Increased fetal epicardial fat thickness: A novel ultrasound marker for altered fetal metabolism in diabetic pregnancies.

OBJECTIVE: To evaluate whether fetal epicardial fat thickness (EFT) differs in diabetic and nondiabetic pregnant women. METHODS: Retrospective case-control study of pregnancies between 24 and 36 weeks complicated by preexisting (PDM) or gestational (GDM) diabetes mellitus, matched one to one with controls for body mass index and gestational age (GA). Epicardial fat was identified as the hypoechogenic area between myocardium and visceral pericardium over the right ventricle and its thickness was measured by a single observer blinded to clinical condition and outcomes. A linear regression analysis was performed to assess the relationship between GA and EFT, and regression lines were compared between diabetics and controls. RESULTS: 53 PDM and 53 GDM pregnant women were matched with controls. With the exception of maternal age, the demographics were similar among groups. EFT increased significantly with advancing gestation in both diabetics and controls (P < 0.0001) and was significantly greater in diabetics than in controls (P < 0.0001). The best fit lines were different between diabetics (EFT = 0.05 × GA + 0.07 mm; R2 = 0.70) and controls (EFT = 0.07 × GA + 0.04 mm; R2 = 0.93) (P < 0.0001). CONCLUSION: Fetal EFT was greater in diabetics than in nondiabetics, and even greater in pregestational diabetics. EFT maybe an additional and/or earlier marker to identify early changes in fetal metabolism before accelerated fetal growth and polyhydramnios is apparent.

microRNA expression profiling and functional annotation analysis of their targets modulated by oxidative stress during embryonic heart development in diabetic mice.

Maternal pregestational diabetes mellitus (PGDM) induces congenital heart defects (CHDs). The molecular mechanism underlying PGDM-induced CHDs is unknown. microRNAs (miRNAs), small non-coding RNAs, repress gene expression at the posttranscriptional level and play important roles in heart development. We performed a global miRNA profiling study to assist in revealing potential miRNAs modulated by PGDM and possible developmental pathways regulated by miRNAs during heart development. A total of 149 mapped miRNAs in the developing heart were significantly altered by PGDM. Bioinformatics analysis showed that the majority of the 2111 potential miRNA target genes were associated with cardiac development-related pathways including STAT3 and IGF-1 and transcription factors (Cited2, Zeb2, Mef2c, Smad4 and Ets1). Overexpression of the antioxidant enzyme, superoxide dismutase 1, reversed PGDM-altered miRNAs, suggesting that oxidative stress is responsible for dysregulation of miRNAs. Thus, our study provides the foundation for further investigation of a miRNA-dependent mechanism underlying PGDM-induced CHDs.

Type 2 diabetes mellitus induces congenital heart defects in murine embryos by increasing oxidative stress, endoplasmic reticulum stress, and apoptosis.

BACKGROUND: Maternal type 1 and 2 diabetes mellitus are strongly associated with high rates of severe structural birth defects, including congenital heart defects. Studies in type 1 diabetic embryopathy animal models have demonstrated that cellular stress-induced apoptosis mediates the teratogenicity of maternal diabetes leading to congenital heart defect formation. However, the mechanisms underlying maternal type 2 diabetes mellitus-induced congenital heart defects remain largely unknown. OBJECTIVE: We aim to determine whether oxidative stress, endoplasmic reticulum stress, and excessive apoptosis are the intracellular molecular mechanisms underlying maternal type 2 diabetes mellitus-induced congenital heart defects. STUDY DESIGN: A mouse model of maternal type 2 diabetes mellitus was established by feeding female mice a high-fat diet (60% fat). After 15 weeks on the high-fat diet, the mice showed characteristics of maternal type 2 diabetes mellitus. Control dams were either fed a normal diet (10% fat) or the high-fat diet during pregnancy only. Female mice from the high-fat diet group and the 2 control groups were mated with male mice that were fed a normal diet. At E12.5, embryonic hearts were harvested to determine the levels of lipid peroxides and superoxide, endoplasmic reticulum stress markers, cleaved caspase 3 and 8, and apoptosis. E17.5 embryonic hearts were harvested for the detection of congenital heart defect formation using India ink vessel patterning and histological examination. RESULTS: Maternal type 2 diabetes mellitus significantly induced ventricular septal defects and persistent truncus arteriosus in the developing heart, along with increasing oxidative stress markers, including superoxide and lipid peroxidation; endoplasmic reticulum stress markers, including protein levels of phosphorylated-protein kinase RNA-like endoplasmic reticulum kinase, phosphorylated-IRE1α, phosphorylated-eIF2α, C/EBP homologous protein, and binding immunoglobulin protein; endoplasmic reticulum chaperone gene expression; and XBP1 messenger RNA splicing, as well as increased cleaved caspase 3 and 8 in embryonic hearts. Furthermore, maternal type 2 diabetes mellitus triggered excessive apoptosis in ventricular myocardium, endocardial cushion, and outflow tract of the embryonic heart. CONCLUSION: Similar to those observations in type 1 diabetic embryopathy, maternal type 2 diabetes mellitus causes heart defects in the developing embryo manifested with oxidative stress, endoplasmic reticulum stress, and excessive apoptosis in heart cells.

