Kakarla V Chalam

University of Florida, Gainesville, Florida, United States

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Publications (141)280.65 Total impact

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    ABSTRACT: The objective of the study was to determine the safety parameters of using brilliant blue green (BBG) for chromovitrectomy by assessing the cytotoxicity of BBG on cultured retinal ganglion cells (RGCs) exposed to illumination. RGCs were exposed to two concentrations of BBG (0.25 and 0.5 mg/mL) under metal halide illumination at varying distances (1 and 2.5 cm), intensities (990 and 2,000 Fc), and durations (1, 5 and 15 minutes). Cell viability was assessed using the WST-1 and CellTiter 96(®) AQueous One solution cell proliferation assays. Using the WST-1 assay, with high-intensity illumination, viability of RGCs ranged from 97.5±16.4% of controls with minimum BBG and light exposure (0.25 mg/mL BBG and illuminated for 1 minute at 2.5 cm distance) to 53.1±11.3% of controls with maximum BBG and light exposure (0.50 mg/mL and illuminated for 15 minutes at 1 cm distance; P < 0.01). With medium-intensity illumination, RGCs showed better viability, ranging from 95.1±7.2% of controls with minimum BBG and light exposure to 72.3±12.8% of controls with maximum BBG and light exposure. CellTiter 96(®) AQueous One assay showed similar results. RGCs seem to safely tolerate up to 5 minutes of exposure to 0.5 mg/mL BBG under diffuse medium-intensity illumination (990 Fc).
    Journal of Clinical Medicine Research 07/2015; 7(7):517-524. DOI:10.14740/jocmr2085e
  • Wenhua Li, Kakarla V Chalam, Sandeep Grover
    Jama Ophthalmology 06/2015; 133(6):715-716. DOI:10.1001/jamaophthalmol.2015.64 · 3.83 Impact Factor
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    ABSTRACT: To evaluate the outcomes of "concurrent vitrectomy" to retrieve dislocated lens fragment during phacoemulsification. In a retrospective, observational case series, data of patients who underwent "concurrent" pars plana vitrectomy (PPV) for dislocated lens fragments between the period 2000 and 2008 were reviewed. Data collected included patient demographics, pre-operative visual acuity, intra-operative occurrence of retinal breaks, duration of follow up, post-operative intraocular pressure, final best-corrected visual acuity (BCVA), presence of cystoid macular edema (CME) and occurrence of rhegmatogenous retinal detachment (RRD). A total of 58 eyes of 58 patients were included in the study. At 12mo the mean postoperative BCVA was logMAR 0.17 (20/30) with a range of logMAR 0 to 0.69 (20/20 to 20/100), with 96.6% (56/58) of patients showing post-operative improvement in visual acuity (P=0.005). None of the patients developed postoperative retinal detachment, endophthalmitis or non-resolving uveitis at 12mo. Our study results suggest concurrent PPV for retained lens fragments after cataract surgery is beneficial and may decrease the risk of glaucoma and prevent development of RRD.
    International Journal of Ophthalmology 02/2015; 8(1):89-93. DOI:10.3980/j.issn.2222-3959.2015.01.16 · 0.50 Impact Factor
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    ABSTRACT: To evaluate the effect of the iPad as a low vision aid in improving the reading ability of low vision patients (LVPs). In this study, 228 consecutive patients that came for their routine eye care examination at the University of Florida, Jacksonville eye clinic, were enrolled. Patients met inclusion criteria if they had best-corrected visual acuity (BCVA) of 20/100 or worse in the best corrected eye and were willing to participate in the study. The patient's reading ability was assessed both with the patient's own spectacles and an iPad. Patients were encouraged to enlarge the reading material as well as change the contrast until they could read comfortably. The number of patients able to read the text comfortably was recorded. Out of the total 228 participants who qualified, 103 (45%) were male and 125 (55%) were female. Only 22% could read standard newsprint-sized text (N8) without the help of an iPad. With the help of an iPad, 94% participants with impaired vision were able to read standard newsprint-sized text (N8) or smaller text (P<0.01). The iPad, a new portable electronic media device, can be adapted by LVPs to improve their reading ability.
