RESUMO
PURPOSE: Phospholipase A2 (PLA2) is a growing family of lipolytic enzymes that play a key role in various biological processes including general lipid metabolism, membrane homeostasis, and in diseases such as atherosclerosis, arthritis, and acute pancreatitis. Oxidative stress as well as inflammation may be associated with glaucoma pathogenesis. Therefore, our aim was to examine the expression of group IIA secretory PLA2 (sPLA2-IIA), group V secretory PLA2 (sPLA2-V), calcium-independent PLA2 (iPLA2), and cytosolic PLA2 (cPLA2) type in the trabecular meshwork (TM) and the canal of Schlemm in normal eyes and in juxtacanalicular tissue samples from patients with primary open angle glaucoma (POAG) or exfoliation glaucoma (ExG). METHODS: TM tissues were isolated from healthy donor eyes for corneal transplantation. Specimens of inner wall of the Schlemm's canal and the juxtacanalicular tissue were collected during deep sclerectomy from the eyes of patients who had POAG or ExG. Antibodies against PLA2s (sPLA2-IIA, sPLA2-V, iPLA2, and cPLA2) and a standard immunohistochemical procedure were used for the analysis. Quantification of immunoreactions was provided using a Photoshop-based image analysis. Double-staining immunofluorescence of macrophages and sPLA2-IIA was performed by using confocal microscopy. RESULTS: sPLA2-IIA was not present in normal TM. In contrast, sPLA2-IIA levels were significantly higher in glaucoma patients than in controls. Furthermore, sPLA2-IIA expression was much higher in POAG when compared to ExG. iPLA2 was found to predominate in normal human TM, and it demonstrated strong labeling in the uveal and corneoscleral meshwork. The staining of juxtacanalicular meshwork was only moderate in density. In contrast, expression of the enzyme was significantly decreased in glaucoma patients, especially in ExG, when compared to normal controls or to POAG. In addition, strong regional differences were detected in sPLA2-IIA and iPLA2 levels in POAG, whereas immunostaining of these enzymes was much lower and rather uniform throughout ExG sample. In POAG, sPLA2-IIA staining was restricted to certain parts of the trabecular samples where sPLA2-IIA positive macrophages were also present. Immunostaining of sPLA2-V or cPLA2 was low, and no significant changes were found in levels of these enzymes between normal and glaucomatous samples. CONCLUSIONS: sPLA2-IIA, an oxidative stress marker in atherosclerosis, is overexpressed especially in POAG. This result supports the hypothesis that oxidative stress may play a significant role in the pathogenesis of POAG. In ExG, a dramatic decrease in the expression level of iPLA2, a housekeeping enzyme in phospholipid remodeling, may indicate imbalance in phospholipid turnover and also inhibition of normal physiological functions in the TM. These findings may contribute to understanding the pathogenesis of POAG and ExG and may be important for the development of novel therapeutic strategies to different glaucomas.
Assuntos
Câmara Anterior/enzimologia , Síndrome de Exfoliação/enzimologia , Glaucoma de Ângulo Aberto/enzimologia , Fosfolipases A/metabolismo , Western Blotting , Citosol/enzimologia , Síndrome de Exfoliação/patologia , Glaucoma de Ângulo Aberto/patologia , Humanos , Imuno-Histoquímica/métodos , Macrófagos/enzimologia , Microscopia Confocal , Fosfolipases A/classificação , Fosfolipases A2 , Coloração e Rotulagem , Distribuição Tecidual , Malha Trabecular/enzimologiaRESUMO
Reversible protein phosphorylation regulates the biological activities of many human proteins involved in crucial cellular processes, e.g., protein-protein interactions, cell signaling, gene transcription, cell growth, and death. A malfunction of cellular homeostasis in retinal pigment epithelial (RPE) cells is involved in the age-related retinal degeneration. In this study, we examined cytotoxicity in human RPE cells subjected to the protein phosphatase inhibitor, okadaic acid (OA). Moreover, the influence of Hsp90 inhibitor geldanamycin (GA), a benzoquinone ansamycin, in cytoprotection was assessed. Hsp70 protein levels were analyzed by Western blot. Cellular viability was determined by LDH and MTT assays. To study apoptotic cell death, caspase-3 enzyme activity was measured by assaying the cleavage of a fluorescent peptide substrate and Hoechst dye was used to visualize nuclear morphology. OA treatment caused morphological changes and induced cytotoxicity by caspase-3-independent manner in the RPE cells. No evidence of nuclear fragmentation was observed in response to OA. Interestingly, GA treatment accumulated Hsp70 protein and attenuated OA-induced cytotoxicity. This study suggests that Hsp70 and Hsp90 are closely related to cytoprotection of RPE cells in response to protein phosphatase inhibition.
