RESUMO
We have previously reported low concentrations of plasma ascorbate and low dietary vitamin C intake in the older Indian population and a strong inverse association of these with cataract. Little is known about ascorbate levels in aqueous humor and lens in populations habitually depleted of ascorbate and no studies in any setting have investigated whether genetic polymorphisms influence ascorbate levels in ocular tissues. Our objectives were to investigate relationships between ascorbate concentrations in plasma, aqueous humor and lens and whether these relationships are influenced by Single Nucleotide Polymorphisms (SNPs) in sodium-dependent vitamin C transporter genes (SLC23A1 and SLC23A2). We enrolled sixty patients (equal numbers of men and women, mean age 63 years) undergoing small incision cataract surgery in southern India. We measured ascorbate concentrations in plasma, aqueous humor and lens nucleus using high performance liquid chromatography. SLC23A1 SNPs (rs4257763, rs6596473) and SLC23A2 SNPs (rs1279683 and rs12479919) were genotyped using a TaqMan assay. Patients were interviewed for lifestyle factors which might influence ascorbate. Plasma vitamin C was normalized by a log10 transformation. Statistical analysis used linear regression with the slope of the within-subject associations estimated using beta (ß) coefficients. The ascorbate concentrations (µmol/L) were: plasma ascorbate, median and inter-quartile range (IQR), 15.2 (7.8, 34.5), mean (SD) of aqueous humor ascorbate, 1074 (545) and lens nucleus ascorbate, 0.42 (0.16) (µmol/g lens nucleus wet weight). Minimum allele frequencies were: rs1279683 (0.28), rs12479919 (0.30), rs659647 (0.48). Decreasing concentrations of ocular ascorbate from the common to the rare genotype were observed for rs6596473 and rs12479919. The per allele difference in aqueous humor ascorbate for rs6596473 was -217 µmol/L, p < 0.04 and a per allele difference in lens nucleus ascorbate of -0.085 µmol/g, p < 0.02 for rs12479919. The ß coefficients for the regression of log10 plasma ascorbate on aqueous humor ascorbate were higher for the GG genotype of rs6596473: GG, ß = 1460 compared to carriage of the C allele, CG, ß = 1059, CC, ß = 1132, p interaction = 0.1. In conclusion we found that compared to studies in well-nourished populations, ascorbate concentrations in the plasma, aqueous humor and lens nucleus were low. We present novel findings that polymorphisms in SLC23A1/2 genes influenced ascorbate concentration in aqueous humor and lens nucleus.
Assuntos
Humor Aquoso/química , Ácido Ascórbico/metabolismo , Catarata/genética , Núcleo do Cristalino/química , Plasma/química , Polimorfismo Genético , Transportadores de Sódio Acoplados à Vitamina C/genética , Adulto , Idoso , Alelos , Catarata/metabolismo , Cromatografia Líquida de Alta Pressão , DNA/genética , Feminino , Frequência do Gene , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Transportadores de Sódio Acoplados à Vitamina C/metabolismoRESUMO
Purpose: The objective of the present study was to investigate the relationship between macular pigment optical density (MPOD) and plasma carotenoids [(L) and (Z)] and serum lipids in South Indian young healthy volunteers and patients with early age-related macular degeneration (AMD). Methods: Two hundred and fourteen (N = 214) study participants (Healthy control group (N) = 178; Early AMD group (N) = 36) were enrolled after getting their written informed consent. The MPOD of the study participants was assessed using MPS II (Electron Technology, UK) after completing their routine ocular examination. Serum lipids were measured by the standard technique. Plasma levels of L, Z, lycopene and beta-carotene were estimated by high performance liquid chromatography with photo diode array detector. Statistical analysis used: Correlations among variables in serum, plasma and the MPOD were established using Spearman's rho correlation coefficient. Results: The overall mean MPOD in healthy control group and early AMD group were found to be 0.47 ± 0.16 (N = 178; 317 eyes) and 0.35 ± 0.22 (N = 36; 38 eyes) at 1° eccentricity respectively and were found to be statistically significant (p < 0.001). A strong positive association was found between plasma L, Z and L + Z and MPOD. Serum HDL showed a strong negative association with MPOD and other lipids showed a very weak association. MPOD was unaffected by body mass index. Conclusions: MPOD is positively associated with plasma L,Z and L + Z, adding further evidence that additional intake of L/Z may be beneficial in delaying the risk of AMD in our population.
