CSH RNA Interference Reduces Global Nutrient Uptake and Umbilical Blood Flow Resulting in Intrauterine Growth Restriction.

Deficiency of the placental hormone chorionic somatomammotropin (CSH) can lead to the development of intrauterine growth restriction (IUGR). To gain insight into the physiological consequences of CSH RNA interference (RNAi), the trophectoderm of hatched blastocysts (nine days of gestational age; dGA) was infected with a lentivirus expressing either a scrambled control or CSH-specific shRNA, prior to transfer into synchronized recipient sheep. At 90 dGA, umbilical hemodynamics and fetal measurements were assessed by Doppler ultrasonography. At 120 dGA, pregnancies were fitted with vascular catheters to undergo steady-state metabolic studies with the 3H2O transplacental diffusion technique at 130 dGA. Nutrient uptake rates were determined and tissues were subsequently harvested at necropsy. CSH RNAi reduced (p ≤ 0.05) both fetal and uterine weights as well as umbilical blood flow (mL/min). This ultimately resulted in reduced (p ≤ 0.01) umbilical IGF1 concentrations, as well as reduced umbilical nutrient uptakes (p ≤ 0.05) in CSH RNAi pregnancies. CSH RNAi also reduced (p ≤ 0.05) uterine nutrient uptakes as well as uteroplacental glucose utilization. These data suggest that CSH is necessary to facilitate adequate blood flow for the uptake of oxygen, oxidative substrates, and hormones essential to support fetal and uterine growth.

Neuroimaging findings associated with the fetal inflammatory response syndrome.

The fetal inflammatory response syndrome (FIRS) is a condition whereby the fetus mounts an inflammatory response to intrauterine infection/inflammation. Clinical consequences include preterm premature rupture of membranes (PPROM), spontaneous preterm delivery, neonatal sepsis, bronchopulmonary dysplasia, and brain and other organ injury. Mechanisms leading to brain injury in FIRS have been investigated in animal and human studies. We review the neuroimaging findings of brain injury in FIRS, which overlap those of hypoxic-ischemic injury, and clinical correlation is necessary for a correct diagnosis. FIRS should be considered the primary diagnosis when neuroimaging findings such as periventricular leukomalacia are identified in preterm children born as a consequence of PPROM and spontaneous preterm labor. Additionally, FIRS should be considered in term infants who do not have the most common features of HIE (e.g. a sentinel event). Systematic histopathologic examination of the placenta and umbilical cord and/or detection of characteristic inflammatory markers in such cases are needed to establish the correct diagnosis.

MHY2233 Attenuates Replicative Cellular Senescence in Human Endothelial Progenitor Cells via SIRT1 Signaling.

Cardiovascular diseases (CVDs) are a major cause of death worldwide. Due to the prevalence of many side effects and incomplete recovery from pharmacotherapies, stem cell therapy is being targeted for the treatment of CVDs. Among the different types of stem cells, endothelial progenitor cells (EPCs) have great potential. However, cellular replicative senescence decreases the proliferation, migration, and overall function of EPCs. Sirtuin 1 (SIRT1) has been mainly studied in the mammalian aging process. MHY2233 is a potent synthetic SIRT1 activator and a novel antiaging compound. We found that MHY2233 increased the expression of SIRT1, and its deacetylase activity thereby decreased expression of the cellular senescence biomarkers, p53, p16, and p21. In addition, MHY2233 decreased senescence-associated beta-galactosidase- (SA-ß-gal-) positive cells and senescence-associated secretory phenotypes (SASPs), such as the secretion of interleukin- (IL-) 6, IL-8, IL-1α, and IL-1ß. MHY2233 treatment protected senescent EPCs from oxidative stress by decreasing cellular reactive oxygen species (ROS) levels, thus enhancing cell survival and function. The angiogenesis, proliferation, and migration of senescent EPCs were enhanced by MHY2233 treatment. Thus, MHY2233 reduces replicative and oxidative stress-induced senescence in EPCs. Therefore, this novel antiaging compound MHY2233 might be considered a potent therapeutic agent for the treatment of age-associated CVDs.

