Modulation of the oscillatory mechanics of lung tissue and the oxidative stress response induced by arginase inhibition in a chronic allergic inflammation model

Abstract\ud \ud \ud \ud Background\ud The importance of the lung parenchyma in the pathophysiology of asthma has previously been demonstrated. Considering that nitric oxide synthases (NOS) and arginases compete for the same substrate, it is worthwhile to elucidate the effects of complex NOS-arginase dysfunction in the pathophysiology of asthma, particularly, related to distal lung tissue. We evaluated the effects of arginase and iNOS inhibition on distal lung mechanics and oxidative stress pathway activation in a model of chronic pulmonary allergic inflammation in guinea pigs.\ud \ud \ud \ud Methods\ud Guinea pigs were exposed to repeated ovalbumin inhalations (twice a week for 4 weeks). The animals received 1400 W (an iNOS-specific inhibitor) for 4 days beginning at the last inhalation. Afterwards, the animals were anesthetized and exsanguinated; then, a slice of the distal lung was evaluated by oscillatory mechanics, and an arginase inhibitor (nor-NOHA) or vehicle was infused in a Krebs solution bath. Tissue resistance (Rt) and elastance (Et) were assessed before and after ovalbumin challenge (0.1%), and lung strips were submitted to histopathological studies.\ud \ud \ud \ud Results\ud Ovalbumin-exposed animals presented an increase in the maximal Rt and Et responses after antigen challenge (p<0.001), in the number of iNOS positive cells (p<0.001) and in the expression of arginase 2, 8-isoprostane and NF-kB (p<0.001) in distal lung tissue. The 1400 W administration reduced all these responses (p<0.001) in alveolar septa. Ovalbumin-exposed animals that received nor-NOHA had a reduction of Rt, Et after antigen challenge, iNOS positive cells and 8-isoprostane and NF-kB (p<0.001) in lung tissue. The activity of arginase 2 was reduced only in the groups treated with nor-NOHA (p <0.05). There was a reduction of 8-isoprostane expression in OVA-NOR-W compared to OVA-NOR (p<0.001).\ud \ud \ud \ud Conclusions\ud In this experimental model, increased arginase content and iNOS-positive cells were associated with the constriction of distal lung parenchyma. This functional alteration may be due to a high expression of 8-isoprostane, which had a procontractile effect. The mechanism involved in this response is likely related to the modulation of NF-kB expression, which contributed to the activation of the arginase and iNOS pathways. The association of both inhibitors potentiated the reduction of 8-isoprostane expression in this animal model.FAPESP and LIM20HCFMUSP.FAPESP and LIM-20-HC-FMUSP

Desenvolvimento e avaliação de massa tipo macarrão a base de milho e quinoa para celiacos

