Ragnhild Bergene Skråstad

Metabolomic Biomarkers in Serum and Urine in Women with Preeclampsia

Objective To explore the potential of magnetic resonance (MR) metabolomics for study of preeclampsia, for improved phenotyping and elucidating potential clues to etiology and pathogenesis. Methods Urine and serum samples from pregnant women with preeclampsia (n = 10), normal pregnancies (n = 10) and non-pregnant women (n = 10) matched by age and gestational age were analyzed with MR spectroscopy and subjected to multivariate analysis. Metabolites were then quantified and compared between groups. Results Urine and serum samples revealed clear differences between women with preeclampsia and both control groups (normal pregnant and non-pregnant women). Nine urine metabolites were significantly different between preeclampsia and the normal pregnant group. Urine samples from women with early onset preeclampsia clustered together in the multivariate analysis. The preeclampsia serum spectra showed higher levels of low and very-low density lipoproteins and lower levels of high-density lipoproteins when compared to both non-pregnant and normal pregnant women. Conclusion The MR determined metabolic profiles in urine and serum from women with preeclampsia are clearly different from normal pregnant women. The observed differences represent a potential to examine mechanisms underlying different preeclampsia phenotypes in urine and serum samples in larger studies. In addition, similarities between preeclampsia and cardiovascular disease in metabolomics are demonstrated.

First Trimester Urine and Serum Metabolomics for Prediction of Preeclampsia and Gestational Hypertension: A Prospective Screening Study

Hypertensive disorders of pregnancy, including preeclampsia, are major contributors to maternal morbidity. The goal of this study was to evaluate the potential of metabolomics to predict preeclampsia and gestational hypertension from urine and serum samples in early pregnancy, and elucidate the metabolic changes related to the diseases. Metabolic profiles were obtained by nuclear magnetic resonance spectroscopy of serum and urine samples from 599 women at medium to high risk of preeclampsia (nulliparous or previous preeclampsia/gestational hypertension). Preeclampsia developed in 26 (4.3%) and gestational hypertension in 21 (3.5%) women. Multivariate analyses of the metabolic profiles were performed to establish prediction models for the hypertensive disorders individually and combined. Urinary metabolomic profiles predicted preeclampsia and gestational hypertension at 51.3% and 40% sensitivity, respectively, at 10% false positive rate, with hippurate as the most important metabolite for the prediction. Serum metabolomic profiles predicted preeclampsia and gestational hypertension at 15% and 33% sensitivity, respectively, with increased lipid levels and an atherogenic lipid profile as most important for the prediction. Combining maternal characteristics with the urinary hippurate/creatinine level improved the prediction rates of preeclampsia in a logistic regression model. The study indicates a potential future role of clinical importance for metabolomic analysis of urine in prediction of preeclampsia.

A randomized controlled trial of third-trimester routine ultrasound in a non-selected population

To compare detection rates of small‐for‐gestational‐age fetuses, large‐for‐gestational‐age fetuses, congenital anomalies and adverse perinatal outcomes in pregnancies randomized to third‐trimester routine ultrasound or ultrasound on clinical indication.

Risk assessment for preeclampsia in nulliparous women at 11–13 weeks gestational age: prospective evaluation of two algorithms

To evaluate two algorithms for prediction of preeclampsia in a population of nulliparous women in Norway.

A prospective study of screening for hypertensive disorders of pregnancy at 11-13 weeks in a Scandinavian population.

To investigate the prediction of preeclampsia and gestational hypertension using maternal characteristics, mean arterial pressure (MAP), uterine artery pulsatility index (UtAPI), pregnancy‐associated plasma protein‐A (PAPP‐A) and placental growth factor (PlGF) at gestational weeks 11–13 in a Scandinavian population with a medium to high prior risk for developing hypertensive disorders of pregnancy.

Distinct First Trimester Cytokine Profiles for Gestational Hypertension and Preeclampsia.

Objective—Gestational hypertension and preeclampsia involve dysregulated maternal inflammatory responses to pregnancy, but whether such responses differ between the disorders has not been determined. We aimed to investigate disease-specific early pregnancy serum cytokine profiles of women subsequently developing gestational hypertension or preeclampsia for new insight into the underlying pathogeneses and differences between the disorders. Approach and Results—The study cohort consisted of 548 pregnant Norwegian women who were either multiparous with previous gestational hypertension or preeclampsia or were nulliparous. Maternal sera at gestational weeks 110–136 were assayed for 27 cytokines, C-reactive protein, total cholesterol, high-density lipoprotein, triglyceride, creatinine, calcium, uric acid, and placental growth factor. Compared with normotensive women, women with both hypertensive conditions presented an atherogenic lipid profile at early gestation, but only those later developing gestational hypertension had significantly higher serum levels of interleukin (IL)-5 and IL-12. Comparing the 2 hypertensive pregnancy disorders, women subsequently developing gestational hypertension had higher serum levels of IL-1&bgr;, IL-5, IL-7, IL-8, IL-13, basic fibroblast growth factor, and vascular endothelial growth factor than the women subsequently developing preeclampsia. Conclusions—This study identifies early pregnancy differences in serum cytokine profiles for gestational hypertension and preeclampsia.