Correlation of fetal blood vessel Doppler measurements with fetal anemia among Rhesus isoimmunized pregnancies after two intrauterine transfusions.

OBJECTIVE: To assess the correlation between fetal blood vessel Doppler measurements and fetal anemia among Rhesus isoimmunized pregnancies after two intrauterine transfusions as a potential guide to therapy. METHODS: A prospective observational study was conducted among 30 women who attended a single hospital in India between April 2, 2015 and October 30, 2016. The participants underwent a third intrauterine transfusion based on a middle cerebral artery (MCA) peak systolic velocity (PSV) of greater than 1.50 multiples of the median (MoM). Cordocentesis was performed before the third intrauterine transfusion and hematocrit values correlated with the blood vessel Doppler measurements. RESULTS: The MCA PSV MoM and fetal hematocrit MoM had a correlation coefficient of -0.43 (95% confidence interval -0.68 to 0.08; P=0.017). The sensitivity, specificity, positive predictive value, and negative predictive value were 68%, 57%, 83%, and 33%, respectively. The descending aorta PSV δ and fetal hematocrit δ had a correlation coefficient of -0.54 (95% confidence interval -0.75 to -0.23; P=0.001). An area under the curve of 0.80 (standard error 0.085; P=0.017) had 87% sensitivity and 57% specificity for diagnosing fetal anemia. CONCLUSION: The descending aorta PSV could offer a useful diagnostic adjunct to MCA PSV after two intrauterine transfusions.

Fetal anemia: Diagnosis and management.

Fetal anemia has been known for many years as a dangerous complication of pregnancy. Its most common causes are maternal alloimmunization and parvovirus B19 infection, although it can be associated with many different pathological conditions including fetal aneuploidies, vascular tumors, and arteriovenous malformations of the fetus or placenta and inherited conditions such as alpha-thalassemia or genetic metabolic disorders. Doppler ultrasonographic assessment of the peak velocity of systolic blood flow in the middle cerebral artery for the diagnosis of fetal anemia and intravascular intrauterine transfusion for its treatment are the current practice standards. Live birth rates as high as 95% have been reported in recent years. The additional role of intravenous immunoglobulin therapy and the long-term consequences of the condition are the subjects of active ongoing research.

Human umbilical cord blood relaxation times and susceptibility at 3 T.

PURPOSE: To characterize the magnetic susceptibility and relaxation times (T1 and T2 ) of fetal blood at 3 T as a function of the hematocrit (Hct) and oxygen saturation (sO2 ). METHODS: Susceptibility and relaxometry measurements were performed on cord blood specimens (N = 90, derived from six caesarean deliveries) with a range of hematocrits and oxygen saturations (0.09 < Hct < 0.82, 7 < sO2 < 100%). To obtain simple, analytic relationships between MRI properties and blood properties, data were fit to established two-compartment (plasma and erythrocytes) models. RESULTS: Two-compartment models effectively described the cord blood data. The root-mean-squared deviation between the model and the data was 6.3, 10.3, and 1.3% for fits to T1 , T2 , and susceptibility measurements. Relaxometry data and estimated T1 and T2 model parameters were generally consistent with those reported in cord blood at 1.5 T and comparable to published values for adult blood. Notably, the measured value of Δχdeo , the susceptibility difference between fully oxygenated (sO2 = 100%) and deoxygenated (sO2 = 0) cord blood was approximately 20% lower than the established adult blood value (Δχdeo,cord = 2.64 ppm, Δχdeo,adult = 3.4 ppm). CONCLUSIONS: The described models and associated parameter values can be used to inform acquisition parameters, and interpret fetal/neonatal blood susceptibility measurements and relaxometry data acquired at 3 T with respect to hematocrit and sO2 . Magn Reson Med 79:3194-3206, 2018. © 2017 International Society for Magnetic Resonance in Medicine.

