Reactivity of mesenteric and aortic rings from trained rats fed with high caloric diet

A strong association between the benefits of physical exercise on the cardiovascular disease with an improvement of the endothelium-derived relaxing factor production has been consistently shown. The purpose of this study was to evaluate the effect of exercise training associated with high caloric diet in the reactivity of rat mesenteric and aortic rings. Experimental protocol consisted of 4 weeks of high caloric diet consumption previous to 4 weeks of run training (1.2 km/h, 0% grade, in sessions of 60 min, 5 days/week). Concentrations of triglycerides, glucose, insulin and nitrite/nitrate levels were measured and atherogenic index was calculated. Concentration-response curves to acetylcholine (10 nM-100 mu M), sodium nitroprusside (100 pM-100 nM) and phenylephrine (1 nM-3 mu M) were obtained. Exercise training reduced body mass (6%) and triglyceride levels (about 54%), without changes in glucose and insulin concentrations. An improvement of endothelium-dependent relaxation responses to acetylcholine in mesenteric and aortic rings was observed in trained group. No changes were seen for sodium nitroprusside and phenylephrine. In conclusion, our study is the first to show clearly that run training promotes an improvement of the endothelium-dependent relaxing response in aorta and mesenteric rings from rats fed with high caloric diet and that is associated with increase of NO production. (c) 2006 Elsevier B.V. All rights reserved.

High-fat diet associated with obesity induces impairment of mouse corpus cavernosum responses

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

Influence of acute pancreatitis on the in vitro responsiveness of rat mesenteric and pulmonary arteries

Background: Acute pancreatitis is an inflammatory disease characterized by local tissue injury and systemic inflammatory response leading to massive nitric oxide (NO) production and haemodynamic disturbances. Therefore, the aim of this work was to evaluate the vascular reactivity of pulmonary and mesenteric artery rings from rats submitted to experimental pancreatitis.Male Wistar rats were divided into three groups: saline (SAL); tauracholate (TAU) and phospholipase A(2) (PLA(2)). Pancreatitis was induced by administration of TAU or PLA(2) from Naja mocambique mocambique into the common bile duct of rats, and after 4 h of duct injection the animals were sacrificed. Concentration-response curves to acetylcholine (ACh), sodium nitroprusside (SNP) and phenylephrine (PHE) in isolated mesenteric and pulmonary arteries were obtained. Potency (pEC(50)) and maximal responses (E(MAX)) were determined. Blood samples were collected for biochemical analysis.Results: In mesenteric rings, the potency for ACh was significantly decreased from animals treated with TAU (about 4.2-fold) or PLA(2) (about 6.9-fold) compared to saline group without changes in the maximal responses. Neither pEC(50) nor E(MAX) values for Ach were altered in pulmonary rings in any group. Similarly, the pEC(50) and the E(MAX) values for SNP were not changed in both preparations in any group. The potency for PHE was significantly decreased in rat mesenteric and pulmonary rings from TAU group compared to SAL group (about 2.2- and 2.69-fold, for mesenteric and pulmonary rings, respectively). No changes were seen in the E(MAX) for PHE. The nitrite/nitrate (NO(x)(-)) levels were markedly increased in animals submitted to acute pancreatitis as compared to SAL group, approximately 76 and 68% in TAU and PLA(2) protocol, respectively.Conclusion: Acute pancreatitis provoked deleterious effects in endothelium-dependent relaxing response for ACh in mesenteric rings that were strongly associated with high plasma NO(x)(-) levels as consequence of intense inflammatory responses. Furthermore, the subsensitivity of contractile response to PHE in both mesenteric and pulmonary rings might be due to the complications of this pathological condition in the early stage of pancreatitis.

