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1.
PLoS One ; 16(2): e0247161, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33596257

RESUMO

Regularly scheduled intravitreal anti-vascular endothelial growth factor (anti-VEGF) injections are essential to maintaining and/or improving many ocular conditions including: neovascular age-related macular degeneration (nAMD), diabetic retinopathy, and retinal vein occlusions with macular edema (RVO). This study aims to assess the effect of unintended delays in anti-VEGF treatment during the first wave of the COVID-19 pandemic. This retrospective case series identified patients receiving regularly scheduled anti-VEGF intravitreal injections based on current procedural terminology (CPT) code at two practices in Minnesota. Diagnoses were limited to nAMD, diabetic macular edema (DME), proliferative diabetic retinopathy, and RVO. Patients were divided into two groups based on whether they maintained or delayed their follow-up visit by more than two weeks beyond the recommended treatment interval during the COVID-19 lockdown. The 'COVID-19 lockdown' was defined as the period after March, 28th, 2020, when a lockdown was declared in Minnesota. We then compared the visual acuity and structural changes to the retina using ocular coherence tomography (OCT) to assess whether delayed treatment resulted in worse visual outcomes. A total of 167 eyes from 117 patients met criteria for inclusion in this study. In the delayed group, the average BCVA at the pre- and post-lockdown visits were 0.614 and 0.715 (logMAR) respectively (p = 0.007). Central subfield thickness (CST) increased from 341 to 447 in the DME delayed group (p = 0.03) while the CST increased from 301 to 314 (p = 0.4) in the nAMD delayed group. The results of this pilot study suggests that treatment delays may have a negative impact on the visual and anatomic outcomes of patients with nAMD and DME. Future studies with larger sample sizes are required for further investigation.


Assuntos
COVID-19/epidemiologia , Doenças Retinianas/tratamento farmacológico , Tempo para o Tratamento/estatística & dados numéricos , Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Adulto , Idoso , Idoso de 80 Anos ou mais , Inibidores da Angiogênese/uso terapêutico , COVID-19/psicologia , Retinopatia Diabética/tratamento farmacológico , Feminino , Humanos , Edema Macular/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Minnesota/epidemiologia , Pandemias/estatística & dados numéricos , Projetos Piloto , Quarentena/métodos , Quarentena/psicologia , Oclusão da Veia Retiniana/tratamento farmacológico , Estudos Retrospectivos , SARS-CoV-2/isolamento & purificação , Acuidade Visual/efeitos dos fármacos
2.
Am J Ophthalmol Case Rep ; 18: 100666, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32258825

RESUMO

PURPOSE: The primary goal of this study was to identify characteristic features of peripheral degenerative retinoschisis (RS), schisis detachment (SD) and retinal detachment (RD) on both fundus autofluorescence (FAF) and infrared (IR) imaging, using spectral domain optical coherence tomography (SD-OCT) imaging of the peripheral retina as the confirmatory imaging tool. METHODS: This is a descriptive case series study. A total of 27 eyes of 22 patients were included. Thirteen eyes of 10 patients diagnosed with RS, 4 eyes of 3 patients diagnosed with SD, and 10 eyes of 9 patients diagnosed with RD were included. Patients with images of poor quality were excluded. Heidelberg Spectralis HRA + OCT machine (Heidelberg Engineering, Heidelberg, Germany) were used to acquire the images. RESULTS: All conditions appeared as areas of hypo-AF on FAF and hypo-reflectance on IR imaging. Accentuated vasculature of the lesion was noted with IR imaging due to elevation of the RS and RD, which was less frequently observed with FAF. On FAF, a hyper-AF leading edge around the RS lesion indicated the presence of intraretinal or subretinal fluid and an extension of the RS. Retinal breaks/holes were best visualized with IR imaging. SD-OCT confirmed the diagnosis in all performed cases. CONCLUSIONS: We were unable to differentiate between RS and RD based solely on findings from FAF and IR imaging. However, the combination of them with SD-OCT can assist in the diagnosis of RS from RD and in the evaluation of RS progression. OCT remains the main modality imaging to differentiate these conditions.