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Glucocorticoids (GCs) are known to regulate several physiological processes and are the mainstay in the management of inflammatory eye diseases. The long-term use of GC causes raised intraocular pressure (IOP) or ocular hypertension (OHT) in about 30-50% of the susceptible individuals depending on the route of administration, and can lead to steroid-induced secondary glaucoma. The present study aims to understand the role of microRNAs (miRNAs) in differential glucocorticoid (GC) responsiveness in human trabecular meshwork (HTM) cells using small RNA sequencing. The human organ-cultured anterior segment (HOCAS) model was used to identify whether donor eyes were from GC-responders (GC-R; n = 4) or GC-non-responders (GC-NR; n = 4) following treatment with either 100 nM dexamethasone (DEX) or ethanol (ETH) for 7 days. The total RNA was extracted from cultured HTM cells with known GC responsiveness, and the differentially expressed miRNAs (DEMIRs) were compared among the following five groups: Group #1: ETH vs. DEX-treated GC-R; #2: ETH vs. DEX-treated GC-NR; #3: overlapping DEGs between Group #1 and #2; #4: Unique DEMIRs of GC-R; #5: Unique DEMIRs of GC-NR; and validated by RT-qPCR. There were 13 and 21 DEMIRs identified in Group #1 and Group #2, respectively. Seven miRNAs were common miRNAs dysregulated in both GC-R and GC-NR (Group #3). This analysis allowed the identification of DEMIRs that were unique to GC-R (6 miRNAs) and GC-NR (14 miRNAs) HTM cells, respectively. Ingenuity Pathway Analysis identified enriched pathways and biological processes associated with differential GC responsiveness in HTM cells. This is the first study to reveal a unique miRNA signature between GC-R and GC-NR HTM cells, which raises the possibility of developing new molecular targets for the management of steroid-OHT/glaucoma.
Assuntos
Glaucoma , MicroRNAs , Hipertensão Ocular , Humanos , Glucocorticoides/farmacologia , Malha Trabecular/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Hipertensão Ocular/induzido quimicamente , Hipertensão Ocular/metabolismo , Glaucoma/genética , Dexametasona/farmacologia , Análise de Sequência de RNA , Esteroides/metabolismoRESUMO
Aim: To investigate genes and pathways involved in differential glucocorticoid (GC) responsiveness in human trabecular meshwork (HTM) cells using RNA sequencing. Methods: Using paired human donor eyes, human organ-cultured anterior segment (HOCAS) was established in one eye to characterize GC responsiveness based on intra ocular pressure (IOP) change and, in the other eye, primary HTM cell culture was established. For RNA sequencing, total RNA extracted from GC-responder (GC-R) and non-responder (GC-NR) cells after dexamethasone (DEX) or ethanol (ETH) treatment for 7d was used. Differentially expressed genes (DEGs) were compared among five groups and validated. Results: In total, 616 and 216 genes were identified as significantly dysregulated in Group #1 and #2 (#1: ETH vs. DEX-treated GC-R; #2: ETH vs. DEX-treated GC-NR), respectively. Around 80 genes were commonly dysregulated in Group #3 (overlapping DEGs between #1 and #2), whereas 536 and 136 genes were uniquely expressed in GC-R (#4) and GC-NR HTM (#5) cells, respectively. Pathway analysis revealed that WNT signaling, drug metabolism cytochrome p450, cell adhesion, TGF-ß signaling, and MAPK signaling were associated with GC responsiveness. Conclusion: This is the first study reporting distinct gene signatures and their associated pathways for GC-R and GC-NR HTM cells. WNT and MAPK signaling are potential therapeutic targets for the management of GC-induced glaucoma.
