Is pycnogenol a double-edged sword? Cataractogenic in vitro, but reduces cataract risk in diabetic rats.

PURPOSE: Pycnogenol was used (a) to study its antioxidant activity, (b) to study its effects on lens integrity in organ culture and (c) in vivo to determine whether it could reduce the damage in model diabetic cataract. METHODS: For (a) our luminescent antioxidant assay was used, (b) lenses were incubated in medium 199, with 55.6 mM glucose. Lenses were stained with 0.014 mM rhodamine 123 for 15 min to stain mitochondria, immobilized in 1% agarose in M199, and the equatorial region examined by a Zeiss confocal microscope. For (c) cataract grades of streptozotocin diabetic rats fed 1% pycnogenol were followed for 12 weeks. RESULTS: (a) Pycnogenol in vitro was an antioxidant when challenged with peroxide. (b) In vitro, when [570 mg/L] pycnogenol in dimethyl sulfoxide (DMSO) was used, lenses turned opaque after 3 d of incubation, in both pycnogenol controls and glucose + pycnogenol. Normal controls (DMSO, n = 4) and controls (n = 4) remained clear after 8 d of incubation. After 3 d of incubation with pycnogenol, cumulative protein leakage was greater than 0.28 mg/mL versus 8 d controls (0.018 mg/mL). Similar damage occurred at pycnogenol concentrations as low as 20 mg/L. The 20 mg/L pycnogenol control showed mitochondrial death, and calcium concentration in the lens equatorial differentiating fiber cells increased. (c) In vivo feeding pycnogenol resulted in similar growth and body condition for diabetic rats, and lower cataract grades at 9 and 11 weeks: final serum glucose levels were not significantly different, but glycohemoglobin A1 levels were significantly lower (83.9% of normal, p < 0.05) in pycnogenol-fed diabetic rats. CONCLUSIONS: Although it appears that pycnogenol has a potential toxic effect on incubated lenses, it appears in vivo to have a marginal protective effect, and also significantly reduces glycation of proteins. Supported by Cognis US (formerly Henkel Chemical Co.) and Horphag Research.

The retinal carotenoids zeaxanthin and lutein scavenge superoxide and hydroxyl radicals: a chemiluminescence and ESR study.

PURPOSE: Carotenoids are present in many biological systems, often decreasing the formation of products of oxidative damage to biological molecules. In the macula their concentration is so high that it has been believed that the yellow color filters out damaging blue light. Recent reports that dietary lutein reduces the risk of cataract in the eye lens suggested that the antioxidant action of carotenoids, which has been inferred from decreased oxidative damage, warranted further direct investigation. METHODS: Superoxide and hydroxyl radical scavenging by lutein and zeaxanthin (retinal carotenoids), beta-carotene, lycopene, lutein esters (from marigolds), and a commercial mixture of soy carotenoids were compared to scavenging by ascorbate and ascorbyl palmitate. Radical scavenging was measured with a chemiluminescent assay (luminol) and by electron spin resonance, ESR. Inhibitory concentrations, IC(50), were determined with the luminescent assay. RESULTS: All of the carotenoids scavenged both superoxide (in ESR 30-50% at 16.7 microM) and hydroxyl radicals (in ESR 50-70% at 16.7 microM, in a luminescent assay 90-99%). CONCLUSIONS: While crocin may be unable to scavenge superoxide, some of the other carotenoids do so quite effectively. The mixtures of 15,15'-cis and all-trans-carotenoids studied by ESR and luminescent assay scavenge both superoxide and hydroxyl radicals. Lycopene and beta-carotene both scavenge superoxide more effectively than the xanthophylls of the retina, zeaxanthin and lutein. All of the carotenoids examined scavenged the hydroxyl radicals more effectively than superoxide radicals. The predominant carotenoid in the fovea of the retina, zeaxanthin, scavenged hydroxyl radicals more effectively than the other retinal carotenoid, lutein. Possible mechanisms of radical scavenging by the carotenoids are discussed.

Model study of AREDS antioxidant supplementation of AMD compared to Visudyne: a dominant strategy?

