Our study was designed using a case–control approach. Sixty pre-eclamptic patients, 60 healthy pregnant women with uncomplicated pregnancies and 59 healthy non-pregnant women were involved in the BMN 673 manufacturer study. The study participants were enrolled from the First Department of Obstetrics and Gynecology and from the Department of Obstetrics and Gynecology of Kútvölgyi Clinical Center, at the Semmelweis University, Budapest, Hungary. All women were Caucasian and resided in the same geographic area in Hungary. Exclusion criteria were multi-fetal gestation, chronic hypertension,

diabetes mellitus, autoimmune disease, angiopathy, renal disorder, maternal or fetal infection and fetal congenital anomaly. The women were fasting; none of the pregnant women were in active labour, and none had rupture of membranes. The healthy non-pregnant women were in the early follicular phase of the menstrual cycle (between cycle days 3 and 5), and none of them received hormonal contraception. Pre-eclampsia was defined by increased blood pressure (≥140 mmHg systolic or ≥90 mmHg diastolic on ≥2 occasions at least 6 h apart) that occurred after 20 weeks of gestation in women with previously normal

blood pressure, accompanied by proteinuria (≥0·3 g/24 h or ≥1 + on dipstick in the absence of urinary tract infection). Protein kinase N1 Blood pressure returned to normal by 12 weeks postpartum in each pre-eclamptic study patient. Pre-eclampsia was regarded as severe if any of the following criteria was present: blood pressure ≥ 160 mmHg LY2606368 supplier systolic or ≥110 mmHg diastolic, or proteinuria ≥ 5 g/24 h

(or ≥3 + on dipstick). Pregnant women with eclampsia or HELLP (haemolysis, elevated liver enzymes and low platelet count) syndrome were not enrolled into this study. Early onset of pre-eclampsia was defined as onset of the disease before 34 weeks of gestation (between 20 and 33 completed gestational weeks). Fetal growth restriction was diagnosed if the fetal birth weight was below the 10th percentile for gestational age and gender, based on Hungarian birth weight percentiles. The study protocol was approved by the Regional and Institutional Committee of Science and Research Ethics of the Semmelweis University, and written informed consent was obtained from each patient. The study was conducted in accordance with the Declaration of Helsinki. Blood samples were taken from an antecubital vein into plain tubes, as well as ethylenediamine tetraacetic acid (EDTA) or sodium citrate anti-coagulated tubes, and then centrifuged at room temperature with a relative centrifugal force of 3000 g for 10 min. The aliquots of serum and plasma were stored at −80°C until the measurements.