Orientador: Jaime Amaya FarfanDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de AlimentosResumo: Doença celíaca é uma intolerância alimentar ao glúten, causada pela ingestão de cereais que contêm essa proteína, como trigo, amido de trigo, centeio, cevada, triticale e, provavelmente, aveia. O único tratamento efetivo contra a doença é a exclusão total da dieta de produtos elaborados a partir dos grãos considerados tóxicos. Essa exclusão pode ser uma tarefa difícil devido à grande quantidade de produtos industrializados e, até mesmo, caseiros preparados a partir desses grãos. No caso do trigo, a exclusão ou substituição desta matéria-prima é particularmente complexa, em função de suas propriedades viscoelásticas únicas. Pseudo cereal isento de glúten, originário da região andina sul americana, a quinoa (Chenopodium quino8, Willd), é uma alternativa para a citada substituição muito atraente para celíacos e pessoas não portadoras de tal doença. Apresenta elevado teor de proteína e pode satisfazer quase completamente, em termos de vitaminas e minerais, o requerimento recomendado pelo "Commitee on Dietary Allowances". O objetivo deste trabalho foi desenvolver um produto tipo macarrão de milho e quinoa para qualquer indivíduo, especialmente os celíacos. Foram avaliados os parâmetros de: qualidade de cozimento (perda de sólidos, aumento de peso, aumento de volume), textura (adesividade e elasticidade), viscosidade (viscosidade de pico e viscosidade final) e umidade da mistura de farinhas pré-gelatinizadas. Dos parâmetros estudados, verificou-se que a perda de sólidos, aumento de peso e volume, adesividade e umidade da farinha tratada termicamente variaram em torno de uma média. Esses valores médios foram: 16,2% de perda de sólidos, aumento de 2,3 vezes para o peso, aumento de 2,6 vezes para o volume, adesividade de 2090,7g e umidade de 38,4% na farinha tratada termicamente. Os resultados de elasticidade e viscosidade de pico forneceram indicações relevantes quanto à formulação e processamento, mas não permitiram chegar a conclusões definitivas, em função do processo ainda não apresentar alta reprodutibilidade. Já a viscosidade final teve modelo significativo (p=O,0657) e, portanto, preditivo. A combinação das variáveis independentes que implicou no maior valor de elasticidade, característica desejável para o produto em questão, foi baixo a médio tempo de tratamento térmico, baixa à média umidade de condicionamento e de baixa à média porcentagem de quinoa. Para a viscosidade de pico, todos os valores obtidos podem ser considerados baixos em relação à farinha inicial não tratada, e, portanto, qualquer combinação de tempo de tratamento térmico, umidade de condicionamento e porcentagem de quinoa pode ser considerada boa, devendo¬se escolher a mais conveniente. Já para a viscosidade final, a melhor combinação foi baixo teor de quinoa (1,6 a 5%) e toda a faixa de tempo de tratamento térmico ou toda faixa de quinoa e tempo de tratamento térmico de aproximadamente 4 minutos. A média obtida para a aceitação global do produto foi 6,83, que se encontra entre "gostei ligeiramente" e "gostei moderadamente". Com relação à intenção de compra, 50% dos provadores "possivelmente comprariam". Isso mostra um considerável potencial de mercado para o produto.Abstract: Celiac disease can be understood as an intolerance to gluten exhibited by certain individuais upon ingestion of such cereais as wheat, wheat starch, rye, barley, triticale and probably oats, which contain this class of proteins. The only effective treatment to date, is the complete exclusion from the diet of products known to be toxic. Exclusion of these grains from a standard diet can be a difficult task for the patient, given the wide spectrum of industrialized and home-made products of the Western diet that include those ingredients. The substitution of wheat requires the use of special technical strategies because of its unique viscoelastic properties. Quinoa (Chenopodium quinoa Willd), a pseudo cereal original of the high Andes that has been adapted to the low lands of the Brazilian "cerrado" is reportedly gluten-free and should be explored as a cereal food substitute for celiac patients. Quinoa products have high contents of protein, and can furnish most of the recommendations for vitamins and minerais. The objective of this work was to assess the potential of quinoa flour in the development of a spaghetti prototype, using corn and quinoa flours as ingredients. Cooking quality (loss of solids, volume and weight increments), texture (adhesiveness, elasticity), viscosity (peak and final viscosities) and moisture contents of the pre-gelatinized flour mixtures were the physical parameters studied. Of these, only texture did not vary, about a mean. Mean values were 16.2%, 2.3 times, 2.6 times, 2090.7g and 38.4% for loss of solids, weight increase, volume increase, adhesiveness and moisture, respectively. The results from elasticity and peak viscosities provided information relevant to both formulation and processing of the product, but did not permit to draw more definite conclusions due to the low reproducibility of the processo The final viscosity, in turn, afforded a significant model (p=0,0657) and was, therefore, predictive. The combination of independent variables that resulted in higher elasticity values, desirable characteristic for the target product, was short-to-medium thermal treatment, low-to-medium conditioning moisture and low to-medium quinoa level of addition. For peak viscosity, all values obtained could be considered low with respect to the unreated, initial flour. Thus, any value could be satisfatory and convenience will influence selection. For the final viscosity, however, the best combinations were low (1,6 a 5%) quinoa content and the entire range of time of thermal treatment or, alternatively, the entire range of quinoa content and a thermal treatment of about 4 minutes. the mean sensory acceptance value was 6,83 placed the product between "I liked it slightly" and "I liked it moderately". Regarding the intention of buying the product, 50% of the tasters declared they "would probably buy it", which shows a substantial potential market for the product.MestradoMestre em Ciência da Nutriçã