3.
Exp Eye Res ; 184: 48-55, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30991052

RESUMO

Increased fundus autofluorescence is directly related to increased RPE lipofuscin deposition in the retina and has been observed in eyes with age-related macular degeneration (AMD). Smoking is the most significant modifiable risk factor for the development and progression of AMD, in which one of the main mechanisms is oxidative damage from smoking leading to RPE cell toxicity. The relationship between smoking and autofluorescence is not established and could provide insight into pathogenic mechanism of AMD. Therefore, our objective was to compare quantitative fundus autofluorescence (qAF) in the retinae of healthy non-smokers to smokers. We conducted a cross-sectional study at the 2016 Minnesota State Fair. Participants self-reported past medical and ocular history and underwent eye examination as well as qAF imaging with Spectralis confocal scanning laser ophthalmoscope (cSLO) equipped with an internal fluorescent reference. Two sets of images were obtained per eye. Stepwise multiple mixed effects regression model was used to examine the relationship between mean qAF values and smoking status. We enrolled 105 individuals (54 smokers, 61 females, mean age 41 years with range 18-78 years old). Fundus autofluorescence images were analyzable for 85 of 105 individuals contributing 161 eyes (80 right, 81 left). The repeatability coefficients between the first set and second set of images were ±21% of their mean qAF values. Older age and female gender were independently associated with higher qAF. Positive smoking history tended to result in higher qAF values after adjusting for age and gender but was not statistically significant (0.118, 95%CI -0.003, 0.240, P = 0.056). Among smokers, the number of pack-years smoked was not significantly associated with higher qAF. Our study's results are consistent with existing literature in which older age is predictive of intensified autofluorescence, while smoking history does not have as important of an impact on autofluorescence as hypothesized. Several large epidemiological studies have shown that smoking is significantly associated with AMD, and qAF is likely not the appropriate modality to clinically assess smoking's impact on retinae.


Assuntos
Lipofuscina/metabolismo , Degeneração Macular/metabolismo , não Fumantes , Imagem Óptica , Epitélio Pigmentado da Retina/metabolismo , Fumantes , Adolescente , Adulto , Idoso , Estudos Transversais , Feminino , Angiofluoresceinografia/métodos , Fundo de Olho , Humanos , Masculino , Pessoa de Meia-Idade , Oftalmoscopia/métodos , Adulto Jovem
4.
Trans Am Ophthalmol Soc ; 115: T3, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29021717

RESUMO

PURPOSE: Understanding the apparent paradoxical role of zinc in the pathogenesis and prevention of age-related macular degeneration (AMD) has been limited by the lack of animal models for its detection in sub-retinal epithelial deposits (drusen), a definitive early hallmark of AMD. In-vitro studies using Zinpyr-1 showed drusen contained high levels of zinc, but the probe was not suitable for in-vivo studies. This study compares Zinpyr-1 to ZPP1, a new fluorescein-based probe for zinc, to assess the potential of ZPP1 for in-vivo detection of zinc in drusen. METHODS: Flat mounts of human sub-RPE tissue using the probes were analyzed by fluorescence and confocal microscopy. Flat mounts of sub-RPE tissue from mice deficient in superoxide dismutase isoform-1 (CuZn-SOD-KO) or isoform-2 (Mn-SOD-RPE-KO) were analyzed with sub-RPE deposits confirmed by histology. RESULTS: Drusen are detected in greater numbers and intensity with ZPP1 compared to Zinpyr-1. Using ZPP1, drusen was detected in a sample from a 46-year old human donor without ocular history, suggesting that ZPP1 might be sensitive enough to detect drusen at an early stage. With CuZn-SOD KO mice, ZPP1 detected sub-RPE deposits at 10 months of age, whereas Zinpyr-1 required 14 months. CONCLUSION: Detection of sub-RPE deposits by ZPP1 was greatly enhanced compared to Zinpyr-1. This enhanced sensitivity will allow for more insightful analysis of zinc in AMD using human specimens and mouse models. This could result in the development of a sensitive in-vivo probe to enhance research on the role zinc in drusen formation and the early clinical diagnosis of AMD.


