The effect of fetal anemia on fetal cardiac troponin T in pregnancies complicated by RhD alloimmunization.

OBJECTIVE: To study the effect of fetal anemia on fetal cardiac troponin T (cTnT) in pregnancies complicated by RhD alloimmunization. METHOD: Twenty pregnant women complicated by RhD alloimmunization who underwent intrauterine transfusion (IUT) for treatment of fetal anemia were studied. Immediately before IUT, fetal blood was obtained for hemoglobin and cTnT measurements. RESULTS: Complete measurements of hemoglobin and cTnT before IUT were obtained in 49 procedures, of which 20 were first-time. The regression analysis between hemoglobin z-score and cTnT values in 49 procedures showed significant negative correlation (r = -0.43, p = 0.002, Regression equation Log(cTnT) = -1.5057 + -0.07563 Hb z-score). Cardiac TnT values before first IUT were significantly associated with perinatal death. In the group with elevated cTnT (n = 7), fetal or neonatal death was more frequent (2 IUD and 2 NND) when compared to normal cTnT group (n = 13, 1 IUD) (57.1 vs. 7.7%, p = 0.031, Fisher's exact test). CONCLUSION: Fetal blood concentration of cTnT before IUT was negatively correlated to hemoglobin z-score, and levels of cTnT help to manage the pregnancies complicated by RhD alloimmunization.

Bilirubin/albumin ratios in fetal blood and in amniotic fluid in rhesus immunization.

OBJECTIVE: To test the hypothesis that unconjugated bilirubin is equally distributed over the albumin molecules present in fetal blood and amniotic fluid in Rhesus (Rh) immunization. METHODS: Molar concentrations of unconjugated bilirubin and albumin were measured in fetal blood and amniotic fluid samples, obtained before the first intrauterine transfusion in 30 nonhydropic, anti-D-alloimmunized fetuses, with gestational ages ranging from 20 to 35 weeks. RESULTS: Bilirubin concentration in amniotic fluid was best predicted by a combination of bilirubin concentration in fetal blood (P<.001), albumin concentration in fetal blood (P=.008), and albumin concentration in amniotic fluid (P<.001) (adjusted R2=0.91). The bilirubin/albumin ratios in fetal blood were linearly correlated with the bilirubin/albumin ratios in amniotic fluid (R2=0.75, P<.001). However, the bilirubin/albumin ratios in fetal blood were always higher than the bilirubin/albumin ratios in amniotic fluid (regression coefficient 1.4, 95% confidence interval 1.1-1.7). In our population, a bilirubin/albumin ratio in amniotic fluid of 0.10 or greater had a better sensitivity and specificity to predict severe anemia (Z hemoglobin -5 standard deviations or less) than the Queenan 4 or the Liley 2c line. CONCLUSION: The relation between fetal hemolysis and amniotic fluid bilirubin concentration is based on the linear correlation between bilirubin/albumin ratios in fetal blood and in amniotic fluid. The slope in Queenan's and Liley's chart follows that of the albumin concentration in amniotic fluid during gestation. LEVEL OF EVIDENCE: III.

On the origin of amniotic fluid bilirubin.

We studied the relationship between bilirubin concentrations in amniotic fluid and fetal blood in 68 non-hydropic rhesus d-alloimmunized anemic fetuses at first blood sampling. In these alloimmunized fetuses, the amniotic fluid/fetal blood ratio for bilirubin decreased from 0.09 at 28 weeks to 0.05 at 33 weeks. In normal fetuses, amniotic fluid/fetal blood ratios for bilirubin, and for albumin, are in the same range and show a similar decrease during gestation. We conclude that amniotic fluid bilirubin concentration is determined, firstly, by fetal blood bilirubin concentration and, secondly, by the amniotic fluid/fetal blood ratio of albumin. Among five possible pathways bilirubin could take to build up a concentration in amniotic fluid (fetal kidneys, lungs, skin, bowel, membranes), the intramembranous pathway is the only one that is compatible with the amniotic fluid/fetal blood ratios for bilirubin that we found and must therefore be the most important.

Fetal bone metabolism in normal and rhesus isoimmunised pregnancies.