Modulation of Bone Morphogenetic Protein-9 Expression and Processing by Insulin, Glucose, and Glucocorticoids: Possible Candidate for Hepatic Insulin-Sensitizing Substance

Bone morphogenetic protein 9 (BMP-9), a member of the TGF-beta superfamily predominantly expressed in nonparenchymal liver cells, has been demonstrated to improve glucose homeostasis in diabetic mice. Along with this therapeutic effect, BMP-9 was proposed as a candidate for the hepatic insulin-sensitizing substance ( HISS). Whether BMP-9 plays a physiological role in glucose homeostasis is still unknown. In the present study, we show that BMP-9 expression and processing is severely reduced in the liver of insulin-resistant rats. BMP-9 expression and processing was directly stimulated by in situ exposition of the liver to the combination of glucose and insulin and oral glucose in overnight fasted rats. Additionally, prolonged fasting ( 72 h) abrogated refeeding-induced BMP-9 expression and processing. Previous exposition to dexamethasone, a known inductor of insulin resistance, reduced BMP-9 processing stimulated by the combination of insulin and glucose. Finally, we show that neutralization of BMP-9 with an anti-BMP-9 antibody induces glucose intolerance and insulin resistance in 12-h fasted rats. Collectively, the present results demonstrate that BMP-9 plays an important role in the control of glucose homeostasis of the normal rat. Additionally, BMP-9 is expressed and processed in an HISS-like fashion, which is impaired in the presence of insulin resistance. BMP-9 regulation according to the feeding status and the presence of diabetogenic factors reinforces the hypothesis that BMP-9 might exert the role of HISS in glucose homeostasis physiology. ( Endocrinology 149: 6326-6335, 2008)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)CNPq Conselho Nacional de Desenvolvimento Cientifico e TecnologicoConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPES

Ginkgo biloba Extract Improves Insulin Signaling and Attenuates Inflammation in Retroperitoneal Adipose Tissue Depot of Obese Rats

Due to the high incidence and severity of obesity and its related disorders, it is highly desirable to develop new strategies to treat or even to prevent its development. We have previously described that Ginkgo biloba extract (GbE) improved insulin resistance and reduced body weight gain of obese rats. In the present study we aimed to evaluate the effect of GbE on both inflammatory cascade and insulin signaling in retroperitoneal fat depot of diet-induced obese rats. Rats were fed with high fat diet for 2 months and thereafter treated for 14 days with 500 mg/kg of GbE. Rats were then euthanized and samples from retroperitoneal fat depot were used for western blotting, RT-PCR, and ELISA experiments. The GbE treatment promoted a significant reduction on both food/energy intake and body weight gain in comparison to the nontreated obese rats. In addition, a significant increase of both Adipo R1 and IL-10 gene expressions and IR and Akt phosphorylation was also observed, while NF-κB p65 phosphorylation and TNF-α levels were significantly reduced. Our data suggest that GbE might have potential as a therapy to treat obesity-related metabolic diseases, with special interest to treat obese subjects resistant to adhere to a nutritional education program

Acute lung injury is reduced by the alpha 7nAChR agonist PNU-282987 through changes in the macrophage profile

