- Email: [email protected]
713: Accuracy and utility of the early fetal anatomy ultrasound
www.AJOG.org
Academic Issues, Antepartum Fetal Assessment, Genetics, Hypertension, Medical-Surgical Complications, Ultrasound-Imaging
ered clinically significant. A total of 88 complications during the 4 weeks’ period after the procedure were register (0,94%). The post procedural loss rate was 0,6% (0,4 % after AC and 4,1% after CVS), 0,3% in singles and 1,6% in twin pregnancies AC group. The overall loss rate was 0,9% (0,8% in AC group, 4,3% in CVS group). There was a significant correlation between post procedure complications and number of fetuses, technical aspects (dark amniotic fluid), indications for referral and operator’s experience. CONCLUSIONS: Invasive prenatal diagnostic practice in our center over a 10-years period, the largest national published series, shows a decrease in number of invasive procedures and a shift towards CVS as a diagnostic test. Despite of the expanding screening policies, maternal anxiety remains one of the main referral indication for invasive test. Monitoring of them is a first step in the process of quality control, recently introduced into medical practice in the area of prenatal diagnosis. This historical analysis enables to define new guidelines, to individualize the prenatal counselling and to implement new indicators of invasive prenatal activity.
712 The effects of exposure to BPA and BBP on the DNA methylation profile of the IGF2/H19 imprinting control region in HTR-8 cells Christian Litton1, Luca Lambertini1, Yula Ma1, Jia Chen1, Yevgeniya Pozharny2, Jennifer Wong1, Men Jean Lee3, Joanne Stone1 1
Mount Sinai School of Medicine, New York, NY, 2North Shore University Hospital, Manhasset, NY, 3Indiana University School of Medicine, Indianapolis, IN
OBJECTIVE: Bisphenol-A (BPA) and Butyl Benzyl Phthalate (BBP) affect a range of endocrine, metabolic and developmental outcomes. While not mutational, these plasticizers may disrupt the placental and fetal epigenetic profile of imprinted genes. Disruption of two of these imprinted genes, IGF2 and H19, may lead to alterations in fetal development and programming of adult disease. Our objective was to measure the effects of exposure to BPA and BBP in first trimester extravillous HTR8 trophoblast cells by: 1.Profiling the DNA methylation of the imprinting control region (ICR) of IGF2 and H19 2.Measuring IGF2 and H19 gene expression STUDY DESIGN: HTR8 cells were treated with two sublethal concentrations of BPA and BBP. 5-Azacitidine (AZA), a demethylating agent, was used as a positive control. Cells were harvested on post-treatment days 1 and 4. Extracted DNA was bisulfite-treated and purified. The methylation profile of six CpG dinucleotides, part of the CTCF6 binding site of the IGF2/H19 ICR, was determined by pyrosequencing. Extracted mRNA was quantified by real-time PCR for changes in IGF2 and H19 gene expression. RESULTS: There were no significant differences in the methylation profiles of the groups at the various time points. The CTCF6 binding site was shown to be insensitive to treatment with AZA in HTR8 cells. The commercially-available AZA-treated “control-unmethylated DNA” from Jurkat cells was shown to be 50% methylated. Finally, no differences were found in IGF2 and H19 gene expression. CONCLUSIONS: This data demonstrates that the epigenetic effects of BPA and BBP are not due to changes in methylation of the CTCF6 binding site of the IGF2/H19 ICR. Additionally, BPA and BBP do not cause dysregulation of IGF2 nor H19 gene expression in HTR8 cells. Interestingly, the CTCF6 binding site appears to be stably methylated and resistant to AZA treatment in the control Jurkat cells. This suggests that a different mechanism exists to control the epigenetic profile of this ICR. This finding is now being explored in the villous 3A first trimester placental cells and freshly prepared dendritic cells.