Assuntos
Epitélio Pigmentado da Retina/diagnóstico por imagem , Degeneração Macular Exsudativa/diagnóstico , Zinco/metabolismo , Animais , Biomarcadores/metabolismo , Modelos Animais de Doenças , Humanos , Camundongos , Camundongos Knockout , Microscopia Confocal , Oftalmologia , Epitélio Pigmentado da Retina/metabolismo , Sociedades Médicas , Estados Unidos , Degeneração Macular Exsudativa/metabolismo
5.
Ophthalmol Ther ; 5(2): 215-222, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27601159

RESUMO

INTRODUCTION: To assess the anatomical changes taking place in the choroid after a scleral buckle (SB) procedure for retinal detachment repair. METHODS: This cross-sectional study looked at 23 adults with a history of unilateral retinal detachment repaired with a SB or other encircling element. The subjects underwent bilateral Enhanced Depth Spectral Domain Optical Coherence Tomography to image the choroid. The choroidal thickness (CT) was measured, and the non-operative eye was used as an internal control. RESULTS: CT was measured to be 170.8 ± 60.9 µm (mean ± SD) in eyes with SBs compared to 175.1 ± 61.9 µm in non-operative eyes. There was no statistically significant difference between the two groups (mean 4.3 µm, 95% CI -8.7, 17.3, p value 0.4973, paired t test). CONCLUSION: Placement of an SB as part of a surgery to repair retinal detachment did not significantly alter CT at the macula.

6.
Ophthalmol Ther ; 5(2): 183-190, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27484365

RESUMO

INTRODUCTION: Many ocular diseases require intravitreal injections of pharmacological agents. Optimizing patients' experiences during injections is important to ensure compliance and maintenance of quality of life. The objective of this study was to identify strategies to help alleviate discomfort during intravitreal injections. METHODS: A cross-sectional study surveying 128 patients during clinic visits between 2014 and 2015 in two outpatient Retina Clinics (one academic and one private). Patients receiving an intravitreal injection(s) for any retinal disorder were given a questionnaire with 10-yes/no responses for various potential strategies. Responses were stratified by sex, age (<30 years, 30-60 years, and >60 years) and total number of prior injections (0-9 injections, 10-20 injections and >20 injections). RESULTS: A total of 128 patients were surveyed: 59 males, 41 females and 28 with no sex specified. Our results identified four favorable strategies as those receiving more than 50% "yes" votes. These included the presence of technician/staff during the procedure, the use of a neck pillow, a verbal warning before the injection and performing injections in both eyes on the same day. Other specific strategies were identified for females, younger patients and those with greatest experience. These included: females preferred having their hand held during injections (P = 0.001) and using a stress ball (P = 0.000) when compared to males. Stratifying by age, patients 30-60 years old preferred having their hand held (P = 0.008) and background music (P = 0.007). Stratifying by prior injections, patients with >20 prior injections preferred having their hand held (P = 0.001), using a stress ball (P = 0.021) and, if necessary, having bilateral injections performed the same day to improve comfort (P = 0.037). CONCLUSIONS: Having an extra staff member present during the injection, having a neck pillow, having a verbal warning prior to injection and having both eyes injected on the same day were indicated as favorable strategies by over half of those surveyed. Further, specific strategies were identified for females, younger patients (30-60 years old) and those with greatest experience (>20 injections).

7.
Ophthalmic Surg Lasers Imaging Retina ; 47(5): 471-3, 2016 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-27183552

RESUMO

Vision loss in giant cell arteritis (GCA) often presents as anterior ischemic optic neuropathy and central retinal artery occlusion. Previous studies have established an acute delay in choroidal perfusion on fluorescein angiography (FA) as a classic sign of GCA. The authors present a unique imaging case report of GCA where ultra wide-field (UWF) indocyanine green angiography (ICGA) offers improved characterization of delayed choroidal perfusion compared to FA. Routine use of ICGA, particularly UWF imaging, in patients with suspected GCA should be studied in a larger cohort to determine whether it may improve detection of choroidal perfusion delay. [Ophthalmic Surg Lasers Imaging Retina. 2016;47:471-473.].


Assuntos
Corioide/irrigação sanguínea , Angiofluoresceinografia/métodos , Arterite de Células Gigantes/complicações , Verde de Indocianina/farmacologia , Neuropatia Óptica Isquêmica/diagnóstico , Vasos Retinianos/patologia , Idoso de 80 Anos ou mais , Feminino , Fundo de Olho , Arterite de Células Gigantes/diagnóstico , Humanos , Neuropatia Óptica Isquêmica/etiologia
8.
Ophthalmic Surg Lasers Imaging Retina ; 47(4): 369-71, 2016 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-27065379

RESUMO

The authors describe the implantation of the Argus II Retinal Prosthesis System (Argus II) (Second Sight Medical Products, Sylmar, CA) into a short axial length (AL) eye. The authors' main modification is the use of endocyclophotocoagulation (endo-CPC) to the ciliary processes in the area that the cable enters through the sclerotomy. This case describes the surgical technique necessary for successful implantation of the Argus II into a short AL eye. The use of endo-PC prevents chafing to the ciliary processes, does not affect postoperative intraocular pressure, and facilitates direct visualization of the structures during the surgery, preventing damage during implantation.