Assuntos
Células Epiteliais/efeitos dos fármacos , Proteínas de Choque Térmico HSP70/efeitos dos fármacos , Ácido Okadáico/antagonistas & inibidores , Fosfoproteínas Fosfatases/antagonistas & inibidores , Epitélio Pigmentado Ocular/efeitos dos fármacos , Quinonas/farmacologia , Benzoquinonas , Caspase 3 , Caspases/efeitos dos fármacos , Caspases/metabolismo , Morte Celular/efeitos dos fármacos , Morte Celular/fisiologia , Linhagem Celular , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Inibidores de Cisteína Proteinase/farmacologia , Citoproteção/efeitos dos fármacos , Citoproteção/fisiologia , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/fisiologia , Inibidores Enzimáticos/toxicidade , Células Epiteliais/metabolismo , Proteínas de Choque Térmico HSP70/metabolismo , Proteínas de Choque Térmico HSP90/efeitos dos fármacos , Proteínas de Choque Térmico HSP90/metabolismo , Humanos , Lactamas Macrocíclicas , Ácido Okadáico/toxicidade , Fosfoproteínas Fosfatases/metabolismo , Fosforilação/efeitos dos fármacos , Epitélio Pigmentado Ocular/metabolismo , Degeneração Retiniana/enzimologia , Degeneração Retiniana/fisiopatologia , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/fisiologiaRESUMO
Development of age-related macular degeneration (AMD) is associated with functional abnormalities and cell death in retinal pigment epithelial (RPE) cells attributable to oxidative stress. To minimize the adverse effects of oxidative stress, cells activate their defence systems, e.g., via increased expression of heat shock protein (Hsp), activation of stress sensitive AP-1 and NF-kappaB transcription factors. In this study, we examined the accumulation of Hsp70 protein, activation of AP-1 and NF-kappaB transcription factors in human ARPE-19 cells subjected to a 4-hydroxynonenal (HNE)-induced oxidative stress. In addition, the influence of Hsp90 inhibitor geldanamycin (GA) was studied in HNE-treated cells. Mitochondrial metabolic activity and apoptosis were determined to evaluate cell death in the ARPE-19 cells. The ARPE-19 cells showed increased accumulation of Hsp70 protein before of the cytotoxic hallmarks appearing in response to HNE. In contrast, increased DNA-binding activities of AP-1 or NF-kappaB transcription factors were not seen under HNE insults. Interestingly, GA significantly increased cell death in the HNE-treated cells, which was involved in caspase-3 independent apoptosis. This study reveals that the Hsps have an important role in the cytoprotection of RPE cells subjected to HNE-derived oxidative stress.
Assuntos
Aldeídos/toxicidade , Inibidores de Cisteína Proteinase/farmacologia , Epitélio Pigmentado Ocular/efeitos dos fármacos , Quinonas/farmacologia , Benzoquinonas , Western Blotting , Caspase 3 , Caspases/fisiologia , Morte Celular/efeitos dos fármacos , Linhagem Celular , Ensaio de Desvio de Mobilidade Eletroforética , Proteínas de Choque Térmico HSP70/biossíntese , Proteínas de Choque Térmico HSP90/fisiologia , Humanos , Lactamas Macrocíclicas , NF-kappa B/fisiologia , Estresse Oxidativo/efeitos dos fármacos , Epitélio Pigmentado Ocular/citologia , Sais de Tetrazólio , Tiazóis , Fator de Transcrição AP-1/fisiologiaRESUMO
OBJECTIVE: To evaluate whether combining ketoprofen, a nonsteroidal anti-inflammatory analgesic, with paracetamol would provide better postoperative analgesia than paracetamol alone in children undergoing strabismus surgery. STUDY DESIGN: A prospective, double-blind, randomised clinical trial. PATIENTS AND METHODS: 56 generally healthy children, aged 3-15 years, undergoing strabismus surgery with standardised endotracheal anaesthesia were randomly assigned to receive either ketoprofen 1 mg/kg intravenously or normal saline at induction of anaesthesia and a second dose 3h later. All children received paracetamol 24 mg/kg solution orally 60 min before surgery and fentanyl 2 mug/kg intravenously during surgery. For rescue analgesia during the first 6h children in pain (pain score >3 on an 11-point scale: 0 = no pain, 10 = worst possible pain) received fentanyl 1 mug/kg intravenously. Any episodes of vomiting and all other adverse events were recorded for the first 24h. RESULTS: 21 of 27 children in the ketoprofen-paracetamol group (78%) and 23 of 29 in the paracetamol group (79%) received fentanyl for rescue analgesia. The mean (SD) number of fentanyl doses during the first 6h was 1.2 (0.9) in the ketoprofen-paracetamol group and 1.7 (1.1) doses in the paracetamol group (mean difference 0.5 doses; 95% CI for difference: -0.1, 1.3; p = 0.11). Eight (30%) vomited in the ketoprofen-paracetamol group and 14 in the paracetamol group (48%) [difference 19%; 95% CI -6, 44; p = 0.15). No serious adverse events occurred. CONCLUSION: Neither paracetamol nor ketoprofen-paracetamol provided sufficient analgesia for children after strabismus surgery because most needed rescue analgesia.