Non-invasive evaluation of blood oxygen saturation and hematocrit from T1 and T2 relaxation times: In-vitro validation in fetal blood.

PURPOSE: We propose an analytical method for calculating blood hematocrit (Hct) and oxygen saturation (sO2 ) from measurements of its T1 and T2 relaxation times. THEORY: Through algebraic substitution, established two-compartment relationships describing R1=T1-1 and R2=T2-1 as a function of hematocrit and oxygen saturation were rearranged to solve for Hct and sO2 in terms of R1 and R2 . Resulting solutions for Hct and sO2 are the roots of cubic polynomials. METHODS: Feasibility of the method was established by comparison of Hct and sO2 estimates obtained from relaxometry measurements (at 1.5 Tesla) in cord blood specimens to ground-truth values obtained by blood gas analysis. Monte Carlo simulations were also conducted to assess the effect of T1 , T2 measurement uncertainty on precision of Hct and sO2 estimates. RESULTS: Good agreement was observed between estimated and ground-truth blood properties (bias = 0.01; 95% limits of agreement = ±0.13 for Hct and sO2 ). Considering the combined effects of biological variability and random measurement noise, we estimate a typical uncertainty of ±0.1 for Hct, sO2 estimates. CONCLUSION: Results demonstrate accurate quantification of Hct and sO2 from T1 and T2 . This method is applicable to noninvasive fetal vessel oximetry-an application where existing oximetry devices are unusable or require risky blood-sampling procedures. Magn Reson Med 78:2352-2359, 2017. © 2017 International Society for Magnetic Resonance in Medicine.

Relaxation properties of human umbilical cord blood at 1.5 Tesla.

PURPOSE: To characterize the MRI relaxation properties of human umbilical cord blood at 1.5 Tesla. METHODS: Relaxometry measurements were performed on cord blood specimens (N = 88, derived from six caesarean deliveries) spanning a broad range of hematocrits (Hct = 0.19-0.76) and oxygen saturations (sO2 = 4-100%), to characterize the dependence of T1 and T2 on these blood properties. Adult blood data (N = 31 specimens, derived from two volunteers) were similarly studied to validate our experimental methods by comparison with existing literature. Using biophysical models previously developed for adult blood, new model parameters were estimated, which relate Hct and sO2 to the observed cord blood relaxation times. RESULTS: Fitted biophysical models explained more than 90% of the variation in T1 and T2 . In general, T2 relaxation times of cord blood were longer (by up to 35%) than those of adult blood, whereas T1 relaxation times were slightly shorter (by up to 10%). CONCLUSIONS: The models and fitted parameters presented here can be used for calibration of future MRI investigations of fetal and neonatal blood physiology. This study is an important step in facilitating accurate, noninvasive assessments of fetal blood oxygen content, a valuable diagnostic parameter in the identification and treatment of fetal hypoxia. Magn Reson Med 77:1678-1690, 2017. © 2016 International Society for Magnetic Resonance in Medicine.

Blood flow Doppler velocimetry measured during active labor.

PURPOSE: To evaluate blood flow Doppler velocimetry during the first and second stages of active labor. METHODS: A prospective observational study was performed. Patients at term (37-42 weeks gestation), with normal fetal heart rate tracing patterns (categorized as category I) were examined during the first and second stages of labor. The sonographic parameters that were measured included the blood flow resistance of the maternal uterine artery (UtA) and umbilical artery (UA). Wilcoxon-matched pair test was used for the comparison of flows between the first and the second stages of labor. RESULTS: UtA and UA Doppler velocimetry measurements were obtained from 31 parturients. The left (LT) and right (RT) UtA pulsatility index (PI) was lower in the second stage of labor as compared with the first stage. However, only the LT side reached a statistically significant difference (0.88 ± 0.32 and 0.73 ± 0.18; P = 0.005). Compared with the first stage of labor, UA PI was significantly higher during the second stage of labor (0.72 ± 0.17 vs. 0.84 ± 0.33; respectively, P = 0.05). CONCLUSION: Significant blood flow resistance changes in maternal as well as in fetal blood vessels occur during the second stage as compared with the first stage of active labor.