Assuntos
Comprimento Axial do Olho/cirurgia , Implantação de Prótese/métodos , Retinose Pigmentar/cirurgia , Transtornos da Visão/reabilitação , Próteses Visuais , Adulto , Comprimento Axial do Olho/patologia , Extração de Catarata , Humanos , Implante de Lente Intraocular , Masculino
9.
Artigo em Inglês | MEDLINE | ID: mdl-23676237

RESUMO

The integrity of macular morphology was examined in a patient with multiple evanescent white dot syndrome (MEWDS) diagnosed through clinical and investigational adaptive optics (AO) retinal imaging techniques. Imaging was performed during the acute and recovery phases to examine changes in retinal morphology, revealing characteristic small multifocal white dots in the perifoveal region and a granular appearance in the fovea. Fluorescein angiography revealed early and intermediate hyperfluorescence, and regions of decreased fundus autofluorescence were observed. Photoreceptor disruption was apparent during the acute phase and recurrence. Conventional multimodal imaging combined with AO imaging offers more insight into the pathology of MEWDS by providing complementary views of the retina throughout the acute phase, recovery, and recurrence.


Assuntos
Angiofluoresceinografia , Imagem Multimodal/métodos , Oftalmoscopia/métodos , Doenças Retinianas/diagnóstico , Adulto , Feminino , Humanos , Lasers
10.
Biofactors ; 37(2): 104-16, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21488133

RESUMO

Carotenoids are known to generate various aldehydes, known as carotenoid-derived aldehydes (CDAs), which could efficiently react with protein or DNA. In this in vitro model study, interaction between CDA and protein has been studied. Various proteins were incubated with CDA, and protein modification and adduct formation were confirmed by using matrix-assisted laser desorption and ionization time-of-flight, amino acid analysis, and measuring enzyme activity on modification with CDA. Using radiolabeled NaB((3) H)H(4) and Raney nickel as well as sulfhydryl assay (Ellman's reagent), we confirmed that CDA could conjugate with cysteine through a thioether linkage. The carbonyl assay using 2,4-dinitrophenylhydrazine revealed the possible involvement of Schiff's base reaction between CDA and lysine. The adducts formed between ß-apo-8-carotenal (BA8C) and N-acetylcysteine and BA8C and N-acetyllysine were confirmed by HPLC and ESI-MS. Our results suggest that CDA could alter protein function by post-translational interaction with cysteine and lysine by thioether linkage and by schiff's based bonds, respectively. Thus, the formation of CDA adducts with proteins could alter functional properties of proteins responsible for maintaining cell homeostasis and thereby cause cellular toxicity. In view of these observations, further studies are required to understand the delicate balance between beneficial and/or harmful effects of carotenoids as a dietary supplement to slow age-related macular degeneration progression.


Assuntos
Carotenoides/química , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Proteínas/química , Proteínas/metabolismo , Aldeídos/química , Aldeídos/farmacologia , Cromatografia Líquida de Alta Pressão , Ligação Proteica , Carbonilação Proteica , Bases de Schiff/química , Espectrometria de Massas por Ionização por Electrospray
11.
Toxicol Lett ; 198(1): 56-62, 2010 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-20435108