Assuntos
Glucocorticoides , Malha Trabecular , Perfilação da Expressão Gênica , Glucocorticoides/metabolismo , Glucocorticoides/farmacologia , Humanos , Pressão Intraocular , Malha Trabecular/metabolismo , Transcriptoma/genéticaRESUMO
Purpose: With age, human retinal pigment epithelium (RPE) accumulates bisretinoid fluorophores that may impact cellular function and contribute to age-related macular degeneration (AMD). Bisretinoids are comprised of a central pyridinium, dihydropyridinium, or cyclohexadiene ring. The pyridinium bisretinoid A2E has been extensively studied, and its quantity in the macula has been questioned. Age-changes and distributions of other bisretinoids are not well characterized. We measured levels of three bisretinoids and oxidized A2E in macula and periphery in human donor eyes of different ages. Methods: Eyes (N = 139 donors, 61 women and 78 men, aged 40-80 years) were dissected into 8 mm diameter macular and temporal periphery punches. Using liquid chromatography - electrospray ionization - mass spectrometry (LC-ESI-MS) and an authentic synthesized standard, we quantified A2E (ng). Using LC-ESI-MS and a 50-eye-extract of A2E, we semiquantified A2E and 3 other compounds (eye extract equivalent units [EEEUs): A2-glycerophosphoethanolamine (A2GPE), dihydropyridine phosphatidyl ethanolamine (A2DHPE), and monofuranA2E (MFA2E). Results: A2E quantities in ng and EEEUs were highly correlated (r = 0.97, P < 0.001). From 262 eyes, 5 to 9-fold higher levels were observed in the peripheral retina than in the macula for all assayed compounds. A2E, A2DHPE, and MFA2E increased with age, whereas A2GPE remained unaffected. No significant right-left or male-female differences were detected. Conclusions: Significantly higher levels were observed in the periphery than in the macula for all assayed compounds signifying biologic differences between these regions. Levels of oxidized A2E parallel native A2E and not the distribution of retinal illuminance. Data will assist with the interpretion of clinical trial outcomes of agents targeting bisretinoid-related pathways.
Assuntos
Degeneração Macular , Epitélio Pigmentado da Retina , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Lipofuscina/metabolismo , Degeneração Macular/metabolismo , Masculino , Pessoa de Meia-Idade , Extratos Vegetais , Compostos de Piridínio/química , Compostos de Piridínio/metabolismo , Epitélio Pigmentado da Retina/metabolismo , Retinoides/metabolismo , Espectrometria de Massas por Ionização por Electrospray/métodosRESUMO
In the quest of identifying newer molecular targets for the management of glucocorticoid-induced ocular hypertension (GC-OHT) and glaucoma (GCG), several microarray studies have attempted to investigate the genome-wide transcriptome profiling of primary human trabecular meshwork (TM) cells in response to dexamethasone (DEX). However, no studies are reported so far to demonstrate the temporal changes in the expression of genes in the cultured human TM cells in response to DEX treatment. Therefore, in the present study, the time-dependent changes in the genome-wide expression of genes in primary human TM cells after short (16 hours: 16 h) and long exposure (7 days: 7 d) of DEX was investigated using RNA sequencing. There were 199 (118 up-regulated; 81 down-regulated) and 525 (119 up-regulated; 406 down-regulated) DEGs in 16 h and 7 d treatment groups respectively. The unique genes identified in 16 h and 7 d treatment groups were 152 and 478 respectively. This study found a distinct gene signature and pathways between two treatment regimes. Longer exposure of DEX treatment showed a dys-regulation of Wnt and Rap1 signaling and so highlighted potential therapeutic targets for pharmacological management of GC-OHT/glaucoma.
Assuntos
Glaucoma , Malha Trabecular , Células Cultivadas , Dexametasona/efeitos adversos , Glaucoma/induzido quimicamente , Glaucoma/tratamento farmacológico , Glaucoma/genética , Glucocorticoides/metabolismo , Humanos , Malha Trabecular/metabolismo , TranscriptomaRESUMO
Purpose: Earlier our group has demonstrated the drug reservoir function of the human amniotic membrane (HAM) using stable moxifloxacin and fortified cefazolin ophthalmic formulations and found it as a suitable tool to deliver drugs for an extended duration. The purpose of this study was to evaluate the extended-release kinetics of voriconazole from the impregnated human amniotic membrane (HAM) in vitro. Methods: HAM buttons were incubated with freshly prepared 1% topical ophthalmic formulation of voriconazole for 5 different exposure time to investigate the ideal exposure time for the extended-release of voriconazole from HAM. The drug release kinetics was studied in simulated tear fluid for 5 weeks and the amount of voriconazole released at different intervals was estimated using high-performance liquid chromatography (HPLC) with photodiode array (PDA) detector. Results: There was a marginal increase in drug entrapment efficiency with increased drug exposure time but neither the drug entrapment nor the drug release was found to be statistically significant (P ≥ 0.5). Voriconazole was detectable even at 5 weeks. Conclusion: A sustained release of voriconazole was achieved up to 5 weeks, when voriconazole was incubated with amniotic membrane for all the studied drug soaking times. Thus, voriconazole impregnated amniotic membrane can be considered for the sustained delivery for its in fungal keratitis.
