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- ItemAcesso aberto (Open Access)Avaliação histopatológica e morfométrica intestinal dos efeitos da administração prolongada de corticosteroide em cães(Sociedade Brasileira para o Desenvolvimento da Pesquisa em Cirurgia, 1999-01-01) Louzada, Murilo; Meneses, Flávia Santana e; Fernandes, Luís César; Kim, Su Bong; Mora, Oswaldo Alves [UNIFESP]; Egami, Mizue Imoto [UNIFESP]; Stávale, João Norberto [UNIFESP]; Silva, Maria Regina Régis [UNIFESP]; Matos, Delcio [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)Corticosteroids are largely used in the medical practice. However, they present side effects, that condition a risk factor for gastro-intestinal surgical procedures. The present study seeks to evaluate the histologycal changes of the mucosa and submucosa in colon of dogs submitted to the extended administration of corticosteroids.Fifteen male dogs with no defined pedigree, weighing 15 kg, randomized, being 6 dogs of the control group and 9 dogs of the experimental group, were submitted to the intramuscular application of hydrocortisone, 35 mg/kg/day for 30 days. The colonic distal segment was resected, fixed in Bouin and the courts were stained by the methods: PAS, alcian blue and HE. In the morphometric analysis, was accomplished the counting of: globet cells (GLO), absorptive cells (ABS), total cells (TOT= GLO+ABS), mitosis (MIT) and lymphocytes (LYMP) in each crypt; mast cells (MAST) in the submucosa. The histopathologycal evaluation revealed lymphocytic and plasma cells infiltrated, disorganization of the crypts, presence of pictotic cells, ulcerations in the mucosa; edema, vascular and lymphatic congestion in the submucosa. Analysis revealed: significant decrease of the number of GLO, ABS, TOT and LYMP, increase of MIT in the experimental group and non alteration of the number of MAST. It was concluded that the extended administration of corticosteroids in dogs induced significant histopathologycal and morphometric alterations in the colonic mucosa, that can be constituted important factors of surgical risk.
- ItemSomente MetadadadosB-1 cell participation in T-cell-mediated alloimmune response(Elsevier B.V., 2010-04-01) Nogueira-Martins, Mauro F. [UNIFESP]; Mariano, Mario [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)B-1 and B cells are important producers of natural antibodies in mice and humans and, therefore, are considered as the first line of defense against pathogens. Because of that, their role in T-cell-mediated immune responses is commonly underrated. However, recent studies have described the participation of B-1 cells in immediate and delayed-type hypersensitivity. the present work assessed the role of B-1 cells in the rejection of allografts in mice, an immune reaction mainly orchestrated by T cells. We have transplanted allogeneic skin and heart to wild-type and B-1-cell-deficient mice, and followed rejection kinetics. Skin graft-infiltrating cells were analyzed by flow cytometry.We observed a delay in rejection kinetics of B-1-cell-deficient mice when compared to wild-type mice. Adoptive transfer of B-1 cells into B-1-cell-deficient mice abrogated this delay. the longer survival observed in the absence of B-I cells correlated with less CD8(+) T cells infiltrating the grafts, as well as with more mast cells. Collectively, our results show the participation of B-1 cells in the allograft rejection process in mice and collaborate to the understanding of B-1 cell biology. (C) 2009 Elsevier GmbH. All rights reserved.
- ItemSomente MetadadadosBeneficial effect of annexin A1 in a model of experimental allergic conjunctivitis(Elsevier B.V., 2015-05-01) Gimenes, Alexandre D. [UNIFESP]; Andrade, Teresa Raquel M. [UNIFESP]; Mello, Claudia B.; Ramos, Lisandra [UNIFESP]; Gil, Cristiane D. [UNIFESP]; Oliani, Sonia M. [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Univ Estadual PaulistaAnnexin A1 (ANXA1), a 37 kDa glucocorticoid-regulated protein, is a potent anti-inflammatory mediator effective in terminating acute inflammatory response, and its role in allergic settings has been poorly studied. the aim of this investigation was to evaluate the mechanism of action of ANXA1 in intraocular inflammation using a classical model of ovalbumin (OVA)-induced allergic conjunctivitis (AC). OVA-immunised Balb/c mice, wild-type (WT) and ANXA1-deficient (AnxA1(-/-)), were challenged with eye drops containing OVA on days 14-16 with a subset of WT animals pretreated intraperitoneally with the peptide AC(2-26) (N-terminal region of ANXA1) or dexamethasone (DEX). After 24 h of the last ocular challenge, WT mice treated with Ac2-26 and DEX had significantly reduced clinical signs of conjunctivitis (chemosis, conjunctival hyperaemia, lid oedema and tearing), plasma IgE levels, leukocyte (eosinophil and neutrophil) influx and mast cell degranulation in the conjunctiva compared to WT controls. These anti-inflammatory effects of DEX were associated with high endogenous levels of ANXA1 in the ocular tissues as detected by immunohistochemistry. Additionally, AC(2-26) administration was effective to reduce IL-2, IL-4, IL-10, IL-13, eotaxin and RANTES in the eye and lymph nodes compared to untreated WT animals. the lack of ANXA1 produced an exacerbated allergic response as detected by the density of the inflammatory cell influx to the conjunctiva and the cytokine/chemokine release. These different effects observed for Ac2-26 were correlated with diminished level of activated ERK at 24 h in the ocular tissues compared to untreated OVA group. Our findings demonstrate the protective effect of ANXA1 during the inflammatory allergic response suggesting this protein as a potential target for new ocular inflammation therapies. (C) 2015 Elsevier B.V. All rights reserved.