RESUMO
PURPOSE: To investigate the conjunctival inflammatory alterations of patients with primary open-angle glaucoma (POAG) and exfoliation glaucoma (ExG) and correlate the findings with the success of deep sclerectomy (DS) surgery and with the patients' medical history. METHODS: Altogether 25 POAG and ExG patients of the prospective DS study were divided, based on the diagnosis and success of the operation, into 4 groups, POAG S (success), POAG F (failure), ExG S, and ExG F. Controls were obtained from other ophthalmologic surgery patients who did not have glaucoma, and their conjunctiva was examined to be normal. Inflammatory cell subtypes in the conjunctiva were identified and quantified by using immunohistochemistry and monoclonal antibodies: CD3 (T-lymphocyte marker), CD4 (T-helper lymphocyte marker), CD8 (T-cytotoxic lymphocyte marker), CD20 (pan-B cell marker), CD38 (plasma cell marker), CD45RA (naïve T-cell marker), and CD68 (macrophage marker). RESULTS: Higher numbers of inflammatory cells were found in the conjunctiva of the glaucoma patients on medical treatment compared with the normal conjunctiva of the controls. Moreover, T-lymphocytes, T-helper lymphocytes, T-cytotoxic lymphocytes, B cells, plasma cells, and macrophages were found in significantly higher numbers in patients in whom DS failed during the follow-up period of 2.5 years than those with surgical success. CONCLUSIONS: High numbers of cytotoxic and helper T-lymphocytes, plasma cells, and macrophages indicate a chronic inflammatory reaction in the conjunctiva of glaucoma patients. The chronic inflammation is most probably owing to the chronic topical treatment of the patients and seems to be a significant risk factor for DS surgery failure.
Assuntos
Linfócitos B/imunologia , Conjuntivite/imunologia , Síndrome de Exfoliação/cirurgia , Glaucoma de Ângulo Aberto/cirurgia , Macrófagos/imunologia , Esclerostomia , Linfócitos T/imunologia , Idoso , Antígenos CD/imunologia , Anti-Hipertensivos/uso terapêutico , Contagem de Células , Síndrome de Exfoliação/tratamento farmacológico , Síndrome de Exfoliação/imunologia , Técnica Indireta de Fluorescência para Anticorpo , Glaucoma de Ângulo Aberto/tratamento farmacológico , Glaucoma de Ângulo Aberto/imunologia , Humanos , Pessoa de Meia-Idade , Estudos ProspectivosRESUMO
PURPOSE: Dacryocystorhinostomy (DCR) is an effective and safe procedure for patients with post-saccal obstruction of the nasolacrimal pathway. The aim of DCR is to relieve symptoms by creating a bypass between the lacrimal sac and the nasal cavity. The most common reason for failure is stenosis caused by a fibrotic process at the rhinostomy site. In this prospective study we assessed the expression of heat shock protein 47 (HSP47), a regulator of fibrosis, in the biopsies of nasal mucosa isolated from patients undergoing primary endoscopic DCR (EN-DCR). METHODS: Thirty consecutive primary EN-DCR procedures in 30 patients were performed using the powered instrumentation technique. The nasal mucosa specimens over the rhinostomy site were collected for histological analysis at the beginning of the operation and the expression of HSP47 was evaluated by immunohistochemistry. The outcome of EN-DCR was estimated in follow-up visits at 1 week, 2 months and 6 months after surgery. RESULTS: At the 6-month follow-up, the overall success rate after primary EN-DCR was 83%. A metaplastic change and strong expression of HSP47 in nasal mucosa were associated with EN-DCR failure (p = 0.009). CONCLUSIONS: HSP47 may be regarded as a novel marker to predict impaired EN-DCR outcome.
Assuntos
Biomarcadores/metabolismo , Dacriocistite/cirurgia , Dacriocistorinostomia , Proteínas de Choque Térmico HSP47/metabolismo , Mucosa Nasal/metabolismo , Mucosa Nasal/patologia , Idoso , Dacriocistite/metabolismo , Endoscopia , Feminino , Seguimentos , Humanos , Técnicas Imunoenzimáticas , Obstrução dos Ductos Lacrimais/metabolismo , Masculino , Metaplasia , Pessoa de Meia-Idade , Estudos Prospectivos , Resultado do TratamentoRESUMO
PURPOSE: To investigate the efficacy and safety of mitomycin C (MMC)-augmented deep sclerectomy with implant (DSCI) in primary open-angle glaucoma (POAG) and exfoliation glaucoma (ExG) patients. METHODS: A total of 68 eyes of 68 patients with POAG and ExG were enrolled consecutively to undergo DSCI with MMC (0.4 mg/ml for 2 min). The intraocular pressure (IOP), number of antiglaucoma medications, neodymium:yttrium-aluminum-garnet (Nd:YAG) laser goniopuncture treatments and complications were compared postoperatively after 36- month follow-up. Surgery was considered as a complete success when IOP was <18 mmHg without antiglaucoma medication. RESULTS: Preoperatively the mean IOPs were 23 ± 6 mmHg and 25 ± 8 mmHg, and 13 ± 4 mmHg and 11 ± 4 mmHg in the POAG and ExG groups, respectively, at 36 months. At 36 months, 74% and 73% of surgeries were a complete success in the POAG and ExG group, respectively [not significant (NS)]. Two patients (8%) of the POAG group and one of the ExG group (3%) were receiving antiglaucoma medication at 36 months (NS). Nd:YAG laser goniopuncture was performed more often in the ExG group (87%) than in the POAG group (61%, p = 0.024). Postoperatively choroidal detachment occurred in 16% of eyes in the POAG group and in 11% of eyes in the ExG group (NS). CONCLUSIONS: DSCI with MMC augmentation appears to be as effective in patients with ExG and POAG in lowering IOP to target levels at medium term with few immediate postoperative complications.