Assuntos
Infecções Oculares Fúngicas , Preparações Farmacêuticas , Âmnio , Antifúngicos/uso terapêutico , Infecções Oculares Fúngicas/tratamento farmacológico , Humanos , Moxifloxacina , VoriconazolRESUMO
The purpose of the present study was to assess the differential intraocular pressure response (IOP) to dexamethasone (DEX) treatment at two dose levels (100 or 500 nM) in perfusion cultured Indian cadaveric eyes to investigate glucocorticoid (GC) responsiveness. In a human organ-cultured anterior segment (HOCAS) set-up, the eye pressure was monitored for every 24 h post DEX infusion (100 or 500 nM) or 0.1% ethanol treatment for 7 days after baseline stabilization. The expression of DEX-inducible proteins such as myocilin and fibronectin in HOCAS-TM tissues was assessed by immunostaining. Elevated IOP was observed in 6/16 eyes [Mean ± SEM (mΔIOP): 15.50 ± 1.96 mmHg; 37.5% responders] and 3/15 eyes (Mean ± SEM mΔIOP: 10 ± 0.84 mmHg; 20% responders) in 100 nM and 500 nM dose groups respectively. Elevated IOP in GC responder eyes was substantiated with a significant increase in myocilin (11.8-fold; p = 0.0002) and fibronectin (eightfold; p = 0.04) expression as compared to vehicle-treated eyes by immunofluorescence analysis. This is the first study reporting the GC responsiveness in Indian cadaveric eyes. The observed GC response rate was comparable with the previous studies and hence, this model will enable us to investigate the relationship between differential gene expression and individual GC responsiveness in our population.
Assuntos
Dexametasona/farmacologia , Olho/fisiopatologia , Glaucoma/fisiopatologia , Glucocorticoides/farmacologia , Malha Trabecular/efeitos dos fármacos , Idoso , Cadáver , Células Cultivadas , Olho/efeitos dos fármacos , Glaucoma/tratamento farmacológico , Humanos , Pressão Intraocular , PerfusãoRESUMO
The intraocular pressure lowering property of a new rho kinase inhibitor, SB772077B (SB77) has been previously demonstrated in perfused human cadaveric eyes. In this study, the efficacy of SB77 in alleviating the aqueous outflow resistance mediated by cyclic mechanical stress in perfused human cadaveric eyes was investigated. A human anterior segment perfusion culture model was used to investigate the effect of cyclic intraocular pressure (IOP) on aqueous outflow facility in presence or absence of SB77. The status of RhoA activation and the downstream effector molecule myosin-light chain phosphorylation (p-MLC) was investigated by Western blot. Cyclic mechanical stress resulted in decrease in aqueous outflow facility (-19.79 ± 4.93%; p = 0.019) in perfused human eyes and treatment with SB77 (50 µM) significantly enhanced outflow facility by 15% (p = 0.05). The increase in outflow facility by SB77 was confirmed with the inactivation of RhoA/ROCK signaling and decreased expression of extracellular matrix markers. SB77 effectively reduced the outflow resistance mediated by cyclic IOP and thus may be a potential clinical candidate for the management of glaucoma.
Assuntos
Humor Aquoso/efeitos dos fármacos , Olho/efeitos dos fármacos , Pressão Intraocular/efeitos dos fármacos , Inibidores de Proteínas Quinases/uso terapêutico , Actinas/metabolismo , Idoso , Animais , Cadáver , Matriz Extracelular/metabolismo , Olho/metabolismo , Humanos , Cadeias Leves de Miosina/metabolismo , Técnicas de Cultura de Órgãos/métodos , Fosforilação/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Estresse Mecânico , Quinases Associadas a rho/metabolismo , Proteína rhoA de Ligação ao GTP/metabolismoRESUMO
Purpose: Our previous study demonstrated the drug reservoir function of human amniotic membrane (HAM) using stable moxifloxacin as a model drug. The purpose of the present study is to evaluate whether HAM can be used as a drug carrier for extended release of extemporaneous preparation of cefazolin. Methods: HAM Buttons (1 Control, 5 Test) were incubated in a freshly prepared (1 ml) sterile topical solution of cefazolin 5% (w/v) for 3 h and 24 h at two different temperatures. The groups were designated as follows: Group IA: Soaking duration 3 h at 4°C; Group IB: Soaking duration 3 h at room temperature; Group IIA: Soaking duration 24 h at 4°C; and Group IIB: Soaking duration 24 h at room temperature. The release kinetics of cefazolin from different groups of drug-laden HAM was studied for a period of 5 days. Samples were assayed for estimation of cefazolin content at different time intervals by High Performance Liquid Chromatography (HPLC) with Photodiode array (PDA) detector. Results: Three-hour cefazolin treatment with HAM at 4°C caused high drug entrapment (24%) compared to room temperature (11%; P < 0.005); however, the release kinetics was not significantly different between Group IA and IB as well as Group IIA and IIB up to the study period. Increase in drug treatment duration did not show increase in entrapment, but caused two-fold (IA Vs IIA) and 1.6-fold (IB Vs IIB) less drug entrapment at 4°C and room temperature, respectively. Conclusion: The results reveal that HAM may be a suitable drug carrier for extended delivery of fortified formulations without compromising stability.