Retrograde diastolic blood flow in the aortic isthmus is not a simple marker of abnormal fetal outcome in pregnancy complicated by IUGR--a pilot study.

AIM: To evaluate the relation between retrograde diastolic blood flow in the aortic isthmus and adverse perinatal outcome in fetuses with IUGR. MATERIALS AND METHODS: The study included 33 fetuses with IUGR defined as the estimated fetal weight and abdominal circumference under the 10th percentile for a given gestational age. The Doppler examination of the blood flow in the aortic isthmus, umbilical artery umbilical vein, middle cerebral artery uterine arteries and ductus venosus was performed regularly The study population was further divided into two subgroups, depending on the aortic isthmus blood flow direction, i.e. with and without retrograde isthmic diastolic flow. Furthermore, the relation between Doppler blood flow parameters and determinants of the perinatal outcome was analyzed. The perinatal outcome was reported as adverse if any of the following occurred: umbilical cord blood pH < 7,2; 5-minute Apgar score < 7; respiratory distress syndrome, intraventricular hemorrhage (/ll/IV grade); necrotizing enterocolitis; sepsis; intrauterine or neonatal death. RESULTS: There was no statistically significant difference in the incidence of adverse perinatal outcome between the antegrade and retrograde isthmic blood flow groups. Moreover; the study showed no statistically significant relationship between the retrograde blood flow in the aortic isthmus and the prevalence of abnormal flow in the analyzed vessels. CONCLUSION: Retrograde diastolic blood flow in the aortic isthmus presents a low sensitivity and low predictive value in predicting the adverse perinatal outcome in pregnancies complicated with IUGR.

Fetal blood flow velocimetry by phase-contrast MRI using a new triggering method and comparison with Doppler ultrasound in a sheep model: a pilot study.

OBJECT: We present the first study demonstrating the feasibility of antenatal blood flow velocimetry performing ECG triggered phase-contrast (PC)-MRI in the fetal aorta by using a newly developed Doppler ultrasound trigger. MATERIALS AND METHODS: Five pregnant sheep carrying singleton fetuses (gestational age 121 days) were anesthetized to undergo fetal 2D PC-MRI in the fetal descending aorta (1.5 T) using a newly developed MR-compatible Doppler ultrasound trigger for fetal cardiac triggering. Inter-operator variability was assessed for PC-MR measurements and reproducibility was tested by repeated scans in one fetus. Inter-modality comparison was performed by Doppler ultrasound velocimetry. RESULTS: Fetal cardiac triggering was possible in all examinations. PC-MR velocimetry revealed a mean inter-operator variability of 3 ± 5%. Average peak systolic flow velocities of 62.5 ± 4.4 cm/s were in good agreement with Doppler ultrasound measurements of 62.0 ± 9.2 cm/s (p (Lord's U test) ≫ 0.05). CONCLUSION: Fetal PC-MR velocimetry was successfully performed using the newly developed MR-compatible Doppler ultrasound trigger for intrauterine fetal cardiac triggering, demonstrating high inter-operator and inter-modality agreement. This new method has the high potential for alternative assessment of hemodynamic decompensation of the fetal circulation.

Relationship between the umbilical cord coiling index and the umbilical blood flow at 11-13 weeks of gestation.