Genetic Variation in the Platelet Endothelial Aggregation Receptor 1 Gene Results in Endothelial Dysfunction.

Platelet Endothelial Aggregation Receptor 1 (PEAR1) is a newly identified membrane protein reported to be involved in multiple vascular and thrombotic processes. While most studies to date have focused on the effects of this receptor in platelets, PEAR1 is located in multiple tissues including the endothelium, where it is most highly expressed. Our first objective was to evaluate the role of PEAR1 in endothelial function by examining flow-mediated dilation of the brachial artery in 641 participants from the Heredity and Phenotype Intervention Heart Study. Our second objective was to further define the impact of PEAR1 on cardiovascular disease computationally through meta-analysis of 75,000 microarrays, yielding insights regarding PEAR1 function, and predictions of phenotypes and diseases affected by PEAR1 dysregulation. Based on the results of this meta-analysis we examined whether genetic variation in PEAR1 influences endothelial function using an ex vivo assay of endothelial cell migration. We observed a significant association between rs12041331 and flow-mediated dilation in participants of the Heredity and Phenotype Intervention Heart Study (P = 0.02). Meta-analysis results revealed that PEAR1 expression is highly correlated with several genes (e.g. ANG2, ACVRL1, ENG) and phenotypes (e.g. endothelial cell migration, angiogenesis) that are integral to endothelial function. Functional validation of these results revealed that PEAR1 rs12041331 is significantly associated with endothelial migration (P = 0.04). Our results suggest for the first time that genetic variation of PEAR1 is a significant determinant of endothelial function through pathways implicated in cardiovascular disease.

The North American Fetal Therapy Network Consensus Statement: prenatal management of uncomplicated monochorionic gestations.

Owing to vascular connections within a single placenta, monochorionic gestations present distinctive prenatal management challenges. Complications that can arise as a result of unbalanced hemodynamic exchange (twin-twin transfusion syndrome and twin anemia polycythemia sequence) and unequal placental sharing (selective fetal growth restriction) should be kept in mind while prenatal management is being planned. Because of unique monochorionic angioarchitecture, what happens to one twin can directly affect the other. Death of one twin can result in death or permanent disability of the co-twin. Early detection of these unique disease processes through frequent ultrasonographic surveillance may allow the opportunity for earlier referral, intervention, or both and potentially better outcomes. Therefore, monochorionic gestations should be managed differently than dichorionic gestations or singletons. The purpose of this document is to present in detail methods for monitoring and management of uncomplicated monochorionic gestations and to review the evidence for the roles of these methods for detection of complications in clinical practice. Finally, we present evidence-based and expert opinion-supported recommendations developed by the North American Fetal Therapy Network for the diagnosis, surveillance, and delivery of uncomplicated monochorionic gestations.

The North American Fetal Therapy Network consensus statement: prenatal surveillance of uncomplicated monochorionic gestations.

Because they share a common placenta, monochorionic gestations are subject to unique pregnancy complications that can threaten the life and health of both fetuses and therefore impose a disproportionate disease burden on overall perinatal morbidity and mortality. Early detection of these unique disease processes may allow for prompt referral to a regional treatment center, comprehensive counseling, and better patient outcomes. The North American Fetal Therapy Network is a consortium of 30 medical institutions in the United States and Canada with established expertise in fetal surgery and other forms of multidisciplinary care for complex fetal disorders. The goal of this publication is to briefly describe complications of monochorionic gestations and to provide multidisciplinary, evidence-based, and consensus-driven recommendations for surveillance of uncomplicated monochorionic gestations.

