Mast cells modulate the inflammatory process in endotoxin-induced uveitis

Purpose: To investigate the role of mast cells and annexin-A1 (Anxa1) in endotoxin-induced uveitis (EIU). Methods: EIU was induced by injection of lipopolysaccharide (LPS) into the paws of rats, which were then sacrificed after 24 and 48 h. To assess EIU in the absence of mast cells, groups of animals were pretreated with compound 48/80 (c48/80) and sacrificed after 24 h after no treatment or EIU induction. The eyes were used for histological studies and the aqueous humor (AqH) pool was used for the analysis of transmigrated cells and Anxa1 levels. In inflammatory cells, Anxa1 expression was monitored by immunohistochemistry. Results: After 24 h, rats with EIU exhibited degranulated mast cells, associated with elevated numbers of infiltrating leukocytes and the high expression of Anxa1 in the AqH and the neutrophils. After 48 h of EIU, the mast cells were intact, indicating granule re-synthesis, and there was a reduction of neutrophil transmigration and an increase in the number of mononuclear phagocytic cells in ocular tissues. Anxa1 expression was decreased in neutrophils but increased in mononuclear phagocytic cells. In the animals pretreated with c48/80 and subjected to EIU, mast cells responded to this secretagogue by degranulating and few transmigrated neutrophils were observed. Conclustions: We report that mast cells are a potential source of pharmacological mediators that are strongly linked to the pathophysiology of EIU, and the endogenous protein Anxa1 is a mediator in the homeostasis of the inflammatory process with anti-migratory effects on leukocytes, which supports further studies of this protein as an innovative therapy for uveitis. © 2011 Molecular Vision

Trivialitat dels coneixements generals sobre l'abandonament i adopció de gossos i gats

Treball presentat a l'assignatura de Deontologia i Veterinària Legal (21223

Análise quantitativa de neutrófilos, mastócitos e eosinófilos no adenocarcinoma prostático humano

Objective: To evaluate the relationship between benign prostatic hyperplasia (BPH) and prostatic adenocarcinoma (AP) through the quantification of neutrophils, eosinophils and mast cells and to correlate with the histopathological grade of the neoplasm. Methodology: This is an observational cross-sectional study. Cataloged patient biopsies were sectioned into 5µm sections, stained and analyzed under a light microscope. In these sections, cells were quantified and compared between HPB and AP conditions by statistical analysis. Results: Biopsies of 47 patients were analyzed, 28 (59.6%) with BPH and 19 (40.4%) with AP. The median age of the BPH group was 69 years (range: 54 – 77 years) and the AP group was 66 years (range: 59 – 92 years). In the statistical analysis, a greater number of extravascular (p <0.001) and total (p <0.05) neutrophils was observed in the AP in relation to the BPH; however, there was no statistical difference between intravascular neutrophils, mast cells and eosinophils between the groups. By correlating the Gleason Score and the influx of inflammatory cells, it was observed that higher scores are associated with a lower influx of neutrophils and intact mast cells. In addition, it was observed that prostatic volume and weight with AP may be associated with inflammatory infiltrate. Conclusion: In this study, it was possible to suggest that prostate cancer is related to the innate immune response by the exacerbated influx of neutrophils in the tumor microenvironment and by the influence of these cells on the Gleason Score and on the values of weight and prostate volume. However, further studies are needed to better illustrate the role of neutrophils in tumorigenesis.Objetivo: Avaliar a relação entre a hiperplasia prostática benigna (HPB) e o adenocarcinoma prostático (AP) por meio da quantificação de neutrófilos, eosinófilos e mastócitos e correlacionar com o grau histopatológico da neoplasia. Metodologia: Este estudo é observacional transversal. Biópsias catalogadas de pacientes foram seccionadas em cortes de 5µm, coradas e analisadas no microscópio de luz. Nessas secções, as células foram quantificadas e comparadas entre as condições HPB e AP por meio de análises estatísticas. Resultados: Foram analisadas biópsias de 47 pacientes, sendo 28 (59,6%) com HPB e 19 (40,4%) com AP. A mediana da idade do grupo com HPB foi 69 anos (intervalo: 54 – 77 anos) e o com AP foi 66 anos (intervalo: 59 – 92 anos). Na análise estatística, foi observado um maior número de neutrófilos extravasculares (p <0,001) e totais (p <0,05) no AP em relação à HPB, porém não houve diferença estatística entre neutrófilos intravasculares, mastócitos e eosinófilos entre os grupos. Ao correlacionar o Escore de Gleason e o influxo de células inflamatórias, foi observado que maiores escores estão associados a menor influxo de neutrófilos e mastócitos intactos. Além disso, foi observado que o volume e o peso prostático com AP podem estar associados com o infiltrado inflamatório. Conclusão: Nesse estudo, foi possível sugerir que o câncer de próstata possui relação com a resposta imune inata pelo influxo exacerbado de neutrófilos no microambiente tumoral e pela influência dessas células no Escore de Gleason e nos valores de peso e volume prostático. Entretanto, novos estudos são necessários para melhor ilustrar o papel dos neutrófilos na tumorigênese