OBJECTIVES: The aim of this study was to clarify the effects of umbilical cord coiling on the umbilical blood flow at 11-13 weeks of gestation. METHODS: A cross-sectional study was conducted among consecutive pregnant females at 11-13 weeks of gestation. Transabdominal ultrasound examinations were performed to obtain the umbilical coiling index (CI), the maximum umbilical arterial peak velocity at the free loop, the venous velocities at the free loop and the umbilical ring, and the umbilical arterial and venous flow volumes. After every measurement was standardized according to the crown-rump length (CRL), correlations between the CI and these measurements were analyzed. RESULTS: A total of 364 subjects were enrolled. The CI significantly decreased in association with advancing gestation. There were significant correlations between the CRLs and the umbilical arterial peak velocities, the venous velocities at the free loop and the umbilical ring, and the umbilical arterial and venous flow volumes. The z-scores of the umbilical arterial and venous velocimetries exhibited no significant correlations with the CI. The umbilical arterial and venous flow volumes were also not found to correlate with the CI. CONCLUSIONS: The CI does not affect either the umbilical arterial or venous blood flow at 11-13 weeks of gestation.

[Doppler blood flow velocimetry in the umbilical artery in uncomplicated pregnancy]. / Ocena przeplywu krwi w tetnicy pepowinowej w ciazy prawldlowej jako predykcia zaburzen czynnosci serca plodu w czasie porodu.

OBJECTIVE: To determine the resistance index (RI) and pulsatility (PI) in the umbilical artery (UA) in prediction of abnormal fetal heart rate during labor and poor fetal outcome in term pregnancy. MATERIAL AND METHODS: The study included 148 patients at term in uncomplicated pregnancy Daily evaluation of blood flow in the UA was performed and PI and RI were calculated. The last value before delivery was taken for the analysis. In turn predictive value of Doppler parameters has been determined in the prediction of abnormal FHR during labor and abnormal newborn condition. Evaluation included fetal CTG parameters and newborn status based on the V.Apgar scale and acid-base equilibrium in the umbilical cord blood. Then selected parameters, characterizing pregnancy and the newborn status, with abnormal Doppler results were compared. The prognostic value of Doppler indices was assessed for selected parameters determining the course of pregnancy and abnormal fetal heart rate. RESULTS: A poor predictive value of UA PI and RI in the prediction of abnormal fetal heart rate during labor and poor fetal outcome was found. The RI in the UA presented the highest predictive value. CONCLUSION: RI in the UA shows higher predictive value than PI in the detection of abnormal fetal outcome and abnormal fetal heart rate in uncomplicated pregnancy at term. However, PI as well as RI in the UA have a low predictive value for the analyzed parameters.

[Doppler blood flow velocimetry in the middle cerebral artery in uncomplicated pregnancy]. / Ocena przeplywu krwi w tetnicy srodkowej mózgu w ciazy niepowiklanej.

OBJECTIVE: To determine the resistance index (RI) and pulsatility (PI) in the middle cerebral artery (MCA) in prediction of abnormal fetal heart rate during labor and poor fetal outcome in term pregnancy. MATERIAL AND METHODS: The study included 148 patients at term in uncomplicated pregnancy Daily evaluation of blood flow in the MCA was performed and PI and RI were calculated. The last value before delivery was taken for the analysis. The predictive value of Doppler parameters has been determined in turn to predict abnormal FHR during labor and abnormal newborn condition. Evaluation included fetal CTG parameters and newborn status based on the V Apgar scale and acid-base equilibrium in the umbilical cord blood. Then selected parameters, characterizing pregnancy and the newborn status, were compared with abnormal Doppler results. The prognostic value of Doppler indices was assessed for selected parameters determining the course of pregnancy and abnormal fetal heart rate. RESULTS: Poor predictive value of UA PI and RI in the prediction of abnormal fetal heart rate during labor and poor fetal outcome was found. The RI in the UA presented the highest predictive value. CONCLUSION: PI in the MCA shows higher predictive value than RI in the detection of abnormal fetal outcome and abnormal fetal heart rate in uncomplicated pregnancy at term. However PI indices, particularly RI in the middle cerebral artery have low predictive value for the analyzed parameters.

Quantitative effects of tobacco smoking exposure on the maternal-fetal circulation.

