Noninvasive prenatal diagnosis. Use of density gradient centrifugation, magnetically activated cell sorting and in situ hybridization.

OBJECTIVE: To develop a noninvasive method suitable for clinical prenatal diagnosis. STUDY DESIGN: Fetal nucleated erythrocytes were separated from peripheral blood of 17 healthy pregnant women using small magnetically activated cell sorting columns (MiniMACS) following density gradient centrifugation and dual antibody labeling methods. The protocol was designed to compare the efficacy of antitransferrin receptor (CD71)/antiglycophorin A (GPA) antibodies with antithrom-bospondin receptor (CD36)/anti-GPA antibodies in identifying nucleated erythrocytes in maternal blood. Cytospin preparations of the isolated cells were subjected to in situ hybridization with specific DNA probes for the Y chromosome and chromosome 21 to confirm the fetal origin. RESULTS: After MiniMACS the enrichment factors for the CD71/GPA- and CD36/GPA-positive cells from maternal blood were similar, and the percentages of fetal cells recovered did not differ. Seven of seven male pregnancies were correctly identified. One case of trisomy 21 was detected. CONCLUSION: The in situ hybridization analysis of fetal nucleated erythrocytes isolated from maternal blood using single density gradient centrifugation, anti-CD71/anti-GPA immunostaining and MiniMACS could be an accurate, sensitive and noninvasive method for prenatal diagnosis.

Role of the pulmonary circulation in the distribution of human fetal cardiac output during the second half of pregnancy.

BACKGROUND: By using Doppler echocardiography, we determined the normal distribution of human fetal combined cardiac output (CCO) from the left and right ventricles. We also established weight-indexed pulmonary and systemic vascular resistances (Rpi and Rsi, respectively) and changes during the second half of pregnancy. METHODS AND RESULTS: Blood flows at the aortic and pulmonary valve annuli (LVCO and RVCO, respectively), right and left pulmonary arteries (QP), and ductus arteriosus (QDA) were calculated in 63 normal fetuses. Foramen ovale blood flow (QFO = LVCO-QP) was estimated. From 20 to 30 weeks of gestation, the proportion of QP of the CCO increased (from 13% to 25%, P < .001), while the proportion of QFO decreased (from 34% to 18%, P < .001). After 30 weeks, the proportions of QP and QFO were unchanged. At 38 weeks, the proportion of RVCO (60%) was higher (P < .05) than that of LVCO (40%). The proportion of QDA did not change significantly. The correlation between RVCO calculated from blood flow through the pulmonary valve and from QDA and QP was good (r = .97, P < .0001). RPi (P < .001) decreased from 20 to 30 weeks of gestation. From 30 to 38 weeks, RPi increased (P < .0001). Rsi increased (P < .001) from 20 to 38 weeks. The ratio of RPi to RSi decreased (P < .01) from 20 to 30 weeks and later remained unchanged. CONCLUSIONS: The human fetal pulmonary circulation has an important role in the distribution of cardiac output.

Volume-rendered, three-dimensional echocardiographic determination of the size, shape, and position of atrial septal defects: validation in an in vitro model.

Accurate evaluation of atrial septal defect (ASD) size and shape is very important for the selection of patients for transcatheter occlusion. The ability of volume-rendered, three-dimensional echocardiography (3DE) in displaying ASDs in a dynamic mode has been demonstrated; however, its accuracy in sizing ASDs is unknown. To assess this, we performed 3DE of 10 explanted pig hearts in which ASDs of various locations, sizes, and shapes had been experimentally created. From en face 3DE views of the atrial septum containing the defects, major and minor diameters of the defect were measured by a blinded observer, and these data were compared to direct anatomic measurements. The correlations between 3DE and anatomy for the major and minor ASD diameters were y = 0.83x + 3.4 (r = 0.97, p < 0.0001) and y = 0.92x + 1.3 (r = 0.92, p < 0.0001) respectively. The correlation between the measures for major and minor axis ratio was y = 1.06 x - 0.052, r = 0.91, p < 0.0002. Good agreement between both methods of measurements was demonstrated for all measurements. In addition, 3DE portrayed the location and shape of the defects accurately. Thus 3DE provides excellent visualization of ASD and is able to accurately define the size of the defects. These qualitative and quantitative capabilities enhance the clinical potential of this technique in the appraisal of ASDs for decisions regarding application of closure devices.