Assuntos
Âmnio , Cefazolina/farmacocinética , Úlcera da Córnea/tratamento farmacológico , Portadores de Fármacos , Lágrimas/química , Antibacterianos/farmacocinética , Células Cultivadas , Cromatografia Líquida de Alta Pressão , Úlcera da Córnea/metabolismo , Úlcera da Córnea/patologia , Estabilidade de Medicamentos , Feminino , Humanos , Soluções OftálmicasRESUMO
PURPOSE: The impact of P-glycoprotein (P-gp) blockade on the intravenous (i.v.) pharmacokinetics of rhodamine-123 (Rho-123), and the subsequent effect on its disposition in ocular and nonocular tissues, was studied by using rabbits. METHODS: Three (3) control rabbits received only an i.v. bolus dose of Rho-123 (1.52 mg/kg). Three (3) blocker-pretreated rabbits received an i.v. dose of GF120918 (3.5 mg/kg) 30 min before the i.v. bolus of Rho-123. The plasma concentration of Rho-123 at different time points was subjected to a pharmacokinetic compartmental analysis, using WinNonlin (Scientific Consultants, Lexington, KY). For tissue-distribution study, a drug treatment similar to the i.v. kinetic study was followed by having 5 rabbits in each group. The animals were sacrificed at 30 min with an excess of anesthesia. Plasma and tissues samples were analyzed by using a validated high-performance liquid chromatographic IV method with a fluorescent detector. RESULTS: The method validated was sensitive enough to estimate Rho-123 up to 1.94 ng/mL in plasma. I.v. Rho-123 data fitted well into the three-compartment model, and P-gp blocker treatment changed it into a two-compartment model. The P-gp blockade significantly increased the mean tissue concentrations in the lungs and spleen, whereas the rise in mean tissue levels in the heart, liver, and kidney and in all ocular tissues were found to be statistically insignificant. CONCLUSIONS: Increasing the ocular concentration of systemically given drugs may not be possible with the degree of P-gp blockade achieved when using GF120918 at the studied concentration after an i.v. administration.
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Membro 1 da Subfamília B de Cassetes de Ligação de ATP/antagonistas & inibidores , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Acridinas/farmacologia , Olho/metabolismo , Preparações Farmacêuticas/metabolismo , Rodamina 123/farmacocinética , Tetra-Hidroisoquinolinas/farmacologia , Animais , Área Sob a Curva , Cromatografia Líquida de Alta Pressão , Interpretação Estatística de Dados , Feminino , Corantes Fluorescentes , Injeções Intravenosas , Masculino , Coelhos , Reprodutibilidade dos Testes , Distribuição TecidualRESUMO
We investigated the effect of a new Rho kinase inhibitor, SB772077B (SB77) on aqueous outflow facility (OF) in human eyes using human organ-cultured anterior segment (HOCAS). IOP was monitored for 24 h post-treatment with either SB77 (0.1/10/50 µM) or vehicle after a stable baseline pressure. The hydrodynamic pattern of aqueous outflow was analysed by labelling outflow pathway with red fluorescent microspheres. The effect of SB77 on cell morphology, actin stress fibers, focal adhesions, ECM, status of RhoA activation and myosin light chain phosphorylation (p-MLC) were evaluated and compared with Y27632, by immunostaining using primary human trabecular meshwork (HTM) cells. Following 24 h treatment, SB77 increased OF by 16% at 0.1 µM (N = 6), 29% at 10 µM (N = 8; p = 0.018) and 39% at 50 µM (N = 8; p = 0.004) in human eyes. There was an overall increase in tracer quantity and in area along inner wall of Schlemm's canal. Treatment with SB77 showed no evidence of cytotoxicity and caused a significant reduction in the expression of fibrotic markers compared to Y27632. The present findings indicate that SB77 treatment was effective in enhancing OF and reducing fibrotic markers in an ex vivo model. Thus SB77 may be a potential clinical candidate for the management of glaucoma.