Assuntos
Alquilantes/administração & dosagem , Síndrome de Exfoliação/cirurgia , Glaucoma de Ângulo Aberto/cirurgia , Mitomicina/administração & dosagem , Esclerostomia/métodos , Idoso , Terapia Combinada , Síndrome de Exfoliação/fisiopatologia , Feminino , Seguimentos , Glaucoma de Ângulo Aberto/fisiopatologia , Humanos , Pressão Intraocular/fisiologia , Complicações Intraoperatórias , Terapia a Laser , Lasers de Estado Sólido , Masculino , Complicações Pós-Operatórias , Estudos Prospectivos , Esclera/efeitos dos fármacos , Esclera/cirurgia , Tonometria Ocular , TrabeculectomiaRESUMO
PURPOSE: To investigate the efficacy and safety of mitomycin C (MMC)-augmented deep sclerectomy with implant (DSCI) in patients with primary open-angle glaucoma (POAG) and exfoliation glaucoma (ExG). METHODS: A total of 68 eyes of 68 patients with POAG and ExG were enrolled consecutively to undergo DSCI with MMC (0.4 mg/ml for 2 min). The intraocular pressure (IOP), number of antiglaucoma medications, neodymium:yttrium-aluminum-garnet (Nd:YAG) laser goniopunctures and complications were compared postoperatively. Surgery was considered as a complete success when IOP was < 18 mmHg without antiglaucoma medication. RESULTS: Preoperatively, the mean IOPs were 23.1 +/- 5.8 and 25.4 +/- 8.3 mmHg, and 13.8 +/- 6.1 and 11.2 +/- 5.6 mmHg in the POAG and ExG groups, respectively, at 12 months. 77.4% and 75.7% of surgeries were a complete success in the POAG and ExG groups, respectively [not significant (NS)]. Five patients (16.1%) in the POAG group but none in the ExG group (0%) were receiving antiglaucoma medication at 12 months (NS). Nd:YAG laser goniopuncture was performed in 29.0% of eyes in the POAG group and in 55.6% of eyes in the ExG group (p = 0.047). Postoperatively, choroidal detachment occurred in 16.1% of eyes in the POAG group and in 10.8% of eyes in the ExG group (NS). We encountered no serious complications related to MMC use. CONCLUSION: DS with MMC augmentation appears to be equally effective in ExG and POAG patients in lowering IOP to target levels, at least in the short term, with few immediate postoperative complications.
Assuntos
Síndrome de Exfoliação/cirurgia , Glaucoma de Ângulo Aberto/cirurgia , Cuidados Intraoperatórios , Mitomicina/administração & dosagem , Esclerostomia/métodos , Implantes Absorvíveis , Idoso , Câmara Anterior/cirurgia , Doenças da Coroide/etiologia , Colágeno/administração & dosagem , Implantes de Medicamento , Síndrome de Exfoliação/fisiopatologia , Feminino , Seguimentos , Glaucoma de Ângulo Aberto/fisiopatologia , Humanos , Estimativa de Kaplan-Meier , Terapia a Laser , Lasers de Estado Sólido/uso terapêutico , Masculino , Pessoa de Meia-Idade , Mitomicina/efeitos adversos , Complicações Pós-Operatórias , Estudos Prospectivos , Punções , Reoperação , Esclerostomia/efeitos adversos , Resultado do TratamentoRESUMO
New straightforward applications of new biopolymers are needed in glaucoma surgery. The aim of this study was to compare biocompatibility of three biomaterials in rabbit eyes after deep sclerectomy; a collagen implant (AquaFlow) represented the "gold standard". A blend of 85:15 poly(L-lactide-co-glycolide) and 70:30 poly(L-lactide-co-1,3-trimethylene carbonate) copolymers in a molar ratio of 70:30 (Bio-1 = Inion GTR membrane) and poly(DL-lactide-co-glycolide with molar compositions of 50:50 (Bio-2) and 85:15 (Bio-3) were inserted into rabbits eyes. Bio-1, Bio-2 or Bio-3 caused very mild eye irritation or tissue response which was comparable to that of the collagen implant. The biodegradation time of Bio-1, Bio-2, and Bio-3 implants was over one year, 3 months and 6 months, respectively. Implant mapping by Fourier transform infrared (FTIR) microscopy revealed a heterogeneous distribution of degradation products throughout Bio-1, Bio-2, and Bio-3. All implants were surrounded by a very fine tissue capsule which was not visible after total degradation of the implants. The FTIR spectrum of tissue capsule around Bio-1 was almost identical to that around Bio-2 whereas significant differences were observed in the spectrum of the tissue capsule around Bio-3. Despite some differences in tissue response, all tested implants represent biologically acceptable materials for drainage devices in glaucoma surgery.