Fetal branch pulmonary arterial vascular impedance during the second half of pregnancy.

OBJECTIVE: Our purpose was to establish normal physiologic parameters in the fetal proximal and distal branch pulmonary arterial vascular impedance during the second half of pregnancy and to analyze relationships between proximal and distal pulmonary arterial blood velocity waveforms. STUDY DESIGN: In this cross-sectional study 100 uncomplicated singleton pregnancies were studied by pulsed color Doppler techniques between 18 and 41 weeks of gestation (median 30 weeks). Both right and left proximal (immediately after the bifurcation of the main pulmonary artery) and distal (beyond the first bifurcation of the branch pulmonary artery) pulmonary artery blood velocity waveforms were recorded and pulsatility index values were calculated. Peak systolic velocities and time-to-peak-velocity intervals were measured. Time-to-peak-velocity intervals were also analyzed at the level of aortic and pulmonary valves and at the ductus arteriosus. Right and left pulmonary artery diameters and right lung length were measured. RESULTS: In both right and left proximal and distal pulmonary arteries pulsatility index values decreased (p < 0.0001) and the peak systolic velocities (p < 0.003) and time-to-peak-velocity intervals (p < 0.0001) increased during the second half of pregnancy. In the proximal pulmonary arteries the pulsatility index values decreased linearly until 34 to 35 weeks of gestation and in the distal pulmonary arteries until 31 weeks of gestation. Thereafter they remained unchanged. In pulmonary arteries time-to-peak-velocity intervals were shorter (p < 0.01) than at the pulmonary valve level. There were no significant differences between the right or left pulmonary arteries in the pulsatility index values, peak systolic velocities, time-to-peak-velocity intervals, or pulmonary artery diameters. In the proximal pulmonary arteries the pulsatility index values (p < 0.02) and peak systolic velocities (p < 0.0001) were higher and time-to-peak-velocity intervals (p < 0.0001) were longer than in the distal pulmonary arteries. There was a 2.5-fold increase in pulmonary artery diameters and right lung length. CONCLUSIONS: Fetal branch pulmonary arterial vascular impedance decreases significantly during the second half of pregnancy. The linear decrease in vascular impedance during the second trimester and in the beginning of the third trimester may be related to the growth of the lung and the increase in the number of resistance vessels. During the latter part of the third trimester pulmonary vascular impedance does not decrease further.

The predictive value of maternal serum testing for detection of fetal anemia in red blood cell alloimmunization.

OBJECTIVE: Current management protocols for pregnancies complicated by red blood cell alloimmunization use the maternal antibody titer to predict the need for invasive testing for detection of fetal anemia. We investigated the use of three maternal serum tests to assess their usefulness in predicting fetal disease: indirect Coombs' titer, Marsh score, and monocyte monolayer assay. STUDY DESIGN: Forty-seven serum samples from pregnant women with red blood cell antibodies associated with fetal anemia were analyzed at cordocentesis. Fetal blood was analyzed for hematocrit (corrected for gestational age) and antigen status. Fetal anemia was defined as a hematocrit value of < 2 SD from the mean value for gestational age. Fetuses were classified into three groups: Antigen positive with anemia (n = 19), antigen positive without anemia (n = 17), antigen negative (n = 11). Statistical methods included Kruskal-Wallis test, Newman-Keuls test, Spearman's rank correlation, and receiver-operator characteristic curves; p < 0.05 was considered significant. RESULTS: The median monocyte monolayer assay (phagocytosis, adherence, and association) did not differ among the three groups. Both maternal titers and Marsh scores were significantly higher in fetuses with anemia compared with the other two groups of fetuses (256 vs 64 vs 64, p < 0.001, and 86 vs 69 vs 64, p = 0.02, respectively). Both titer and Marsh score exhibited significant correlations with corrected fetal hematocrit (r = -0.70, p < 0.001; r = -0.63, p < 0.001, respectively). Comparison of the overall receiver-operator characteristic curves for titer and Marsh score revealed no statistical difference; however, a Marsh score of 57 was noted to have a superior specificity than a titer of 16 (p = 0.02). CONCLUSION: The maternal Marsh score can be performed in conjunction with standard indirect Coombs' titers to enhance the predictability of fetal anemia.