Reference ranges for ductus venosus velocity ratios in pregnancies with normal outcomes.

OBJECTIVES: The purpose of this study was to establish reference ranges for ductus venosus velocity ratios. METHODS: Singleton pregnancies from 11 to 38 weeks with exactly established gestational ages (GAs) were recruited for the study. Pregnancies with fetal anomalies, growth abnormalities, maternal medical complications, stillbirth, birth weight below the 10th or above the 90th percentile, and neonatal anomalies were excluded. The ductus venosus pulsatility index for veins (PIV) and velocity ratios (S/v, S/D, v/D, S/a, v/a, and D/a, where S indicates ventricular systole [s-wave], v, ventricular end-systolic relaxation [v-descent], D, passive diastolic ventricular filling [D-wave], and a, active ventricular filling during atrial systole [a-wave]) were calculated. Separate regression models were fitted to estimate the mean and standard deviation at each GA for each ratio. RESULTS: A total of 902 velocity wave ratios and ductus venosus PIVs were used for reference ranges. The S/v, S/D, and v/D ratios were not changed with GA (P > .05 for all). The PIV and S/a, v/a, and D/a ratios were reduced with GA (P < .0001 for all). Significant reductions in the means and standard deviations of the PIV and S/a, v/a, and D/a ratios were observed between 17 and 18 weeks' gestation. Therefore, nomograms were separately created between 11 and 17 weeks and 18 and 38 weeks. CONCLUSIONS: We created reference ranges for ductus venosus velocity ratios between 11 and 38 weeks' gestation in normal pregnancies. These reference ranges may prove beneficial for evaluation of fetal conditions that are associated with cardiovascular abnormalities.

First trimester maternal characteristics, Doppler parameters and serum analytes after preeclampsia.

OBJECTIVE: To evaluate the impact of prior preeclampsia on first trimester assessment in subsequent pregnancy. METHODS: A total of 1283 parous patients were prospectively enrolled at 9-14 weeks of gestation. Maternal biophysical characteristics, ultrasound parameters and placental analytes were compared between women with and without prior preeclampsia. RESULTS: There is no association between prior preeclampsia and the first trimester ultrasound parameters or placental analytes studied. The effects of prior preeclampsia in subsequent pregnancy are exaggerated by increasing parity and are predominantly blood pressure-related, independent of other cardiovascular risk factors. CONCLUSION: There is a potential role for lifestyle modification and stricter pregnancy blood pressure control in patients with prior preeclampsia.

First-trimester detection of fetal anomalies in pregestational diabetes using nuchal translucency, ductus venosus Doppler, and maternal glycosylated hemoglobin.

OBJECTIVE: The frequency of fetal anomalies in women with pregestational diabetes correlates with their glycemic control. This study aimed to assess the predictive performance of first-trimester fetal nuchal translucency (NT), ductus venosus (DV) Doppler, and hemoglobin A1c (HbA1c) to predict fetal anomalies in women with pregestational diabetes. STUDY DESIGN: This was a prospective observational study of patients undergoing first-trimester NT with DV Doppler. Screening performance was tested for first-trimester parameters to detect fetal anomalies. RESULTS: Of 293 patients, 17 had fetal anomalies (11 cardiac, 7 major, 3 multisystem). All anomalous fetuses were suspected prenatally. One had NT >95th centile, 2 had reversed DV a-wave, and 13 had HbA1c >7.0%. The HbA1c was the primary determinant of anomalies (r(2), 0.15; P < .001) and >8.35% was the optimal cutoff for prediction of anomalies with an area under the curve of 0.72 (95% confidence interval, 0.57-0.88). Therefore, first-trimester prediction of anomalies was best in women with increased NT or HbA1c >8.3% (sensitivity 70.6%, specificity 77.4%, positive predictive value 16.2%, negative predictive value 97.7%, P < .001). CONCLUSION: In women with pregestational diabetes and poor glycemic control, an increased NT increases risks for major fetal anomalies. Second-trimester follow-up is required to achieve accurate prenatal diagnosis.