Assuntos
Implantes Absorvíveis , Biopolímeros/metabolismo , Olho/metabolismo , Olho/patologia , Animais , Biopolímeros/química , Movimento Celular , Olho/efeitos da radiação , Macrófagos/citologia , Coelhos , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
PURPOSE: To evaluate the effects of 5-fluorouracil (5-FU) on ocular cells in vitro and the effects of degradable 5-FU-loaded poly(DL-lactide-co-glycolide; PDLGA) 50:50 implant in the rabbit eye in vivo. METHODS: Cytotoxicity was assessed with a tetrazolium salt WST-1 cell proliferation/viability test and a lactate dehydrogenase (LDH) leakage test in rabbit corneal stromal fibroblasts (SIRCs), bovine corneal endothelial cells (BCECs), human conjunctival epithelial cells (IOBA-NHCs), human retinal pigment epithelial cells (ARPE-19), and human corneal epithelial cells (HCECs). The 5-FU-loaded PDLGA implants were surgically placed in rabbit eyes with a deep sclerectomy technique and the histopathology of the eyes was examined. RESULTS: In vitro, 5-FU affected cell proliferation and survival in a time- and dose-dependent manner. In the WST-1 test, adverse effects in serum-free conditions started from 0.0005 mg/mL 5-FU in SIRCS and HCECs, whereas in other cell types, 0.005 mg/mL 5-FU hindered cell proliferation. In serum-free conditions 72-hour 5 mg/mL 5-FU treatment decreased cell viability to 40% in BCECs and to 10% to 15% in other cell types. 5-FU had no or very minor effects on LDH leakage. In vivo, the 5-FU implant showed no signs of toxicity in cornea and retina, whereas in the conjunctival stroma near the implantation site, some inflammatory cells and a marked subepithelial condensation of stromal connective tissue was observed during the postoperative period of 4 weeks. CONCLUSIONS: 5-FU had a broad therapeutic range, and the 5-FU implant showed only minor tissue reactions in conjunctiva at the surgical site. 5-FU is a possible candidate for controlled drug release.
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Alquilantes/farmacologia , Implantes de Medicamento , Olho/efeitos dos fármacos , Fluoruracila/farmacologia , Poliglactina 910 , Animais , Bovinos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Túnica Conjuntiva/efeitos dos fármacos , Túnica Conjuntiva/patologia , Córnea/efeitos dos fármacos , Córnea/patologia , Substância Própria/efeitos dos fármacos , Substância Própria/patologia , Relação Dose-Resposta a Droga , Endotélio Corneano/efeitos dos fármacos , Endotélio Corneano/patologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/patologia , Olho/patologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/patologia , Humanos , Coelhos , Retina/efeitos dos fármacos , Retina/patologia , Epitélio Pigmentado da Retina/efeitos dos fármacos , Epitélio Pigmentado da Retina/patologia , Esclerostomia , Fatores de TempoRESUMO
BACKGROUND: Chronic situations like long-term use of topical medications induces conjunctival inflammation and is also a significant risk factor for failure of filtering surgery. We evaluated conjunctival expression of group IIA secretory PLA(2) (sPLA(2)-IIA), group V secretory PLA(2) (sPLA(2)-V), calcium-independent PLA(2) (iPLA(2)) and cytosolic PLA(2) (cPLA(2)). METHODS: Samples were obtained from non-glaucomatous patients (control subjects), and patients with either primary open-angle glaucoma (POAG) or exfoliation glaucoma (ExG). All the glaucoma patients had been treated with antiglaucomatous medication, and underwent deep sclerectomy surgery. Antibodies against sPLA(2)-IIA, sPLA(2)-V, iPLA(2) and cPLA(2) were used for immunohistochemical staining of frozen tissue sections. RESULTS: In the human conjunctiva of non-glaucomatous patients, immunostaining of sPLA(2)-IIA, sPLA(2)-V or cPLA(2) was low and positively stained cells were mainly localized in the surface of the epithelium. In contrast, iPLA(2) was found to predominate in human normal conjunctiva and it demonstrated strong labeling throughout the epithelium. The stromal staining of iPLA(2) was weak. Expression of sPLA(2)-IIA was significantly increased in stromal fibers of patients with POAG or ExG. No changes were found in levels of sPLA(2)-V, iPLA(2) or cPLA(2) between the patient groups and controls. CONCLUSIONS: These findings demonstrate that sPLA(2)-IIA, sPLA(2)-V, iPLA(2) and cPLA(2) are expressed in the conjunctiva of non-glaucomatous patients. In the epithelium, sPLA(2)-IIA, sPLA(2)-V, and cPLA(2) may participate in protection against risks caused by mechanical wear and tear stress whereas iPLA(2) may regulate remodeling and maintenance of membrane phospholipids. sPLA(2)-IIA may also have the important role in the degradation of bacteria. In conjunctival stroma of POAG and ExG patients, sPLA(2)-IIA may play a role in the development of scar tissue after glaucoma filtration surgery.
