RESUMO
Objetivos: Este artigo visa exemplificar uma parceria internacional técnico cientifica por meio da utilização de técnica idealizada e patenteada por pesquisadores brasileiros. Materiais e Métodos: Um grupo de pesquisadores brasileiros idealizou, pesquisou e patenteou uma técnica de obtenção e utilização de íris digitalizada na reabilitação protética ocular. A internacionalização do método foi decorrente do intercâmbio estabelecido entre professores do Brasil e do México.A permanência do pesquisador mexicano, por um período em que especializou e concluiu o curso de mestrado, junto aos colegas brasileiros, pôde oferecer conhecimento e treinamento na aplicação da técnica proposta. Resultados: São apresentados casos clínicos em que a reabilitação protética ocular foi realizada usando a técnica brasileira de íris digitalizada. Conclusão:Pesquisas brasileiras, principalmente na área da saúde, vêm tendo destaque internacional. A preocupação com a melhora da qualidade de vida torna relevantes nossos estudos e técnica sem âmbito mundial.
Objectives: This article aims to illustrate a scientific technical international relation ship through the use of technique created and patented by Brazilian researchers. Materials and Methods: A group of Brazilian researchers devised, researched and patented a technique of obtaining and using scanned iris in ocular prosthetic rehabilitation. The internationalization of the technique was due to technical and scientific partner ship between Brazilian and Mexican researchers. A Mexican researcher remained for a period in which he specialized and completed his masters course with the Brazilian team that was able to transfer knowledge and offer atraining in the application of the technique. Results: Three cases are shown, two Brazilian an done Mexican. Ocular Prosthetic rehabilitation of these cases was performed using the Brazilian technique of scanned iris. Conclusion: Brazilian research, especially in health, comes with international relevance. The concern with improving life quality makes our technical studies relevant world wide.
Assuntos
Humanos , Masculino , Feminino , Iris/metabolismo , Olho Artificial/efeitos adversos , Olho Artificial/normas , Olho Artificial , Reabilitação/ética , Reabilitação/instrumentação , Reabilitação/métodos , Reabilitação , Reabilitação/tendênciasRESUMO
PURPOSE: This study evaluates the effects of the gold nanoparticle in endotoxin-induced uveitis in rats. METHODS: Adult male Wistar rats were divided into five groups: saline + saline, lipopolysaccharide (LPS) + saline, LPS + prednisolone, LPS + gold salt (GS) and LPS + gold nanoparticle (GNP). Two hours after LPS administration, prednisolone acetate 1%, GS, and GNP were topically applied to both eyes of rats and repeated every 6 hours for 24 hours. After 24 hours, rats were anesthetized and aqueous humor was sampled and the irides were removed. Aqueous humor TNF-α, myeloperoxidase activity were determined. Irides oxidative damage and content of toll-like receptor 4 (TLR4) and nuclear factor-κB (NF-κB) were determined. RESULTS: The administration of LPS-induced eye inflammatory response characterized by an increase in aqueous humor TNF-α, myeloperoxidase, and by irides oxidative damage. All these parameters were decreased by the administration of GNP. Since the inflammatory response secondary to LPS administration depends, in part, to the activation of the TLR4-NF-κB pathway we demonstrated here that a potential mechanism to explain the GNP effects was the decrease on TLR4 content and NF-κB activation. CONCLUSIONS: These findings suggest that topical GNP decreases intraocular inflammation and oxidative damage by interfering in the TLR4-NF-κB pathway.