Cardiovascular transition to extrauterine life in growth-restricted neonates: relationship with prenatal Doppler findings.

OBJECTIVE: Cardiovascular status in fetal growth restriction (FGR) can be classified by the severity of individual Doppler abnormalities (early and late) or by the rate of clinical progression. We tested the hypothesis that aspects of the fetal cardiovascular status in FGR affect neonatal cardiovascular findings. STUDY DESIGN: FGR cases [abdominal circumference <5th percentile and an elevated umbilical (MCA) artery (UA) pulsatility index] had UA, middle cerebral artery and ductus venosus (DV) Doppler. Positive UA end-diastolic velocity and/or a low MCA pulsatility index denoted early and absent/reversed UA end-diastolic velocity, whereas an increased DV pulsatility index for veins denoted late responses. The rate of progression was classified into mild, progressive and severe. After delivery, shunt dynamics and blood flow across the patent ductus arteriosus (PDA), foramen ovale and atriaventricular valves, myocardial contractility and pharmacologic pressor requirement were noted at neonatal echocardiography. These findings were related to prenatal Doppler parameters. RESULTS: In 94 patients, only individual Doppler parameters related to neonatal echocardiographic findings. Absent/reversed UA DV significantly predicted PDA with right to left shunt (p = 0.016). The pressor need for cardiovascular instability was observed in neonates with abnormal prenatal DV Doppler and with lower birth weights delivered at earlier gestational age (p < 0.0001 for both). Pressor need was significantly related to neonatal death (Nagelkerke R² = 0.35, p = 0.002). CONCLUSION: A markedly abnormal UA Doppler predisposes growth-restricted neonates to persistence of fetal circulation associated with right to left shunting. Abnormal venous Doppler is a risk factor for cardiovascular instability which in turn significantly contributes to neonatal mortality. Further clarification of the neonatal cardiovascular transition may be helpful in guiding early neonatal assessment and management.

Serum pentraxin-3 levels at 11 to 14 weeks' gestation: association with maternal and placental characteristics.

OBJECTIVE: Pentraxin (PTX)-3 is an inflammatory molecule that may be increased in the first trimester in pregnancies with subsequent preeclampsia. We measured first-trimester serum PTX-3 and correlated levels with maternal/placental factors related to placental development. STUDY DESIGN: Prospectively enrolled women had ultrasound, physical examination, and blood draw at 11-14 weeks. PTX-3 determined by enzyme-linked immunosorbent assay was related to maternal age, parity, race, body mass index (BMI), mean arterial blood pressure (MAP), smoking/caffeine, and uterine/umbilical artery Doppler pulsatility index (PI). RESULTS: In 111 patients PTX-3 levels ranged from 0.2-13.8 ng/mL. Spearman correlation between PTX-3 and gestational age (rho = 0.096), maternal age (rho = -0.049), BMI (rho = -0.07), MAP (rho = -0.085), mean uterine artery PI (rho = 0.150), and umbilical artery PI (rho = -0.021) was nonsignificant (all P > .05). Similarly, PTX-3 distribution was unaffected by smoking/caffeine use, BMI >30, MAP >100 mm Hg, or uterine artery notching (P > .05 for all). CONCLUSION: First-trimester PTX-3 is unrelated to maternal characteristics and placental Doppler.

Amniotic fluid abnormalities.

The complex nature of amniotic fluid reflects contributions from many fetal systems, many functional roles, and multiple interactions with fetal maturation, obstetric, and maternal factors. Simple ultrasound measurement, probably done best with the maximum vertical pocket method, has a clinical role in fetal surveillance, substantiated by extensive level II and some level I evidence. Interventions (amnioinfusion for oligohydramnios, amnioreduction for polyhydramnios) have not been studied effectively in controlled fashion, with the exception of intrapartum applications, where effective reduction of cesarean delivery for fetal distress and perinatal impacts of meconium aspiration may follow amnioinfusion. A wealth of research opportunities exists into regulation of amniotic fluid constituents and their relation to preterm delivery.

Predictors of necrotizing enterocolitis in preterm growth-restricted neonates.

