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1.  Phototransformations of Advanced Glycation End Products in the Human Eye Lens due to Ultraviolet A Light Irradiation 
Previous studies from this laboratory have shown that ultraviolet A (UVA) light can bleach the yellow advanced glycation end products (AGEs) of aged and cataractous human lenses. The AGEs OP-lysine and argpyrimidine are two UVA-absorbing posttranslational modifications that are abundant in the eye lens. The purpose of this study was to outline the changes in these two AGEs due to UVA irradiation. The changes of OP-lysine, OP-phenethylamine (a phenethylamine analogue of OP-lysine), and argpyrimidine due to irradiation with UVA light in the presence or absence of air and ascorbic acid were followed by different spectral methods. Aged human lenses were similarly irradiated in artificial aqueous humor. The amounts of OP-lysine in the irradiated lenses and in the corresponding dark controls were determined by HPLC. Both OP-lysine and argpyrimidine decreased 20% when irradiated with UVA light in the absence of ascorbic acid. Under the same conditions, OP-lysine was bleached 80% in the presence of ascorbic acid during irradiation experiments. In contrast, argpyrimidine UVA light bleaching was not affected by the presence of ascorbic acid. Interestingly the major product of OP-phenethylamine after UVA irradiation in the presence of ascorbic acid was phenethylamine, which indicates that the entire heterocycle of this AGE was cleaved and the initial amino group was restored. Some AGEs in the human eye lens can be transformed by UVA light.
PMCID: PMC1564128  PMID: 16037236
ascorbic acid; OP-lysine; UVA light; eye lens; glycation
2.  Measurement of Lens Protein Aggregation in Vivo Using Dynamic Light Scattering in a Guinea Pig/UVA Model for Nuclear Cataract 
Photochemistry and photobiology  2008;84(6):1589-1595.
The role of UVA radiation in the formation of human nuclear cataract is not well understood. We have previously shown that exposing guinea pigs for 5 months to a chronic low level of UVA light produces increased lens nuclear light scattering and elevated levels of protein disulfide. Here we have used the technique of dynamic light scattering (DLS) to investigate lens protein aggregation in vivo in the guinea pig/UVA model. DLS size distribution analysis conducted at the same location in the lens nucleus of control and UVA-irradiated animals showed a 28% reduction in intensity of small diameter proteins in experimental lenses compared with controls (P < 0.05). In addition, large diameter proteins in UVA-exposed lens nuclei increased five-fold in intensity compared to controls (P < 0.05). The UVA-induced increase in apparent size of lens nuclear small diameter proteins was three-fold (P < 0.01), and the size of large diameter aggregates was more than four-fold in experimental lenses compared with controls. The diameter of crystallin aggregates in the UVA-irradiated lens nucleus was estimated to be 350 nm, a size able to scatter light. No significant changes in protein size were detected in the anterior cortex of UVA-irradiated lenses. It is presumed that the presence of a UVA chromophore in the guinea pig lens (NADPH bound to zeta crystallin), as well as traces of oxygen, contributed to UVA-induced crystallin aggregation. The results indicate a potentially harmful role for UVA light in the lens nucleus. A similar process of UVA-irradiated protein aggregation may take place in the older human lens nucleus, accelerating the formation of human nuclear cataract.
PMCID: PMC2702991  PMID: 18627516
3.  Violet and blue light blocking intraocular lenses: photoprotection versus photoreception 
To analyse how intraocular lens (IOL) chromophores affect retinal photoprotection and the sensitivity of scotopic vision, melanopsin photoreception, and melatonin suppression.
Transmittance spectra of IOLs, high pass spectral filters, human crystalline lenses, and sunglasses are used with spectral data for acute ultraviolet (UV)‐blue photic retinopathy (“blue light hazard” phototoxicity), aphakic scotopic luminous efficiency, melanopsin sensitivity, and melatonin suppression to compute the effect of spectral filters on retinal photoprotection, scotopic sensitivity, and circadian photoentrainment.
Retinal photoprotection increases and photoreception decreases as high pass filters progressively attenuate additional short wavelength light. Violet blocking IOLs reduce retinal exposure to UV (200–400 nm) radiation and violet (400–440 nm) light. Blue blocking IOLs attenuate blue (440–500 nm) and shorter wavelength optical radiation. Blue blocking IOLs theoretically provide better photoprotection but worse photoreception than conventional UV only blocking IOLs. Violet blocking IOLs offer similar UV‐blue photoprotection but better scotopic and melanopsin photoreception than blue blocking IOLs. Sunglasses provide roughly 50% more UV‐blue photoprotection than either violet or blue blocking IOLs.