    Clinical ophthalmology (Auckland, N.Z.) 01/2015; 9:17-20. DOI:10.2147/OPTH.S73193
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    ABSTRACT: Central corneal thickness (CCT) can be measured by using contact and non-contact methods. Ultrasound pachymetry (US pachymetry) is a contact method for measuring CCT and is perhaps the most commonly used method. However, non-contact methods like scanning slit topography (Orbscan II), slit-lamp optical coherence tomography (SL-OCT), and specular microscopy are also used. Not many studies have correlated the measurement of CCT with all four modalities. The purpose of this study was to compare and correlate the CCT measurements obtained by US pachymetry with SL-OCT, specular microscopy, and Orbscan. This is a prospective, comparative study done in an institutional setting. Thirty-two eyes of 32 subjects with no known ocular disease and best-corrected visual acuity of 20/20 were enrolled. CCT measurements were obtained using SL-OCT, specular microscopy, scanning slit topography (Orbscan), and US pachymetry. Three measurements were made with each instrument by the same operator. Mean, standard deviation, and coefficient of variation were calculated for CCT measurements acquired by the four measurement devices. Bland-Altman plot was constructed to determine the agreements between the CCT measurements obtained by different equipment. The mean CCT was 548.16±48.68 μm by US pachymetry. In comparison, CCT averaged 546.36±44.17 μm by SL-OCT, 557.61±49.92 μm by specular microscopy, and 551.03±48.96 μm by Orbscan for all subjects. Measurements by the various modalities were strongly correlated. Correlations (r (2)) of CCT, as measured by US pachymetry compared with other modalities, were: SL-OCT (r (2)=0.98, P<0.0001), specular microscopy (r (2)=0.98, P<0.0001), and Orbscan (r (2)=0.96, P<0.0001). All modalities had a linear correlation with US pachymetry measurements. In subjects with healthy corneas, SL-OCT, specular microscopy, and Orbscan (with correction factor) can be used interchangeably with US pachymetry in certain clinical settings. The four modalities showed significant linear correlations with one another.
    Clinical ophthalmology (Auckland, N.Z.) 01/2015; 9:1065-70. DOI:10.2147/OPTH.S81376
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    ABSTRACT: Background This study determines ‘correlation constants’ between the gold standard histological measurement of retinal thickness and the newer spectral-domain optical coherence tomography (SD-OCT) technology in adult C57BL/6 mice. Methods Forty-eight eyes from adult mice underwent SD-OCT imaging and then were histologically prepared for frozen sectioning with H&E staining. Retinal thickness was measured via 10x light microscopy. SD-OCT images and histological sections were standardized to three anatomical sites relative to the optic nerve head (ONH) location. The ratios between SD-OCT to histological thickness for total retinal thickness (TRT) and six sublayers were defined as ‘correlation constants’. Results Mean (± SE) TRT for SD-OCT and histological sections was 210.95 µm (±1.09) and 219.58 µm (±2.67), respectively. The mean ‘correlation constant’ for TRT between the SD-OCT and histological sections was 0.96. The retinal thickness for all sublayers measured by SD-OCT vs. histology were also similar, the ‘correlation constant’ values ranged from 0.70 to 1.17. All SD-OCT and histological measurements demonstrated highly significant (p<0.01) strong positive correlations. Conclusion This study establishes conversion factors for the translation of ex vivo data into in vivo information; thus enhancing the applicability of SD-OCT in translational research.
    PLoS ONE 10/2014; 9(10):e111203. DOI:10.1371/journal.pone.0111203 · 3.53 Impact Factor
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    ABSTRACT: Ocular ischemic microenvironment plays a critical role in the progression of diabetic retinopathy (DR). In this study, human CD34(+) cells were incubated with vitreous or aqueous of subjects with PDR and evaluated for cell migration with CXCL12. Intracellular levels of nitric oxide (NO) was measured with DAF-FM. Tube formation assay used to evaluate the effect of treated-CD34(+) cells on in vitro angiogenesis. Angiogenic protein array and mass spectrometry (MS) was performed to ascertain factors secreted by healthy non diabetic CD34(+) cells exposed to diabetic vitreous or aqueous. PDR vitreous/aqueous reduced migration of CD34(+) cells (672.45±42.1 AFU / 736.75±101.7 AFU; p<0.01) and attenuated intracellular NO levels (182±1.4 AFU / 184.5±6.3 AFU; p=0.002). Pretreatment with PDR vitreous suppressed tube formation of human retinal endothelial cells (64±1.6 Vs 80±2.5). CD34(+) exposed to PDR vitreous resulted in the increased expression of CXCL4 and serpin F1 whereas CD34(+) exposed to PDR aqueous showed increased expression of CXCL4, serpin F1 and endothelin-1 (ET-1). MS analysis of CD34(+) (exposed to PDR vitreous) expressed J56 gene segment, isoform 2 of SPARC-related modular calcium-binding protein 2, isoform 1 of uncharacterized protein c1 orf167, integrin alpha-M, and 40s ribosomal protein s21. Exposure of healthy non diabetic CD34(+) cells to PDR vitreous and aqueous resulted in decreased migration, reduced generation of NO and altered paracrine secretory function. Our results suggest that the contribution of CD34(+) cells to the aberrant neovascularization observed in PDR is driven more by the pro-angiogenic effects of the retinal cells rather than the influence of the vitreous.