Poster Session V
713 Accuracy and utility of the early fetal anatomy ultrasound Christian Litton1, Tina Nguyen1, Nok Cchun1, Erin Moshier1, Jennifer Wong1, Keith Eddleman1, Joanne Stone1 1
Mount Sinai School of Medicine, New York, NY
OBJECTIVE: To assess the concordance rates for fetal anomalies when
comparing an early anatomy ultrasound at 13-16 weeks (EAUS) to a routine anatomy ultrasound at 18-22 weeks (RAUS). In addition, to determine the rate and accuracy with which the fetal anatomic structures were identified at the EAUS vs. RAUS. STUDY DESIGN: This was a retrospective cohort of 461 unselected patients identified to have an EAUS followed by RAUS. Forty-five anatomic structures were examined during each ultrasound. We compared the rate of detection and concordance of fetal anomalies at the EAUS and RAUS. In addition, the rate of visualization of each of the fetal structures was compared for each of the sonograms. The concordance of the estimated fetal weight (EFW) at each sonogram was also assessed. RESULTS: The complete anatomy was visualized in patients in 60.43% at 16 weeks and 87.42% at 20 weeks. The detection and concordance of anomalies is noted in Table 1. For the majority of patients, the findings of normal fetal anatomy at EAUS were highly correlated with findings at the RAUS. However, certain structural defects were less easily visualized at the EAUS, particularly cardiac anatomy and nuchal fold. Choroid plexus cysts (CPC’s) were the only anomaly found to be abnormal at EAUS but normal at the RAUS. In addition, 89.4% of patients had an EFW in the normal range at both the EAUS and RAUS. However, 5.2% of patients had a normal EFW at the EAUS but an abnormal EFW at the RAUS, whereas 3.5% of patients had an abnormal EFW at EAUS that became normal at the later anatomy scan. CONCLUSIONS: This study confirms the feasibility of performing an early ultrasound exam in a large series of patients. For the majority of patients, a normal EAUS correlated highly with a normal RAUS. The EAUS was less likely to detect a cardiac defect or an increased nuchal fold, and more likely to overpredict the presence of CPC’s. This study suggests that the EAUS is useful, but it clearly reaffirms the importance of performing the 18-22 week anatomy scan for confirmation. (-) Anomaly 16w (-) Anomaly 20w
(-) Anomaly 16w (ⴙ) Anomaly 20w
(ⴙ) Anomaly 16w (-) Anomaly 20w
(ⴙ) Anomaly 16w (ⴙ) Anomaly 20w
Nuchal Fold
451(97.61%)
11(2.39%)
0
0
Umbilical Cord
455(98.70%)
2(0.43%)
0
4(0.87%)
Heart
456(98.90%)
5(1.10%)
0
0
Choroid Plexus
458(99.35%)
1(0.22%)
2(0.43%)
0
Lips
459(99.57%)
2(0.43%)
0
0
Cisterna Mag
460(99.78%)
1(0.22%)
0
0
Intracranial
460(99.78%)
1(0.22%)
0
0
Stomach
460(99.78%)
1(0.22%)
0
0
Kidneys
460(99.78%)
1(0.22%)
0
0
Hands
460(99.78%)
1(0.22%)
0
0
Lat Ventricle
460(99.78%)
0
0
1(0.22%)
Feet
460(99.78%)
0
0
1(0.22%)
.......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... ..........................................................................................................................................................................................