RESUMO

Cadmium is reported to accumulate in human eye tissues suggesting its implication in diverse ocular pathology. Using an in vitro cell culture model we investigated the effects of cadmium on human lens epithelial cells (HLECs) (HLE-B3). We observed cadmium-induced dose- as well as time-dependent decline in HLECs viability which was exacerbated significantly upon reduction of intracellular glutathione levels by buthionine sulfoximine (BSO). There was a dose-dependent significant increase in lactate dehydrogenase (LDH) release from HLECs suggesting cadmium-induced alteration of membrane integrity as well as necrotic cell death. The decline in cell viability was also due to apoptosis of the HLECs as determined by quantifying % apoptotic cells as well as PARP cleavage. Moreover, release of apoptosis inducing factor (AIF) into the cytosol was also detected. Cadmium was also observed to increase oxidative stress, lipid peroxidation and activation of MAPK pathway in HLECs. Antioxidants like N-acetylcysteine (NAC) and alpha-Tocopherol significantly prevented cadmium-induced toxicity in HLECs. Our findings suggest that cadmium-induced elevated oxidative stress as well as activation of MAPK signaling cascade eventually led to cell death of HLECs through apoptosis as well as necrosis. The loss of HLECs by cadmium could possibly explain its implication in cataract development particularly associated with smoking.


Assuntos
Apoptose/efeitos dos fármacos , Cádmio/toxicidade , Células Epiteliais/efeitos dos fármacos , Cristalino/efeitos dos fármacos , Catarata/induzido quimicamente , Catarata/etiologia , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Células Epiteliais/metabolismo , Glutationa/metabolismo , Humanos , Cristalino/citologia , Cristalino/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Estresse Oxidativo , Transdução de Sinais/efeitos dos fármacos , Fumar/efeitos adversos
12.
Invest Ophthalmol Vis Sci ; 51(10): 5105-13, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20435582

RESUMO

PURPOSE: To investigate the anti-inflammatory effects of guggulsterone, an antioxidant and antitumor agent, in endotoxin-induced uveitis (EIU) in rats and to elucidate the underlying molecular mechanism or mechanisms related to ocular inflammation. METHODS: EIU was induced by subcutaneous injection of lipopolysaccharide (LPS; 150 µg) into Lewis rats treated with guggulsterone (30 mg/kg body weight, intraperitoneally) or its carrier. After 24 hours the rats were killed, eyes were enucleated, and aqueous humor (AqH) was collected. Numbers of infiltrating cells and levels of matrix metalloproteinase-2 (MMP-2), nitric oxide (NO), and prostaglandin E(2) (PGE(2)) were determined in AqH by specific ELISAs. An antibody array was used to measure the expression of various inflammatory cytokines in AqH. The expression of MMP-2, iNOS, Cox-2, phospho-IκB, and phospho-NF-κB was determined immunohistochemically. Human primary nonpigment ciliary epithelial cells (HNPECs) were used to determine the in vitro efficacy of guggulsterone on the LPS-induced inflammatory response. RESULTS: Compared with control, the EIU rat eye AqH had a significantly higher number of infiltrating cells, total protein, and inflammatory markers, such as MMP-2, NO, and PGE(2), and the treatment of guggulsterone prevented EIU-induced increases. Guggulsterone also prevented the expression of MMP-2, iNOS, and Cox-2 proteins and of IκB and NF-κB in various eye tissues. Moreover, in cultured HNPECs, guggulsterone inhibited LPS-induced expression of inflammatory proteins. CONCLUSIONS: These results for the first time demonstrate that the plant sterol guggulsterone suppresses ocular inflammation in EIU, suggesting that the supplementation of guggulsterone could be a novel approach for the treatment of ocular inflammation.


Assuntos
Antineoplásicos/uso terapêutico , Antioxidantes/uso terapêutico , Commiphora , Modelos Animais de Doenças , Lipopolissacarídeos , Pregnenodionas/uso terapêutico , Uveíte/prevenção & controle , Animais , Humor Aquoso/citologia , Humor Aquoso/metabolismo , Western Blotting , Células Cultivadas , Corpo Ciliar/efeitos dos fármacos , Citocinas/metabolismo , Dinoprostona/metabolismo , Ensaio de Imunoadsorção Enzimática , Escherichia coli , Humanos , Proteínas I-kappa B/metabolismo , Injeções Intraperitoneais , Masculino , Metaloproteinase 2 da Matriz/metabolismo , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Ratos , Ratos Endogâmicos Lew , Uveíte/induzido quimicamente , Uveíte/metabolismo
13.
Curr Eye Res ; 34(9): 737-47, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19839867

