Receptores envolvidos na produção de citocinas e eicosanóides por monócitos e neutrófilos humanos em resposta ao Paracoccidioides brasiliensis

Pós-graduação em Doenças Tropicais - FMB

Resveratrol na apoptose e produção de superóxido em neutrófilos de cães

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Leucograma e metabolismo oxidativo de neutrófilos em cabras da raça Saanen nos períodos de gestação, parto e pós-parto

The purpose of this study was to evaluate the leukocyte count and the oxidative metabolism of neutrophil in Saanen goats during periods of pregnancy, parturition and postpartum. Were used 20 Saanen goats, clinically healthy and serologically negative for caprine arthritis encephalitis virus (CAEV). Blood samples were collected by jugular venipuncture 49 (M1), 42 (M2), 35 (M3), 28 (M4), 21 (M5), 14 (M6), seven (M7), three (M8) days before the parturition, on the day of birth (M9), three (M10) and seven (M11) days postpartum, for the leukocyte count, and serum for cortisol, estradiol and progesterone determination. From 28 days (M4) before parturition until seven days postpartum (M11) blood samples were collected for evaluation of oxidative metabolism of neutrophils by the nitroblue tetrazolium reduction test (NBT). The results showed that at parturition day there were an increase in cortisol and estradiol levels and a decrease in progesterone serum, neutrophilic leukocytosis and left shift slight, decrease of lymphocytes, increase in the neutrophil: lymphocyte, eosinopenia, monocytosis and basophilia. There was a neutrophilic leukocytosis and an increase in the neutrophil: lymphocyte on the seventh day postpartum. There were not significant alterations in oxidative metabolism of neutrophils during pregnancy, parturition and postpartum. It was concluded that parturition causes an elevation in cortisol and estradiol levels and a decrease in progesterone serum determining a neutrophilic leukocytosis and left shift slight, with a reduction of lymphocytes, increase in the neutrophil:lymphocyte, eosinopenia, monocytosis and basophilia. Neutrophilic leukocytosis, increase in the neutrophil: lymphocyte and fibrinogen are detected on the seventh day postpartum. Pregnancy, parturition and the postpartum do not change the oxidative metabolism of neutrophils evaluated by NBT reduction test.

Papel de neutrófilos humanos no direcionamento da resposta imune adaptativa do tipo Th1 contra o Paracoccidioides brasiliensis

Pós-graduação em Doenças Tropicais - FMB

Participação das NETs (neutrophil extracellular traps) na atividade fungicida de neutrófilos humanos contra o Paracoccidioides brasiliensis

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

Síntese e estudo da diapocinina como um inibidor da NADPH Oxidase em neutrófilos e células mononucleares de sangue periférico

Apocynin, a methoxy-catechol originally extracted from the root of Picrorhiza kurroa, has been used as an inhibitor of the NADPH oxidase complex in phagocytic and nonphagocytic cells. Its mechanism of inhibition is linked to their prior activation through the action of peroxidases leading to oxidation of the dimeric product, diapocynin. In this study, dipocinina was synthesized and investigated its effect as an inhibitor of activation NADPH oxidase in neutrophils (PMN) and peripheral blood mononuclear cells (PBMC). The synthesis of diapocinina was performed by oxidation of apocinina by potassium persulphate in the midst of water for 5 minutes at room temperature. The precipitate was filtered and washed with water and methanol. Diapocinina was characterized by mass spectrometry. PMN and PBMC were obtained from peripheral blood of healthy donors and purified for gelatin sedimentation, or centrifugation with Histopaque ®, the red cells were lysed with ice water or ammonium chloride. Diapocinina or apocinina were incubated with opsonized zymosan, activation of PMNs and release of superoxide anion, these monitored by chemiluminescent assay dependent lucigenina. We found that diapocinina inhibitor was no better than the apocinina in PMN. However, diapocinina was more efficient than apocinina as an inhibitor of NADPH oxidase in PBMC. In conclusion, whereas PBMC are relatively poor compared with peroxidases PMN, our results are consistent with the need for oxidation apocinina for its effect as an inhibitor of NADPH oxidase

Apocinina versus diapocinina como inibidor da produção de peróxido de hidrogênio e ácido hipocloroso por neutrófilos ativados

Apocynin has been used as an efficient inhibitor of the multi-enzymatic complex NADPH oxidase in many experimental models involving phagocytic and nonphagocytic cells. The mechanism of inhibition has been linked with the previous activation of apocynin through the action of cellular peroxidases leading to the formation of a dimeric oxidation product, diapocynin. In this study we compared apocynin with pure diapocynin regarding their effects as scavenger of hydrogen peroxide and hypochlorous generated by glucose/glucose oxidase and myeloperoxidase respectively, and as inhibitors of the production of hydrogen peroxide and hypochlorous acid by activated neutrophils. The production of hydrogen peroxide was measured by the oxidation of the fluorescent substance Amplex Red and the production of hypochlorous acid by was measured as taurine-chloramine derivative using the chromogenic substrate 3,3’,5,5’- tetramethylbenzidine (TMB). Neutrophils (1 x106 cells/mL) were pre-incubated in PBS buffer supplemented with 1 mM calcium chloride, 0.5 mM magnesium chloride, 1 mg/mL glucose and 5 mM taurine in the presence or absence of inhibitors. The reactions were triggered by adding the soluble stimulus Forbol Miristate Acetate PMA or zymosan and incubated by additional 30 minutes. We found that pure diapocynin was not better than apocynin regarding its scavenger and inhibitory properties. These results suggest that the formation of diapocynin is not essential for the action of apocynin as inhibitor of NADPH oxidase activation

Avaliação do envolvimento do receptor dectina-1 na atividade fungicida e na produção de metabólitos do oxigênio por neutrófilos e monócitos humanos desafiados com Paracoccidioides brasiliensis

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

Identificação de Neutrophil Extracellular Traps em lesões tegumentares de pacientes com Paracoccidiodomicose e análise da formação de NETs in vitro por neutrófilos humanos desafiados com Paracoccidioides brasiliensis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Avaliação da lipocalina associada à gelatinase de neutrófilos (NGAL) em idosos após osteossíntese de fêmur

Pós-graduação em Anestesiologia - FMB

Avaliação in vitro do P-cresol sobre o metabolismo oxidativo e apoptose dos neutrófilos de cães

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

Efeito das toxinas urêmicas guanidina e ácido guanidinoácetico sobre o metabolismo oxidativo e apoptose em neutrófilos de cães

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

Efeito do plasma rico em uréia na apoptose de neutrófilos de cães

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

Efeito do plasma rico em glicose na apoptose de neutrófilos de cães

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

Efeito do edta sobre o teste de redução do tetrazólio nitroazul (NBT) para avaliação do metabolismo oxidativo dos neutrófilos de equinos

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