    AJP Endocrinology and Metabolism 08/2014; 307(8). DOI:10.1152/ajpendo.00253.2014 · 4.09 Impact Factor
  • Kakarla V Chalam, Vikram S Brar, Ravi K Murthy
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    ABSTRACT: Importance Retinal ischemia–induced upregulation of vascular endothelial growth factor (VEGF) leads to endothelial proliferation of the anterior segment, resulting in neovascular glaucoma.Objective To investigate the ciliary epithelium as a possible source of VEGF in human eyes enucleated for intractable neovascular glaucoma.Design, Setting, and Participants In this proof-of-concept, laboratory-based study, 16 human enucleated eyes (8 with neovascular glaucoma and 8 as controls) were investigated.Main Outcomes and Measures Presence of VEGF by immunohistochemical analysis (VEGF protein) and in situ hybridization (VEGF messenger RNA).Results In eyes with neovascular glaucoma, strong VEGF immunoreaction in the nonpigmented epithelial cells of the ciliary processes and in the retina was noted. In situ hybridization for VEGF messenger RNA revealed a similar pattern, with positive stain results only in eyes with neovascular glaucoma. A minimal amount of VEGF immunostaining was seen in control eyes.Conclusions and Relevance The nonpigmented ciliary epithelium is an important site of VEGF synthesis in patients with neovascular glaucoma. The ciliary epithelium may represent an additional focus of treatment in the management of neovascular glaucoma, especially in eyes that are nonresponsive to panretinal photocoagulation.
    Jama Ophthalmology 07/2014; 132(11). DOI:10.1001/jamaophthalmol.2014.2356 · 3.83 Impact Factor
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    ABSTRACT: Understanding measurement variability and relationships between measurements obtained on different optical coherence tomography (OCT) machines is critical for clinical trials and clinical settings.
    Jama Ophthalmology 07/2014; 4(1). DOI:10.1001/jamaophthalmol.2014.1698 · 3.83 Impact Factor
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    ABSTRACT: Objective. To prospectively evaluate the effect of intravitreal bevacizumab on aqueous levels of interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF) in patients with exudative age-related macular degeneration (AMD) and correlate clinical outcomes with cytokine levels. Methods. 30 eyes of 30 patients with exudative AMD underwent intravitreal injection of bevacizumab three times at monthly intervals. The aqueous samples prior to the 1st injection (baseline) and 3rd injection were analyzed for VEGF and IL-6 levels. Subjects were subgrouped based upon change in the central subfield (CSF) macular thickness on SD-OCT at 8 weeks. Group 1 included patients (n = 14) with a decrease in CSF thickness greater than 10% from the baseline (improved group). Group 2 included patients (n = 16) who had a decrease in CSF thickness 10% or less (treatment-resistant). Results. In subgroup analysis, in both groups 1 and 2 patients, compared to aqueous VEGF, aqueous IL-6 levels showed a better correlation with CSF thickness on SD-OCT (r = 0.72 and 0.71, resp.). Conclusions. Aqueous IL-6 may be an important marker of treatment response or resistance in wet macular degeneration. Future therapeutic strategies may include targeted treatment against both VEGF and IL-6, in patients who do not respond to anti-VEGF treatment alone.