- ItemAcesso aberto (Open Access)Caracterização morfológica do intestino do robalo peva (Centropomus parallelus) e avaliação do efeito do Bacillus subtilis sobre mastócitos com grânulos eosinofílicos (MCs/EGC) e células caliciformes intestinais(Universidade Federal de São Paulo (UNIFESP), 2016-10-26) Silva, Wémeson Ferreira da [UNIFESP]; Simões, Manuel de Jesus [UNIFESP]; http://lattes.cnpq.br/5987164343458678; http://lattes.cnpq.br/6658420589724525; Universidade Federal de São Paulo (UNIFESP)Characterize anatomical and morphologically different sites and layers of the intestine of fat snook (Centropomus parallelus). Quantify mast cells containing eosinophilic granules (MCs/EGC) and goblet cells after study specimens being fed with probiotic Bacillus subtilis. Material and Methods: A total of 50 animals were used. The morphologic, histochemical and ultra-structural study was carried out on 23 individuals, the histochemical and immune-histochemical was performed on 12 animals and 15 specimens was used as controls for the number of MCs/EGC and goblet cells, and for any variation in number of them after the supplementation of probiotic at the doses of 5g and 10g per kg of regular fish treat. Histological sections for light microscopic study were 5?m thick, were stained with HE dye, toluidine blue (semithin sections of 1?m), Nilceu M. Castro trichrome staining method and silver staining. For histochemical analysis, staining methods comprised PAS, pH-2.5 and pH-0.5 Alcian blue (AB), pH-2.5 AB plus PAS. For MCs/EGC morphology study the methods used were ferric hematoxylin eosin, Congo red, pH-6 acridine orange and toluidine blue. The histochemical analysis was carried out on sections treated with PAS, pH-2.5 and pH-0.5 AB, pH-2.5 AB plus PAS. Antibodies anti-tryptase, chimase and serotonine were used for immunohistochemical study. Tissue fragments 0.5mm-sized, fixed in Karnovsky solution were conventionally processed for Transmission Electron Microscope imaging. MCs/EGC and goblet cells were quantified in one-square millimeter areas of serial sections from sites of the intestine of animals from all 3 study groups. Results: Anatomically, the intestine comprised pyloric caeca, proximal and distal intestine, and rectum. Morphologically, the layer comprised the mucosa, submucosa, muscular layer and serosa. The mucosa presented an epithelium containing absorptive cells (enterocytes), endocrine cells, intraepithelial limphocytes, rodlet cells, goblet cells positive for neutral glycoproteins, acidic carboxylated and sulfated glycosaminoglycans, lamina propria consisted of connective tissue, blood vessels, MCs / EGC, leukocytes and leukocyte infiltrate, and a muscular mucosa between the mucosa and the submucosa. Connective tissue, blood vessels and leucocytic infiltrate formed the submucosa. Their granules of MCs/EGC showed an acidic and basic character and were also metachromatic, presence of neutral glycoproteins, carboxylated and sulfated glycosaminoglycans and positive for tryptase, chymase and serotonine were finding. The muscular layer showed an inner circular and an outer longitudinal arrangement and it?s connective tissue contained nerve plexuses. The serosa was formed by a thin layer of connective tissue, containing vessels and the subserosa nerve plexus. Ultrastructurally, the mucosa showed enterocytes showing an euchromatic nucleus within a cytoplasm containing mitochondria and electrondense spherical granules, goblet cells showing euchromatic nucleus and cytoplasm plentiful of granules of varied size and electrondensity, four types of enteroendocrine cells, intraepithelial phagocytes, rodlet cells showing euchromatic nucleus and rod-shaped cytoplasmic granules of varying electron density and MCs/EGC with euchromatic nucleus and spherical cytoplasmic granules. The quantification of MCs/EGC showed statistically significant difference only for animals in the group 5-grams, which had an increased mean number of these cells when compared with that showed by the control group. Conclusions: The intestine of the fat snook has pyloric caeca, proximal and distal intestines and rectum. Its mucosa contains enterocytes, goblet cells and enteroendocrine cells, intraepithelial lymphocytes, rodlet cells, phagocytes and MCs/EGC. The granules of MCs/EGC had basic and acidic content as well as tryptase, chymase and serotonine. Ultra-structurally, four different types of endocrine cells were identified and the supplementation of probiotic increased the number of MCs/EGC in the intestinal mucosa and submucosa.