Transthoracic real-time three-dimensional echocardiography using a fan-like scanning approach for data acquisition: methods, strengths, problems, and initial clinical experience.

Three-dimensional echocardiography is an emerging clinical method to assess cardiovascular disorders. The feasibility of using a linear mode scanning (parallel slicing) for transthoracic data acquisition has been demonstrated. In this study, we evaluated the feasibility of real-time transthoracic three-dimensional imaging of the heart using a fan-like scanning mode of echocardiographic data acquisition. We used a computer-driven motor to sequentially angulate transthoracic transducers over a fan-like arc up to 90 degrees. With careful ECG and respiratory gating, we acquired basic two-dimensional data set via parasternal and subcostal windows and performed dynamic three-dimensional reconstructions. The problems encountered included the need to repeat data acquisition sequences because of transducer movement or inappropriate gain and gray scale settings. From 15 scanning sequences in four patients, we were able to use ten sets of data. These yielded good quality three-dimensional studies projecting normal valves, a stenotic mitral valve, and an atrial septal defect, in a number of novel views. The valves could be visualized from above and from below as well as in other orientations, and the detailed anatomy appraised. Spatial relationships of the atrial septal defect with inferior and superior vena cava, coronary sinus, or tricuspid annulus could be uniquely displayed through views from the right side of the heart. This technique provided adequate new imaging planes not available from two-dimensional echocardiography. This experience demonstrates for the first time that transthoracic three-dimensional echocardiography using a fan-like scanning mode of data acquisition is feasible, and that it provides adequate visualization of intracardiac structures in unique projections.(ABSTRACT TRUNCATED AT 250 WORDS)

Diagnosis, pathophysiology, and treatment of chronic twin-to-twin transfusion syndrome.

The discovery of a 'stuck twin' during an ultrasound examination has often been equated with twin-to-twin transfusion based to a great extent on postnatal confirmation of discordant fetal size and hemoglobin concentration. However, the diagnosis of twin-to-twin transfusion cannot be made with certainty after birth since virtually all monochorionic gestations have placental anastomoses and there are many causes of growth deficiency and abnormal hemoglobin concentration. The purpose of this study is to investigate the pathophysiology of chronic twin-to-twin transfusion syndrome and apply the findings to the development of a management algorithm. In 42 twin gestations with stuck twin associated with acute hydramnios, we performed targeted ultrasound cordocentesis in each fetus and therapeutic amniocentesis. The diagnosis of chronic twin-to-twin transfusion syndrome required: sonographic evidence of monochorionicity; rapid reaccumulation of fluid after amniocentesis; discordant fetal size, and divergent fetal hematocrit measurements with at least one above or below the 95% confidence interval for gestational age. These criteria were met in 20 of 42 (48%) pregnancies. The mean gestation was 23.8 +/- 2 weeks (range 21-27 weeks). In 4 pregnancies, the transfer of adult RBCs from the donor to the recipient was documented. Monochorionicity was confirmed in all postnatally. All recipients had polycythemia and hyperproteinemia. Hydrops developed only in the recipient twin (6 of 20) and was associated with an elevated umbilical venous pressure. All pregnancies were treated with aggressive serial therapeutic amniocenteses. There was no objective evidence that amniocentesis altered the magnitude of the shunt.(ABSTRACT TRUNCATED AT 250 WORDS)

Dynamic three-dimensional echocardiography: methods and clinical potential.