BACKGROUND: Despite the existence of various published studies regarding the effects of tobacco smoking on pregnancy, and especially in regards to placental blood flow and vascular resistance, some points still require clarification. In addition, the amount of damage due to tobacco smoking exposure that occurs has not been quantified by objective means. In this study, we looked for a possible association between flow resistance indices of several arteries and the levels of urinary cotinine and the concentration of carbon monoxide in the exhaled air (COex) of both smoking and non-smoking pregnant women. We also looked for a relationship between those findings and fetal growth and birth weight. METHODS: In a prospective design, thirty pregnant smokers and thirty-four pregnant non-smokers were studied. The volunteers signed consent forms, completed a self-applied questionnaire and were subjected to Doppler velocimetry. Tobacco smoking exposure was quantified by subject provided information and confirmed by the measurement of urinary cotinine levels and by the concentration of carbon monoxide in the exhaled air (COex). The weight of newborns was evaluated immediately after birth. RESULTS: Comparing smoking to non-smoking pregnant women, a significant increase in the resistance index was observed in the uterine arteries (P = 0.001) and umbilical artery (P = 0.001), and a decrease in the middle cerebral artery (P = 0.450). These findings were associated with progressively higher concentrations of COex and urinary cotinine. A decrease in the birth weight was also detected (P < 0.001) in association with a progressive increase in the tobacco exposure of the pregnant woman. CONCLUSIONS: In pregnant women who smoke, higher arterial resistance indices and lower birth weights were observed, and these findings were associated with increasing levels of tobacco smoking exposure. The values were significantly different when compared to those found in non-smoking pregnant women. This study contributes to the findings that smoking damage during pregnancy is dose-dependent, as demonstrated by the objective methods for measuring tobacco smoking exposure.

Normal reference ranges of fetal regional cerebral blood perfusion as measured by fractional moving blood volume.

OBJECTIVES: To establish normal reference intervals of fetal regional brain blood perfusion using power Doppler ultrasound as measured by fractional moving blood volume (FMBV). METHODS: A cohort of consecutive singleton normally grown fetuses was selected including at least 12 fetuses for each completed week of gestation between 24 and 41 weeks. Cerebral blood perfusion was estimated using conventional power Doppler ultrasound in the following brain regions: frontal area, basal ganglia and posterior brain. Five consecutive good-quality images were recorded in each area and the region of interest was delineated offline. The FMBV was quantified as the average of all images and expressed as a percentage. Normal reference ranges were constructed by means of the LMS (lambda-mu-sigma) method. RESULTS: A total of 230 fetuses were included. The median gestational age at evaluation and at delivery was 33.1 (range, 24.0-41.0) and 39.7 (range, 34.9-42.3) weeks, respectively. From 24 to 41 weeks' gestation, the mean FMBV increased from 13.21 to 14.97% in the frontal area, 11.17 to 14.86% in the basal ganglia and 4.83 to 6.70% in the posterior brain. CONCLUSIONS: Normal data of fetal cerebral blood perfusion in the frontal area, basal ganglia and posterior brain are provided, which could be of clinical use in the assessment of fetal brain circulation.

Human fetal amino acid metabolism at term gestation.