OBJECTIVES: In Ontario, Canada, in a cohort of all people initially aged 50-54 years, modeling whether the Age-Related Eye Disease Study (AREDS) antioxidant supplementation for stage 3 and 4 AMD would decrease the costs of photodynamic treatment with Visudyne. PERSPECTIVE: Third party payer, the Ontario Health Insurance Plan. METHODS: Using reported risk reductions, prevalence data by age and sex from the Beaver Dam studies, and yearly costs: AREDS 182.50 Canadian dollars, potential savings were calculated as the difference or incremental cost between the estimated medical costs for the untreated cohort of 17,000 Canadian dollars for Visudyne treatment of individuals with neovascularization and the same cohort if stage 3 and 4 AMD patients were treated with antioxidants, decreasing progression to neovascularization. Different scenarios were explored for sensitivity analysis of direct cost savings. RESULTS: For the Ontario cohort of approximately 788,000 aged 51-55 years in 2001, for photodynamic therapy of the untreated cohort, modeled costs were 1.7 billion Canadian dollars. AREDS treatment costs would be 513 million Canadian dollars. AREDS would reduce photodynamic therapy costs, a net saving of 431 million Canadian dollars, a saving of 547 Canadian dollars per person in the total cohort, or 6,753 Canadian dollars per stage 3 and 4 patient treated. To explore the sensitivity of this model to AMD incidence rather than prevalence data, Framingham incidence data were incorporated in the model: net savings of 70.3 million Canadian dollars were modeled using Framingham incidence data. CONCLUSION: Under reasonable assumptions, if the case progresses to wet AMD (1) AREDS with Visudyne is less expensive than Visudyne alone in every five-year time period for the cohort that is age 50-54 right now until they become 75-79; thus, the lifetime cost is lower; (2) AREDS with Visudyne yields more QALYs than Visudyne alone in every five-year interval; (3) under all but the most extreme assumptions, the conclusions reached are robust. Even when AREDS costs a little more, it yields more QALYs at a reasonable cost per QALY. Thus, AREDS antioxidant supplementation appears to be a dominant strategy for macular degeneration. Applied to the whole Canadian population, the potential medical cost savings for Visudyne treatment of neovascular AMD are 5.6 billion Canadian dollars in direct costs. These values would be tenfold higher for the USA, because of the currency and population size differences.

Effect of hyperbaric oxygen on guinea pig lens optical quality and on the refractive state of the eye.

The main objective of this study was to investigate the effect of in vivo hyperbaric oxygen (HBO) treatment of albino guinea pigs on ocular refractive state and optical properties of the lens in vitro, as well as on the integrity of the mitochondria of the lens. The animals were treated 30-35 times (2.5-3 months) or 70 times (6 months) with HBO. An increased level of lens nuclear light scattering was evident by slit-lamp at 30 treatments, and this increased at 70 treatments. After 30-35 HBO treatments a myopic shift in refractive state of the eye was seen in two separate studies with two different refractionists. Also, the average back vertex distance of the lens was significantly shorter after 35 HBO treatments while spherical aberration (focal variability) increased after 70 treatments. No difference in refractive state was noted after 70 HBO treatments (a reversal of the initial myopic effect). The mitochondrial distribution and morphology of the lens epithelium and the superficial cortical fibre cells were normal after both 35 and 70 HBO treatments, highlighting that HBO treatment does not affect the superficial cortex of the lens. The results of the in vitro lens optical analysis carried out in this study correlate with the myopia observed after 30-35 HBO in vivo treatments. A similar reversible myopia and increase in lens nuclear light scattering is known to occur in humans treated with HBO for extended periods and the results suggest that the myopia was caused by a change in the refractive index of the lens. The significant loss of sharp focus after 70 HBO treatments can be correlated with previous reports of biochemical and morphological changes associated with HBO-induced loss of lens nuclear transparency in mature guinea pigs. The guinea pig HBO model may be a useful approach for the study of lens development and refractive error.

Focal length variability and protein leakage as tools for measuring photooxidative damage to the lens.