OBJECTIVE: To construct gestation-specific reference intervals for fetal concentrations of biochemical markers of bone metabolism and assess the effect of rhesus isoimmunisation on these. METHODS: Fetal blood samples were obtained by cordocentesis from 175 pregnancies (43 complicated by rhesus isoimmunisation) and assayed for carboxy terminal pro-peptide of type I pro-collagen (PICP) and cross-linked carboxyterminal telopeptide of type I collagen (ICTP) which directly monitor the rate of bone formation and resorption respectively. RESULTS: Both plasma PICP and ICTP were negatively correlated with gestational age (r = -0.351 and -0.472 for PICP and ICTP, respectively, and P < 0.001 for both). In fetuses affected by rhesus isoimmunisation PICP levels were lower (P=0.030) and more variable (P <0.001) than expected, compared with normal unaffected fetuses. However, no such differences were found in the ICTP levels. In the fetuses affected by rhesus isoimmunisation there was a significant correlation between haemoglobin concentration and both PICP (r = 0.504, P = 0.001) and ICTP (r = 0.343, P = 0.030). CONCLUSIONS: Fetal bone turnover declines from early second trimester to term, and may be deranged in fetuses affected by rhesus isoimmunisation.

[Relationship between erythropoietin activity in fetal blood and Liley's curve and bilirubin concentration in amniotic fluid in serologic conflict]. / Zwiazek miedzy aktywnoscia erytropoetyny we krwi plodowej a krzywa Lileya i stezeniem bilirubiny w plynie owodniowym w konflikcie serologicznym.

Authors analysed concentration of erythropoietin (EPO) in fetal blood from pregnancies complicated by serologic conflict. There was conducted the trial of settle correlation between EPO activity in fetal blood and bilirubin concentration in amniotic fluid and its optical density ratio obtained by amniopuncture in the same pregnancies.

Cytokines in fetal blood and amniotic fluid in Rh-immunized pregnancies.

OBJECTIVE: To determine fetal serum and amniotic fluid (AF) levels of interleukin (IL)-3, IL-6, granulocyte-macrophage colony-stimulating factor, stem cell factor, and erythropoietin, and to explore the relationship between cytokines and hemoglobin concentration, white blood cell count (WBC), and platelet count in fetuses affected by Rh immunization. METHODS: Thirty-four consecutive Rh-immunized patients in gestational weeks 19-33 were included. All patients were investigated by funipuncture and 13 by amniocentesis. The levels of IL-3, IL-6, granulocyte-macrophage colony-stimulating factor, stem cell factor, and erythropoietin were estimated using commercially available immunoassays. RESULTS: There was a significant correlation between erythropoietin concentrations in fetal serum and AF (r = 0.54, P < .05), whereas none of the other cytokines showed a positive correlation between these two compartments. Fetal serum contained higher concentrations of IL-3, granulocyte-macrophage colony-stimulating factor, stem cell factor, and erythropoietin compared with AF. In contrast, the IL-6 level was significantly higher in AF compared with fetal serum (P = .002). Erythropoietin and IL-3 levels were both negatively correlated with fetal hemoglobin concentrations (r = -0.75, P = .02, and r = -0.67, P = .045). The fetal WBC correlated significantly with the fetal serum concentration of granulocyte-macrophage colony-stimulating factor (r = 0.38, P = .04). CONCLUSION: Human fetuses with anemia due to erythrocyte immunization exhibit an increased production of erythropoietin and IL-3. Other studied cytokines (such as stem cell factor, granulocyte-macrophage colony-stimulating factor, and IL-6) did not correlate with the degree of fetal anemia. Among the studied cytokines, only erythropoietin showed a positive correlation between fetal serum and AF.

Oxygenation of the human fetus as a function of hemoglobulin concentration.

We studied the relationships between fetal hemoglobin concentration, acid base status, and lactate concentration in umbilical venous and fetal heart blood in 157 fetuses affected by blood group incompatibility who had been exposed for 214 fetal blood sampling procedures (cordocentesis in 153 and intracardiac puncture in 61 cases). All blood samplings were obtained before fetal blood transfusions were administered. The results indicate that the human fetus can maintain a normal acid-base status until a 50% reduction of the hemoglobin concentration. A further reduction of hemoglobulin is associated with an accumulation of lactate both in umbilical venous and fetal heart blood. The partial oxygen tension and the oxygen saturation in umbilical venous blood remained virtually unchanged with decreasing hemoglobin concentration (r = -0.11, P = 0.21; r = 0.09, P = 0.31, respectively), whereas these parameters decreased significantly (r = 0.33, P = 0.02; r = 34, P = 0.02) in blood obtained from the fetal heart. The partial carbon dioxide tension of umbilical vein blood decreases significantly with a reduced hemoglobin concentration (r = 0.25 P = 0.008). We speculate that these alterations in acid-base status in umbilical vein and fetal heart blood reflect a circulatory transition from a high to a low cardiac output as the hemoglobin concentration decreases.