RESUMO

PURPOSE: To investigate the genotoxic effects of lutein (LBP) and beta -carotene breakdown products (beta -apo-8-carotenal, BA8C) and the preventive role of GSH in human retinal pigment epithelial cells (ARPE-19). METHODS: LBP- and BA8C-induced DNA damage in human retinal pigment epithelial cells (ARPE-19) was determined by comet assay. The DNA damage was quantified by the image analysis system using Comet Score software. ARPE-19 cell viability was determined by CellTiter 96 AQ(ueous) one-solution cell proliferation assay kit. Intracellular GSH levels were measured by Ellman's reagent. RESULTS: Incubation of serum-starved ARPE-19 cells with LBP and BA8C caused significant DNA damage in a dose- and time-dependent manner. The DNA damage and cell death incurred by LBP and BA8C were significantly prevented by N-acetylcysteine (NAC) but not by alpha -tocopherol + ascorbic acid (T + AA). Furthermore, BSO-induced GSH depletion in ARPE-19 cells caused a significant elevation in LBP- and BA8C-induced DNA damage, whereas increased GSH levels in ARPE-19 cells prevented it. CONCLUSIONS: Our results suggest that breakdown products of dietary carotenoids could be genotoxic in ARPE-19 cells. LBP-induced genotoxic effects could worsen oxidative stress. The intracellular GSH pool in ARPE-19 cells might play a critical role in carotenoid breakdown products-induced genotoxicity.


Assuntos
Carotenoides/toxicidade , Dano ao DNA , DNA/efeitos dos fármacos , Luteína/toxicidade , Epitélio Pigmentado da Retina/efeitos dos fármacos , Acetilcisteína/farmacologia , Apoptose/efeitos dos fármacos , Ácido Ascórbico/farmacologia , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Ensaio Cometa , Relação Dose-Resposta a Droga , Glutationa/metabolismo , Glutationa/farmacologia , Humanos , Microscopia de Fluorescência , Fatores de Tempo , alfa-Tocoferol/farmacologia
14.
Exp Eye Res ; 89(4): 494-502, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19524565

RESUMO

Cadmium (Cd), released from cigarette smoke and metal industrial activities, is known to accumulate in human body organs including retina and is particularly higher in retinal tissues of age-related macular degeneration (AMD) eyes compared to non-AMD eyes. We have determined the cytotoxic effects of Cd on human retinal pigment epithelial (RPE) cells. Upon Cd treatment, there was a dose- and time-dependent decline in ARPE-19 cell viability as well as early apoptotic changes such as altered mitochondrial membrane potential (MMP) and Cytochrome C release in cytosol. Depletion of GSH by buthionine-[S,R]-sulfoximine (BSO) resulted in increased Cd toxicity in ARPE-19 cells. Cadmium also caused reactive oxygen species (ROS) generation and activation of mitogen-activated protein kinases (MAPKs) pathway including c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase 1/2 (Erk1/2), and p38 in ARPE-19 cells. Antioxidants such as N-acetylcysteine (NAC) significantly reduced Cd-induced toxicity. These results indicate that elevated ROS-induced activation of the MAPK signaling pathway could be associated with Cd-induced RPE cell apoptosis, one of the major contributing factors in AMD. The toxic effects of Cd on ARPE-19 cells indicate that environmental heavy metals such as Cd could be important potential factors in RPE cells death associated retinal diseases particularly related to smoking.


Assuntos
Apoptose/efeitos dos fármacos , Cloreto de Cádmio/toxicidade , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Epitélio Pigmentado da Retina/patologia , Acetilcisteína/farmacologia , Western Blotting , Butionina Sulfoximina/farmacologia , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Citocromos c/metabolismo , Citosol/enzimologia , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Citometria de Fluxo , Glutationa/metabolismo , Humanos , Peróxido de Hidrogênio/toxicidade , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Epitélio Pigmentado da Retina/metabolismo , Fatores de Tempo
15.
Invest Ophthalmol Vis Sci ; 50(2): 752-9, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19011011

