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793 Characterization of discriminatory urinaryroteomic biomarkers for severe preeclampsiasing seldi-TOF mass spectrometry

Seung Mi Lee1, Joong Shin Park1, Errol Norwitz2,Sun Min Kim1, Byoung Jae Kim1, Chan-WookPark1, Jong Kwan Jun1, Hee Chul Syn1

1Seoul National University College of Medicine, Seoul,2Tufts University School of Medicine, Boston, MAOBJECTIVE: To identify and characterize novel urinary biomarkershat are differentially expressed in women with severe preeclampsiasing surface-enhanced laser desorption ionization time-of-flightass spectrometry (SELDI-TOF-MS).

STUDY DESIGN: Urine samples were collected from women with severepreeclampsia (n�11 [sPE]), mild preeclampsia (n�7 [mPE]), andnormotensive controls (n�8) and analyzed individually usingSELDI-TOF-MS to identify discriminatory protein peaks in the sPEcohort. A scoring system was constructed— designated PreeclampsiaProteomic Score of Urine (PPSU)—to differentiate sPE from mPEand normotensive controls: the presence of a discriminatory peak wasassigned a score of 1, the absence of the peak was scored as 0, and thetotal number of discriminatory protein peaks present in each urinesample was converted to a numeric score, which was defined as PPSU.RESULTS: Four discriminatory protein peaks were identified (m/z ra-io: 4155, 6044, 6663, and 7971), all of which were down-regulated inomen with sPE. PPSU scores in women with sPE were significantly

ower than that in both mPE and controls (sPE 0 [0-4] vs mPE 3 [0-4]s controls 4 [2-4]; median [range]; P�0.05 for both). PPSU�2 had aensitivity of 90.9% and specificity of 93.3% in discriminating patientsith sPE from mPE and controls (Figure).

CONCLUSIONS: Proteomic analysis of urine can accurately distinguishPE from mPE and normotensive controls. Additional studies are un-erway to identify and characterize these proteomic biomarkers, ando determine the role of these differentially expressed proteins in theathogenesis of preeclampsia. (Legend of the figure: ROC curve show-

ng performance of PPSU score in discriminating patients with sPErom mPE and controls)

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794 A longitudinal study of plasma beta-carotene leveluring pregnancy in relation to the risk of preeclampsia

Shu Qin Wei1, Pierre Julien2, Franois Audibert1, Amelieagne2, Anne-Monique Nuyt1, Hairong Xu1, Yuquanu1, Zhong-Cheng Luo1, William Fraser1

1University of Montreal, Montreal, QC, 2Laval University, Laval, QCOBJECTIVE: A longitudinal study of plasma beta-carotene level during

regnancy in relation to the risk of preeclampsia.STUDY DESIGN: A nested case-control study using a prospective preg-

ancy cohort from a trial of antioxidant supplementation for the pre-ention of PE (INTAPP). Plasma beta-carotene levels were measuredongitudinally at 12-18 weeks (prior to intervention), 24-26 weeks,nd 32-34 weeks of gestation using high-performance liquid chroma-ography (HPLC) with coulometric electrochemical detection. A totalf 115 women with PE and 229 matched controls were included. Wesed logistic regression to calculate odds ratios (ORs) and 95% con-dence intervals (95% CI).

RESULTS: Longitudal analyses of maternal plasma beta-carotene con-entrations demonstrate no change across gestation (12-14 weeks vs.4-26 weeks vs. 32-34 weeks) (p� 0.05). The plasma beta-caroteneoncentrations were significant lower in the patients with PE com-ared to the normontensive controls at the above different gestationalges (�g/ml) (mean�SD:0.65�0.60 vs. 0.83�0.52; 0.62�0.62 vs.

0.89�0.60; 0.69�0.62 vs. 0.90�0.69; respectively, p�0.05). Aftermultivariate adjustment, women with beta-carotene concentrationsin the lowest quartiles, experienced a 3-6 fold increased risk of pre-eclampsia (using the highest quartile as referent group; 12-14 weeks:OR 3.62, 95%CI 1.88 �6.97; 24-26 weeks: OR 6.03, 95% CI 3.07�11.85; at 32-34 weeks: OR 2.88, 95%CI 1.42- 5.83).CONCLUSIONS: Although plasma beta-carotene concentration re-

ained unchanged across gestation, lower beta-carotene levels weressociated with an increased risk of PE.

795 Seven eclampsia candidate genes differentiallyethylated in preeclampsia

Wendy White1, Brian Brost1, Joshua Nitsche1, Jonathan’Brien1, William Watson1, Carl Rose1, Norman Davies1,ent Bailey1, Stephen Turner1, Vesna Garovic1

1Mayo Clinic College of Medicine, Rochester, MNOBJECTIVE: Altered gene expression in biomarkers associated with

reeclampsia/eclampsia (PE) could be explained in part by epigenetichenomena such as variable methylation.