Action spectra for most retinal photosensitisers increase or peak in the violet part of the spectrum. Melanopsin, melatonin suppression, and rhodopsin sensitivities are all maximal in the blue part of the spectrum. Scotopic sensitivity and circadian photoentrainment decline with ageing. UV blocking IOLs provide older adults with the best possible rhodopsin and melanopsin sensitivity. Blue and violet blocking IOLs provide less photoprotection than middle aged crystalline lenses, which do not prevent age related macular degeneration (AMD). Thus, pseudophakes should wear sunglasses in bright environments if the unproved phototoxicity‐AMD hypothesis is valid.
PMCID: PMC1860240  PMID: 16714268
intraocular lens; phototoxicity; scotopic; macular degeneration; melanopsin
4.  A Class I UV-Blocking (senofilcon A) Soft Contact Lens Prevents UVA-induced Yellow Fluorescence and NADH loss in the Rabbit Lens Nucleus in vivo 
Experimental eye research  2012;102C:17-27.
It is known that fluorescence, much of it caused by UVA light excitation, increases in the aging human lens, resulting in loss of sharp vision. This study used an in vivo animal model to investigate UVA-excited fluorescence in the rabbit lens, which contains a high level of the UVA chromophore NADH, existing both free and bound to λ-crystallin. Also, the ability of a Class I (senofilcon A) soft contact lens to protect against UVA-induced effects on the rabbit lens was tested. Rabbit eyes were irradiated with UVA light in vivo (100 mW/cm2 on the cornea) for 1 hour using monochromatic 365 nm light. Irradiation was conducted in the presence of either a senofilcon A contact lens, a minimally UV-absorbing lotrafilcon A contact lens, or no contact lens at all. Eyes irradiated without a contact lens showed blue 365 nm-excited fluorescence initially, but this changed to intense yellow fluorescence after 1 hour. Isolated, previously irradiated lenses exhibited yellow fluorescence originating from the lens nucleus when viewed under 365 nm light, but showed normal blue fluorescence arising from the cortex. Previously irradiated lenses also exhibited a faint yellow color when observed under visible light. The senofilcon A contact lens protected completely against the UVA-induced effects on fluorescence and lens yellowing, whereas the lotrafilcon A lens showed no protection. The UVA-exposure also produced a 53% loss of total NADH (free plus bound) in the lens nucleus, with only a 13% drop in the anterior cortex. NADH loss in the nucleus was completely prevented with use of a senofilcon A contact lens, but no significant protection was observed with a lotrafilcon A lens. Overall, the senofilcon A lens provided an average of 67% protection against UVA-induced loss of four pyridine nucleotides in four different regions of the lens. HPLC analysis with fluorescence detection indicated a nearly six-fold increase in 365 nm-excited yellow fluorescence arising from lens nuclear λ-crystallin after the in vivo UVA exposure. It is concluded that UVA-induced loss of free NADH (which fluoresces blue) may have allowed the natural yellow fluorescence of λ-crystallin and other proteins in the lens nucleus to become visible. Increased fluorescence exhibited by UVA-exposed λ-crystallin may have been the result of a UVA-induced change in the conformation of the protein occurring during the initial UVA-exposure in vivo. The results demonstrate the greater susceptibility of the lens nucleus to UVA-induced stress, and may relate to the formation of human nuclear cataract. The senofilcon A contact lens was shown to be beneficial in protecting the rabbit lens against effects of UVA light, including changes in fluorescence, increased yellowing and loss of pyridine nucleotides.
PMCID: PMC3432665  PMID: 22766154
UVA light; rabbit; in vivo; lens; yellowing; fluorescence; pyridine nucleotides; nuclear cataract
5.  Sunlight and age-related eye disease. 
Within 50 years, if current trends continue, 50 million elderly Americans will suffer visual impairment from macular degeneration or cataract. However, available evidence indicates that this impending crisis of visual health can be minimized by a simple, safe, inexpensive, and practical means of prevention. Cataract and macular degeneration are the ultimate consequences of normal aging, a lifelong process of deterioration. Three major causes of ocular deterioration have been identified: oxygen, heat, and solar radiation. Among these, the radiation hazard is readily accessible to human intervention. The lens is damaged by ultraviolet radiation in sunlight, whereas the retina can be harmed by high-energy visible radiation (the "violet and blue"). Use of sunglasses that block all ultraviolet radiation and severely attenuate high-energy visible radiation will slow the pace of ocular deterioration and delay the onset of age-related disease, thereby reducing its prevalence. A 20-year delay would practically eliminate these diseases as significant causes of visual impairment in the United States.
PMCID: PMC2637688  PMID: 1507250
6.  Retardation of Regeneration and Division of Blepharisma by Ultraviolet Radiation and Its Photoreversal 
The Journal of General Physiology  1961;44(3):543-554.