Subcicatricial fat grafting associated with rigotomy: a clinical and microstructural analysis

INTRODUCTION: The increase in survival of large burn patients during the acute phase has also increased the prevalence of esthetic-functional scarring sequelae. With regard to the clinical relevance of subcicatricial fat grafting, the actual changes after the procedure need to be understood and microscopically evaluated. METHODS: Eight patients with alcohol burns, with an average time after burn of 12 months (10-14 months), who underwent subcicatricial fat grafting associated with rigotomy were selected. Skin biopsies were performed in before and 14 weeks after operation. The following issues were assessed: 1) esthetic-functional improvement of the scar, by using the Vancouver scar scale; 2) quantitative and qualitative analyses of cicatricial collagen; 3) immunohistochemical analysis of scar vascularization with anti-vascular endothelial growth factor (anti-VEGF) antibody. RESULTS: When comparing the pre- and post-fat grafting period, a significant esthetic-functional improvement and, microscopically, a redefinition of the boundaries of the papillary and reticular dermis were observed, as well as quantitative reduction and reorganization of collagen, in addition to the decrease of the vascularization of the tissue through immunohistochemical analysis. CONCLUSION: The basic principle of the whole physiological healing process is to reestablish local homeostasis, that is, excessively intensified steps that lead to severe clinical changes. When subcicatricial fat grafting associated with rigotomy was performed, qualitative and quantitative improvements of the tissue were verified in this study. Thus, it becomes evident that this procedure can complement the treatment of cicatricial pathologies

Annexin A1 Mimetic Peptide and Piperlongumine: Anti-Inflammatory Profiles in Endotoxin-Induced Uveitis

Uveitis is one of the main causes of blindness worldwide, and therapeutic alternatives are worthy of study. We investigated the effects of piperlongumine (PL) and/or annexin A1 (AnxA1) mimetic peptide Ac2-26 on endotoxin-induced uveitis (EIU). Rats were inoculated with lipopolysaccharide (LPS) and intraperitoneally treated with Ac2-26 (200 µg), PL (200 and 400 µg), or Ac2-26 + PL after 15 min. Then, 24 h after LPS inoculation, leukocytes in aqueous humor, mononuclear cells, AnxA1, formyl peptide receptor (fpr)1, fpr2, and cyclooxygenase (COX)-2 were evaluated in the ocular tissues, along with inflammatory mediators in the blood and macerated supernatant. Decreased leukocyte influx, levels of inflammatory mediators, and COX-2 expression confirmed the anti-inflammatory actions of the peptide and pointed to the protective effects of PL at higher dosage. However, when PL and Ac2-26 were administered in combination, the inflammatory potential was lost. AnxA1 expression was elevated among groups treated with PL or Ac2-26 + PL but reduced after treatment with Ac2-26. Fpr2 expression was increased only in untreated EIU and Ac2-26 groups. The interaction between Ac2-26 and PL negatively affected the anti-inflammatory action of Ac2-26 or PL. We emphasize that the anti-inflammatory effects of PL can be used as a therapeutic strategy to protect against uveitis

Efeito anti-inflamatórios e mecanismo de ação da proteína anexina A1 em modelo de uveíte induzida por endotoxina