Assuntos
Modelos Animais de Doenças , Compostos de Ouro/farmacologia , Uveíte Anterior/tratamento farmacológico , Administração Tópica , Animais , Humor Aquoso/metabolismo , Western Blotting , Endotoxinas , Ensaio de Imunoadsorção Enzimática , Compostos de Ouro/administração & dosagem , Iris/metabolismo , Lipopolissacarídeos , Masculino , NF-kappa B/metabolismo , Nanopartículas/química , Estresse Oxidativo/efeitos dos fármacos , Peroxidase/metabolismo , Prednisolona/farmacologia , Ratos , Ratos Wistar , Receptor 4 Toll-Like/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Uveíte Anterior/induzido quimicamente , Uveíte Anterior/metabolismoRESUMO
PURPOSE: Some studies have shown the role of gastrin-releasing peptide (GRP) on the production and release of cytokines both in animal models and in humans with inflammatory diseases, but there are no reports on the effects of GRP in ocular inflammatory disease, mainly uveitis. The authors report on the effects of the GRP receptor (GRPR) antagonist RC-3095 in a well-established model for uveitis induced by the administration of lipopolysaccharide (LPS), comparing its effects with those of glucocorticoids. METHODS: Adult male Wistar rats (weight range, 250-300 g; n = 6 per group) were randomly divided into four groups: saline, LPS + saline, LPS + dexamethasone, LPS + RC-3095. Two hours after LPS administration, RC-3095 (0.3 mg/kg, single dose, subcutaneously) or dexamethasone (1 mg/kg, each 6 hours, subcutaneously) was administered. After 24 and 48 hours, rats were anesthetized, aqueous humor was sampled, and the irides were removed. Aqueous humor tumor necrosis factor-alpha, monocyte chemoattractant protein-1 concentration, myeloperoxidase activity were determined. In addition, oxidative damage to the irides was determined by the measure of thiobarbituric acid reactive substances and protein carbonyl content. RESULTS: The acute administration of RC-3095 exhibited anti-inflammatory actions, characterized by a reduction of myeloperoxidase activity and a decrease in tumor necrosis factor-alpha and monocyte chemoattractant protein-1 levels, to a greater extent than dexamethasone. In addition, RC-3095 elicits important action against irides oxidative damage. CONCLUSIONS: These findings suggest that GRP participates in the inflammatory response in an animal model of uveitis, making GRPR a target for new therapeutic options in the treatment of uveitis.
Assuntos
Anticarcinógenos/uso terapêutico , Bombesina/análogos & derivados , Modelos Animais de Doenças , Fragmentos de Peptídeos/uso terapêutico , Receptores da Bombesina/antagonistas & inibidores , Uveíte/tratamento farmacológico , Animais , Humor Aquoso/metabolismo , Bombesina/uso terapêutico , Quimiocina CCL2/metabolismo , Dexametasona/uso terapêutico , Ensaio de Imunoadsorção Enzimática , Escherichia coli , Peptídeo Liberador de Gastrina/fisiologia , Glucocorticoides/uso terapêutico , Iris/metabolismo , Lipopolissacarídeos/farmacologia , Masculino , Peroxidase/metabolismo , Carbonilação Proteica , Ratos , Ratos Wistar , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Uveíte/induzido quimicamente , Uveíte/metabolismoRESUMO
We investigated the expression of the functional endotoxin receptor proteins Toll-like receptor-4 and CD14 in human eyes. Toll-like receptor-4 and CD14 proteins were detected by immunohistochemical analysis of sections of whole human eyes embedded in paraffin with monoclonal antibodies against human toll-like receptor-4 (HTA-125), human CD14 (RPA-M1), or as a control, an irrelevant mouse IgG1k (MOPC-21). Incubation of explants with a neutralizing anti-toll-like receptor-4 monoclonal antibody was used to determine if lipopolysaccharide stimulation of tumor necrosis factor or interleukin-6 secretion was dependent on Toll-like receptor-4 activity. Reverse transcription-polymerase chain reaction was used to detect mRNAs for toll-like receptor-4, tumor necrosis factor-alpha, interleukin-1beta, interleukin-6 and interleukin-8, 3 hr after stimulation of cultured iris microvascular endothelial cells. By immunohistochemistry, human ciliary body non-pigmented epithelial cells showed strong expression of the endotoxin receptor proteins, toll-like receptor-4 and CD14. Toll-like receptor-4 antibodies significantly inhibited lipopolysaccharide-stimulated tumor necrosis factor secretion by the ciliary body. Toll-like receptor-4 mRNA was constitutively expressed in iris endothelial cells and slightly down-regulated by endotoxin. mRNA levels for tumor necrosis factor-alpha, interleukin-1beta, interleukin-6 and interleukin-8 were all increased by endotoxin treatment. This is the first report that shows intraocular (ciliary body and iris) expression of toll-like receptor-4, other than in cornea. Our results show that the ciliary body also expresses CD14, which is anatomically colocalized with toll-like receptor-4. This suggests a potential interaction between both molecules during endotoxin activation of ciliary body cells. The juxtaposition of toll-like receptor-4 and CD14 in the anterior uveal tract helps to explain the sensitivity of the iris/ciliary body to bacterial endotoxin as seen in the standard animal model of endotoxin-induced uveitis.