RESUMO

PURPOSE: The purpose of this study was to evaluate the effect of aldose reductase (AR) inhibition on posterior capsular opacification (PCO) with the use of a pig eye capsular bag model. METHODS: Pig eye capsular bags were prepared by capsulorhexis and cultured in medium without or with AR inhibitors for 7 days. Immunostaining was performed in paraformaldehyde-fixed capsular bags to determine the expression of proliferating cell nuclear antigen (PCNA), alpha-smooth muscle actin (SMA), beta-crystallin, and intercellular adhesion molecule (ICAM)-1. The effect of AR inhibition on basic fibroblast growth factor (BFGF)-induced mitogenic signaling in cultured human lens epithelial cells (HLECs) was examined. Cell growth was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and cell counting, the expression of alpha-SMA, beta-crystallin, and ICAM-1 by Western blot and immunocytochemical analysis, protein kinases by Western blot analysis, and NF-kappaB activation by gel shift and reporter assays. RESULTS: During culture of pig eye capsular bags, residual cells on both the anterior and the posterior capsule showed vigorous growth. Treatment with AR inhibitors significantly prevented the lens epithelial cell growth in capsular bags and expression of alpha-SMA, beta-crystallin, and ICAM-1. HLECs showed a dose-dependent response to BFGF, proliferation at lower concentrations (<20 ng/mL) and differentiation/transdifferentiation at higher concentrations (>50 ng/mL). Inhibition of AR also prevented the BFGF-induced activation of ERK1/2, JNK, and NF-kappaB in HLECs. CONCLUSIONS: Results suggest that AR is required for lens epithelial cell growth and differentiation/transdifferentiation in the capsular bags, indicating that inhibition of AR could be a potential therapeutic target in the prevention of PCO.


Assuntos
Aldeído Redutase/antagonistas & inibidores , Catarata/prevenção & controle , Inibidores Enzimáticos/farmacologia , Células Epiteliais/patologia , Cápsula do Cristalino/patologia , Actinas/metabolismo , Animais , Western Blotting , Capsulorrexe , Catarata/metabolismo , Catarata/patologia , Contagem de Células , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Ensaio de Desvio de Mobilidade Eletroforética , Células Epiteliais/metabolismo , Fator 2 de Crescimento de Fibroblastos/farmacologia , Imidazolidinas/farmacologia , Técnicas Imunoenzimáticas , Molécula 1 de Adesão Intercelular/metabolismo , Cápsula do Cristalino/metabolismo , Microscopia de Fluorescência , NF-kappa B/metabolismo , Técnicas de Cultura de Órgãos , Antígeno Nuclear de Célula em Proliferação/metabolismo , Transdução de Sinais/fisiologia , Suínos , beta-Cristalinas/metabolismo
17.
Exp Eye Res ; 86(1): 70-80, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17977529

RESUMO

Carotenoids have been advocated as potential therapeutic agents in treating age-related macular degeneration (AMD). In ocular tissues carotenoids may undergo oxidation and form carotenoid-derived aldehydes (CDA), which would be toxic to tissues. We have investigated the cytotoxic effects of CDA from beta-carotene, Lutein and Zeaxanthin on human retinal pigment epithelial cells (ARPE-19). The serum-starved ARPE-19 cells were treated with CDA without or with antioxidant, N-acetylcysteine (NAC) and cell viability, apoptosis, reactive oxygen species (ROS) levels, nuclear chromatin condensation as well as fragmentation, change in mitochondrial membrane potential (MMP) and activation of transcription factors NF-kappaB and AP-1 were determined. We observed a dose and time-dependent decline in cell viability upon incubation of ARPE-19 cells with CDA. The CDA treatment also led to elevation in ROS levels in a dose-dependent manner. Upon CDA treatment a significant number of apoptotic cells were observed. Also early apoptotic changes in ARPE-19 cells induced by CDA were associated with change in MMP. Increased nuclear chromatin condensation and fragmentation were also observed in cells treated with CDA. The cytotoxicity of CDA in ARPE-19 cells was significantly ameliorated by the antioxidant, NAC. Furthermore, CDA induced the activation of NF-kappaB and AP-1 which was significantly inhibited by NAC. Thus our results demonstrate that CDA could increase the oxidative stress in ARPE-19 cells by elevating ROS levels that would cause imbalance in cellular redox status, which could lead to cell death. This would suggest that high carotenoid supplementation for treatment of AMD should be used cautiously.


