Acompanhamento da construção de obras de arte

No âmbito do Trabalho Final de Mestrado (TFM), propus a realização do presente relatório de estágio para obtenção do grau de Mestre em Engenharia Civil, inserido no perfil de estruturas do mestrado de engenharia civil, do Instituto Superior de Engenharia de Lisboa. Para realização deste relatório, realizei um estágio com o período de quatro meses, na Mota-Engil, Engenharia e Construção, SA, onde estive integrado na obra de construção do sublanço da estrada denominada “EN238 – Sertã/ Oleiros” integrado na subconcessão do Pinhal Interior (Lote 10). A obra localiza-se na província da Beira Baixa, no distrito de Castelo Branco e abrange os concelhos de Sertã e Oleiros. Neste trabalho desenvolveram-se essencialmente duas vertentes da área da Engenharia: “Coordenação” e “Construção”.

Cholesterol 24S-Hydroxylase overexpression inhibits the liver X receptor (LXR) pathway by activating small guanosine triphosphate-binding proteins (sGTPases) in neuronal cells

The neuronal-specific cholesterol 24S-hydroxylase (CYP46A1) is important for brain cholesterol elimination. Cyp46a1 null mice exhibit severe deficiencies in learning and hippocampal long-term potentiation, suggested to be caused by a decrease in isoprenoid intermediates of the mevalonate pathway. Conversely, transgenic mice overexpressing CYP46A1 show an improved cognitive function. These results raised the question of whether CYP46A1 expression can modulate the activity of proteins that are crucial for neuronal function, namely of isoprenylated small guanosine triphosphate-binding proteins (sGTPases). Our results show that CYP46A1 overexpression in SH-SY5Y neuroblastoma cells and in primary cultures of rat cortical neurons leads to an increase in 3-hydroxy-3-methyl-glutaryl-CoA reductase activity and to an overall increase in membrane levels of RhoA, Rac1, Cdc42 and Rab8. This increase is accompanied by a specific increase in RhoA activation. Interestingly, treatment with lovastatin or a geranylgeranyltransferase-I inhibitor abolished the CYP46A1 effect. The CYP46A1-mediated increase in sGTPases membrane abundance was confirmed in vivo, in membrane fractions obtained from transgenic mice overexpressing this enzyme. Moreover, CYP46A1 overexpression leads to a decrease in the liver X receptor (LXR) transcriptional activity and in the mRNA levels of ATP-binding cassette transporter 1, sub-family A, member 1 and apolipoprotein E. This effect was abolished by inhibition of prenylation or by co-transfection of a RhoA dominant-negative mutant. Our results suggest a novel regulatory axis in neurons; under conditions of membrane cholesterol reduction by increased CYP46A1 expression, neurons increase isoprenoid synthesis and sGTPase prenylation. This leads to a reduction in LXR activity, and consequently to a decrease in the expression of LXR target genes.