714 Invasive placentation: an investigation into the gene expression profile of pregnancies complicated by placenta previa, accreta, increta and percreta Dennis McWeeney1, Jeremy Chien2, Amy Clayton2, Brian Brost3 1
Vanderbilt University Medical Center, Nashville, TN, 2Mayo Clinic, Rochester, MN, 3Mayo Clinic College of Medicine, Rochester, MN
OBJECTIVE: To determine the gene expression profile of extravillous trophoblasts among pregnancies complicated by invasive placentation. STUDY DESIGN: Obstetrics database isolated 34 patients requiring cesarean hysterectomy for placenta previa (n⫽31) and non-obstetric indications “control” (n⫽3) between 01/1993-06/2009 that were his-
Supplement to JANUARY 2011 American Journal of Obstetrics & Gynecology
S281
Academic Issues, Antepartum Fetal Assessment, Genetics, Hypertension, Medical-Surgical Complications, Ultrasound-Imaging
ered clinically significant. A total of 88 complications during the 4 weeks’ period after the procedure were register (0,94%). The post procedural loss rate was 0,6% (0,4 % after AC and 4,1% after CVS), 0,3% in singles and 1,6% in twin pregnancies AC group. The overall loss rate was 0,9% (0,8% in AC group, 4,3% in CVS group). There was a significant correlation between post procedure complications and number of fetuses, technical aspects (dark amniotic fluid), indications for referral and operator’s experience. CONCLUSIONS: Invasive prenatal diagnostic practice in our center over a 10-years period, the largest national published series, shows a decrease in number of invasive procedures and a shift towards CVS as a diagnostic test. Despite of the expanding screening policies, maternal anxiety remains one of the main referral indication for invasive test. Monitoring of them is a first step in the process of quality control, recently introduced into medical practice in the area of prenatal diagnosis. This historical analysis enables to define new guidelines, to individualize the prenatal counselling and to implement new indicators of invasive prenatal activity.
712 The effects of exposure to BPA and BBP on the DNA methylation profile of the IGF2/H19 imprinting control region in HTR-8 cells Christian Litton1, Luca Lambertini1, Yula Ma1, Jia Chen1, Yevgeniya Pozharny2, Jennifer Wong1, Men Jean Lee3, Joanne Stone1 1
Mount Sinai School of Medicine, New York, NY, 2North Shore University Hospital, Manhasset, NY, 3Indiana University School of Medicine, Indianapolis, IN
OBJECTIVE: Bisphenol-A (BPA) and Butyl Benzyl Phthalate (BBP) affect a range of endocrine, metabolic and developmental outcomes. While not mutational, these plasticizers may disrupt the placental and fetal epigenetic profile of imprinted genes. Disruption of two of these imprinted genes, IGF2 and H19, may lead to alterations in fetal development and programming of adult disease. Our objective was to measure the effects of exposure to BPA and BBP in first trimester extravillous HTR8 trophoblast cells by: 1.Profiling the DNA methylation of the imprinting control region (ICR) of IGF2 and H19 2.Measuring IGF2 and H19 gene expression STUDY DESIGN: HTR8 cells were treated with two sublethal concentrations of BPA and BBP. 5-Azacitidine (AZA), a demethylating agent, was used as a positive control. Cells were harvested on post-treatment days 1 and 4. Extracted DNA was bisulfite-treated and purified. The methylation profile of six CpG dinucleotides, part of the CTCF6 binding site of the IGF2/H19 ICR, was determined by pyrosequencing. Extracted mRNA was quantified by real-time PCR for changes in IGF2 and H19 gene expression. RESULTS: There were no significant differences in the methylation profiles of the groups at the various time points. The CTCF6 binding site was shown to be insensitive to treatment with AZA in HTR8 cells. The commercially-available AZA-treated “control-unmethylated DNA” from Jurkat cells was shown to be 50% methylated. Finally, no differences were found in IGF2 and H19 gene expression. CONCLUSIONS: This data demonstrates that the epigenetic effects of BPA and BBP are not due to changes in methylation of the CTCF6 binding site of the IGF2/H19 ICR. Additionally, BPA and BBP do not cause dysregulation of IGF2 nor H19 gene expression in HTR8 cells. Interestingly, the CTCF6 binding site appears to be stably methylated and resistant to AZA treatment in the control Jurkat cells. This suggests that a different mechanism exists to control the epigenetic profile of this ICR. This finding is now being explored in the villous 3A first trimester placental cells and freshly prepared dendritic cells.