Hypericin is the ingredient used to standardize the popular over-the-counter antidepressant medication St. John's Wort. Because hypericin readily produces singlet oxygen and other excited state intermediates, it is a very efficient phototoxic agent in the eye that can potentially induce the development of the cataract photooxidative mechanism. Hypericin absorbs in the UV and visible ranges, binds to the lens crystallins (alpha, beta and gamma) and damages these proteins through a photooxidative mechanism. Effects were measured previously using fluorescence, UV and mass spectrometry. We report here two additional methods to monitor lens damage: (1) measuring focal length variability using a ScanTox instrument and (2) measuring protein leakage from the damaged lens. Because nonenzymic glycation results in free radical production, we chose to use elevated glucose concentrations as a convenient model for studying oxidative stress. To compare and contrast photooxidative damage against oxidative damage to the lens, we also measured the focal length variability and protein leakage induced by the presence of elevated glucose concentrations. We found that the total accumulated protein leakage was positively correlated (r = 0.9) with variability in focal length. Lenses treated with hypericin and irradiated with UVB had an increase in focal length variability as compared with the lenses that were only UVB-irradiated. Lenses without UVB irradiation had much lower focal length variability than irradiated lenses. For non-hypericin-treated lenses, UVB-irradiated lenses had a larger variability (4.58 mm) than the unirradiated lenses (1.78 mm). The lenses incubated in elevated glucose concentrations had a focal length variability (3.23 mm) equivalent to that of the unirradiated hypericin-treated lenses (3.54 mm). We conclude that photooxidative damage by hypericin results in changes in the optical properties of the lens, protein leakage and finally cataract formation. In contrast to this, high concentrations of glucose induced protein leakage but not changes in optical properties or the opacity associated with a cataract. This work provides further evidence that people should protect their eyes from intense sunlight when taking St. John's Wort.

Phenol antioxidant quantity and quality in foods: beers and the effect of two types of beer on an animal model of atherosclerosis.

The free phenols have been measured in 15 lagers, 6 porters and ales, and 11 light and nonalcoholic beers. Phenols were measured colorimetrically using an oxidation-reduction reaction with Folin-Ciocalteu reagent and catechin as the standard. The order of phenol concentration was ales > lagers > low calorie > nonalcoholic. The quality of antioxidants of the major phenols in beers and the quality of beer antioxidants were measured by (1) dose-response inhibition of lower density lipoprotein oxidation and (2) concentration of phenols in the beers at which 50% of the peroxide was destroyed in a luminescent assay for antioxidant activity. The beers' lipoprotein antioxidant quality was clearly superior to that of vitamin antioxidants and to that of the phenol ingredients, suggesting synergism among the antioxidants in the mixture. The average per capita consumption of beer in the United States in 2000 was 225 mL/day, equivalent to 42 mg/day of catechin equivalents. Beer provides more antioxidants per day than wine in the U.S. diet. A dark beer and a lager beer were given at two concentrations to cholesterol-fed hamsters, an animal model of atherosclerosis. At the high dose ((1)/(2)-diluted beer) both lager and dark beer significantly inhibited atherosclerosis compared to a control of 2% alcohol. At the high dose, lager significantly decreased cholesterol and triglycerides, and both beers acted as in vivo antioxidants by decreasing the oxidizability of lower density lipoproteins. At the low dose ((1)/(10)-diluted beer) only the lager beer significantly decreased atherosclerosis compared to the 0.4% alcohol control. The polyphenols in the beers appear to be responsible for the benefits of beer in this model. Lager beer inhibited atherosclerosis at a human equivalent dose in this hamster model of atherosclerosis.

Mechanisms of ocular toxicity using the in vitro bovine lens and sodium dodecyl sulfate as a chemical model.

Previous work using the in vitro bovine lens as a model has shown a correlation between toxicity and lens optical function and showed much higher sensitivity in detecting irritancy of several surfactants at much lower concentrations than the Draize score. In the current study, cultured bovine lenses were used to study the effects of the surfactant sodium dodecyl sulfate (SDS) on lens optical properties and mitochondrial integrity. Bovine lenses were exposed to SDS (0.1 to 0.00625%) for 30 min and cultured for 24 h. Compared to controls (n = 17), loss of sharp focus was evident immediately following exposure to 0.1% SDS (n = 14, p < 0.0001). At 24 h loss of sharp focus became evident in all groups. Loss of lens transparency, significant increase in lens wet weight, and axial length were seen 24 h postexposure in lenses treated with 0.1 to 0.025% SDS. Confocal analysis 24 h postexposure showed SDS concentration-dependent decrease in number and length of the mitochondria in lens epithelial and superficial cortical fiber cells. The results of this study show a correlation between lens optical properties and metabolic function and together provide a sensitive in vitro model of ocular chemical toxicity. Results of confocal analysis suggest that the mitochondrial integrity of the superficial cortical fiber cells is most sensitive to damage caused by SDS. The results further suggest that recovery of lens metabolic function is necessary for the recovery of lens optical properties.