Pulmonary arterial hypertension in heroin users.

endothelial cells (HAECs) subject to normal pulsatile flow or to flow associated with continuous-flow ventricular assist device (CFVAD) support. (B) Quantitative real-time polymerase chain reaction analysis of anti-oxidant gene expression in HAECs cultured under normal pulsatile flow or flow associated with CFVAD support maintained for 96 hours. Total messenger RNA (mRNA) from HAECs was analyzed and mRNA levels of antioxidant genes Gclc, Gclm, GSR, GPX1, Catalase (CAT), SOD1, SOD2, and G6PD were determined. The relative mRNA levels were determined by normalizing the Ct values of control cells (normal flow) with mRNA levels of reference gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The data represent mean standard error of the mean of relative mRNA levels of respective genes. *p o 0.05 for a sample size of n 1⁄4 4. (C) Immunoblots of representative anti-oxidant proteins glutamate–cysteine ligase catalytic sub-unit (GCLC), catalase, glucose-6-phosphate dehydrogenase (G6PD), catalase, NAD(P)H-quinone oxidoreductase 1 (NQO1), glutathione peroxidase 1 (GPx1), superoxide dismutase (SOD1 and SOD2) in HAECs cultured under normal pulsatile flow or flow associated with CFVAD support. Bar graphs represent the mean SEM of relative density units of band intensity of immunoreactive signal of antioxidant gene products with respect to band intensity of the housekeeping gene, GAPDH. *p o 0.05. for a sample size of n 1⁄4 4. The Journal of Heart and Lung Transplantation, Vol 35, No 7, July 2016 932