In this article, we will attempt to review basic requirements for three-dimensional reconstruction, methods of cavity reconstruction, approaches to gray scale tissue-depiction displays, and current clinical experience, and also present some directions for future development.

Maternal and fetal plasma vitamin E to total lipid ratio and fetal RBC antioxidant function during gestational development.

Placental transfer of vitamin E was investigated from 19 to 35 weeks of gestation by analysis of fetal and maternal blood samples for total tocopherol, total lipids, and fetal red blood cell antioxidant reserves. Fifty-two fetal blood samples were obtained under ultrasonographic guide by percutaneous umbilical blood sampling. Thirteen were from fetuses with gestational age less than or equal to 22 weeks (x serum vitamin E = 0.4 +/- 0.14 mg/dl), 12 were from fetuses at 23-27 weeks gestation (x serum vitamin E = 0.4 +/- 0.21 mg/dl), and 27 were from fetuses with gestational age 28-38 weeks (x serum level = 0.37 +/- 0.18). Total lipid levels ranged from 140 to 216 mg/dl. Maternal plasma vitamin E concentrations correlated significantly with concurrent values in the fetus. There were no significant differences in serum vitamin E levels or vitamin E to total lipid ratio in samples from early, mid, or late gestation in either the mother or fetus. Red blood cell antioxidant reserve on samples from 18 fetuses were grossly abnormal by three different functional assays. On the basis of these data, placental transfer of vitamin E appears to be relatively constant through advancing gestation. Red blood cell antioxidant reserve is uniformly low.

Care of the neonate with erythroblastosis fetalis.

Erythroblastosis fetalis, hemolytic disease of the newborn, occurs when an isoimmunized mother produces antibodies that cross the placenta and cause hemolysis of fetal red blood cells. This hemolysis can be accompanied by severe anemia, ascites, pleural and pericardial effusions, congestive heart failure, and neurological damage with resultant perinatal mortality. Rh isoimmunization in pregnancy still occurs in spite of the advent of Rh immune globulin. This article describes the complex management and nursing implications associated with caring for the neonate with erythroblastosis fetalis.

Percutaneous umbilical blood sampling.

Percutaneous umbilical blood sampling (PUBS) provides a new and exciting method for assessment and management of certain fetal disorders. This procedure offers direct access to the fetal circulation for obtaining blood samples or for transfusing the fetus in utero. Although investigational, PUBS offers treatment approaches that were not previously available. This ability to treat the fetus in utero can prolong pregnancy, resulting in decreased prematurity and mortality rates for infants with erythroblastosis fetalis. This article describes the PUBS procedure and the indications for PUBS and discusses the nursing implications associated with the care of women undergoing PUBS.

Percutaneous umbilical blood sampling. A new technique for prenatal diagnosis.

Direct access to fetal blood during the second and third trimesters of pregnancy opens new fields of prenatal diagnosis and in utero fetal treatment. Percutaneous umbilical blood sampling, a method that involves ultrasonographically guided needle insertion into the umbilical vein, appears to have an acceptable complication rate. This technique has tremendous potential for use in evaluating fetal status.

Detection of fetal risk in postmaturity.

Ninety-seven postmature pregnancies were monitored by amnioscopy or amniocentesis (to determine presence or absence of meconium), oxytocin challenge tests (OCT), 24-hour urinary oestriol estimations and fetal movement counts. The colour of the amniotic fluid and the result of the OCT predicted almost all cases of fetal distress in labour and infants with low Apgar scores. Oestriol estimations and fetal movement counts predicted fetal distress only when combined with other positive tests. Of 50 patients with no abnormal test results, 49 had uneventful labours. The Caesarean section rate was not above average and all babies were liveborn.