Nicotinic alpha-7 acetylcholine receptor (nAChR alpha 7) is a critical regulator of cholinergic anti-inflammatory actions in several diseases, including acute respiratory distress syndrome (ARDS). Given the potential importance of alpha 7nAChR as a therapeutic target, we evaluated whether PNU-282987, an alpha 7nAChR agonist, is effective in protecting the lung against inflammation. We performed intratracheal instillation of LPS to generate acute lung injury (ALI) in C57BL/6mice. PNU-282987 treatment, either before or after ALI induction, reduced neutrophil recruitment and IL1 beta, TNF-alpha, IL-6, keratinocyte chemoattractant (KC), and IL-10 cytokine levels in the bronchoalveolar lavage fluid (P<0.05). In addition, lung NF-kappa B phosphorylation decreased, along with collagen fiber deposition and the number of matrix metalloproteinase-9(+) and -2(+) cells, whereas the number of tissue inhibitor of metalloproteinase-1(+) cells increased (P < 0.05). PNU-282987 treatment also reduced lung mRNA levels and the frequency of M1 macrophages, whereas cells expressing the M2-related markers CD206 and IL-10 increased, suggesting changes in the macrophage profile. Finally, PNU-282987 improved lung function in LPS-treated animals. The collective results suggest that PNU-282987, anagonist of alpha 7nAChR, reduces LPS-induced experimental ALI, thus supporting the notion that drugs that act on alpha 7nAChRs should be explored for ARDS treatment in humans.-Pinheiro, N. M., Santana, F. P. R., Almeida, R. R., Guerreiro, M., Martins, M. A., Caperuto, L. C., Camara, N. O. S., Wensing, L. A., Prado, V. F., Tiberio, I. F. L. C., Prado, M. A. M., Prado, C. M. Acute lung injury is reduced by the alpha 7nAChR agonist PNU-282987 through changes in the macrophage profile.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Univ Sao Paulo, Dept Med, Sch Med, Sao Paulo, BrazilUniv Sao Paulo, Dept Immunol, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Biol Sci, Diadema, BrazilUniv Western Ontario, Dept Physiol & Pharmacol, London, ON, CanadaUniv Western Ontario, Dept Anat & Cell Biol, London, ON, CanadaUniv Fed Sao Paulo, Dept Biosci, Santos, BrazilDepartment of Biological Science, Universidade Federal de São Paulo, Diadema, BrazilDepartment of Bioscience, Universidade Federal de São Paulo (UNIFESP), Rua Silva Jardim, 136 Vila Mathias, Santos SP, Brazil, 11015-020FAPESP: 2008/55359-5FAPESP: 2012/02270-2FAPESP: 2013/02881-4FAPESP: 2014/25689-4CNPq: 471224/2009-0CNPq: 476877/2012-1CNPq: 304465/2012-7Web of Scienc

Congress of Chemical Biology and Symposium of Biological Sciences Department

Essa publicação é a compilação dos resumos científicos apresentados no "III Congresso do Programa de Pós-Graduação em Biologia Química e IV Simpósio do Departamento de Ciências Biológicas"Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Effects of VAChT reduction and α7nAChR stimulation by PNU-282987 in lung inflammation in a model of chronic allergic airway inflammation

The cholinergic anti-inflammatory pathway has been shown to regulate lung inflammation and cytokine release in acute models of inflammation, mainly via α7 nicotinic receptor (α7nAChR). We aimed to evaluate the role of endogenous acetylcholine in chronic allergic airway inflammation in mice and the effects of therapeutic nAChR stimulation in this model. We first evaluated lung inflammation and remodeling on knock-down mice with 65% of vesicular acetylcholine transport (VAChT) gene reduction (KDVAChT) and wild-type(WT) controls that were subcutaneously sensitized and then inhaled with ovalbumin(OVA). We then evaluated the effects of PNU-282987(0.5-to-2mg/kg),(α7nAChR agonist) treatment in BALB/c male mice intraperitoneal sensitized and then inhaled with OVA. Another OVA-sensitized-group was treated with PNU-282987 plus Methyllycaconitine (MLA,1 mg/kg, α7nAChR antagonist) to confirm that the effects observed by PNU were due to α7nAChR. We showed that KDVAChT-OVA mice exhibit exacerbated airway inflammation when compared to WT-OVA mice. In BALB/c, PNU-282987 treatment reduced the number of eosinophils in the blood, BAL fluid, and around airways, and also decreased pulmonary levels of IL-4,IL-13,IL-17, and IgE in the serum of OVA-exposed mice. MLA pre-treatment abolished all the effects of PNU-282987. Additionally, we showed that PNU-282987 inhibited STAT3-phosphorylation and reduced SOCS3 expression in the lung. These data indicate that endogenous cholinergic tone is important to control allergic airway inflammation in a murine model. Moreover, α7nAChR is involved in the control of eosinophilic inflammation and airway remodeling, possibly via inhibition of STAT3/SOCS3 pathways. Together these data suggest that cholinergic anti-inflammatory system mainly α7nAChR should be further considered as a therapeutic target in asthma.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)FAPESP: 2018/15738-9FAPESP: 08/55359-5FAPESP: 14/25689-4CNPq: 476877/2012-