- ItemSomente MetadadadosHarmful effects of carbamazepine on the postnatal development of the rat ventral prostate(Biomed Central Ltd, 2012-03-25) Oliva, Samara Urban de [UNIFESP]; Scarano, Wellerson Rodrigo; Okada, Fatima Kazue [UNIFESP]; Miraglia, Sandra Maria [UNIFESP]; Universidade de São Paulo (USP); Universidade Federal de São Paulo (UNIFESP)Background: Carbamazepine (CBZ) is a first-line antiepileptic drug (AED), although it is also used for the treatments of psychiatric disorders and neuropathic pain. the CBZ utilization has been associated with male reproductive damage, including hormonal alterations, sexual dysfunction and reduction of sperm quality. the wide and long-term use of the CBZ is a common schedule in children and adolescents and alters the testosterone level in adult rats and humans. the objective of this work was to evaluate the CBZ side effects on the ventral prostate of rats from pre-puberty to sexual maturation, since the prostate is an androgen-dependent organ.Methods: Twenty three day-old male albino Wistar rats received CBZ diluted in propylene glycol (20 mg/Kg/i.p via). the treatment lasted 20, 40 and 70 days, according to the different stages of the rat sexual maturation. At the end of each treatment period, ventral prostates were removed and histologically processed. the prostate sections were submitted to the histopathological, morphological and stereological analyses using image analysis system.Results: Reductions of the glandular epithelium, glandular lumen and fibromuscular stroma volume of the ventral prostate were observed in adult rats treated with CBZ since the weaning. Triggering and degranulation of mast cells were observed in the fibromuscular stroma of prepubertal and pubertal CBZ treated rats.Conclusions: the results suggest a direct effect of the CBZ on rat ventral prostate, evidenced by increase of mast cell and macrophage populations during pre-puberty and puberty causing a ventral prostate accentuated damage in the adult phase.
- ItemSomente MetadadadosInflammatory events induced by brown spider venom and its recombinant dermonecrotic toxin: A pharmacological investigation(Elsevier B.V., 2009-04-01) Paludo, Katia Sabrina [UNIFESP]; Petris Biscaia, Stellee Marcela; Chaim, Olga Meiri [UNIFESP]; Otuki, Michel Fleith; Naliwaiko, Katya; Dombrowski, Patricia Andreia; Franco, Celia Regina Cavichiolo [UNIFESP]; Veiga, Silvio Sanches; Universidade Federal de São Paulo (UNIFESP); Univ Fed ParanaAccidents involving Brown spider (Loxosceles sp.) venom produce a massive inflammatory response in injured region. This venom has a complex mixture of different toxins, and the dermonecrotic toxin is the major contributor to toxic effects. the ability of Loxosceles intermedia venom and a recombinant isoform of dermonecrotic toxin to induce edema and increase in vascular permeability was investigated. These toxins were injected into hind paws and caused a marked dose and time-dependent edema and increase in vascular permeability in mice. Furthermore, the enzymatic activity of venom toxins may be primal for these effects. A mutated recombinant isoform of dermonecrotic toxin, that has only residual enzymatic activity, was not able to induce these inflammatory events. Besides the previous heating of toxins markedly reduced the paw edema and vascular permeability showing that thermolabile constituents can trigger these effects. in addition, the ability of these venom toxins to evoke inflammatory events was partially reduced in compound 48/80-pretreated animals, suggesting that mast cells may be involved in these responses. Pretreating mice with histamine (prometazine and cetirizine) and serotonin (methysergide) receptor antagonists significantly attenuated toxins induced edema and vascular permeability. Moreover, HPLC analysis of whole venom showed the presence of histamine sufficient to induce inflammatory responses. in conclusion, these inflammatory events may result from the activation of mast cells, which in turn release bioamines and may be related to intrinsic histamine content of venom. (C) 2008 Published by Elsevier Inc.