Regeneration of Blepharisma undulans variety japonicus from which the hypostome has been removed is retarded by dosages of 3000 to 4600 ergs/mm.2 at wavelength 2654A most strongly when the fragment is exposed soon after cutting. Dosages greater than 4600 ergs/mm.2 prevent regeneration. Regeneration is also retarded strongly when the Blepharisma are cut soon after irradiation. Starvation retards regeneration and potentiates the effect of ultraviolet radiations. Division after regeneration of Blepharisma is also retarded by ultraviolet radiations about equally, regardless of when the Blepharisma are cut indicating a more lasting effect of the radiations upon the cells. Blepharisma cut after irradiation usually recover from the effects of the radiations sooner than uncut individuals given the same dosage. Retardation of division by ultraviolet radiation is subject to photoreversal by visible light, especially in a nitrogen atmosphere, provided the ultraviolet dose is not excessive. Visible light alone if prolonged, retards regeneration or may even kill the cut fragments of Blepharisma.
PMCID: PMC2195106  PMID: 13705175
7.  Non-Invasive Bleaching of the Human Lens by Femtosecond Laser Photolysis 
PLoS ONE  2010;5(3):e9711.
Globally, cataract is the leading cause of blindness and impaired vision. Cataract surgery is an attractive treatment option but it remains unavailable in sufficient quantity for the vast majority of the world population living in areas without access to specialized health care. Reducing blindness from cataract requires solutions that can be applied outside operating theatres. Cataract is a protein conformational disease characterized by accumulation of light absorbing, fluorescent and scattering protein aggregates. The aim of the study was to investigate whether these compounds were susceptible to photobleaching by a non-invasive procedure and whether this would lead to optical rejuvenation of the lens.
Methodology/Principal Findings
Nine human donor lenses were treated with an 800 nm infra-red femtosecond pulsed laser in a treatment zone measuring 1×1×0.52 mm. After laser treatment the age-induced yellow discoloration of the lens was markedly reduced and the transmission of light was increased corresponding to an optical rejuvenation of 3 to 7 years.
The results demonstrate that the age-induced yellowing of the human lens can be bleached by a non-invasive procedure based on femtosecond laser photolysis. Cataract is a disease associated with old age. At the current technological stage, lens aging is delayed but with a treatment covering the entire lens volume complete optical rejuvenation is expected. Thus, femtosecond photolysis has the potential clinical value of replacing invasive cataract surgery by a non-invasive treatment modality that can be placed in mobile units, thus breaking down many of the barriers impeding access to treatment in remote and poor regions of the world.
PMCID: PMC2838787  PMID: 20300521
8.  Tryptophan Cluster Protects Human γD-Crystallin from Ultraviolet Radiation-Induced Photoaggregation In Vitro 
Photochemistry and Photobiology  2013;89(5):1106-1115.
Exposure to ultraviolet radiation (UVR) is a significant risk factor for age-related cataract, a disease of the human lens and the most prevalent cause of blindness in the world. Cataract pathology involves protein misfolding and aggregation of the primary proteins of the lens, the crystallins. Human γD-crystallin (HγD-Crys) is a major γ-crystallin in the nucleus of the human lens. We report here analysis of UVR-induced damage to HγD-Crys in vitro. Irradiation of solutions of recombinant HγD-Crys with UVA/UVB light produced a rise in solution turbidity due to polymerization of the monomeric crystallins into higher molecular weight aggregates. A significant fraction of this polymerized protein was covalently linked. Photoaggregation of HγD-Crys required oxygen and its rate was protein concentration and UVR dose dependent. To investigate the potential roles of individual tryptophan residues in photoaggregation, triple W:F mutants of HγD-Crys were irradiated. Surprisingly, despite reducing UVR absorbing capacity, multiple W:F HγD-Crys mutant proteins photoaggregated more quickly and extensively than wild type. The results reported here are consistent with previous studies that postulated that an energy transfer mechanism between the highly conserved pairs of tryptophan residues in HγD-Crys could be protective against UVR-induced photodamage.
PMCID: PMC3823069  PMID: 23683003
9.  In vitro ultraviolet–induced damage in human corneal, lens, and retinal pigment epithelial cells 
Molecular Vision  2011;17:237-246.
The purpose was to develop suitable in vitro methods to detect ocular epithelial cell damage when exposed to UV radiation, in an effort to evaluate UV-absorbing ophthalmic biomaterials.
Human corneal epithelial cells (HCEC), lens epithelial cells (HLEC), and retinal pigment epithelial cells (ARPE-19) were cultured and Ultraviolet A/Ultraviolet B (UVA/UVB) blocking filters and UVB-only blocking filters were placed between the cells and a UV light source. Cells were irradiated with UV radiations at various energy levels with and without filter protections. Cell viability after exposure was determined using the metabolic dye alamarBlue and by evaluating for changes in the nuclei, mitochondria, membrane permeability, and cell membranes of the cells using the fluorescent dyes Hoechst 33342, rhodamine 123, calcein AM, ethidium homodimer-1, and annexin V. High-resolution images of the cells were taken with a Zeiss 510 confocal laser scanning microscope.