Interaction between Advanced Glycation End Products Formation and Vascular Responses in Femoral and Coronary Arteries from Exercised Diabetic Rats

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

Regeneration of the Corneal Epithelium after Debridement of its Central Region: An Autoradiographic Study on Rabbits

Purpose: To investigate the proliferative behavior of the corneal and limbal epithelia after debridement on the central region of the rabbit cornea. Methods: After scraping a circular epithelial area, 5 mm in diameter, in the center of the cornea, (3)H-thymidine ((3)H-TdR) was injected intravitreally, and the rabbits killed from 1 to 49 days afterward. The cornea, together with the adjacent conjunctiva, was processed for autoradiography. Results: The regenerating epithelium at the center of the cornea exhibited high frequencies of labeled nuclei when compared to controls. The mitotic indexes for the limbus were comparable in experimental and control eyes. The unique basal stratum of the limbal epithelium exhibited quick proliferation and vertical migration in all eyes. Cells that remained labeled for four weeks or more were observed throughout the corneal epithelium, including its basal stratum, and this did not depend on epithelial damage. Conclusion: Corneal epithelium wounds are healed by sliding and proliferation of cells surrounding the epithelial gap without any evidence for the participation of the limbal epithelium. Daughter cells labeled with (3)H-TdR were visualized in all layers of the corneal epithelium up to 7 weeks after the DNA precursor injection. However, at this long interval, the only labeled cells in the limbus were in the suprabasal layers.

Morphological and autoradiographic studies on the corneal and limbal epithelium of rabbits

The investigation was centered on the morphological features of the conjunctiva-cornea transition (limbus) of the rabbit eye and the proliferative behavior of its epithelium. The eyes were processed for examination with light and electron microscopy, as well as for autoradiography after intravitreal injection of [H-3]thymidine ([H-3]TdR). At the sites of extraocular muscle insertion, the vascularization of the stroma extended to the peripheral cornea, and the limbal epithelium was thin with its basal stratum made up by clear cuboidal cells. In between the muscle insertions, the cuboidal clear cells, as well as the stroma blood vessels; were scarce. At the light microscope level, the basement membrane was distinct in the cornea but not in the limbus or the conjunctiva. Autoradiographs demonstrated that, at the limbus, the basal cells migrated very quickly to the suprabasal region and remained there up to the 28-day interval. Labeled cells were identified in all epithelial layers of the cornea, including the basal one, at 21 and 28 days but not in the limbal basal clear cells. The rate of renewal of conjunctival epithelium was similar to that observed for the transition with scarce clear cells. The high-resolution autoradiographs demonstrated that the basal cuboidal clear limbal cells exhibit a quick renewal and that they are not label-retaining cells. These latter ones were detected all over the corneal epithelium and in the suprabasal layers of the limbus up to 28 days, in physiological conditions, without the need of stimulation by damage to the corneal epithelium.

Crystallization and preliminary X-ray crystallographic studies of Protac (R), a commercial protein C activator isolated from Agkistrodon contortrix contortrix venom

The protein C pathway plays an important role in the control and regulation of the blood coagulation cascade and prevents the propagation of the clotting process on the endothelium surface. In physiological systems, protein C activation is catalyzed by thrombin, which requires thrombomodulin as a cofactor. The protein C activator from Agkistrodon contortrix contortrix acts directly on the zymogen of protein C converting it into the active form, independently of thrombomodulin. Suitable crystals of the protein C activator from Agkistrodon contortrix contortrix were obtained from a solution containing 2 M ammonium sulfate as the precipitant and these crystals diffracted to 1.95 angstrom resolution at a synchrotron beamline. The crystalline array belongs to the monoclinic space group C2 with unit cell dimensions a=80.4, b = 63.3 and c = 48.2 angstrom, alpha = gamma = 90.0 degrees and beta = 90.8 degrees. (C) 2005 Elsevier B.V. All rights reserved.