BACKGROUND: Knowledge on human fetal amino acid (AA) metabolism, largely lacking thus far, is pivotal in improving nutritional strategies for prematurely born infants. Phenylalanine kinetics is of special interest as is debate as to whether neonates will adequately hydroxylate phenylalanine to the semiessential AA tyrosine. OBJECTIVE: Our aim was to quantify human fetal phenylalanine and tyrosine metabolism. DESIGN: Eight fasted, healthy, pregnant women undergoing elective cesarean delivery at term received primed continuous stable-isotope infusions of [1-(13)C]phenylalanine and [ring-D(4)]tyrosine starting before surgery. Umbilical blood flow was measured by ultrasound. Maternal and umbilical cord blood was collected and analyzed by gas chromatography-mass spectrometry for phenylalanine and tyrosine enrichments and concentrations. Data are expressed as medians (25th-75th percentile). RESULTS: Women were in a catabolic state for which net fetal AA uptake was responsible for > or = 25%. Maternal and fetal hydroxylation rates were 2.6 (2.2-2.9) and 7.5 (6.2-15.5) micromol phenylalanine/(kg . h), respectively. Fetal protein synthesis rates were higher than breakdown rates: 92 (84-116) and 73 (68-87) micromol phenylalanine/(kg . h), respectively, which indicated an anabolic state. The median metabolized fraction of available phenylalanine and tyrosine in the fetus was <20% for both AAs. CONCLUSIONS: At term gestation, fetuses still show considerable net AA uptake and AA accretion [converted to tissue approximately 12 g/(kg . d)]. The low metabolic uptake (AA usage) implies a very large nutritional reserve capacity of nutrients delivered through the umbilical cord. Fetuses at term are quite capable of hydroxylating phenylalanine to tyrosine.

Doppler vascular changes in intrauterine growth restriction.

Intrauterine growth restriction (IUGR) secondary to placental insufficiency is a major cause of perinatal morbidity and mortality in the United States. Historically, Doppler changes occurring in IUGR fetuses play an important role in the diagnosis and management of these fetuses, and now, based on these changes, we have proposed a staging system for IUGR fetuses that demonstrates prognostic value. This manuscript also summarizes a practical classification for IUGR fetuses. We believe that future studies should differentiate among the different types of IUGR fetuses.

Doppler assessment of the aortic isthmus and perinatal outcome in preterm fetuses with severe intrauterine growth restriction.

OBJECTIVES: To evaluate the characteristics and association with perinatal outcome of the aortic isthmus (AoI) circulation as assessed by Doppler imaging in preterm growth-restricted fetuses with placental insufficiency. METHODS: This was a prospective cross-sectional study. Fifty-one fetuses with intrauterine growth restriction (IUGR) and either an umbilical artery (UA) pulsatility index (PI) > 95(th) centile or a cerebroplacental ratio < 5(th) centile were examined at 24-36 weeks' gestation. AoI impedance indices (PI and resistance index) and absolute velocities (peak systolic (PSV), end-diastolic and time-averaged maximum (TAMXV) velocities), were measured in all cases and compared with reference ranges by gestational age. Furthermore, fetuses were stratified into two groups according to the direction of the diastolic blood flow in the AoI: those with antegrade flow (n = 41) and those with retrograde flow (n = 10). Clinical surveillance was based on gestational age and Doppler assessment of the UA, middle cerebral artery and ductus venosus (DV). Adverse perinatal outcome was defined as stillbirth, neonatal death and severe morbidity (respiratory distress syndrome, bronchopulmonary dysplasia, Grade III/IV intraventricular hemorrhage, necrotizing enterocolitis and a neonatal intensive care unit stay > 14 days). RESULTS: Adverse perinatal outcome was significantly associated with an increased AoI-PI (area under the curve 0.77; 95% CI, 0.63-0.92; P < 0.005). A significant correlation (P < 0.001) was found between retrograde blood flow in the AoI and adverse perinatal outcome, the overall perinatal mortality being higher in the retrograde group (70% vs. 4.8%, P < 0.001). In 4/5 (80%) fetuses the reversal of flow in the AoI preceded that in the DV by 24-48 h. AoI-PSV and AoI-TAMXV were < 5(th) centile in 40/51 (78%) and 48/51 (94%) cases, respectively, whereas AoI-PI was > 95(th) centile in 21/51 (41%) cases. CONCLUSIONS: Retrograde flow in the AoI in growth-restricted fetuses correlates strongly with adverse perinatal outcome. Absolute velocities in the AoI are decreased in growth-restricted fetuses. The data suggest a potential role for Doppler imaging of the AoI in the clinical surveillance of fetuses with severe IUGR, which should be confirmed in larger prospective studies.