STUDY DESIGN: Genome wide methylation profiles of maternal DNAwere evaluated in 14 PE cases and 14 normotensive controls. Subjectswere nulliparous, non-smokers, age and BMI matched. GenomicDNA was run on the Illumina Methylation Assay 27,578 CpG sitesgenome wide. Mean methylation at sites in genes from a Metacore-defined eclampsia gene set present on our platform were comparedusing a t-test.RESULTS: QC confirmed high correlation of replicates and detection palues �95%. Of the 39 genes in the “eclampsia gene set”, 34 wereresent on our platform with 73 CpG sites. Seven out of 34 tested inhis gene set had differential methylation with p value �0.05. Twoenes were found to be less methylated in PE which may result in morexpression. AGT (-3%; p� 0.027), angiotensin, is a potent vasocon-trictor with exaggerated effect in PE. DDAH1 (-6%; p�0.031) isnvolved in nitric oxide generation, via asymmetric dimethylarginineADMA), levels of which are known to be altered in PE. Five genesere more methylated and therefore may correlate with reduced tran-

cription. CALCA (�4%; p�0.001) forms calcitonin-gene relatedeptide, a potent vasodilator decreased in the PE. F5 (�1%;�0.016), coagulation Factor V, is a target of activated protein C, and

ncreased resistance related to genetic variants (Factor V Leiden) orregnancy have been associated with PE. MTHFR (�3%; p�0.041)egulates homocysteine; high levels are associated with a 20x increase

n risk for PE. POMC (�4%; p�0.014) produces beta endorphin and

ent to JANUARY 2011 American Journal of Obstetrics & Gynecology S311

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Poster Session V Academic Issues, Antepartum Fetal Assessment, Genetics, Hypertension, Medical-Surgical Complications, Ultrasound-Imaging www.AJOG.org

through ACTH stimulates aldosterone, both decreased in PE. PTGS2(�3%; p�0.03) is part of the COX 2 prostaglandin pathway involvedin inflammation.CONCLUSIONS: This is the first study that we know of that describes

ethylation differences in eclampsia candidate genes from maternalNA. The differential methylation of these 7 genes may affect tran-

cription and explain known alterations in gene product associatedith PE.

796 History of hypertension in pregnancy predictsyperhomocysteinemia decades later

Wendy White1, Brian Brost1, Vesna Garovic1, Joshua Nitsche1,onathan O’Brien1, William Watson1, Carl Rose1, Normanavies1, Kent Bailey1, Heather Wiste1, Stephen Turner1

1Mayo Clinic College of Medicine, Rochester, MNOBJECTIVE: Hypertension in pregnancy is increasingly recognized as a

redictor of future cardiovascular disease. Elevated plasma homocys-eine has been noted in preeclampsia and is also a recognized bio-

arker for cardiovascular morbidity later in life. We sought to deter-ine if a history of hypertension in pregnancy was an independent

isk factor for hyperhomocysteinemia later in life.STUDY DESIGN: This was a cohort of 2429 women in the GENOA net-work of the Family Blood Pressure Program, who were assessed at amedian age of 61 years old. A validated questionnaire was used todetermine history of nulliparity, normotensive, or hypertensive preg-nancy. Other covariates including family history, age at assessment,education level, race, BMI, diabetes, tobacco, hypertension, and use ofstatins were determined by structured interview and physical exami-nation. Serum homocysteine was measured by the Mayo Clinic HPLCmethod and elevated homocysteine was defined as �13 micromol/L.Multiple linear and logistic regression models were used to assess re-lationships between history of hypertension during pregnancy andhomocysteine levels measured at the time of assessment.RESULTS: A history of hypertension in pregnancy, when compared

ith normotensive pregnancy, predicted a 4.5% higher serum homo-ysteine level (p�0.011) and a 1.64-fold increased odds of havinglevated homocysteine (95% CI, 1.18-2.28, p�0.003) after adjust-ent for potentially confounding covariates. In contrast, a history of

ormotensive pregnancy, as compared with nulliparity, predicted a.49-fold reduced odds of elevated homocysteine (95% CI, 0.32-.74,p �0.001).

CONCLUSIONS: A history of hypertension in pregnancy predicts higheromocysteine levels decades later in life and may be a mediator in theathway that links a history of hypertension in pregnancy with futureardiovascular risk. Normotensive pregnancy, in contrast, indicates anegative stress test” associated with lower biomarkers of CVD riskater in life. Pregnancy history should become part of CVD risk assess-

ent and factor into decisions about screening and primary preven-ion.