Assuntos
Corpo Ciliar/metabolismo , Iris/metabolismo , Receptores de Lipopolissacarídeos/metabolismo , Glicoproteínas de Membrana/metabolismo , Receptores de Superfície Celular/metabolismo , Adulto , Anticorpos Monoclonais/imunologia , Células Cultivadas , Citocinas/biossíntese , Células Endoteliais/metabolismo , Regulação da Expressão Gênica , Humanos , Lipopolissacarídeos , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/imunologia , Pessoa de Meia-Idade , RNA Mensageiro/genética , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/imunologia , Receptor 4 Toll-Like , Receptores Toll-LikeRESUMO
PURPOSE: We have previously reported that transferrin is one of several glycoproteins synthesized within the eye and secreted into the vitreous. The present investigation was designed to determine the role of the ciliary body in the production of this vitreous transferrin. METHODS: Isolated ciliary body-iris were incubated with 3H-fucose, 3H-tyrosine or 35S-methionine and afterwards the culture media were processed for affinity chromatography using columns of Sepharose conjugated with antibody to rabbit plasma transferrin. Reverse transcription-polymerase chain reaction (RT-PCR) was carried out using total RNA extracted from fresh ciliary body-iris and primers constructed on the basis of the known sequence of transferrin mRNA from rabbit liver. The fragment obtained was employed as a probe in northern-blots of total RNA of ciliary body-iris. Furthermore, paraffin sections of eyes were treated for immunocytochemical visualization of transferrin. RESULTS: A labeled polypeptide, specifically eluted from the antitransferrin columns, was detected in the incubation medium, transferrin mRNA was found in extracts of whole ciliary body-iris, and transferrin antigenicity was identified in the ciliary and iridial epithelial cells by immunocytochemistry. CONCLUSIONS: These results demonstrate the ciliary epithelium as one of the sources of the vitreous transferrin.
Assuntos
Corpo Ciliar/metabolismo , Transferrina/biossíntese , Animais , Northern Blotting , Cromatografia de Afinidade , Eletroforese em Gel de Poliacrilamida , Fluorometria , Imuno-Histoquímica , Técnicas In Vitro , Iris/metabolismo , Masculino , Reação em Cadeia da Polimerase , RNA Mensageiro/metabolismo , Coelhos , Transcrição Gênica , Transferrina/genéticaRESUMO
The contribution of the ciliary body to the origin of vitreous proteins was investigated in rabbits by incubating explants of this eye component under novel conditions. At the end of incubations for up to 21 h, the tissues were processed histologically and were shown to be in an excellent state of morphological preservation. When radioactive amino acids and fucose were added to the culture medium, protein and glycoprotein synthesis and secretion were detected using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) plus fluorography. The origin of these secretory products was traced by autoradiography to the ciliary epithelium. When samples of vitreous bodies - injected intravitreally with the same radioactive precursors - were run beside samples of the tissue culture media, comigration of at least 8 radioactively labelled bands including the one previously identified as transferrin was detected. This indicates that some vitreous proteins may be secreted by the ciliary body and that cultures of explants of ciliary body-iris are useful tools for studies on vitreous protein secretion.
Assuntos
Corpo Ciliar/metabolismo , Proteínas do Olho/metabolismo , Glicoproteínas/metabolismo , Epitélio Pigmentado Ocular/metabolismo , Aminoácidos/metabolismo , Animais , Autorradiografia , Corpo Ciliar/efeitos dos fármacos , Meios de Cultura , Cicloeximida/farmacologia , Eletroforese em Gel de Poliacrilamida , Proteínas do Olho/biossíntese , Fucose/metabolismo , Glicoproteínas/biossíntese , Iris/efeitos dos fármacos , Iris/metabolismo , Masculino , Técnicas de Cultura de Órgãos , Epitélio Pigmentado Ocular/efeitos dos fármacos , Inibidores da Síntese de Proteínas/farmacologia , CoelhosRESUMO
L-[3H]fucose was injected either intravitreally or intra-aqueously into adult rabbits which were killed at several time points after injection. SDS-polyacrylamide gel electrophoresis and fluorography of iris extracts revealed that most of the proteins are glycoproteins containing fucose residues. Autoradiography of semi-thin histologic sections demonstrated that glycoprotein synthesis was most prominent in the epithelium of the iris, while little protein synthesis was evident in the stroma of the iris. The results of these experiments indicated that the glycoproteins of the iris undergo renewal. The protein band pattern of the iris extracts was very similar to that of extracts of the ciliary body. The high-molecular-weight cartilage matrix glycoprotein (CMGP), an intrinsic component of the ciliary body, vitreous, and aqueous humor, was detected by immunohistologic studies only in the stroma of the iris. The results of immunohistochemical analyses of the eyes of young rabbits (1-21 days old), in addition to the autoradiographic findings, strongly suggest that CMGP is not an intrinsic glycoprotein of the iris stroma, at least in this species.