Role of phospholipase A(2) and tyrosine kinase in Clostridium difficile toxin A-induced disruption of epithelial integrity, histologic inflammatory damage and intestinal secretion

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Aorticopulmonary septal defect in a dog: case report

Na avaliação da dispneia pós-exercício em uma cadela de 10 meses de idade, não castrada e sem raça definida, foi detectado sopro sistólico leve grau II/VI. A comunicação entre a aorta ascendente e o tronco pulmonar, observada pela presença de fluxo contínuo logo abaixo das valvas semilunares, à ecoDopplercardiografia, foi confirmada pela cirurgia. Após o procedimento cirúrgico, a cadela apresentou boa condição clínica e ausência de dispneia mesmo ao exercício. Ressalta-se a importância do diagnóstico precoce e preciso para o sucesso terapêutico. Este é o primeiro relato brasileiro dessa rara doença e a única cirurgia, bem sucedida, descrita na literatura veterinária consultada.

Invasion of epithelial mammalian cells by Paracoccidioides brasiliensis leads to cytoskeletal rearrangement and apoptosis of the host cell

Paracoccidioides brasiliensis (Pb) yeast cells can enter mammalian cells and probably manipulate the host cell environment to favor their own growth and survival. We studied the uptake of strain Pb 18 into A549 lung and Vero epithelial cells, with an emphasis on the repercussions in the cytoskeleton and the apoptosis of host cells. Cytoskeleton components of the host cells, such as actin and tubulin, were involved in the P. brasiliensis invasion process. Cytochalasin D and colchicine treatment substantially reduced invasion, indicating the functional participation of microfilaments (MFs) and microtubules (MTs) in this mechanism. Cytokeratin could also play a role in the P. brasiliensis interaction with the host. Gp43 was recognized by anti-actin and anti-cytokeratin antibodies, but not by anti-tubulin. The apoptosis induced by this fungus in infected epithelial cells was demonstrated by various techniques: TUNEL, DNA fragmentation and Bak and Bcl-2 immunocytochemical expression. DNA fragmentation was observed in infected cells but not in uninfected ones, by both TUNEL and gel electrophoresis methods. Moreover, Bcl-2 and Bak did not show any differences until 24 h after infection of cells, suggesting a competitive mechanism that allows persistence of infection. Overexpression of Bak was observed after 48 h, indicating the loss of competition between death and survival signals. In conclusion, the mechanisms of invasion of host cells, persistence within them, and the subsequent induction of apoptosis of such cells may explain the efficient dissemination of P. brasiliensis. (C) 2004 Published by Elsevier SAS.

Epithelial cells treated with genistein inhibit adhesion and endocytosis of Paracoccidioides brasiliensis

Paracoccidioidomycosis is caused by Paracoccidioides brasiliensis, which although not formally considered an intracellular pathogen, can be internalized by epithelial cells in vitro and in vivo. The mechanisms used by P. brasiliensis to adhere to and invade non-professional phagocytes have not been identified. The signal-transduction networks, involving protein tyrosine kinase (PTK) and protein phosphatase activities, can modulate crucial events during fungal infections. In this study, the involvement of PTK has been investigated in P. brasiliensis adherence and invasion in mammalian epithelial cells. A significant inhibition of the fungal invasion occurred after the pre-treatment of the epithelial cells with genistein, a specific tyrosine kinase inhibitor, indicating that the tyrosine kinase pathway is involved in P. brasiliensis internalization. In contrast, when the fungus was treated, a slight (not significant) inhibition of PTK was observed, suggesting that PTK might not be the fungus' transduction signal pathway during the invasion process of epithelial cells. An intense PTK immunofluorescence labeling was observed in the periphery of the P. brasiliensis infected cells, little PTK labeling was found in both uninfected cells and yeast cells, at later infection times (8 and 24 h). Moreover, when the epithelial cells were treated with genistein and infected with P. brasiliensis, no labeling was observed, suggesting the importance of the PTK in the infectious process. These results suggest that PTK pathway participates in the transduction signal during the initial events of the adhesion and invasion processes of P. brasiliensis to mammalian epithelial cells.

Expression of cell cycle regulatory proteins in epithelial components of dental follicles

Dental follicle is a component of tooth germs, which remain adjacent to the crown of unerupted or impacted teeth. Under the influence of pathologic changes, however, dental follicles that possess reduced epithelium can proliferate into stratified squamous epithelium as far as originate dental cysts. In order to clarify the role of apoptosis and cellular proliferation herein, expression of p53 and PCNA was examined in epithelial components of dental follicles associated with impacted third molars by means of immunohistochemistry. A total of 40 cases was included in this study being 22 cases with reduced epithelium and 18 cases with stratified epithelium. Expression of p53 expression was weak or not detected in dental follicles with reduced and stratified squamous epithelium. By contrast, PCNA positive cells were evidenced in basal and supra basal layers of the stratified squamous epithelium and in reduced epithelium of dental follicles, but without any significant statistically differences between them (P > 0.05). In conclusion, these data suggest that dental follicles possess proliferative activity as depicted by PCNA-positive nuclei in some epithelial cells. However, the biological behavior of dental follicles during the late stage of dental eruptive process may not be associated with deregulation of death and/or cell proliferation.