Assuntos
Aldeídos/farmacologia , Apoptose/efeitos dos fármacos , Carotenoides/química , Estresse Oxidativo/efeitos dos fármacos , Epitélio Pigmentado Ocular/efeitos dos fármacos , Retina/efeitos dos fármacos , Acetilcisteína/farmacologia , Aldeídos/antagonistas & inibidores , Células Cultivadas , Relação Dose-Resposta a Droga , Sequestradores de Radicais Livres/farmacologia , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Potencial da Membrana Mitocondrial/fisiologia , NF-kappa B/metabolismo , Estresse Oxidativo/fisiologia , Epitélio Pigmentado Ocular/citologia , Espécies Reativas de Oxigênio/metabolismo , Retina/citologia , Fator de Transcrição AP-1/metabolismo
18.
Exp Eye Res ; 84(4): 772-80, 2007 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17313944

RESUMO

One of the hallmarks of age-related macular degeneration (AMD), the leading cause of blindness in the elderly in Western societies, is the accumulation of sub-retinal pigment epithelial deposits (sub-RPE deposits), including drusen and basal laminar deposits, in Bruch's membrane (BM). The nature and the underlying mechanisms of this deposit formation are not fully understood. Because we know that zinc contributes to deposit formation in neurodegenerative diseases, we tested the hypothesis that zinc might be involved in deposit formation in AMD. Using zinc specific fluorescent probes and microprobe synchrotron X-ray fluorescence we showed that sub-RPE deposits in post-mortem human tissues contain unexpectedly high concentrations of zinc, including abundant bio-available (ionic and/or loosely protein bound) ions. Zinc accumulation was especially high in the maculae of eyes with AMD. Internal deposit structures are especially enriched in bio-available zinc. Based on the evidence provided here we suggest that zinc plays a role in sub-RPE deposit formation in the aging human eye and possibly also in the development and/or progression of AMD.


Assuntos
Degeneração Macular/metabolismo , Zinco/análise , Idoso , Idoso de 80 Anos ou mais , Disponibilidade Biológica , Lâmina Basilar da Corioide/metabolismo , Humanos , Macula Lutea/metabolismo , Pessoa de Meia-Idade , Epitélio Pigmentado Ocular/metabolismo , Drusas Retinianas/metabolismo , Espectrometria por Raios X/métodos , Síncrotrons
19.
Arch Biochem Biophys ; 458(2): 128-35, 2007 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-17084803

RESUMO

The xanthophylls lutein (L) and zeaxanthin (Z) form the macular pigment with the highest density in the macula lutea. We investigated Macular Pigment Optical Density (MPOD) responses to supplementation with identically formulated (Actilease) L or Z (OPTISHARP) or L+Z over 6-12 months using doses of 10 or 20mg/day. MPOD as well as blue light sensitivity in fovea and parafovea were measured monthly by heterochromatic flicker photometry. Average xanthophyll plasma concentrations, analysed monthly by HPLC, increased up to 27-fold. MPOD increased by 15% upon L or L+Z supplementation. Supplementation of Z alone produced similar pigment accumulation in fovea and parafovea, which confounded MPOD measurements. After correction for this, a 14% MPOD increase resulted for Z. Thus, during supplementation with xanthophylls, L is predominantly deposited in the fovea while Z deposition appears to cover a wider retinal area. This may be relevant to health and disease of the retina.


Assuntos
Luteína/administração & dosagem , Luteína/metabolismo , Retina/metabolismo , Xantofilas/administração & dosagem , Xantofilas/metabolismo , Adolescente , Adulto , Suplementos Nutricionais , Método Duplo-Cego , Humanos , Luteína/sangue , Macula Lutea/química , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Xantofilas/sangue , Zeaxantinas
20.
Perception ; 34(8): 1029-34, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-16178159

RESUMO

The optical density of macular pigment was measured at twelve retinal locations in ten subjects by minimum motion photometry, comparing 460 nm with 580 nm or 550 nm. Fundus autofluorescence images were obtained for the same subjects with a scanning laser ophthalmoscope. Optical density was computed from mean calibrated grey-scale values for a central circular field and for annular segments, identical to areas tested psychophysically, and for complete annuli. Psychophysical assessments of optical density were similar irrespective of whether 550 nm or 580 nm was used. Optical density values derived psychophysically showed a linear correlation with assessments based on identical sampled areas of annular segments (slope = 0.98, r2 = 0.97) or complete annuli (slope = 0.89, r2 = 0.96) in autofluorescence images.


Assuntos
Angiofluoresceinografia , Percepção de Movimento/fisiologia , Pigmentos da Retina/fisiologia , Humanos , Degeneração Macular , Fotometria , Psicofísica , Retina/fisiologia
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