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Túnica Conjuntiva/enzimologia , Síndrome de Exfoliação/enzimologia , Glaucoma de Ângulo Aberto/enzimologia , Fosfolipases A2/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Anti-Hipertensivos/uso terapêutico , Síndrome de Exfoliação/tratamento farmacológico , Síndrome de Exfoliação/cirurgia , Glaucoma de Ângulo Aberto/tratamento farmacológico , Glaucoma de Ângulo Aberto/cirurgia , Fosfolipases A2 do Grupo II/metabolismo , Fosfolipases A2 do Grupo V/metabolismo , Fosfolipases A2 do Grupo VI/metabolismo , Humanos , Técnicas Imunoenzimáticas , Pessoa de Meia-Idade , Fosfolipases A2 Citosólicas/metabolismo , Fosfolipases A2 Secretórias/metabolismo , EsclerostomiaRESUMO
BACKGROUND: Endoscopic dacryocystorhinostomy (EN-DCR) is an effective procedure when treating saccal and postsaccal obstructions of the nasolacrimal pathway. The benefit of silicone tubing after DCR is still controversial. We conducted a prospective, randomized study to evaluate the necessity of bicanalicular silicone tubes after primary EN-DCR. METHODS: Forty-six consecutive primary EN-DCR procedures were performed in 42 patients during 2004-2007. The patients were randomized into two study groups according to whether silicone tubing was used or not. RESULTS: The overall success rate after primary EN-DCR was 89%: with silicone tubes it was 78%, and without silicone tubes it was 100%. The difference between these two groups was statistically significant (p<0.049). The follow-up period was 6 months and included three follow-up visits: 1 week, 2 months, and 6 months postoperatively. If inserted, the silicone tubes were removed at the 2-month visit. CONCLUSION: The results of our prospective, randomized study showed that the use of silicone tubes after primary EN-DCR is not necessary.
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Dacriocistorinostomia/instrumentação , Endoscopia , Intubação , Ducto Nasolacrimal/cirurgia , Idoso , Materiais Biocompatíveis , Feminino , Humanos , Intubação/instrumentação , Masculino , Pessoa de Meia-Idade , Silicones , Resultado do TratamentoRESUMO
BACKGROUND: Hyperhomocysteinemia (HH), oxidative stress and endothelial dysfunction are all implicated as possible pathogenetic factors in exfoliation syndrome (XFS) and exfoliative glaucoma (XFG). Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of endothelial nitric oxide and plasma level of ADMA is often elevated in HH. Thus the present study was undertaken to study plasma levels of ADMA with concomitant measurement of symmetric dimethylarginine (SDMA) and L-arginine (L-Arg) in XFS and XFG. METHODS: This cross-sectional, prospective study involved 36 XFS patients, 11 of them having XFG, and 36 age- and gender-matched controls. Fasting plasma ADMA, SDMA and L-Arg levels of participants were determined. A special view was created how plasma L-Arg, ADMA and SDMA correlate to plasma homocysteine (P-Hcy). In addition, the influence of P-Hey values derived from our previous study on the above mentioned parameters were evaluated by cut-off values of P-Hcy, 12 micromol/l for women and 14.5 micromol/l for men. RESULTS The mean plasma ADMA, SDMA and L-Arg levels were 0.41, 0.49 and 62.9 micromol/l in the XFS/XFG group, and 0.41, 0.44 and 69.7 micromol/l in the control group, respectively. As all parameters within the XFS and control group were compared, no statistical significance was stated. On the other hand, a positive correlation was observed between plasma SDMA and P-Hcy in XFGs (P = 0.002), and additionally, also a statistically significant difference was in plasma SDMA between the two groups sorted by cut-off levels of P-Hcy 0.49 +/- 0.15 vs. 0.36 +/- 0.04 micromol/l, above and below cut-off levels, respectively (P = 0.001), but not between ADMA in a respective assay. The mean values of L-Arg were 64.6 +/- 17.2 vs. 74.8 +/- 13.3 microg/l, respectively (P = 0.031). In the XFS subgroup, on the contrary, there was no positive correlation between P-Hcy and plasma SDMA. CONCLUSIONS: A positive correlation of plasma SDMA in respect to P-Hcy in XFGs and increase of SDMA in mild or intermediate hyperhomocysteinemia may indicate SDMA as a marker of developing XFG in hyperhomocysteinemic
Assuntos
Arginina/análogos & derivados , Síndrome de Exfoliação/sangue , Glaucoma/sangue , Idoso , Arginina/sangue , Cromatografia Líquida de Alta Pressão , Estudos Transversais , Feminino , Homocisteína/sangue , Humanos , Masculino , Estresse Oxidativo , Estudos ProspectivosRESUMO