- ItemSomente MetadadadosMyenteric Denervation Downregulates Galectin-1 and-3 Expression in Gastric Carcinogenesis(Springer, 2011-06-01) Estofolete, Cassia F.; Zucoloto, Sergio; Oliani, Sonia M.; Polli-Lopes, Ana Claudia; Gil, Cristiane D. [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Sao Jose do Rio Preto Sch Med FAMERP; Universidade de São Paulo (USP); Univ Estadual PaulistaBackground This study evaluated the galectin-1 and -3 expression during N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced gastric carcinogenesis in denervated rat stomachs using benzalkonium chloride.Method Four experimental situations were evaluated: nondenervated and denervated stomachs without lesions and nondenervated and denervated stomachs with lesions. Sections of the pyloric region were stained with toluidine blue and incubated with mouse monoclonal anti-Gal-1 and rabbit polyclonal anti-Gal-3 for histopathological and immunohistochemical analysis, respectively.Result MNNG caused the development of benign and malignant epithelial lesions, which were more pronounced in nondenervated stomachs with lesions and accompanied by inflammatory cell-enriched stroma. By immunostaining, the epithelial cells, blood vessels, muscle layer, and myenteric plexus were Gal-1 and -3 positive. Gal-3 was also detected in the gastric crypts, mucus secretion, and fibroblasts of pyloric fragments. Development of lesions in denervated stomachs was associated with a significant decrease in Gal-1 and -3 expression in epithelial cells, mast cells, and neutrophil cytoplasm, compared with that of nondenervated stomach lesions (P < 0.01; P < 0.001; P < 0.001, respectively).Conclusion These results demonstrate that myenteric denervation downregulates endogenous Gal-1 and -3 expression, which might inhibit tumor development in this experimental model.
- ItemSomente MetadadadosSpecial dyeing, histochemistry, immunohistochemistry and ultrastructure: A study of mast cells/eosinophilic granules cells (MCs/EGC) from Centropomus parallelus intestine(Academic Press Ltd- Elsevier Science Ltd, 2017) Silva, Wemeson Ferreira da [UNIFESP]; Simões, Manuel de Jesus [UNIFESP]; Gutierre, Robson Campos [UNIFESP]; Egami, Mizue Imoto [UNIFESP]; Santos, Antenor Aguiar [UNIFESP]; Antoniazzi, Marta M.; Sasso, Gisela Rodrigues da Silva [UNIFESP]; Ranzani-Paiva, Maria Jose T.Intestine mast cells/eosinophilic granule cells (MCs/EGC) of the marine species Centropomus parallelus (fat snook) were first studied using light and electron microscopy techniques. Mast cells are cells from the connective tissue found in almost all organs and tissues of vertebrates. In fish, they appear in greater numbers in parts of their bodies that are exposed to their environment, such as skin, gills and intestine. The granules in fat snook's mast cell contain a variety of substances, such as histamine, heparin, chondroitin sulfate, serotonin, proteases and cytokines. The present study of intestine MCs/EGC was carried out in 20 specimens of fat snook. Samples of tissue were fixed in Bouin solution and in buffered formalin. Ferric hematoxylin - Congo red, pH6 acridine orange, pH2.5 and pH0,5 Alcian Blue (AB), toluidine blue, PAS, AB + PAS and immunohistochemistry protocols were used. In the mucosa and submucosa layers, MCs/EGCs granules with basic contents were evidenced by Congo red staining, and with acid contents granules were identified through pH 2.5 and 0,5 AB, and acridine orange. Basic and acid contents were simultaneously evidenced using ferric hematoxylin - Congo red stain. Metachromasia was observed in both mucosal and submucosal mast cells. Neutral glycoproteins were evidenced by using PAS protocol, glycosaminoglycan through AB and both simultaneously through AB + PAS. In immunohistochemistry assays, MCs/EGC were positive for tryptase, chymase and serotonin. As in mammals, the study of samples fixed in modified Karnovsky for transmission electron microscopy evidenced that most of the MCs granules were spherical and showed varying electron density, as described in previous reports on other teleost fish species. The metachromasia observed and the identification of tryptase, chymase and serotonin suggest a great similarity between fat snook's MCs/EGC and those described in the mucosa of mammals. (C) 2016 Elsevier Ltd. All rights reserved.