The alamarBlue assay results of UV-exposed cells without filters showed energy level-dependent decreases in cellular viability. However, UV treated cells with 400 nm LP filter protection showed the equivalent viability to untreated control cells at all energy levels. Also, UV irradiated cells with 320 nm LP filter showed lower cell viability than the unexposed control cells, yet higher viability than UV-exposed cells without filters in an energy level-dependent manner. The confocal microscopy results also showed that UV radiation can cause significant dose-dependent degradations of nuclei and mitochondria in ocular cells. The annexin V staining also showed an increased number of apoptotic cells after UV irradiation.
The findings suggest that UV-induced HCEC, HLEC, and ARPE-19 cell damage can be evaluated by bioassays that measure changes in the cell nuclei, mitochondria, cell membranes, and cell metabolism, and these assay methods provide a valuable in vitro model for evaluating the effectiveness of UV-absorbing ophthalmic biomaterials, including contact lenses and intraocular lenses.
PMCID: PMC3025821  PMID: 21270970
10.  Mechanical Tissue Optical Clearing Devices: Enhancement of Light Penetration in Ex-Vivo Porcine Skin and Adipose Tissue 
Lasers in surgery and medicine  2008;40(10):688-694.
Background and Objective
The complex morphological structure of tissue and associated variations in the indices of refraction of components therein, provides a highly scattering medium for visible and near-infrared wavelengths of light. Tissue optical clearing permits delivery of light deeper into tissue, potentially improving the capabilities of various light-based therapeutic techniques, such as adipose tissue removal or reshaping.
Study Design/ Materials and Methods
We report results of a study to evaluate effectiveness of novel mechanical tissue optical clearing devices (TOCD) using white light photography and infrared imaging radiometry (IIR). The TOCD consists of a pin array and vacuum pressure source applied directly to the skin surface. IIR images recorded light absorption and temperature increase of ex vivo porcine skin and adipose during laser irradiation (980 and 1210 nm) before and after TOCD application.
White light photographic images of in vivo human skin demonstrated localized compression and altered visual appearance, indicative of water and blood movement in skin. White light photographic images also showed increased visible light transport through regions of ex vivo porcine skin compressed by TOCD pins. Rate of heating in sub-dermal adipose regions beneath TOCD pins was two-fold higher following TOCD application.
Results of our study suggest that mechanical optical clearing may provide a means to deliver increased light fluence to dermal and adipose tissues.
PMCID: PMC2605205  PMID: 19065559
Vacuum; compression; fat; cellulite; reshaping; water transport; refractive index; scattering; absorption; thickness; radiometry; dermis
11.  Photochemotherapy and the eye: photoprotective factors. 
The photosensitizer, 8-Methoxypsoralen, binds reversibly to a greater extent to human serum albumin than to human lens in the absence of UVA irradiation. Since experimental and clinical evidence suggests that peak levels in skin exceed those in plasma, which in turn exceed those in the lens, 5 to 10 fold more UVA is required to induce lenticular injury than to produce a threshold skin effect. To prevent the potential photoactivation of residual of 8-MOP in the lens, UVA opaque wraparound sunglasses should be worn for 24 hours after drug ingestion. Glasses with limited UVA transmission but good visible transmission should be worn after the 24 hour period following drug ingestion or indoors during the first 24 hours as a substitute for the wraparound. Every effort should be made to insure complete compliance in the wearing of UVA photoprotective lenses by all psoralen photosensitized patients.
PMCID: PMC1312142  PMID: 7257058
12.  The Role of Optical Radiations in Skin Cancer 
ISRN Dermatology  2013;2013:842359.
Purpose. Electromagnetic radiation with wavelength in the range 100 nm to 1 mm is known as optical radiation and includes ultraviolet radiation, the visible spectrum, and infrared radiation. The deleterious short- and long-term biological effects of ultraviolet radiation, including melanoma and other skin cancers, are well recognized. Infrared radiation may also have damaging biological effects. Methods. The objective of this review was to assess the literature over the last 15 years and to summarize correlations between exposure to optical radiation and the risk of melanoma and other cancers. Results. There is a clear correlation between exposure to UV radiation and the development of skin cancer. Most importantly, a strong association between artificial UV radiation exposure, for example, tanning devices, and the risk of melanoma and squamous cell carcinoma has been clearly demonstrated. There is no clear evidence that exposure to IR and laser radiation may increase the risk of skin cancer, although negative health effects have been observed. Conclusions. Preventative strategies that involve provision of public information highlighting the risks associated with exposure to sunlight remain important. In addition, precautionary measures that discourage exposure to tanning appliances are required, as is legislation to prevent their use during childhood.