OBJECTIVE: The purpose of this study was to test the hypothesis that multivessel fetal Doppler imaging provides enhanced prediction of necrotizing enterocolitis (NEC) in preterm placental insufficiency. STUDY DESIGN: Placental-based growth-restricted fetuses (abdominal circumference <5%, abnormal umbilical artery [UA] Doppler imaging) were examined. UA, middle cerebral artery, ductus venosus, and umbilical vein (UV) were evaluated prenatally and were assessed for their ability to predict NEC in neonates who were delivered at <37 weeks of gestation. RESULTS: Thirty-nine of 404 neonates (9.7%) experienced NEC. Among these, the mortality rate was 15.4% (6/39 neonates; odds ratio, 2.7; 95% CI, 1.03-7.11). NEC cases had higher UA Doppler indices prenatally (P = .023), lower gestational ages and birthweight at delivery (P < .0001, respectively), 5-minute Apgar scores of <7, and higher umbilical cord artery base deficit (P < .01, respectively). NEC was more likely after prenatal UV pulsations (odds ratio, 2.4; 95% CI, 1.13-5.14; P = .028) and severe cardiovascular abnormality (composite variable incorporating UA- absent or reversed end diastolic velocity, absent or reversed ductus venosus a-wave, or UV pulsations; odds ratio, 2.1; 95% CI, 1.06-4.05; P = .029) Logistic regression revealed birthweight and base deficit as the main contributors of NEC (r(2) = 0.20; P < .0001). Receiver operating characteristic analyses revealed birthweight of <790 g (sensitivity, 74.4%; specificity, 72.9%; P < .0001) and gestational age of < or =32.2 weeks (sensitivity, 94.9%; specificity, 45.8%; P < .0001) as optimal cut-offs that provide an odds ratio for NEC of 8.2 (95% CI, 3.9-17.6; P < .0001). CONCLUSION: Placental disease predisposes the severely growth-restricted neonate to necrotizing enterocolitis. Even when arterial and venous Doppler variables are taken into consideration, birthweight remains the predominant risk factor for NEC. Further research should focus on the critical transition to neonatal life to identify relevant triggers in predisposed neonates.

Determinants of the middle cerebral artery peak systolic velocity in the human fetus.

OBJECTIVE: The purpose of this study was to identify physiologic determinants of the peak systolic blood flow velocity (PSV) of the middle cerebral artery (MCA) in the human fetus. STUDY DESIGN: MCA PSV was measured with pulsed wave Doppler ultrasound in human fetuses who underwent cordocentesis. Hemoglobin, hematocrit, and blood gas values were analyzed from umbilical venous blood, and the data were normalized for gestational age. Total oxygen content of fetal venous blood was calculated from oxygen saturation, hemoglobin value, and pO2. Correlation and logistic regression analyses were performed to identify primary physiologic determinants of MCA PSV. RESULTS: In 136 fetuses who underwent cordocentesis (predominantly for alloimmune disease), hematocrit, hemoglobin, and blood oxygen content correlated significantly with the MCA PSV (P < .01). Logistic regression modeling demonstrated that fetal hemoglobin content (odds ratio, 7.1; 95% CI, 3.71-13.7) and pCO2, but not pO2 or fetal blood oxygen content, accounted for increases in MCA PSV. CONCLUSION: Under physiologic circumstances, fetal hemoglobin, and not fetal oxygenation, primarily determines the middle cerebral artery peak systolic velocity.

Nucleated red blood cell counts in the first week of life: a critical appraisal of relationships with perinatal outcome in preterm growth-restricted neonates.