Poster Session V
713 Accuracy and utility of the early fetal anatomy ultrasound Christian Litton1, Tina Nguyen1, Nok Cchun1, Erin Moshier1, Jennifer Wong1, Keith Eddleman1, Joanne Stone1 1
Mount Sinai School of Medicine, New York, NY
OBJECTIVE: To assess the concordance rates for fetal anomalies when
comparing an early anatomy ultrasound at 13-16 weeks (EAUS) to a routine anatomy ultrasound at 18-22 weeks (RAUS). In addition, to determine the rate and accuracy with which the fetal anatomic structures were identified at the EAUS vs. RAUS. STUDY DESIGN: This was a retrospective cohort of 461 unselected patients identified to have an EAUS followed by RAUS. Forty-five anatomic structures were examined during each ultrasound. We compared the rate of detection and concordance of fetal anomalies at the EAUS and RAUS. In addition, the rate of visualization of each of the fetal structures was compared for each of the sonograms. The concordance of the estimated fetal weight (EFW) at each sonogram was also assessed. RESULTS: The complete anatomy was visualized in patients in 60.43% at 16 weeks and 87.42% at 20 weeks. The detection and concordance of anomalies is noted in Table 1. For the majority of patients, the findings of normal fetal anatomy at EAUS were highly correlated with findings at the RAUS. However, certain structural defects were less easily visualized at the EAUS, particularly cardiac anatomy and nuchal fold. Choroid plexus cysts (CPC’s) were the only anomaly found to be abnormal at EAUS but normal at the RAUS. In addition, 89.4% of patients had an EFW in the normal range at both the EAUS and RAUS. However, 5.2% of patients had a normal EFW at the EAUS but an abnormal EFW at the RAUS, whereas 3.5% of patients had an abnormal EFW at EAUS that became normal at the later anatomy scan. CONCLUSIONS: This study confirms the feasibility of performing an early ultrasound exam in a large series of patients. For the majority of patients, a normal EAUS correlated highly with a normal RAUS. The EAUS was less likely to detect a cardiac defect or an increased nuchal fold, and more likely to overpredict the presence of CPC’s. This study suggests that the EAUS is useful, but it clearly reaffirms the importance of performing the 18-22 week anatomy scan for confirmation. (-) Anomaly 16w (-) Anomaly 20w
(-) Anomaly 16w (ⴙ) Anomaly 20w
(ⴙ) Anomaly 16w (-) Anomaly 20w
(ⴙ) Anomaly 16w (ⴙ) Anomaly 20w
Nuchal Fold
451(97.61%)
11(2.39%)
0
0
Umbilical Cord
455(98.70%)
2(0.43%)
0
4(0.87%)
Heart
456(98.90%)
5(1.10%)
0
0
Choroid Plexus
458(99.35%)
1(0.22%)
2(0.43%)
0
Lips
459(99.57%)
2(0.43%)
0
0
Cisterna Mag
460(99.78%)
1(0.22%)
0
0
Intracranial
460(99.78%)
1(0.22%)
0
0
Stomach
460(99.78%)
1(0.22%)
0
0
Kidneys
460(99.78%)
1(0.22%)
0
0
Hands
460(99.78%)
1(0.22%)
0
0
Lat Ventricle
460(99.78%)
0
0
1(0.22%)
Feet
460(99.78%)
0
0
1(0.22%)
.......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... .......................................................................................................................................................................................... ..........................................................................................................................................................................................
714 Invasive placentation: an investigation into the gene expression profile of pregnancies complicated by placenta previa, accreta, increta and percreta Dennis McWeeney1, Jeremy Chien2, Amy Clayton2, Brian Brost3 1
Vanderbilt University Medical Center, Nashville, TN, 2Mayo Clinic, Rochester, MN, 3Mayo Clinic College of Medicine, Rochester, MN
OBJECTIVE: To determine the gene expression profile of extravillous trophoblasts among pregnancies complicated by invasive placentation. STUDY DESIGN: Obstetrics database isolated 34 patients requiring cesarean hysterectomy for placenta previa (n⫽31) and non-obstetric indications “control” (n⫽3) between 01/1993-06/2009 that were his-
Supplement to JANUARY 2011 American Journal of Obstetrics & Gynecology
S281