    Journal of Ophthalmology 07/2014; 2014:502174. DOI:10.1155/2014/502174 · 1.94 Impact Factor
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    ABSTRACT: Abstract Purpose: To assess the cytotoxicity of varying concentrations of Brilliant Blue Green (BBG) on Human Retinal Pigment Epithelial cells (HRPE) exposed to metal halide surgical endoilluminator (SE) at varying distances of illumination. Methods: HRPE (ARPE-19) were exposed to 2 concentrations (0.25 and 0.5 mg/mL) of BBG and illuminated with SE for 1, 5, and 15 min. Illumination (measured with light meter) was positioned at varying distances (1 and 2.5 cm) to mimic surgical distance of illumination. Cell viability (WST-1 assay) as well as structural changes in cells was quantified. Results: At 1 cm distance of illumination, 0.25 mg/mL BBG decreased viability of HRPE to 89.6%±4.3%, 83.9%±10.9%, and 38.9%±5.1% of controls after 1, 5, and 15 min of exposure, respectively. Similarly, 0.5 mg/mL BBG at 1 cm distance of illumination reduced the viability of HRPE to 93.7%±2.8%, 59.6%±16%, and 34.7%±3.5% of controls. At the distance of 2.5 cm, HRPE showed improved viability; cells exposed to 0.25 mg/mL BBG maintained 98.85%±3.3%, 95.31%±7.12%, and 62.07%±3.0% of viability compared with controls after 1, 5, and 15 min of exposure. Morphometric evaluation of RPE cells showed increased width (swelling) of HRPE. Conclusion: BBG at its clinically used concentration (0.25 mg/mL) during vitreoretinal surgery is safe and not toxic to HRPE for up to 5 min under focal illumination (1 cm) and up to 15 min under diffuse illumination (2.5 cm) from the commonly used SE. Translational Relevance: Results of our study are useful in establishing safety parameters for the use of BBG dye in vitreoretinal surgery.
    Journal of Ocular Pharmacology and Therapeutics 07/2014; 30(8). DOI:10.1089/jop.2013.0225 · 1.42 Impact Factor
  • Open Journal of Ophthalmology 01/2014; 04(01):24-30. DOI:10.4236/ojoph.2014.41005
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    Sankarathi Balaiya, Ravi K Murthy, Kakarla V Chalam
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    ABSTRACT: Resveratrol, a polyphenolic phytoalexin present in red wine, has a protective role against tumor-induced angiogenesis. Exudative age-related macular degeneration is characterized by hypoxia-induced choroidal vascular endothelial cell (CVEC) proliferation. In this study, we evaluated the effect of resveratrol on hypoxic CVECs and the underlying signaling pathways involved. CVECs (RF/6A) after induction of hypoxia with cobalt chloride (CoCl2, 200 μM) were exposed to increasing doses of resveratrol (2, 4, 6, 8, 10, and 12 μg/ml). Cell viability was measured with 4-[3-(4Iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1, 3-benzene disulfonate (WST-1) colorimetric assay. The effect of resveratrol on hypoxia-induced vascular endothelial growth factor (VEGF) release was analyzed with enzyme-linked immunosorbent assay. The mechanistic pathway was further evaluated by analyzing phosphorylated stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) using immunoblot and cleaved caspase-3 with In-Cell enzyme-linked immunosorbent assay. Resveratrol inhibited hypoxic CVEC proliferation. Hypoxia-induced VEGF release (30.9±2.6 pg/ml) was inhibited in a dose-dependent fashion by 2, 4, 6, 8, 10, and 12 μg/ml resveratrol to 12.4±2.1, 11.0±1.9, 10.3±3.0, 7.5±1.9, 5.5±2.0, and 5.5±2.3 pg/ml, respectively. SAPK/JNK increased by 1.8-fold and 3.9-fold after treatment with 4 and 12 μg/ml resveratrol, respectively. Significant increase in caspase-3 levels was observed with 12 μg/ml resveratrol. Our study demonstrates that resveratrol suppresses hypoxic CVEC proliferation through activation of the SAPK/JNK pathway. Resveratrol, a nutritional supplement and inhibitor of CVECs, may be a useful adjunct to current anti-VEGF therapy in wet age-related macular degeneration.