Diuron-Induced Rat Bladder Epithelial Cytotoxicity

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Comparative Study Between Porcine Small Intestinal Submucosa and Buccal Mucosa in a Partial Urethra Substitution in Rabbits

Purpose: Several urethral conditions may require tissue substitution. One collagen-base biomaterial that recently emerged as an option is small intestinal submucosa (SIS). The aim of this study was to compare the results of SIS and buccal mucosa for urethral substitution in rabbits.Materials and Methods: Thirty-six North Folk male rabbits were randomized into three groups. In all animals, a 10 x 5 mm urethral segment was excised, and the urethral defect was repaired using a one-layer SIS patch (group I [GI]); four-layer SIS (group II [GII]); or buccal mucosa (group III [GIII]). Urethrography was performed preoperatively and after 12 weeks. After sacrifice, graft retraction was objectively measured using Scion Image (R) computer analysis and by calculation of ellipse area. The grade of fibrosis, inflammatory reaction, vascular/epithelial regeneration, and collagen III/I ratio were analyzed by hematoxylin/eosin and Picrosirius red staining.Results: Urethrography confirmed a wide urethral caliber without any signs of strictures after surgery. Urethral fistulae was diagnosed in 8.3% of cases (1 animal each group). Average graft shrinkage was 55.2% in GI; 44.2% in GII; and 57.2% in GIII (p < 0.05). The intensity of chronic inflammation, fibrosis, epithelium regeneration, and neovascularization was similar in all groups (p > 0.05). Collagen III/I ratio was higher in GII (GI: 119.6; GII: 257.2 and GIII: 115.0); p < 0.01.Conclusions: The four-layer SIS is more advantageous than the one-layer SIS and buccal mucosa for urethral substitution in rabbits.

Large segmental radius and ulna defect treated by bone transportation with the Ilizarov technique

An 8-year-old male Boxer with a severely contaminated open fracture of the left radius and ulna fracture, produced by a helicopter propeller, was treated using bone transport by the Ilizarov method. Extensive diaphyseal bone loss and soft-tissue vascular damage were present. The radius and ulna were stabilised with an Ilizarov ring external fixator. The bone defect was partially shortened and restored by gradual transport of a bone segment created from proximal segments of the radius and ulna. The external fixator was removed 4fi months after the beginning of the latency period, due to instability caused by osteolysis around the wires. A cast was placed for 3 weeks. Although the bone transport had resulted in formation of approximately 4 cm of bone, the antebrachium showed approximately 50% shortening when compared to the contralateral limb. The infection was eradicated, and the dog was able to bear weight on the operated limb when walking.

Alveolar osseous defect in rat for cell therapy. Preliminary report

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Periodontal tissue response to coverage of root cavities restored with resin materials: A histomorphometric study in dogs

Background: the purpose of this study was to histomorphometrically evaluate the response of periodontal tissues covering Class V resin restorations in dogs.Methods: After raising a mucoperiosteal flap, bony defects measuring 5 x 5 mm were created on the buccal aspect of the canines of five dogs followed by cavity preparations on the root surface measuring 3 x 3 x 1 mm. Before repositioning the flap to cover the bone defect, the cavities were restored with composite resin (CR) or resin-modified glass ionomer cement (RMGIC) or were left unrestored as control (C). The dogs were euthanized 90 days after surgery. Specimens comprising the tooth and periodontal tissues were removed, processed routinely, cut into longitudinal serial sections in the bucco-lingual direction, and stained with hematoxylin and eosin (H&E) or Masson's trichrome. The most central sections were selected for histomorphometric analysis.Results: Histomorphometric analysis revealed apical migration of epithelial tissue onto the restorative materials (RMGIC and CR). The C group presented significantly longer connective tissue attachment (P < 0.05) than the RMGIC and CR groups and significantly higher bone regeneration (P < 0.05) compared to the RMGIC group. Histologically, the cervical third (CT) of all groups had the most marked chronic inflammatory infiltrate.Conclusions: Within the limits of this study, it can be concluded that the restorative materials used exhibit biocompatibility; however, both materials interfered with the development of new bone and the connective tissue attachment process.

Maxillary Bone Defect Reconstruction Using Porous Polyethylene Implants

Purpose: The aim of this study was to evaluate the bone repair process in the maxillary sinus in monkeys treated with high-density porous polyethylene (Medpor)Methods: Four capuchin monkeys (Cebus apella) were submitted to bilateral horizontal osteotomies in the anterior wall of the maxillary sinus and divided into 2 groups: control group, left side with no implants, and porous polyethylene group, right side with Medpor. After a period of 145 days after implant placement, the maxillae were removed for histologic and histometric analyses.Results: Bone repair in osteotomized areas took place by connective tissue in 58.5% and 58.7% in the control group and the porous polyethylene group, respectively. In the contact surface with Medpor, bone repair occurred in 41.3%.Conclusions: Medpor was not reabsorbed within the period of this study and allowed bone repair surrounding it. The porous polyethylene constitutes a feasible alternative for bone defect reconstruction.

Correction of a Mucogingival Defect Using an Implant-Retained Postoperative Stent in a Severely Resorbed Mandible: Case Report

This case report describes a technique that uses a palatal mucosal graft and an acrylic resin postoperative stent in an attempt to increase the layer of keratinized tissue around osseointegrated implants in an atrophic mandible. During second-stage surgery a split-thickness labial flap is reflected and apically repositioned by being sutured onto the periosteum and connective tissue. A palatal mucosal graft is then sutured onto the recipient site. The stent is worn for at least 4 weeks following surgery. This procedure helps to eliminate mobility of the mucosa in the area, deepen the vestibule, and prevent muscle reinsertion. (Int J Periodontics Restorative Dent 2008;28:617-623.)