PMCID: PMC3655639  PMID: 23710365
13.  Luteolin Prevents Solar Radiation-Induced Matrix Metalloproteinase-1 Activation in Human Fibroblasts: A Role for p38 Mitogen-Activated Protein Kinase and Interleukin-20 Released from Keratinocytes 
Rejuvenation Research  2012;15(5):466-475.
Human skin is continuously exposed to solar radiation, which can result in photoaging, a process involving both dermal and, to a lesser extent, epidermal structures. Previously, we have shown that the flavonoid luteolin protects the epidermis from ultraviolet (UV)-induced damage by a combination of UV-absorbing, antioxidant, and antiinflammatory properties. The aim of the present study was to determine direct and indirect effects of luteolin on dermal fibroblasts as major targets of photoaging. Stimulation of fibroblasts with UVA light or the proinflammatory cytokine interleukin-20 (IL-20) is associated with wrinkled skin, increased IL-6 secretion, matrix metalloproteinase (MMP-1) expression, and hyaluronidase activity. All of these targets were inhibited by luteolin via interference with the p38 mitogen-activated protein kinase (MAPK) pathway. Next, we assessed the role of conditioned supernatants from keratinocytes irradiated with solar-simulated radiation (SSR) on nonirradiated dermal fibroblasts. In keratinocytes, luteolin inhibited SSR-induced production of IL-20, also via interference with the p38 MAPK pathway. Similarly, keratinocyte supernatant-induced IL-6 and MMP-1 expression in fibroblasts was reduced by pretreatment of keratinocytes with luteolin. Finally, these results were confirmed ex vivo on skin explants treated with luteolin before UV irradiation. Our results suggest that SSR-mediated production of soluble factors in keratinocytes is modulated by luteolin and may attenuate photoaging in dermal fibroblasts.
PMCID: PMC3482844  PMID: 23004935
14.  Glycation by Ascorbic Acid Oxidation Products Leads to the Aggregation of Lens Proteins 
Biochimica et biophysica acta  2007;1782(1):22-34.
Previous studies from this laboratory have shown that there are striking similarities between the yellow chromophores, fluorophores and modified amino acids released by proteolytic digestion from calf lens proteins ascorbylated in vitro and their counterparts isolated from aged and cataractous lens proteins. The studies reported in this communication were conducted to further investigate whether ascorbic acid-mediated modification of lens proteins could lead to the formation of lens protein aggregates capable of scattering visible light, similar to the high molecular aggregates found in aged human lenses. Ascorbic acid, but not glucose, fructose, ribose or erythrulose, caused the aggregation of calf lens proteins to proteins ranging from 2.2 × 106 up to 3.0 × 108 Da. This compared to proteins ranging from 1.8 × 106 up to 3.6 × 108 Da for the water-soluble (WS) proteins isolated from aged human lenses. This aggregation was likely due to the glycation of lens crystallins because [U-14C] ascorbate was incorporated into the aggregate fraction and because CNBH3, which reduces the initial Schiff base, prevented any protein aggregation.
Reactions of ascorbate with purified crystallin fractions showed little or no aggregation of α-crystallin, significant aggregation of βH-crystallin, but rapid precipitation of purified βL- and γ-crystallin. The aggregation of lens proteins can be prevented by the binding of damaged crystallins to alpha-crystallin due to its chaperone activity. Depending upon the ratios between the components of the incubation mixtures, α-crystallin prevented the precipitation of the purified βL- and γ-crystallin fractions during ascorbylation. The addition of at least 20% of alpha-crystallin by weight into glycation mixtures with βL-, or γ-crystallins completely inhibited protein precipitation, and increased the amount of the high molecular weight aggregates in solution. Static and dynamic light scattering measurements of the supernatants from the ascorbic acid-modified mixtures of α- and βL-, or γ-crystallins showed similar molar masses (up to 108 Da) and hydrodynamic diameter (up to 80 nm). These data support the hypothesis, that if the lens reducing environment is compromised, the ascorbylation of lens crystallins can significantly change the short range interactions between different classes of crystallins leading to protein aggregation, light scattering and eventually to senile cataract formation.
PMCID: PMC2274899  PMID: 18023423
ascorbic acid; glycation; lens proteins; protein aggregation; light scattering
15.  A look into the invisible: ultraviolet-B sensitivity in an insect (Caliothrips phaseoli) revealed through a behavioural action spectrum 
Caliothrips phaseoli, a phytophagous insect, detects and responds to solar ultraviolet-B radiation (UV-B; λ ≤ 315 nm) under field conditions. A highly specific mechanism must be present in the thrips visual system in order to detect this narrow band of solar radiation, which is at least 30 times less abundant than the UV-A (315–400 nm), to which many insects are sensitive. We constructed an action spectrum of thrips responses to light by studying their behavioural reactions to monochromatic irradiation under confinement conditions. Thrips were maximally sensitive to wavelengths between 290 and 330 nm; human-visible wavelengths (λ ≥ 400 nm) failed to elicit any response. All but six ommatidia of the thrips compound eye were highly fluorescent when exposed to UV-A of wavelengths longer than 330 nm. We hypothesized that the fluorescent compound acts as an internal filter, preventing radiation with λ > 330 nm from reaching the photoreceptor cells. Calculations based on the putative filter transmittance and a visual pigment template of λmax = 360 nm produced a sensitivity spectrum that was strikingly similar to the action spectrum of UV-induced behavioural response. These results suggest that specific UV-B vision in thrips is achieved by a standard UV-A photoreceptor and a sharp cut-off internal filter that blocks longer UV wavelengths in the majority of the ommatidia.