Assuntos
Humor Aquoso/metabolismo , Inibidores Enzimáticos/metabolismo , Olho/efeitos dos fármacos , Quinases Associadas a rho/antagonistas & inibidores , Humanos , Hidrodinâmica , Modelos Biológicos , Técnicas de Cultura de ÓrgãosRESUMO
PURPOSE: Aldose reductase (ALR), the first and rate-limiting enzyme involved in polyol pathway plays a central role in diabetes and its related complications, including diabetic retinopathy (DR). Inhibition of ALR may also be an ideal target for reducing the deleterious effects of DR. Therefore, the purpose of the present study was to investigate the protective effect of epalrestat (EPL), ALR inhibitor on glucose-induced toxicity in ARPE-19 cells. METHODS: ARPE-19 cells were challenged with normal glucose (NG, 5 mM) and high glucose (HG1, 25 mM and HG2, 50 mM) in the presence or absence of EPL. ALR and VEGF165 expression in retinal pigment epithelial (RPE) cells under experimental conditions were quantified by real-time polymerase chain reaction using SYBR Green chemistry. Vascular endothelial growth factor (VEGF) secretion in the cell supernatant was measured by Sandwich ELISA. Cytotoxicity of EPL was assessed by MTT assay. ALR inhibitory activity, apoptosis, and sorbitol accumulation were also investigated. RESULTS: EPL at studied concentration did not show any toxicity to RPE cells and showed as maximum as 65% ALR inhibition under high glucose condition (HG1). The presence of EPL significantly reduced ALR expression and VEGF levels as induced by high glucose in ARPE-19 cells. CONCLUSION: Inhibition of ALR appeared to be beneficial in reducing diabetes-related complications in RPE cells under high glucose condition.
Assuntos
Aldeído Redutase/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Glucose/antagonistas & inibidores , Epitélio Pigmentado da Retina/efeitos dos fármacos , Rodanina/análogos & derivados , Tiazolidinas/farmacologia , Aldeído Redutase/genética , Aldeído Redutase/metabolismo , Apoptose/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/administração & dosagem , Glucose/toxicidade , Humanos , Epitélio Pigmentado da Retina/patologia , Rodanina/administração & dosagem , Rodanina/farmacologia , Sorbitol/antagonistas & inibidores , Sorbitol/metabolismo , Relação Estrutura-Atividade , Tiazolidinas/administração & dosagem , Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Fatores de Crescimento do Endotélio Vascular/metabolismoRESUMO
PURPOSE: The purpose of this study is to develop methods to identify glaucoma by examining the optic nerve head (ONH) of donor's eyes when information on the preexisting ocular disease is unavailable. MATERIALS AND METHODS: The ONH of the donor's eyes was evaluated under a stereomicroscope for the cup-disc ratio (CDR) and focal retinal rim thinning. The vertical diameter of the cup and disc was also measured using a precalibrated eyepiece micrometer. The suspect eyes were subjected to histological analysis to confirm the presence of specific glaucomatous changes. RESULTS: A total of 202 eyes from 119 donors (68 males and 51 females, aged 42-96) were evaluated for glaucoma. Among them, 190 (94%) eyes showing vertical CDR in the of 0.0-0.6 range were considered nonglaucomatous and the remaining eyes with >0.6 as glaucoma suspect. The calculated mean CDR of the two groups (0.3 ± 0.16, 0.62 ± 0.27) was highly significant (P = 0.0003). Of 12 eyes suspected of glaucoma, 7 eyes from 5 donors showed specific glaucomatous changes by histology. The prevalence of glaucoma was 4.2% among the donors studied. CONCLUSIONS: A simple method of screening fresh donor eyes for selecting those with glaucoma features using CDR and histological analysis was reported. This method helps to obtain biologically active human ocular tissue for glaucoma research on gene expression, ultrastructural/proteome changes, and outflow mechanism.