BACKGROUND: In glaucoma, extensive pathological changes occur in the trabecular meshwork (TM) and juxtacanalicular tissue of the chamber angle. Aqueous humor drainage is disturbed due to the accumulation of extracellular matrix (ECM) material in the outflow system. Matrix metalloproteinases (MMPs) remodel ECM material and, thus, they may have a role in regulating outflow facility and intraocular pressure (IOP). This study examined the expression of MMPs and tissue inhibitors of MMPs (TIMPs) in the chamber angle of normal eyes and in primary open-angle glaucoma (POAG) and in exfoliation glaucoma (ExG). METHODS: TM tissues were isolated from healthy donor eyes for corneal transplantation. Specimens of the inner wall of Schlemm's canal and the juxtacanalicular tissue were collected from patients with POAG or ExG during deep sclerectomy operation. Monoclonal antibodies against MMPs (MMP-1, -2, -3, and -9) and antibodies against TIMPs (TIMP-1, -2, and -3) were used for immunohistochemical staining. RESULTS: Immunoreactivity for MMP-2, TIMP-2, or TIMP-3 was observed in human normal TM and in the inner wall of Schlemm's canal. In general, immunoreactions for all of the tested MMPs were more intense in POAG samples than in ExG samples or in the control group. The only exception was the MMP-2 level, which was the highest in the control group. The staining intensity of MMP-1 or MMP-3 was significantly higher in POAG when compared to ExG. TIMP-1 was significantly increased in POAG compared with ExG and there were no marked differences in the levels of TIMP-2 or TIMP-3 between POAG and ExG. The ratios of MMP-1/TIMP-1 and MMP(1+2+3+9) and TIMP(1+2+3) were significantly higher in samples from POAG compared to those of ExG. CONCLUSIONS: Our results reveal an expression imbalance between MMPs and their endogenous tissue inhibitors in tissue samples from patients with POAG and ExG. Differences in immunohistochemical reactions reflect discrete local pathogenic mechanisms involved in POAG and ExG. With respect to the proposed role of MMPs in the remodeling of ECM material, this may point to a weaker reactivity to the accumulation of ECM material in TM in ExG than POAG eyes.
Assuntos
Câmara Anterior/enzimologia , Síndrome de Exfoliação/enzimologia , Glaucoma de Ângulo Aberto/enzimologia , Metaloproteinases da Matriz/metabolismo , Inibidores Teciduais de Metaloproteinases/metabolismo , Malha Trabecular/enzimologia , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Processamento de Imagem Assistida por Computador , Técnicas Imunoenzimáticas , Masculino , Pessoa de Meia-IdadeRESUMO
PURPOSE: Because of the regulatory role of neuropeptide Y (NPY) in angiogenesis, we set out to determine the presence of the leucine 7-proline 7 (Leu7Pro) polymorphism in exudative age-related macular degeneration (AMD) patients and to analyse its implications. METHODS: Genotype analysis of the Leu7Pro polymorphism in the signal peptide region of the human prepro-NPY was performed in blood samples from exudative AMD patients (n = 240) and control subjects (n = 79). RESULTS: In all, 11% of exudative AMD patients and 14% of control subjects exhibited the NPY signal peptide Leu7Pro polymorphism. There were no statistically significant differences in Leu7Pro polymorphism frequency between the exudative AMD and control cases, as analysed by Fisher's exact two-sided test. CONCLUSIONS: Leu7Pro polymorphism in the signal peptide region of the human prepro-NPY is not a risk factor for exudative AMD.
Assuntos
Leucina , Degeneração Macular/genética , Neuropeptídeo Y/genética , Polimorfismo Genético , Prolina , Sinais Direcionadores de Proteínas/genética , Idoso , Idoso de 80 Anos ou mais , Substituição de Aminoácidos , Exsudatos e Transudatos , Feminino , Genótipo , Humanos , Masculino , Reação em Cadeia da Polimerase , Fatores de RiscoRESUMO
BACKGROUND: The aim of the study was to evaluate the vitreous penetration of two commercially available ophthalmic fluoroquinolones: ofloxacin and levofloxacin. METHODS: This prospective, double-blind, randomized clinical trial comprised 16 patients scheduled for vitrectomy surgery of one eye for macular hole or macular pucker. The patients were randomly assigned to receive topical ofloxacin 0.3% (n=9) or levofloxacin 0.5% (n=7) the day before, one drop at noon, 4 p.m., 8 p.m. and midnight. The next morning, patients were given their assigned masked antibiotic every 5 min for four doses starting 1 h before surgery. The vitreous humour samples, at least 0.3 ml each, were collected 1 h after the administration of the last dose, at the beginning of the pars plana vitrectomy with infusion disconnected. Samples were assayed for ofloxacin and levofloxacin concentrations by a method using high-performance liquid chromatography (HPLC) coupled with single mass spectrometry with electrospray ionization RESULTS: Equal topical administration of levofloxacin yielded 2.5 times higher vitreal concentration than ofloxacin. The mean vitreous concentrations of ofloxacin and levofloxacin were 5.30+/-3.04 (SD) ng/ml and 13.09+/-5.24 ng/ml, respectively (P=0.002). CONCLUSIONS: Equal dosing with topical administration of levofloxacin 0.5% and ofloxacin 0.3% allows better penetration into the vitreous for levofloxacin, but the levels of mean concentrations of each drug did not exceed the MIC(90) or MIC(50) for most ocular pathogenic bacteria in terms of conventional endophthalmitis therapy.