PMCID: PMC2842652  PMID: 19846453
UV-B; insect vision; insect behaviour
To determine whether digital spatial intensity patterns can be developed to effect precise in vitro correction of myopic, hyperopic, and astigmatic refractive errors in a silicone light-adjustable lens (LAL). Also, to determine whether a new spatial intensity pattern for “lock-in” is effective in vitro.
A digital interferometer/irradiation system was developed to irradiate LALs and measure the power change following irradiation. Light-adjustable lenses were mounted into a wet cell maintained at 35.0 ± 0.5°C (simulated ocular temperature) and allowed to equilibrate for a minimum of 2 hours. Ultraviolet light was then applied with spatial light intensity patterns to correct hyperopia, myopia, and astigmatism. Light-adjustable lenses were also treated to effect lock-in with a separate spatial light intensity pattern. Treated lenses were characterized for power change and optical quality. In the case of lock-in, exhaustive chemical extraction was also performed to determine the percentage of remaining macromer.
Appropriate digital irradiation spatial intensity patterns were created to develop nomograms for in vitro correction of myopia, hyperopia, and astigmatism in approximate 0.25 D steps. Power changes were reproducible and did not alter optical quality of the LALs. Further, lock-in dosing of the LALs did not alter optical quality or significantly change LAL power.
In vitro nomograms have been developed for a silicone LAL that permit precise correction of myopia, hyperopia, and astigmatism. Furthermore, a spatial light intensity pattern has been devised that effects lock-in without significantly altering LAL power or optical quality.
PMCID: PMC1280088  PMID: 15747746
17.  Experimental Coronary Angioplasty Using a UV-Excimer Laser 
Texas Heart Institute Journal  1985;12(4):339-343.
Both conventional Argon- and NdYAG-lasers in continuous wave or pulsed application and far ultraviolet laser radiation are able to cause a loss of substance of biologic tissue. The thermic and ablative effects of NdYAG-lasers and UV-Excimer lasers at the wavelengths of 193 nm (ArF) and 248 mm (KrF) on inconspicuous and atherosclerotic human and animal coronary vessels were compared by histologic and, in some cases, by scanning electron microscopic examinations. Whereas common lasers generally produce thermal injuries of the surroundings, pulsed far ultra-violet radiation is characterized by a lack of thermic damage. The UV-radiation in vitro cleaned precise defects of substance, and assessable tissue-removing effects were found. These results were influenced by the wavelength used. In general, the removing effect was good in normal and atherosclerotic tissue, whereas massive calcification was very resistant. Excimer lasers seem to be preferable for ablation of atherosclerotic tissue, but still there is a great number of technical problems to be solved until use in the clinical setting can be justified.
PMCID: PMC341885  PMID: 15226990
18.  UV-B-Induced Damage to the Lens In Vitro: Prevention by Caffeine 
Ultraviolet (UV) irradiation is one of the significant risk factors in the genesis of cataracts. Pathogenetically, the process can be triggered by the intraocular generation of various reactive species of oxygen that are well known to be initiated by the penetration of light, especially of the UV frequencies. The contribution of UV exposure in the etiology of this disease is likely to increase further due to ozone depletion in the upper atmosphere. The present studies were undertaken to examine if the UV effects can be attenuated with the xanthine-based alkaloids primarily present in tea and coffee. We have examined this possibility by in vitro lens culture studies with caffeine. As expected, mice lenses incubated in Tyrode solution exposed to UV at 302 nm are physiologically damaged, as evidenced by the inhibition of the active transport of 86Rb+, an ion acting as a surrogate of the K+. There was a simultaneous decrease in the levels of adenosine triphosphate and glutathione. The addition of caffeine to the medium prevented such deleterious effects. That caffeine and perhaps other xanthinoids have a protective effect against cataract formation induced by UV has hence been demonstrated for the first time.
PMCID: PMC2958437  PMID: 18788993
The Journal of General Physiology  1955;38(3):295-305.