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Bancos de Olhos , Glaucoma de Ângulo Aberto/complicações , Pressão Intraocular , Disco Óptico/patologia , Doenças do Nervo Óptico/diagnóstico , Doadores de Tecidos , Campos Visuais/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Glaucoma de Ângulo Aberto/diagnóstico , Glaucoma de Ângulo Aberto/epidemiologia , Humanos , Índia/epidemiologia , Masculino , Pessoa de Meia-Idade , Doenças do Nervo Óptico/etiologia , Prevalência , Tomografia de Coerência Óptica , Testes de Campo VisualRESUMO
Purpose: The purpose of this study is to develop methods to identify glaucoma by examining the optic nerve head (ONH) of donor’s eyes when information on the preexisting ocular disease is unavailable. Materials and Methods: The ONH of the donor’s eyes was evaluated under a stereomicroscope for the cup‑disc ratio (CDR) and focal retinal rim thinning. The vertical diameter of the cup and disc was also measured using a precalibrated eyepiece micrometer. The suspect eyes were subjected to histological analysis to confirm the presence of specific glaucomatous changes. Results: A total of 202 eyes from 119 donors (68 males and 51 females, aged 42–96) were evaluated for glaucoma. Among them, 190 (94%) eyes showing vertical CDR in the of 0.0–0.6 range were considered nonglaucomatous and the remaining eyes with >0.6 as glaucoma suspect. The calculated mean CDR of the two groups (0.3 ± 0.16, 0.62 ± 0.27) was highly significant (P = 0.0003). Of 12 eyes suspected of glaucoma, 7 eyes from 5 donors showed specific glaucomatous changes by histology. The prevalence of glaucoma was 4.2% among the donors studied. Conclusions: A simple method of screening fresh donor eyes for selecting those with glaucoma features using CDR and histological analysis was reported. This method helps to obtain biologically active human ocular tissue for glaucoma research on gene expression, ultrastructural/proteome changes, and outflow mechanism.
RESUMO
PURPOSE: The purpose of the present study was to evaluate the kinetics of single and multiple doses of topical, non-preserved voriconazole (VZ) in human eyes. METHODS: For single dose kinetics, 119 patients undergoing cataract surgery were divided into group I and group II and each group received a single drop (30 µl) of either 1% or 0.1% VZ formulation. Aqueous humor was collected at designated time intervals. For multidose kinetics, a single drop of 1% VZ was instilled 5 times either hourly or every 2 hr. The aqueous humor was tested for VZ at the 5th hr and 9th hr, respectively, after initial instillation. The stability and efficacy of the reconstituted VZ formulations were also evaluated after 30 days. RESULTS: Single dose ocular kinetics of 1% VZ resulted in a maximum mean aqueous concentration of 3.333 ± 1.61 µg/ml in 30 min whereas 0.1% showed a maximum mean aqueous concentration of 0.817 ±.36 µg/ml. In the multidose kinetic study, hourly and bi-hourly dosing resulted in mean aqueous concentrations of 7.47 ± 2.14 µg/ml and 4.69 ± 2.7 µg/ml, respectively. The reconstituted VZ formulations were stable at all studied temperatures, and their efficacy was maintained throughout the study period. CONCLUSION: The present study showed that the achieved mean concentration of VZ in both single dose and multi dose kinetic studies satisfactorily met the MIC(90) for almost all causative fungal organisms. The frequency of instillation may be designed for an "every 2 hr regimen" to maintain a therapeutic concentration for successful therapy.
Assuntos
Antifúngicos/administração & dosagem , Antifúngicos/farmacocinética , Humor Aquoso/metabolismo , Pirimidinas/administração & dosagem , Pirimidinas/farmacocinética , Triazóis/administração & dosagem , Triazóis/farmacocinética , Administração Tópica , Idoso , Idoso de 80 Anos ou mais , Área Sob a Curva , Extração de Catarata , Cromatografia Líquida de Alta Pressão , Úlcera da Córnea/prevenção & controle , Estabilidade de Medicamentos , Infecções Oculares Fúngicas/prevenção & controle , Feminino , Meia-Vida , Humanos , Masculino , Pessoa de Meia-Idade , Conservantes Farmacêuticos/administração & dosagem , Espectrometria de Massas em Tandem , Distribuição Tecidual , VoriconazolRESUMO
Multiple epidemiological studies have emphasized the intake of dark green leafy vegetables rich in xanthophylls in reducing the risk of developing age-related macular degeneration (AMD). Therefore, the present study was undertaken to quantify the levels of major carotenoids in commonly consumed fruits and vegetables of Indian origin and of xanthophylls in the macula of Indian human donor eyes. Fresh fruits (n=20) and vegetables (n=51) collected from two zones of India were tested for the estimation of xanthophyll, lycopene and ß-carotene by using HPLC with Photodiode Array Detection. Lutein and zeaxanthin were quantified from macula and in selected vegetables collected from both southern (SI) and northern (NI) regions of India. Xanthophylls, ß-carotene and lycopene were found in many affordable vegetables commonly available for consumption in India. Higher content of lutein and zeaxanthin was confirmed in many economical leafy vegetables and fruits. Surprisingly, the mean macular levels of lutein and zeaxanthin of SI donor eyes (n=13) were found to be significantly (p<0.001) four times less than in NI donor eyes (n=15) and the macular levels of Northern India were comparable with reported levels in western populations. The present study showed considerable levels of xanthophylls in many of the commonly consumed fruit and vegetable sources in both parts of India. However, SI donor eyes showed lower levels as compared to NI donors and this warrants further investigation about the bioavailability of xanthophylls in their blood and food intake. The relevance of these findings with prevalence of AMD in South India needs to be explored.