Assuntos
Antibacterianos/farmacocinética , Levofloxacino , Ofloxacino/farmacocinética , Corpo Vítreo/metabolismo , Administração Tópica , Idoso , Cromatografia Líquida de Alta Pressão , Método Duplo-Cego , Feminino , Humanos , Masculino , Estudos Prospectivos , Perfurações Retinianas/metabolismo , Perfurações Retinianas/cirurgia , Espectrometria de Massas por Ionização por Electrospray , VitrectomiaRESUMO
PURPOSE: To retrospectively compare the efficacy of deep sclerectomy in the treatment of primary open-angle glaucoma (POAG) and exfoliation glaucoma (ExG). METHODS: Deep sclerectomy with either collagen or hyaluronate implants was performed in 31 eyes (45%) with POAG and 38 eyes (55%) with ExG. Pre- and postoperative intraocular pressure (IOP) was recorded, as was the number of glaucoma medications used pre- and postoperatively in each group. The follow-up data referred to a mean period of 18 months (range: 2 weeks to 36 months). RESULTS: At 18 months, complete success had been achieved in 56.3% of POAG eyes and 44.9% of ExG eyes. Qualified success had been achieved in 83.1% and 71.6% of POAG and ExG eyes, respectively. The mean IOP was 18.6 mmHg in POAG eyes and 16.3 mmHg in ExG eyes. YAG-descemetotomies were performed in nine eyes in each group. There were no statistically significant differences between the groups in IOP (except at 1 week postoperatively in favour of POAG; p = 0.05), success rates, need for postoperative glaucoma medication or number of complications. Reoperations were required in three (10%) POAG eyes and seven (18%) ExG eyes. CONCLUSIONS: Deep sclerectomy is equally effective in controlling IOP in both POAG and ExG and has low rates of serious complications, even when the surgeon is inexperienced in the technique. Both survival rates and IOP control were similar between the groups, and there were no serious intra- or postoperative complications.
Assuntos
Síndrome de Exfoliação/cirurgia , Glaucoma de Ângulo Aberto/cirurgia , Esclerostomia/métodos , Idoso , Síndrome de Exfoliação/fisiopatologia , Feminino , Seguimentos , Glaucoma de Ângulo Aberto/fisiopatologia , Humanos , Pressão Intraocular/fisiologia , Complicações Intraoperatórias , Masculino , Complicações Pós-Operatórias , Estudos Retrospectivos , Resultado do Tratamento , Acuidade Visual/fisiologiaRESUMO
BACKGROUND: Endoscopic dacryocystorhinostomy (EN-DCR) is increasing in popularity as a treatment of nasolacrimal obstructions because it has proven to be an effective and safe procedure. In this study the success of EN-DCR combined with or without bicanalicular silicone stents was evaluated in patients with nasolacrimal obstructions. METHODS: Forty-two consecutive EN-DCR procedures were performed in 36 patients during 2000-2004. The surgery was primary in 23 cases and revision in 19 cases. Bicanalicular silicone stents were inserted in 18 cases and in 24 cases stenting was avoided. RESULTS: The overall success rate after EN-DCR was 81%. The success rate of EN-DCR was 89% with silicone tubing and 75% without silicone tubing. The mean duration of postoperative follow-up was 4 months. CONCLUSION: In our retrospective study no significant statistical differences were seen between the patients with stenting compared with patients without stenting. However, prospective, randomized studies are clearly needed to answer the question of whether the use of stents is advisable.
Assuntos
Dacriocistorinostomia/métodos , Endoscopia , Intubação/instrumentação , Ducto Nasolacrimal , Elastômeros de Silicone , Stents , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Desenho de Prótese , Reoperação , Estudos Retrospectivos , Resultado do TratamentoRESUMO
PURPOSE: Cross-sectional analysis of iris transluminance in type 2 diabetic patients compared with control subjects, all followed in a cohort study. METHODS: A cohort consisting of a well-characterized group of 82 type 2 diabetic patients were followed for 10 years after diagnosis, as were 125 control subjects. The prevalence of iris transluminance was determined by transscleral transillumination and by grading of black-and-white positive prints at the 10-year examination. The frequency of diabetic retinopathy was prospectively determined by grading of fundus photographs at baseline and after five and 10 years. RESULTS: Iris transluminance was found in 27% of diabetic patients and 8.0% of control subjects (P = 0.001). The diabetic patients with more severe and short-term retinopathy more commonly had abnormal iris transluminance than those with no or only mild retinopathy (P < 0.05). Iris transluminance was not associated with intraocular pressure or glycemic control. CONCLUSIONS: In type 2 diabetic patients, abnormal iris transluminance tended to associate with more severe and short-term retinopathy. Therefore we assume that hypoxia is responsible for the defects in the pigment layer of the iris. Thus, abnormal iris transluminance may serve as a marker for rapidly progressive retinopathy.