High Throughput UPLC®-MSMS Method for the Analysis of Phosphatidylethanol (PEth) 16:0/18:1, a Specific Biomarker for Alcohol Consumption, in Whole Blood

Phosphatidylethanol (PEth) is an alcohol biomarker formed in the presence of ethanol in the body. Both due to its specificity and because it has a detection window of up to several weeks after alcohol intake, its application potential is broader than for other ethanol biomarkers. The aim of this study was to develop and validate a robust method for PEth in whole blood with fast and efficient sample extraction and a short analytical runtime, suitable for high throughput routine purposes. A validated ultra-performance liquid chromatography tandem mass spectrometry (UPLC®-MSMS) method for quantification of PEth 16:0/18:1 in the range 0.05-4.00 μM (R2 ≥ 0.999) is presented. PEth 16:0/18:1 and the internal standard (IS) PEth-d5 (0.55 μM), were extracted from whole blood (150 μL) by simple protein precipitation with 2-propanol (450 μL). Chromatography was achieved using a BEH-phenyl (2.1 × 30 mm, 1.7 μm) column and a gradient elution combining ammonium formate (5 mM, pH 10.1) and acetonitrile at a flow rate of 0.5 mL/min. Runtime was 2.3 min. The mass spectrometer was monitored in negative mode with multiple reaction monitoring (MRM). The m/z 701.7 > 255.2 and 701.7 > 281.3 transitions were monitored for PEth 16:0/18:1 and the m/z 706.7 > 255.3 for PEth-d5. Limit of quantification was 0.03 μM (coefficient of variation, CV = 6.7%, accuracy = 99.3%). Within-assay and between-assay imprecision were 0.4-3.3% (CV ≤ 7.1%). Recoveries were 95-102% (CV ≤ 4.9%). Matrix effects after IS correction ranged from 107% to 112%. PEth 16:0/18:1 in patient samples were stable for several days at 30°C. Repeated freezing (-80°C) and thawing did not affect the concentration. After thawing and analysis patient samples were stable at 4-8°C for at least 4 weeks. Results from a proficiency test program, showing |Z| values ≤1.2, confirm the validity of the method. Analysis of the first 3,169 samples sent to our laboratory for routine use has demonstrated its properties as a robust method suitable for high throughput purposes.

A Randomized Controlled Trial of Third-Trimester Routine Ultrasound in a Nonselected Population

Fetuses who are small for gestational age (SGA) and with intrauterine growth restriction have increased risk for perinatal morbidity and mortality. This study was undertaken to compare detection rates of SGA after 2 ultrasound (US) examinations and to evaluate detection rates of fetuses who are large for gestational age (LGA), congenital anomalies, and adverse perinatal outcomes. This trial (1989–1992) included 6780 pregnant women to whom routine US examinations at 18 weeks of gestation were offered. In the study group, 1 additional routine US examination was offered at 33/0 wk 3 d. The fetal head, abdomen, growth deviation, and birth weight were assessed. The cutoff value for SGA was birth weight for gestational age of greater than 22% weight deviation, and that for suspected intrauterine growth restriction was growth deviation greater than 5%. The cutoff value for suspected LGA in utero was growth deviation greater than +10%, and that at birth was defined as weight for gestational age of more than +22%. Data from a study group of 3175 women and a control group of 3224 women were analyzed. The groups did not differ in size or gestational age at birth. In the study group, 77 of 467 fetuses thought to be SGA in utero were SGA at birth; 19 of 2689 fetuses thought to be non-SGA in utero were SGA at birth. Sensitivity and specificity for detection of SGA infants in the study group were 80% (95% confidence interval [CI], 71%–88%) and 87% (95% CI, 86%–88%), respectively. In controls, 44 of 217 fetuses suspected to be SGA in utero were SGA at birth; 52 of 2984 fetuses without suspected SGA in utero were SGA. The sensitivity and specificity for detection of SGA in the control group were 46% (95% CI, 36%–56%) and 94% (95% CI, 94%–95%), respectively. Preconception and Prenatal Care 185 Copyright

PP002. Metabolomic biomarkers in serum and urine of preeclamptic women

INTRODUCTION Preeclampsia (PE) affects about 3% of pregnancies. The syndrome cannot be accurately predicted, and large variation complicates the search for early biomarkers. Metabolites are components of the metabolism; the chemical interactions in the body necessary for life. Metabolomics, the study of metabolism, has been used to characterize diabetes, cancer and cardiovascular disease (CVD). OBJECTIVES Explore the use of magnetic resonance (MR) metabolomics on PE, and to elucidate potential clues to PE etiology and pathogenesis. METHODS Serum and urine from non-pregnant women (n=10) and pregnant women with PE (n=10) or normal pregnancies (n=10), was analyzed with MR spectroscopy and subjected to multivariate analysis (MVA). Metabolites were quantified and compared between groups. RESULTS Urine and serum samples revealed differences between PE and both control groups. Ten urine metabolites were significantly different between the three groups. Urine samples from women with early-onset PE clustered together in MVA. PE serum spectra had higher levels of low and very-low density lipoproteins, and lower high-density lipoproteins compared to control groups. CONCLUSION PE and control samples were effectively discriminated using MR metabolomics, suggesting that MR metabolomics is a useful method for improved sub-phenotyping of PE in larger studies. Information relevant to the disease was found both for serum and urine samples, and indicated similarities between PE and CVD.