Assuntos
Carotenoides/análise , Dieta , Frutas/química , Macula Lutea/química , Degeneração Macular/prevenção & controle , Verduras/química , Xantofilas/análise , Cromatografia Líquida de Alta Pressão , Bancos de Olhos , Humanos , ÍndiaRESUMO
PURPOSE: To evaluate the effect of P-glycoprotein modulation at blood-ocular barriers using gamma scintigraphy. METHODS: Ofloxacin, a fluoroquinolone, was selected as a substrate to study the drug efflux transporter (P-glycoprotein) modulation after labeling with Technetium ((99m)Tc) in rabbits. New Zealand albino rabbits were randomized into two groups of 4 each. Group I received labeled ofloxacin intravitreally (100 micro Ci) and Group II animals were given verapamil intravitreally 15 min before the labeled ofloxacin. Static imaging was done at predetermined time, and dynamic images were also taken after 30 min of intravitreal injection. RESULTS: The radio-chemical purity of labeled ofloxacin was found to be 90-95% with the labeling efficiency of 90%. The static anterior planar images of verapamil pre-treated group showed marginal increase in the uptake of labeled ofloxacin, and dynamic images showed less systemic pool as compared to its control. CONCLUSION: This study further confirms the findings of our laboratory regarding the involvement of P-glycoprotein in the intraocular disposition of susceptible drugs.
Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Barreira Hematoaquosa/fisiologia , Barreira Hematorretiniana/fisiologia , Animais , Antibacterianos/farmacocinética , Barreira Hematoaquosa/diagnóstico por imagem , Barreira Hematorretiniana/diagnóstico por imagem , Câmaras gama , Masculino , Ofloxacino/farmacocinética , Coelhos , Tecnécio , Tomografia Computadorizada de Emissão de Fóton Único , Vasodilatadores/administração & dosagem , Verapamil/administração & dosagem , Corpo Vítreo/diagnóstico por imagem , Corpo Vítreo/metabolismoRESUMO
PURPOSE: To evaluate the functional role of P-gp and ocular tissue distribution of intravitreally injected Rhodamine-123 (Rho-123) in the presence of P-gp specific blocker (GF 120918) in normal as well as rifampicin-fed rabbits using microdialysis and direct sampling technique. METHODS: Intravitreal pharmacokinetics of Rho-123 were conducted in male New Zealand albino rabbits. Direct sampling and microdialysis were employed to study the disposition of Rho-123 in normal as well as rifampicin-fed conditions. Control animals received Rho-123 at the concentration of 350 ng in PBS (0.05 ml) intravitreally, and the blocker-treated group received GF 120918 intravenously at the dose of 3.5 mg/kg 30 min prior to intravitreal injection of Rho-123. In case of direct sampling, four eyes were enucleated at different time points, and ocular tissues and humors were stored at -86 degrees C until analysis by HPLC with fluorescence detection. RESULTS: In direct sampling, the blocker group showed significant increase (2.6 fold) in the mean vitreous concentration of Rho-123. Other tissues like ret-choroid, iris, and cornea also showed significant increase in their mean concentration. Microdialysis did not significantly predict the changes observed with direct sampling. Rifampicin-fed rabbits showed a vitreous pharmacokinetic profile comparable with non-fed (control) animals, and the pharmacokinetic parameters were unaffected by the blocker pretreatment. CONCLUSION: Intravenously injected blocker significantly altered the ocular disposition of intravitreally injected P-gp substrate. Rifampicin pretreatment did not upregulate P-gp transporters of the retina to the extent to affect the intravitreal kinetics of Rho-123 significantly.