A proteína anexina A1 (AnxA1) apresenta importantes propriedades anti-inflamatórias e estudos sugerem que suas ações podem ser mediadas por receptores para peptídeos formilados (FPR). Embora os efeitos anti-inflamatórios da AnxA1 e de seus peptídeos miméticos, especialmente o Ac2-26, tenham sido explorados em diversas investigações, são raros os estudos da AnxA1 nas inflamações oculares. Em razão dos graves efeitos colaterais dos tratamentos atuais para a uveíte, uma importante causa de cegueira, investigamos, in vivo, os efeitos e o mecanismo de ação da AnxA1 nos tecidos oculares de roedores na uveíte induzida por endotoxina (EIU). Ratos machos (Rattus novergicus) foram anestesiados e inoculados, por via subcutânea, na pata direita com lipopolissacarídeo (LPS) (100µg) para o desenvolvimento da uveíte. Após, foram divididos em grupos experimentais (n=10/grupo): EIU por 24 e 48h; EIU por 24h e tratados farmacologicamente por administrações tópica e sistêmica do peptídeo Ac2-26 (100µg) e EIU 24h tratado sistemicamente com peptídeo e Boc2, antagonista do FPR (50µg/animal). Para confirmar a importância da AnxA1 endógena na resolução da inflamação ocular, camundongos selvagens e deficientes para AnxA1 (AnxA1-/-) foram induzidos à uveíte por 24h sem tratamento. Nesses animais AnxA1-/- a resposta inflamatória foi exacerbada em comparação com os selvagens. Enquanto, nos olhos dos ratos, as análises quantitativas dos leucócitos, dosagens de interleucina (IL)-1β, IL-6, fator de necrose tumoral (TNF)-α, óxido nítrico (NO) e expressão da ciclo-oxigenase (COX)-2 nos tecidos e/ou no humor aquoso indicaram os efeitos anti-inflamatórios do peptídeo. Efeitos que foram revertidos na presença do Boc2. As análises imuno-histoquímicas das proteínas AnxA1, AnxA1 fosforilada em...Annexin A1 (AnxA1) is a protein that displays anti-inflammatory properties and some studies suggest that its effects may be mediated by formyl peptide receptors (FPR). Although the anti-inflammatory activities of AnxA1 and its mimetic peptides, including Ac2-26, have been explored in several investigations, the role of AnxA1 in ocular inflammatory processes has not yet been elucidated. Given the common side effects of the current therapies used to treat uveitis, an important cause of blindness worldwide, we investigated, in vivo, the AnxA1 effects and mechanism of action in endotoxin-induced uveitis (EIU). Rattus norvegicus were induced to uveitis (lipopolysaccharide - 100 µg) and divided into experimental groups (n=10/group): EIU untreated for 24 and 48h, EIU treated with topical applications (4/4h) or a single intravenous injection of Ac2-26 (100µg) and EIU systemically treated with the peptide and Boc2, the FPR antagonist (50µg/animal). To confirm the importance of endogenous AnxA1 in the resolution of ocular inflammation, wild-type and AnxA1 deficient (AnxA1-/-) mice were also induced to uveitis without treatment for 24h. AnxA1-/- mice showed exacerbated inflammation compared to wild-type animals. As, quantitative analyses of leukocytes, interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, nitric oxide (NO) levels and cyclooxigenase (COX)-2 expression in ocular tissues and/or in aqueous humor of rats eyes revealed the anti-inflammatory effects of the peptide, which were abrogated in the Boc2 presence. Immunohistochemical analysis of AnxA1, serine-or-tyrosine-phosphorylated AnxA1 (AnxA-S27-PO4 - AnxA-Y21-PO4) in the ocular tissues showed AnxA1 and AnxA1-S27-PO4 expression in epithelial (cornea, iris and ciliary processes) and nervous cells. These expressions were increased in... (Complete abstract click electronic access below

Mast cells in the developing avian eye

Mast cells are present in the eye of Gallus domesticus, appearing in the anterior uvea in embryos at stage 39 HH (13th day). In hatching and adult birds, they are present in the sclera, uvea, pectinate Ligament, and conjunctiva. Mast cells are absent in the cornea, retina, and pecten oculi.Maturing mast cells in the anterior eye segment appear as round cells having eccentric nuclei and a few cytoplasmic metachromatic granules, whose fluorescence increases during development. Mature cells are more numerous in late development, and their cytoplasm is rich in metachromatic and intensely fluorescent granules. Ultrastructurally, maturing mast cells display progranules and a few electron dense and homogeneous granules on one side of the cell. Mast cells of adult birds possess homogeneous cytoplasmic granules, some of which display protuberances that penetrate hollows of adjoining granules. Heterogeneous granules exhibiting latticed and mottled patterns are also present. The existence of mast cells in the anterior eye segment indicates that these cells might perform a physiological role during development and in aqueous humor outflow. They might modulate exchanges between blood and aqueous humor through chemical mediators present in their granules. (C) 1996 Wiley-Liss, Inc.STATE UNIV SAO PAULO,DEPT BIOL,IBILCE,BR-15054000 S JOSE RIO PR,BRAZILWeb of Scienc