OBJECTIVE: Nucleated red blood cells (NRBC) are fetal hematologic markers for placental dysfunction, hypoxemia, and asphyxia. NRBC count elevation at birth or persistence is linked statistically to adverse outcome, but clinical predictive value is variable. We studied novel indices to better define overall magnitude of NRBC response. STUDY DESIGN: Peripheral NRBC count was obtained from preterm (<34 weeks of gestation) growth-restricted neonates within 2 hours of life. Daily counts and duration of NRBC count >30/100 white blood cells were determined. Mean counts (NRBC-mean), area under the curve (NRBC-AUC), and declination (NRBC-slope) were analyzed over week 1. NRBC parameters were related to major morbidity (bronchopulmonary dysplasia, grade III/IV intraventricular hemorrhage, necrotizing enterocolitis included) and neonatal death (NND). RESULTS: Twenty-two of 176 patients (12.5%) had acidosis. Complications included bronchopulmonary dysplasia (n = 36; 20.5%), intraventricular hemorrhage (n = 18; 10.2%), necrotizing enterocolitis (n = 18; 10.2%), NND (n = 18; 10.2%). NRBC-AUC and NRBC-mean correlated most strongly with pH, birthweight, and gestational age (Pearson coefficient -0.45 to -0.18; all P < .001). NRBC-AUC varied most between nonmorbid and morbid; NRBC-mean varied most between survivors and NND (all P < .001). NRBC persistence strongly predicted NND: clearance by day 4 was achieved by 80% of survivors and only 35% of NNDs. Logistic regression identified prematurity and persistent NRBC counts as primary morbidity determinants (r2 = 0.56; P < .01). Although the importance of individual NRBC counts varied, day-4 NRBC counts of >70 predicted morbidity best (sensitivity, 82%; specificity, 96%). Presence of morbidity and birthweight were prime determinants of death (r2 = 0.42; P < .01). CONCLUSION: Simple daily NRBC counts provide clinical information that is equivalent to more complicated methods. The importance of prematurity and growth are emphasized, but elevated NRBC counts beyond day 3 are relevant independent predictors of adverse outcome.

Acute intrapartum twin-to-twin transfusion following successful fetoscopic laser ablation.

Fetoscopic laser ablation of placental vascular anastomoses is an intrauterine therapy targeting the source of twin-to-twin transfusion syndrome (TTTS). Our patient had successful laser treatment, with resolution of stage II TTTS, suggesting closure of all significant anastomoses. After an uneventful pregnancy course, she elected a trial of labor. Following spontaneous onset of labor, acute intrapartum twin-to-twin transfusion was suspected, suggested by sinusoidal heart rate changes, confirmed postdelivery by twins' complete blood counts. The potential risks of labor with prior fetoscopic laser ablation are reviewed in the context of current knowledge about placental architecture in TTTS.

Sustained hematological consequences in the first week of neonatal life secondary to placental dysfunction.

OBJECTIVE: To evaluate the relationship between umbilical artery end diastolic velocity in growth restricted fetuses and neonatal hematologic parameters. STUDY DESIGN: Growth restricted fetuses were studied with ultrasound and Doppler evaluations. Neonates were analyzed in two groups based on umbilical artery Doppler status: positive end-diastolic velocities (PEDV) and absent or reversed end-diastolic velocities (AEDV). At birth and throughout the first week of life, groups were compared for anemia and thrombocytopenia; transfusion of red blood cells, platelets, and fresh frozen plasma; and intraventricular hemorrhage (IVH). RESULTS: Seventy-three neonates met inclusion criteria, 38 with PEDV, 35 with AEDV. Those with AEDV were delivered 3 weeks earlier, were 450 g smaller, had lower cord arterial pH values, and greater cord artery base deficits (p<0.05, respectively). AEDV neonates were twice as likely to be anemic and thrombocytopenic at birth and remain so during the first week, requiring more red blood cell and platelet transfusions. There was no difference in occurrence of severe IVH between groups. CONCLUSION: Hematological alterations associated with intrauterine growth restriction appear to continue into the first week of neonatal life. These are proportional to the degree of placental dysfunction and are predicted by fetal Doppler status. SUMMARY: Abnormal development of the placental vascular tree is the primary step in a cascade of fetal compromises leading to intrauterine growth restriction (IUGR). Doppler ultrasound evaluation of fetal and placental blood flows provides a non-invasive assessment of the fetal condition which reflects the impact of placental vascular abnormalities. The degree of placental dysfunction determines the severity of fetal disease, which can affect many fetal organ systems. In addition to disturbances in placental respiratory function, abnormal umbilical artery Doppler status is also indicative of hematologic abnormalities during fetal life and at birth. Neonates who had more severe placental dysfunction, as depicted by absent umbilical artery end diastolic velocity, were more likely to be anemic and thrombocytopenic at birth and remain so during the first week of life, and required more transfusions than those with positive end diastolic velocities. The severity of hematologic alterations during the first week of life in growth restricted neonates was proportional to and predicted by the antenatal umbilical artery end diastolic velocity Doppler status.

Persistent fetal hemoglobin in maternal circulation complicating the diagnosis of fetomaternal hemorrhage.