Transplante de membrana amniótica canina criopreservada para cicatrização de córnea com deficiência de células límbicas em coelhos

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

Excision of sclerocorneal limbus in dogs and resulting clinical events: study of an experimental model

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Aspectos clínicos e imunopatológicos da ceratoplastia com membrana amniótica xenógena fresca e conservada em glicerina: estudo experimental em coelhos

Avaliaram-se, por meio da análise clínica, histopatologia e imunoistoquímica, os efeitos da aplicação da membrana amniótica xenógena fresca e conservada em glicerina, sobre os mecanismos imunológicos da superfície ocular. Para tal, utilizaram-se 40 coelhos, distribuídos em dois grupos experimentais, os quais foram avaliados por 21 dias. A avaliação clínica revelou que a membrana amniótica xenógena conservada em glicerina estimulou uma resposta inflamatória aguda maior que a membrana aplicada fresca. A análise histopatológica indicou que ambas se comportaram de forma semelhante a partir da primeira semana de pós-operatório, apresentando as alterações clássicas da resposta inflamatória da córnea, com o predomínio de infiltrado do tipo polimorfonuclear. A análise imunoistoquímica indicou que, ainda aos 21 dias, a resposta imune local é inespecífica, permitindo concluir que a resposta imune específica na córnea é tardia e que a córnea é um sítio privilegiado para aplicação de enxertos com características imunológicas diferentes, visto que não houve o estímulo para o desenvolvimento de uma resposta mais específica nos grupos avaliados durante toda a execução do experimento.

Superficial keratectomy and 360º conjunctival flap for bullous keratopathy in a dog: a case report

Relata-se o caso ocorrido em um cão, da raça Pinscher, com dois anos de idade e histórico de desconforto no olho direito. O olho esquerdo havia sido enucleado por outro profissional, por apresentar os mesmos sinais, cujo tratamento clínico instituído não lograra êxito. O valor do teste da lágrima de Schirmer encontrava-se aumentado e identificou-se diminuição da pressão intraocular à tonometria de aplanação. Observaram-se, à biomicroscopia, edema corneal profuso e ceratocone, e o teste da fluoresceína foi negativo. Gonioscopia e oftalmoscopia não lograram fornecer dados relevantes dadas as condições da córnea. Diagnosticou-se ceratite bolhosa. Optou-se pelo tratamento cirúrgico, que fora realizado em duas etapas: 1- ceratectomia superficial e flap conjuntival de 360º; 2- ceratectomia superficial para devolver transparência à córnea. Transcorridos 30 dias da segunda ceratectomia superficial, o flap de terceira pálpebra foi desfeito. Observou-se conjuntivalização do quadrante nasal superior da córnea, córnea clara no eixo visual e retorno da visão.

Microbiological profile of donor corneas stored for tectonic transplantation purposes in rabbits

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

Experimental lamellar keratoplasty in rabbits using microfibrilar cellulose membrane: clinical, morphological and immunohistochemical findings

Avaliaram-se aspectos clínicos, histopatógicos e imunoistoquímicos da córnes de coelhos da raça Nova Zelândia adultos e machos em ceratoplastias lamelares com membrana de celulose microfibrilar. Trinta animais distribuídos em cinco grupos (n=6) foram estudados por até 60 dias de pós-operatório. A avaliação clínica revelou manifestações moderadas de edema, blefaroespasmo e fotofobia ao segundo dia, evoluindo para formas discretas ou ausentes a partir do sétimo dia, período em que se observou, clinicamente, reparo do defeito corneal. A histopatologia revelou uma fina camada de células escamosas, recobrindo a área lesada já aos sete dias, com discreto infiltrado de células polimorfonucleares. Observaram-se vasos no epitélio a partir do 15o dia, com regressão ao 48o dia. A marcação com o anticorpo Ki67 mostrou aumento de células em proliferação aos 15 dias no epitélio e aos 30 dias no estroma. Nesse período, ocorreram remodelamento e adesão epitealial. Considerando a boa integração do implante, admite-se a membrana de celulose como um bom material a ser utilizado em ceratoplastia lamellar.

Topical 1% Nalbuphine on corneal sensivity and epitheilization after experimental lamellar keratectomy in rabbits

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

Use of fresh autogenous vaginal tunic in the experimental lamellar keratoplasty in dogs (Canis familiaris, LINNAEUS, 1758)

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