Increased Glutathione Levels Contribute To The Beneficial Effects Of Hydrogen Sulfide And Inducible Nitric Oxide Inhibition In Allergic Lung Inflammation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)The interaction between nitric oxide (NO) and hydrogen sulfide (H2S) in the airways could have significant implications for the pathogenesis and therapeutic effects of both on lung diseases. In this study we investigated whether the beneficial effects of H2S on asthma could be comparable to that inhibition of inducible NO synthase (iNOS). Methods: Female BALB/C mice sensitized with ovalbumin (OVA) received either the H2S donor sodium hydrosulfide (NaHS, 14 mu mol/kg) or the iNOS inhibitor 1400W (1 mg/kg), 30 min before each OVA challenge during six days. On the first, second and sixth days, the leucocyte infiltration in lung parenchyma and bronchoalveolar lavage was evaluated. The aconitase activity (a sensor of O-2(center dot-) formation) and lipid peroxidation, as well as levels of reduced glutathione (GSH) and oxidized glutathione (GSSG) were determined in the lung tissues. Results: OVA-challenge caused a significant and time-dependent increase in the eosinophil number in the airways, which was accompanied by a significant decrease of aconitase activity and GSH/GSSG ratio along with enhanced lipid peroxidation in the lungs. Treatment with NaHS or 1400 W significantly attenuated the airways eosinophilia that was paralleled by an increase in aconitase activity and decrease of lipid peroxidation. NaHS or 1400 W treatments also reversed the decreased GSH/GSSG ratio seen after OVA-challenge. Conclusions: The present study shows for the first time that the increased GSH/GSSG ratio caused by either H2S supplementation or iNOS-inhibition is a potential mechanism protecting airways against oxidative stress and inflammatory lung diseases. (C) 2016 Elsevier B.V. All rights reserved.395762Sao Paulo Research Foundation (FAPESP), Brazil [2012/02145-3]National Council for Scientific and Technological Development (CNPq) [2013/04151-3, 2014/24518-1]Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Markers of vascular differentiation, proliferation and tissue remodeling in juvenile nasopharyngeal angiofibromas

Juvenile nasopharingeal angiofibroma (JNA) is a histologically benign locally aggressive tumor characterized by irregular vessels embedded. in a fibrous stroma. Excessive vascularity results in bleeding complications, and the inhibition of angiogenesis is a promising strategy for managing extensive JNA tumors. To better characterize the endothelial components of JNA, we aimed to evaluate markers of vascular differentiation and proliferation, such as friend leukemia integration-1 (FLI-1) and endoglin, lymphatic markers, including podoplanin and vascular endothelial growth factor receptor 3 (VEGFR3) and its cognate ligand VEGFC, GLUT-1, a diagnostic marker that discriminates between hemangiomas and vascular malformations, and two markers of tissue remodeling, stromelysin 3 (ST3) and secreted acid protein rich in cysteine (SPARC). Antigens were assessed immunohistochemically in vessels and stromal cells of JNA archival cases (n=22). JNA endothelial cells were positive for endoglin, VEGFC and FLI-1, whereas podoplanin and VEGFR3 were negative in all cases. Both endothelial cells and fibroblasts stained for ST3 and SPARC. GLUT-1 was investigated in JNA cases, in infantile hemangiomas (n=123) and in vascular malformations (n=135) as controls. JNAs and vascular malformations were GLUT-1-negative, while hemangiomas showed positive staining. The presence of markers of endothelial differentiation and proliferation highlighted the hyper-proliferative state of JNA vessels. The absence of podoplanin and VEGFR3 underscores their blood endothelial cell characteristic. The absence of GLUT-1 discriminates JNAs from hemangiomas. ST3 and SPARC up-regulation in endothelial cells and fibroblasts may contribute to a compensatory signaling for controlling angiogenesis. Some of these markers may eventually serve as therapeutic targets. Our results may aid in the understanding of JNA pathophysiology.CNPqFAPES

Increased glutathione levels contribute to the beneficial effects of hydrogen sulfide and inducible nitric oxide inhibition in allergic lung inflammation

The interaction between nitric oxide (NO) and hydrogen sulfide (H2S) in the airways could have significant implications for the pathogenesis and therapeutic effects of both on lung diseases. In this study we investigated whether the beneficial effects of H2S on asthma could be comparable to that inhibition of inducible NO synthase (iNOS). Female BALB/C mice sensitized with ovalbumin (OVA) received either the H2S donor sodium hydrosulfide (NaHS, 14 μmol/kg) or the iNOS inhibitor 1400 W (1 mg/kg), 30 min before each OVA challenge during six days. On the first, second and sixth days, the leucocyte infiltration in lung parenchyma and bronchoalveolar lavage was evaluated. The aconitase activity (a sensor of O2radical dotsingle bond formation) and lipid peroxidation, as well as levels of reduced glutathione (GSH) and oxidized glutathione (GSSG) were determined in the lung tissues. OVA-challenge caused a significant and time-dependent increase in the eosinophil number in the airways, which was accompanied by a significant decrease of aconitase activity and GSH/GSSG ratio along with enhanced lipid peroxidation in the lungs. Treatment with NaHS or 1400 W significantly attenuated the airways eosinophilia that was paralleled by an increase in aconitase activity and decrease of lipid peroxidation. NaHS or 1400 W treatments also reversed the decreased GSH/GSSG ratio seen after OVA-challenge. The present study shows for the first time that the increased GSH/GSSG ratio caused by either H2S supplementation or iNOS-inhibition is a potential mechanism protecting airways against oxidative stress and inflammatory lung diseases395762CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2013/04151-3; 2014/24518-12012/02145-

Padronização de metodologia para determinação de ácido propiônico em pães de forma

Alguns tipos de pães para sanduíches são sensíveis à contaminação por bolores e leveduras. Por isso a legislação brasileira de alimentos permite a adição de propionato de cálcio no limite máximo de 0,2%. A cromatografia gasosa é um método utilizado para quantificação do ácido propiônico, sendo necessária a etapa de extração desse ácido do pão que comumente se encontra sob a forma de propionato de cálcio. Foram feitos vários testes utilizando-se métodos de extração por arraste à vapor, extração etérea contendo 3% de ácido ortofosfórico e extração direta com solução aquosa de ácido fórmico 0,05 M. Foram também testadas várias colunas cromatográficas e verificado o comportamento do ácido isovalérico, ácido butírico e metil isobutil cetona (MIBK) como padrões internos. Em função dos melhores resultados obtidos, optou-se por extrair o ácido propiônico com solução etérea de ácido fórmico 3%, usando ácido butírico como padrão interno e análise cromatográfica na coluna DEGS. A recuperação média de 97,4% foi obtida com propionato de cálcio padrão e de 85,0% com amostras de pão adicionadas de propionato de cálcio. O método mostrou-se preciso, rápido e de fácil execução

Simulation of the diffusion process of NaCl and KCl in olive pulp using the finite element method

The simultaneous diffusion of inorganic components in the olive pulp in wet brine was modeled based on Fick’s generalized 2nd Law and simulated using the finite element method. The main and crossed diffusion coefficients, the film coefficient and the Biot number were determined, with the application of the simplex optimization method, through the minimization of the percentage errors. The errors between the simulated and experimental data were 5.35% for NaCl and 4.77% for KCl and the adjusted main diffusion coefficients were 0.4358 × 10-12 m2 s-1 for NaCl and 0.5408 × 10-12 m2 s-1 for KCl. The system developed to simulate diffusion allows the control and modulation of the salts content that diffuses through the olive pulp