BACKGROUND: Transplacental hemorrhage can be life threatening to a fetus and has important maternal treatment implications. In contrast, hereditary persistence of fetal hemoglobin is a condition that has little consequence. The Kleihauer-Betke test, which is routinely used to document transplacental hemorrhage, will be positive in either case. CASES: We report two cases in which maternal persistence of fetal hemoglobin was unknown and led to the erroneous diagnosis of fetomaternal hemorrhage. These cases highlight both the limitations of the Kleihauer-Betke test and the role of flow cytometry in diagnosing fetomaternal hemorrhage. CONCLUSION: The use of flow cytometry can clarify Kleihauer-Betke test results when there is known maternal persistence of fetal hemoglobin and can more precisely quantify a fetomaternal hemorrhage for accurate Rh immune globulin dosing.

Detection of viral deoxyribonucleic acid in amniotic fluid: association with fetal malformation and pregnancy abnormalities.

OBJECTIVE: To first test the hypothesis that the presence of viral nucleic acid in amniotic fluid (AF) is associated with an abnormal pregnancy outcome, and second, to determine if the overall rate of polymerase chain reaction (PCR) positivity and the distribution of virus types vary geographically. STUDY DESIGN: Cytomegalovirus (CMV), parvovirus B19, adenovirus, enterovirus, herpes simplex virus, Epstein-Barr virus, and respiratory syncytial virus nucleic acids were sought in 423 AF samples obtained for clinical indications: 284 from the East Coast (EC) and 139 from the Midwest (MW). RESULTS: Gestational age at sampling was 19.1 weeks for EC and 20.1 weeks for MW. 13.5% of karyotypically normal singleton pregnancies (57/423) had a positive AF PCR. 11% of AF PCR from the EC while 18% of AF PCR from the MW were positive (p = 0.06). The most commonly detected viruses were adenovirus (77%), enterovirus (12%), and CMV and parvovirus B19 (5% each). Twenty-four percent of sonographically abnormal pregnancies (33/136) had a positive AF PCR compared to only 8.4% (24/287) of normal pregnancies (p < 0.001). CONCLUSION: A positive AF PCR is associated with an increased rate of fetal structural malformations, intrauterine growth restriction, hydrops and other fetal abnormalities. There were no significant geographic differences in the incidence of AF viral PCR positivity.

Kleihauer-betke testing is important in all cases of maternal trauma.

BACKGROUND: In maternal trauma, the Kleihauer-Betke (KB) test has traditionally been used to detect transplacental hemorrhage (TPH), so that Rh-negative women could receive appropriate Rh immune prophylaxis. Reasoning that the magnitude of TPH would reflect uterine injury, we evaluated Kleihauer-Betke testing as an independent predictor of preterm labor (PTL) after maternal trauma. METHODS: Admissions to the Shock Trauma Center, University of Maryland, from January 1996 to January 2002, were reviewed. Of 30,362 trauma patients admitted, 166 were pregnant, and 93 of these underwent electronic fetal monitoring. Their records were abstracted for demographics, injury type, three separate trauma scores, documented uterine contractions, PTL (contractions with progressive cervical change), and serious perinatal complications. In 71 cases, transplacental hemorrhage was assessed by maternal KB test. RESULTS: TPH, defined as KB-positive for greater than 0.01 mL of fetal blood in the maternal circulation, occurred in 46 women. Forty-four had documented contractions (25 had overt PTL) and 2 had no contractions. In 25 women with a negative KB test, none had uterine contractions. All patients with contractions or PTL had positive KB tests. By logistic regression, KB test result was the single risk factor associated with PTL (p < 0.001; likelihood ratio, 20.8 for positive KB test). Compared with other sites, abdominal trauma was associated more often with uterine contractions (p < 0.001), PTL (p = 0.001), and a positive KB test (p < 0.001, chi). None of the trauma scoring systems predicted PTL. CONCLUSION: Kleihauer-Betke testing accurately predicts the risk of preterm labor after maternal trauma. Clinical assessment does not. With a negative KB test, posttrauma electronic fetal monitoring duration may be limited safely. With a positive KB test, the significant risk of PTL mandates detailed monitoring. KB testing has important advantages to all maternal trauma victims, regardless of Rh status.