NOX2, p22phox and p47phox are targeted to the nuclear pore complex in ischemic cardiomyocytes colocalizing with local reactive oxygen species.

BACKGROUND: NADPH oxidases play an essential role in reactive oxygen species (ROS)-based signaling in the heart. Previously, we have demonstrated that (peri)nuclear expression of the catalytic NADPH oxidase subunit NOX2 in stressed cardiomyocytes, e.g. under ischemia or high concentrations of homocysteine, is an important step in the induction of apoptosis in these cells. Here this ischemia-induced nuclear targeting and activation of NOX2 was specified in cardiomyocytes. METHODS: The effect of ischemia, mimicked by metabolic inhibition, on nuclear localization of NOX2 and the NADPH oxidase subunits p22(phox) and p47(phox), was analyzed in rat neonatal cardiomyoblasts (H9c2 cells) using Western blot, immuno-electron microscopy and digital-imaging microscopy. RESULTS: NOX2 expression significantly increased in nuclear fractions of ischemic H9c2 cells. In addition, in these cells NOX2 was found to colocalize in the nuclear envelope with nuclear pore complexes, p22(phox), p47(phox) and nitrotyrosine residues, a marker for the generation of ROS. Inhibition of NADPH oxidase activity, with apocynin and DPI, significantly reduced (peri)nuclear expression of nitrotyrosine. CONCLUSION: We for the first time show that NOX2, p22(phox) and p47(phox) are targeted to and produce ROS at the nuclear pore complex in ischemic cardiomyocytes

Involvement of PPAR-γ in the neuroprotective and anti-inflammatory effects of angiotensin type 1 receptor inhibition: effects of the receptor antagonist telmisartan and receptor deletion in a mouse MPTP model of Parkinson's disease

<p>Abstract</p> <p>Background</p> <p>Several recent studies have shown that angiotensin type 1 receptor (AT1) antagonists such as candesartan inhibit the microglial inflammatory response and dopaminergic cell loss in animal models of Parkinson's disease. However, the mechanisms involved in the neuroprotective and anti-inflammatory effects of AT1 blockers in the brain have not been clarified. A number of studies have reported that AT1 blockers activate peroxisome proliferator-activated receptor gamma (PPAR γ). PPAR-γ activation inhibits inflammation, and may be responsible for neuroprotective effects, independently of AT1 blocking actions.</p> <p>Methods</p> <p>We have investigated whether oral treatment with telmisartan (the most potent PPAR-γ activator among AT1 blockers) provides neuroprotection against dopaminergic cell death and neuroinflammation, and the possible role of PPAR-γ activation in any such neuroprotection. We used a mouse model of parkinsonism induced by the dopaminergic neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and co-administration of the PPAR-γ antagonist GW9662 to study the role of PPAR-γ activation. In addition, we used AT1a-null mice lesioned with MPTP to study whether deletion of AT1 in the absence of any pharmacological effect of AT1 blockers provides neuroprotection, and investigated whether PPAR-γ activation may also be involved in any such effect of AT1 deletion by co-administration of the PPAR-γ antagonist GW9662.</p> <p>Results</p> <p>We observed that telmisartan protects mouse dopaminergic neurons and inhibits the microglial response induced by administration of MPTP. The protective effects of telmisartan on dopaminergic cell death and microglial activation were inhibited by co-administration of GW9662. Dopaminergic cell death and microglial activation were significantly lower in AT1a-null mice treated with MPTP than in mice not subjected to AT1a deletion. Interestingly, the protective effects of AT1 deletion were also inhibited by co-administration of GW9662.</p> <p>Conclusion</p> <p>The results suggest that telmisartan provides effective neuroprotection against dopaminergic cell death and that the neuroprotective effect is mediated by PPAR-γ activation. However, the results in AT1-deficient mice show that blockage of AT1, unrelated to the pharmacological properties of AT1 blockers, also protects against dopaminergic cell death and neuroinflammation. Furthermore, the results show that PPAR-γ activation is involved in the anti-inflammatory and neuroprotective effects of AT1 deletion.</p

Hidrolisado proteico de resíduo de sardinha como atrativo alimentar para juvenis de jundiá

RESUMO O objetivo deste estudo foi avaliar a utilização do hidrolisado proteico de resíduo de sardinha como atrativo na alimentação do Rhamdia quelen. No experimento 1, foram utilizados os seguintes atrativos alimentares: 1. extrato aquoso de músculo de tilápia-do-Nilo (controle positivo); 2. hidrolisado proteico de resíduo de sardinha com baixo grau de hidrólise (GH); 3. hidrolisado proteico de resíduo de sardinha com alto GH; 4. hidrolisado proteico de resíduo de sardinha com alto GH diluído (10% da concentração) e 5. controle usando somente água destilada. Após jejum de 48 horas, o comportamento foi registrado em vídeo por um período basal de dois minutos e por mais 18 minutos após a inoculação do atrativo. O delineamento foi inteiramente ao acaso, com três tratamentos e 20 repetições. O experimento 2 foi realizado para avaliar a capacidade do hidrolisado proteico de estimular a ingestão de alimento em juvenis de jundiá. Para isso, foram confeccionados pellets de ágar contendo ou não hidrolisado proteico de resíduo de sardinha. Os peixes foram avaliados individualmente e tiveram um período de adaptação de sete dias. Os resultados foram analisados por meio do teste de proporção de Goodman (1964). A inoculação dos hidrolisados com alto e baixo GH aumentou o tempo de movimentação dos barbilhões. O hidrolisado com alto GH diluído proporcionou os mesmos resultados que o hidrolisado com baixo GH , mas as médias não diferiram das obtidas para a água destilada (controle negativo) e do extrato de músculo. O incremento na movimentação de um lado para outro do aquário foi maior (P<0,05) para os hidrolisados com alto e baixo GH. No experimento 2, a proporção de peixes que ingeriu os pellets contendo hidrolisado proteico de resíduo de sardinha com alto GH foi maior (P<0,05) em relação aos que ingeriram os pellets contendo água destilada. O hidrolisado proteico foi eficiente para estimular o comportamento associado à alimentação em juvenis de Rhamdia quelen

Determinação do fluxo gênico em Brachiaria sp. usando marcadores microssatélites.

O objetivo deste estudo é estimar a taxa de fluxo gênico em Brachiaria por dispersão de pólen de B. brizantha cv. Marandu para auxiliar no estabelecimento de padrões de biossegurança para futuros ensaios de campo com Brachiaria geneticamente modificada