1. The effect of the nutritional state of Didinium nasutum on its resistance to short ultraviolet (UV) radiation (2654 A) and its recovery from the injury following illumination with visible light (4350 A, blue) was studied. 2. The resistance of a didinium to UV is considerably increased by feeding it a paramecium 15 to 60 minutes before exposure to UV. If fed just before exposure to UV, the resistance is less than that of an unfed control. 3. Photoreversal is only slightly greater in didinia fed after irradiation with UV but before exposure to visible light as compared to those fed after exposure to visible light. 4. Irradiated paramecia are eaten by didinia, provided they have not started to cytolyze. Didinia fed on irradiated paramecia divide at about the same rate as controls or slightly faster. 5. The available stock of Didinium declines in vigor with lapse of time after excystment, as measured by the time required for division. The sensitivity of Didinium to UV did not change essentially during the 5 month period over which tests were made. 6. The theoretical implications of the results are considered.
PMCID: PMC2147485  PMID: 13221774
20.  A clinical study of radiation cataract formation in adult life following γ irradiation of the lens in early childhood 
AIMS—To analyse long term effects on the lens of radium irradiation during infancy.
METHODS—An infant cohort (n = 20, median age 6 months) treated for skin haemangioma with one or two radium-226 needles located at or within the orbital rim was examined 30 to 45 years after γ radiation. Detailed information about the treatment procedure was available for all cases. Subcapsular opacities were graded semiquantitatively according to a scale based on extent and density of the opacities.
RESULTS—A high prevalence of light to moderate posterior, subcapsular, and cortical cataract formation was found in the lenses on the treated side irradiated with a mean dose ranging from approximately 1 to 8 Gy. The cataract formation increased as a function of dose. The presence of subcapsular punctate opacities and vacuoles in the lenses on the untreated side receiving irradiation of an estimated dose varying around 0.1 Gy indicates a higher sensitivity than expected.
CONCLUSION—The growing lens during infancy is sensitive to radium irradiation at doses lower than those previously stated. The eye lens seems suitable for studies of effects of low dose radiation since damaged cells are retained in the lens for a lifetime.

PMCID: PMC1722161  PMID: 9215051
21.  Pigmentation effects of solar simulated radiation as compared with UVA and UVB radiation 
Pigment cell & melanoma research  2008;21(4):487-491.
Different wavelengths of ultraviolet (UV) radiation elicit different responses in the skin. UVA induces immediate tanning and persistent pigment darkening through oxidation of pre-existing melanin or melanogenic precursors, while UVB induces delayed tanning which takes several days or longer to develop and requires activation of melanocytes. We compared the effects of a two-week repetitive exposure of human skin to solar-simulated radiation (SSR), UVA or UVB at doses eliciting comparable levels of visible tanning and measured levels of melanins and melanin-related metabolites. Levels of eumelanin and pheomelanin were significantly higher in the order of SSR, UVB, UVA or unexposed control skin. Levels of free 5-S-cysteinyldopa (5SCD) were elevated about four-fold in SSR- or UVB-exposed skin compared with UVA-exposed or control skin. Levels of protein-bound form of 5SCD tended to be higher in SSR- or UVB-exposed skin than in UVA-exposed or control skin. Total levels of 5-hydroxy-6-methoxyindole-2-carboxylic acid (5H6MI2C) and 6H5MI2C were higher in SSR- than in UVB-exposed or control skin. These results show that SSR is more effective in promoting delayed tanning than UVB radiation alone, suggesting a synergistic effect of UVA radiation. Furthermore, free 5SCD may serve as a good marker of the effect of SSR and UVB.
PMCID: PMC2593890  PMID: 18627527
ultraviolet; solar radiation; melanin; eumelanin; pheomelanin; 5-S-cysteinyldopa; 5(6)-hydroxy-6(5)-methoxyindole-2-carboxylic acid
22.  A Class I (Senofilcon A) Soft Contact Lens Prevents UVB-Induced Ocular Effects, Including Cataract, in the Rabbit In Vivo 
This is the first study to show that a class I UV-blocking soft contact lens can prevent UVB-induced cataract in an animal model.
UVB radiation from sunlight is known to be a risk factor for human cataract. The purpose in this study was to investigate the ability of a class I UV-blocking soft contact lens to protect against UVB-induced effects on the ocular tissues of the rabbit in vivo.
Eyes of rabbits were exposed to UVB light for 30 minutes (270–360 nm, peak at 310 nm, 1.7 mW/cm2 on the cornea). Eyes were irradiated in the presence of either a UV-blocking senofilcon A contact lens, a minimally UV-blocking lotrafilcon A contact lens, or no contact lens at all. Effects on the cornea and lens were evaluated at various times after exposure.
Eyes irradiated with no contact lens protection showed corneal epithelial cell loss plus lens epithelial cell swelling, vacuole formation, and DNA single-strand breaks, as well as lens anterior subcapsular opacification. The senofilcon A lens protected nearly completely against the UVB-induced effects, whereas the lotrafilcon A lens showed no protection.