797 Genome wide methylation profiling in preeclampticregnancies yields novel neuronal candidate geneshat may explain seizure susceptibility orhe protective effect of magnesium

Wendy White1, Brian Brost1, Joshua Nitsche1, Jonathan’Brien1, William Watson1, Carl Rose1, Norman Davies1,ent Bailey1, Stephen Turner1, Vesna Garovic1

1Mayo Clinic College of Medicine, Rochester, MNOBJECTIVE: Preeclampsia (PE) is a major source of maternal and fetal

orbidity and mortality worldwide. Seizures complicate up to 3% ofases of preeclampsia. It is not well understood why this occurs or whos most at risk. This study sought to determine if a genome wide as-essment of DNA methylation in PE could identify novel biomarkersssociated with the disease state.

STUDY DESIGN: Genome wide methylation profiles of maternal DNA

from blood were obtained at the time of delivery in 14 PE cases and 14 t

S312 American Journal of Obstetrics & Gynecology Supplement to JANUARY 2

normotensive controls. The patients were nulliparous, non-smokersand were matched for age and BMI. Genomic DNA was derived frombuffy coat, purified, and bisulfite modified then run on the IlluminaMethylation Assay platform looking at 27,578 CpG sites in 14,495genes. Mean methylation levels at each CpG site were compared usinga t-test.RESULTS: Magnesium prophylaxis was used in 9 of 14 PE cases due to

severe phenotype; none of the controls were exposed to magnesium.here did not appear to be an association between methylation andagnesium exposure. When ranked by p value, 4 of the top 15 genes

ound to be differentially methylated in PE compared with controlsere surprisingly highly expressed in the brain. GRIN2b, 6% moreethylated in PE (p�0.00006), produces a NMDA excitatory neu-

otransmitter receptor which has voltage dependent sensitivity toagnesium. PCDHB7, 6% more methylated in PE (p � 0.00007), is

involved in the establishment and function of specific cell-cell neuralconnections. BEX1, 3% more methylated in PE (p�0.0003), plays arole in cell cycle progression and neuronal differentiation. GABRA1,5% more methylated in PE (p�0.00066), produces a major inhibitoryneurotransmitter and is associated with epilepsy.CONCLUSIONS: Genome wide methylation profiling of preeclampticases suggests novel eclampsia candidate genes that are highly ex-ressed in the brain. Further investigation of these genes in terms ofNP frequencies, gene expression and protein levels may lead to aetter understanding of both the physiology of seizure activity in PEnd the mechanism of action of magnesium prophylaxis.

798 Improving patient understanding of preeclamspsia:randomized controlled trial

Whitney You1, Michael Wolf2, Stacyailey2, William A. Grobman3

1Naval Medical Center San Diego, San Diego, CA, 2Institute for HealthcareStudies, Northwestern University, Chicago, IL, 3NorthwesternUniversity, Feinberg School of Medicine, Chicago, ILOBJECTIVE: Ninety million people have low health literacy, however,here is a paucity of information regarding how to enhance patientnderstanding in obstetrics. Recently, our group has shown that manyomen have a poor understanding of preeclampsia. We developed an

ducational tool to inform women about the syndrome, the symp-oms associated with it, and what to do should they experience theseymptoms. The objective of this study was to assess whether this toolould improve understanding of preeclampsia.

STUDY DESIGN: This was a randomized controlled trial that took placen a tertiary care, urban, university hospital. One hundred and twentyomen were randomly assigned to: 1) the preeclampsia educational

ool 2) a standard ACOG pamphlet or 3) no additional information.he preeclampsia educational tool was a graphics-based card withinimum text created with the input of obstetricians, health literacy

esearchers, and patient focus groups. Preeclampsia knowledge wasssessed using a questionnaire that addressed the symptoms, conse-uences, and proper patient actions associated with preeclampsia.dditional information about demographics, medical history, andealth literacy was obtained. Health literacy was assessed using theewest Vital Sign.

RESULTS: The mean age of the participants was 27.2 (� 5.9) years, 62%ere multiparous, approximately half were African American, and2% were deemed to have adequate literacy. There were no differencesetween the groups according to demographic characteristics. Pa-ients who received the educational tool scored significantly better ontest of preeclampsia knowledge than those who received the ACOGamphlet or no additional information (71%, 63%, 49%, respectively,� 0.05). This improved understanding was equally evident amongomen with adequate and inadequate health literacy (interaction �

05).CONCLUSIONS: A graphics-based preeclampsia educational tool has

he ability to significantly improve patient understanding of pre-

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