    Molecular vision 11/2013; 19:2385-2392. · 2.25 Impact Factor
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    ABSTRACT: IMPORTANCE Sickle cell disease (SCD) is characterized by vaso-occlusive crisis. In the eye, central retinal artery occlusion (CRAO) is a rare complication in SCD, with only 1 previous report of bilateral, concurrent CRAO. We report a case of bilateral, concurrent CRAO in a patient with SCD, possibly precipitated by the use of phosphodiesterase 5 inhibitors. OBSERVATIONS A 37-year-old African American woman with a known medical history significant for SCD and pulmonary arterial hypertension who was receiving treatment with tadalafil, a phosphodiesterase 5 inhibitor, developed bilateral, concurrent CRAO that persisted after exchange transfusion. CONCLUSIONS AND RELEVANCE Bilateral CRAO secondary to SCD is extremely rare, with only 1 previous case report in the literature. The use of phosphodiesterase 5 inhibitors is an additional risk factor and may have contributed to the development of concurrent CRAO in this patient.
    Jama Ophthalmology 09/2013; 131(11). DOI:10.1001/jamaophthalmol.2013.5047 · 3.83 Impact Factor
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    ABSTRACT: Abstract Context: Lutein (LUT) and zeaxanthin (ZEA) are currently under investigation in clinical trials as prophylactic nutritional agents for age-related macular degeneration (AMD). However, dose used in these trials is empirical and not been investigated in in vitro studies. Objective: In this study, we investigated the dose-response effect of LUT and ZEA in protecting retinal pigment epithelium (RPE) from oxidative stress, a common underlying pathology in AMD. Methods: Three thousand cultured human retinal pigment epithelial cells (ARPE-19) were plated in 72-well plate and after 24 h were exposed to increasing concentrations of hydrogen peroxide (H2O2). ARPE-19 cells were exposed to four different concentrations of LUT (0.5, 1, 2 and 4 µg/mL) and ZEA (0.1, 0.2, 0.4 and 0.8 µg/mL). After 24 h incubation, cells were subjected to oxidative stress induced with H2O2. Cultures containing saline solution and dichloromethane served as controls. Cell viability was assessed using the WST-1 assay. Pathophysiological pathways were evaluated by measuring caspase-3 levels as an indicator of apoptosis induction. Reactive oxygen species (ROS) levels were measured using dihydrorhodamine-123. Results: Cell viability as a percentage of control was 81.3%, 81.1%, and 88.8% at 0.5, 1, and 2 µg/ml, respectively of LUT (p < 0.001). The maximum cytoprotective effect was seen with LUT at 2 μg/mL. ZEA did not show any cytoprotective effect at all concentrations used in the study. Caspase-3 showed a corresponding decrease in levels with LUT (1 and 2 µg/ml). Significant decrease in ROS levels were measured only with LUT at 4 µg/ml (p = 0.02). Discussion and conclusions: Results from our study provide in vitro data to support the epidemiologic studies, which are currently underway to provide evidence that lutein may act as cofactor that modulates processes implicated in AMD pathogenesis.
    Cutaneous and Ocular Toxicology 07/2013; 33(2). DOI:10.3109/15569527.2013.812108 · 0.92 Impact Factor
  • Florida Society of Ophthalmology; 06/2013
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    ABSTRACT: To report normative data for retinal thickness in wild-type C57BL/6 mouse utilizing a miniature SD-OCT system. THIRTY ADULT MICE (RANGE: 3-5 months) were anesthetized and secured into the Bioptigen Spectral Domain Ophthalmic Imaging System. Right eye SD-OCT images were standardized by centralizing the optic nerve head (ONH) prior to image acquisition. Global and quadrant total retinal thickness (TRT) values were measured from retinal nerve fiber layer to retinal pigment epithelial layer. Posterior segment analyses also included the outer retinal layer (ORL) and inner retinal layer (IRL). Further sublayer analyses of four layers from the ORL and three layers comprising the IRL were also performed. The overall mean±SD global TRT in a C57BL/6 mouse model was 204.41±5.19 µm. Quadrant mean TRT values were 204.85±5.81 µm inferiorly, 204.97±6.71 µm nasally, 205.08±5.44 µm temporally, and 202.74±4.85 µm superiorly. Mean±SD thickness for ORL, and IRL were 126.37±10.01 µm, and 107.03±10.98 µm respectively. The mean±SD estimates for the four layers of the ORL were 18.23±2.73 µm, 26.04±4.21 µm, 63.8±6.23 µm, and 19.22±4.34 µm. Mean±SD values for the three IRL sublayers were 27.82±4.04 µm, 59.62±6.66 µm and 19.12±3.71 µm. This study established normative values for the total retinal thickness and sublayer thickness for the wild-type C57BL/6 mice. Moreover, it provides a standard of retinal morphology, in a commonly used animal model, for evaluating therapeutic interventions and retinal disease pathophysiology.