The results indicate that use of a senofilcon A contact lens is beneficial in protecting ocular tissues of the rabbit against the harmful effects of UVB light, including photokeratitis and cataract.
PMCID: PMC3109047  PMID: 21421866
23.  The ageing lens and cataract: a model of normal and pathological ageing 
Cataract is a visible opacity in the lens substance, which, when located on the visual axis, leads to visual loss. Age-related cataract is a cause of blindness on a global scale involving genetic and environmental influences. With ageing, lens proteins undergo non-enzymatic, post-translational modification and the accumulation of fluorescent chromophores, increasing susceptibility to oxidation and cross-linking and increased light-scatter. Because the human lens grows throughout life, the lens core is exposed for a longer period to such influences and the risk of oxidative damage increases in the fourth decade when a barrier to the transport of glutathione forms around the lens nucleus. Consequently, as the lens ages, its transparency falls and the nucleus becomes more rigid, resisting the change in shape necessary for accommodation. This is the basis of presbyopia. In some individuals, the steady accumulation of chromophores and complex, insoluble crystallin aggregates in the lens nucleus leads to the formation of a brown nuclear cataract. The process is homogeneous and the affected lens fibres retain their gross morphology. Cortical opacities are due to changes in membrane permeability and enzyme function and shear-stress damage to lens fibres with continued accommodative effort. Unlike nuclear cataract, progression is intermittent, stepwise and non-uniform.
PMCID: PMC3061107  PMID: 21402586
cortical cataract; nuclear cataract; lens morphology; light-scattering
24.  The Cytotoxic Effects of Low Intensity Visible and Infrared Light on Human Breast Cancer (MCF7) cells 
A concept of using low intensity light therapy (LILT) as an alternative approach to cancer treatment is at early stages of development; while the therapeutic effects of LILT as a non-invasive treatment modality for localized joint and soft tissue wound healing are widely corroborated. The LEDs-based exposure system was designed and constructed to irradiate the selected cancer and normal cells and evaluate the biological effects induced by light exposures in visible and infrared light range. In this study, human breast cancer (MCF7) cells and human epidermal melanocytes (HEM) cells (control) were exposed to selected far infrared light (3400nm, 3600nm, 3800nm, 3900nm, 4100nm and 4300nm) and visible and near infrared wavelengths (466nm, 585nm, 626nm, 810nm, 850nm and 950nm). The optical intensities of LEDs used for exposures were in the range of 15µW to 30µW. Cellular morphological changes of exposed and sham-exposed cells were evaluated using light microscopy. The cytotoxic effects of these low intensity light exposures on human cancer and normal cell lines were quantitatively determined by Lactate dehydrogenase (LDH) cytotoxic activity and PrestoBlue™ cell viability assays. Findings reveal that far-infrared exposures were able to reduce cell viability of MCF7 cells as measured by increased LDH release activity and PrestoBlue™ assays. Further investigation of the effects of light irradiation on different types of cancer cells, study of possible signaling pathways affected by electromagnetic radiation (EMR) and in vivo experimentation are required in order to draw a firm conclusion about the efficacy of low intensity light as an alternative non-invasive cancer treatment.
PMCID: PMC3962108
Electromagnetic Radiation (EMR); Lactate dehydrogenase (LDH) Assay; PrestoBlue™ Assay; Light microscopy; Far Infrared wavelength exposure; Visible light exposure
25.  Phototoxicity in Human Retinal Pigment Epithelial Cells Promoted by Hypericin, a Component of St. John’s Wort† 
Photochemistry and photobiology  2007;83(3):706-713.
St. John’s Wort (SJW), an over-the-counter antidepressant, contains hypericin, which absorbs light in the UV and visible ranges. In vivo studies have determined that hypericin is phototoxic to skin and our previous in vitro studies with lens tissues have determined that it is potentially phototoxic to the human lens. To determine if hypericin might also be phototoxic to the human retina, we exposed human retinal pigment epithelial cells to 10−7 to 10−5 M hypericin. Fluorescence emission detected from the cells (λexc = 488 nm; λem = 505 nm) confirmed hypericin uptake by human RPE. Neither hypericin exposure alone nor visible light exposure alone reduced cell viability. However when irradiated with 0.7 J/cm2 of visible light (λ> 400 nm) there was loss of cell viability as measured by MTS and LDH assays. The presence of hypericin in irradiated hRPE cells significantly changed the redox equilibrium of glutathione and a decrease in the activity of glutathione reductase. Increased lipid peroxidation as measured by the TBARS assay correlated to hypericin concentration in hRPE cells and visible light radiation. Thus, ingested SJW is potentially phototoxic to retina and could contribute to retinal or early macular degeneration.
PMCID: PMC2092452  PMID: 17576381

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