    PLoS ONE 06/2013; 8(6):e67265. DOI:10.1371/journal.pone.0067265 · 3.53 Impact Factor
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    ABSTRACT: PURPOSE: To evaluate the differential sensitivity of choroidal endothelial, retinal pigment epithelial, and retinal ganglion cells to escalating doses of proton beam radiation and to establish a safe dose range for the management of choroidal neovascularization associated with age-related macular degeneration (AMD). DESIGN: Laboratory investigation. METHODS: Proliferating simian choroidal endothelial cells (RF/6A), differentiated rat retinal ganglion cells (RGC-5), and serum-starved human retinal pigment epithelial cells (ARPE-19) were exposed to 2, 4, 8, and 12 cobalt gray equivalent of proton beam radiation and cell viability was quantified on day 9. Reactive oxygen species levels were analyzed. RESULTS: Significant decline of choroidal endothelial cell viability was noted as dose escalated from 4 to 8 cobalt gray equivalent with maximum effect observed at 12 cobalt gray equivalent. RGC-5 and ARPE-19 cell count decreased to 95% and 62.7% at 8 cobalt gray equivalent, respectively. Sub-analysis between 4 and 8 cobalt gray equivalent radiation revealed significant decrease in choroidal endothelial cell viability (43.1% at 7 cobalt gray equivalent and 32.3% at 8 cobalt gray equivalent of radiation). Correspondingly, RGC-5 and ARPE-19 cells did not show decrease in cell count or viability. Reactive oxygen species levels significantly increased in radiation-treated choroidal endothelial cells (8.3%-11.9%). CONCLUSIONS: At 6-8 cobalt gray equivalent proton beam radiation, retinal ganglion and retinal pigment epithelial cells are preserved while choroidal endothelial cells are completely inhibited. This dosage offers optimum therapeutic safety window for treatment using proton beam radiation for exudative AMD.
    American Journal of Ophthalmology 06/2013; 156(3). DOI:10.1016/j.ajo.2013.04.036 · 4.02 Impact Factor
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    ABSTRACT: The lens and retina of the human eye are exposed constantly to light and oxygen. In situ phototransduction and oxidative phosphorylation within photoreceptors produces a high level of phototoxic and oxidative related stress. Within the eye, the carotenoids lutein and zeaxanthin are present in high concentrations in contrast to other human tissues. We discuss the role of lutein and zeaxanthin in ameliorating light and oxygen damage, and preventing age-related cellular and tissue deterioration in the eye. Epidemiologic research shows an inverse association between levels of lutein and zeaxanthin in eye tissues and age related degenerative diseases such as macular degeneration (AMD) and cataracts. We examine the role of these carotenoids as blockers of blue-light damage and quenchers of oxygen free radicals. This article provides a review of possible mechanisms of lutein action at a cellular and molecular level. Our review offers insight into current clinical trials and experimental animal studies involving lutein, and possible role of nutritional intervention in common ocular diseases that cause blindness.
    Nutrients 05/2013; 5(5):1823-1839. DOI:10.3390/nu5051823 · 3.15 Impact Factor

Publication Stats

1k Citations
280.65 Total Impact Points

Institutions

  • 2002–2015
    • University of Florida
      • Department of Ophthalmology
      Gainesville, Florida, United States
  • 2007–2010
    • Jacksonville University
      Jacksonville, Florida, United States
  • 2004–2010
    • Florida State College at Jacksonville
      Jacksonville, Florida, United States
  • 2008
    • University of Florida Health Science Center-Jacksonville
      Jacksonville, Florida, United States
    • University of Missouri - Kansas City
      Kansas City, Missouri, United States
  • 1996–2004
    • University of South Carolina
      • Department of Ophthalmology
      Columbia, South Carolina, United States