Erythropoietin concentrations in amniotic fluid and umbilical venous blood from Rh-immunized pregnancies.

We set out to investigate prospectively the levels of erythropoietin in amniotic fluid and umbilical venous blood, and to attempt to relate these to fetal haemoglobin and lactate concentrations and to pCO2 and PO2 in Rh immunised patients studied before the onset of labor. Fetal blood was obtained by cordocentesis, and amniotic fluid by amniocentesis from a consecutive series of 36 Rh immunized patients at the time of fetal blood sampling. There was a close correlation (tau = 0.357, P = 0.0001) between the concentrations of erythropoietin in umbilical venous blood and those in amniotic fluid. Erythropoietin in umbilical venous blood correlated inversely with hemoglobin (tau = 0.453, P = 0.0001), and directly with lactate concentrations (tau = 0.450, P = 0.0005). When all other variables were considered, multiple regression analysis demonstrated hemoglobin concentration to be the only variable to be related to the level of erythropoietin in umbilical venous blood taken before transfusion. When the same analysis was performed on the same variables, adding erythropoietin concentration in amniotic fluid as the dependent variable, only erythropoietin in umbilical venous blood was found to be related to the level of erythropoietin in amniotic fluid. We conclude that the erythropoietin concentration in umbilical venous blood from Rh-immunized patients before the onset of labor, is related to fetal anemia. We also conclude that erythropoietin concentration in amniotic fluid is related to that in fetal blood, thereby indicating that the fetus is an important source of amniotic fluid erythropoietin in non laboring patients.

Pregnancy-associated major basic protein in amniotic fluid.

The pregnancy-associated major basic protein, a protein elevated in the sera of all pregnant women, is virtually identical to the eosinophil granule major basic protein. To determine whether pregnancy-associated major basic protein is present in amniotic fluid, we examined samples from both early and late gestation by a double antibody radioimmunoassay. A total of 112 amniotic fluids were tested and all but three contained levels of pregnancy-associated major basic protein greater than 400 ng/ml. Amniotic fluid pregnancy-associated major basic protein antigenic activity was immunochemically identical to that of the eosinophil granule major basic protein and also had identical physicochemical properties such as heat stability and the need for reduction and alkylation. Although the majority of amniotic fluid samples (90 of 112) were obtained from healthy women with normal gestations, the remaining 21 amniotic fluid samples were from women with Rh sensitization and from one gestation complicated by intrauterine growth retardation.

[Characteristics of the renin-aldosterone system of the fetoplacental complex in fetal erythroblastosis]. / Osobennosti renin-al'dosteronovoi sistemy plodovo-platsentarnogo kompleksa pri gemoliticheskoi bolezni ploda.

Renin activity and aldosterone concentrations have been determined in amniocentesis samples and fetal plasma obtained by intrauterine umbilical vein puncture in 23 women at 25 to 34 weeks' gestation, normal or complicated by hemolytic disease of the fetus. Prior to the intrauterine interventions, Doppler studies of umbilical arterial circulation were done and 1-hour fetal diuresis was measured. Fetal plasma pH and fetoplacental flood volume were determined. Hemolytic disease was shown to increase renin activity and aldosterone concentration in the amniotic fluid and fetal plasma in parallel with reduction in the fetoplacental blood flow and 1-hour fetal diuresis, increase in vascular resistance of fetal placenta and hypoxia. The increase in renin-aldosterone activity of the fetoplacental unit may be a compensatory fetal and chorionic response to hemodynamic and water-mineral balance in the presence of impaired uteroplacental circulation.

Amniotic fluid prolactin levels in rhesus isoimmunized patients.

The concentration of prolactin in the amniotic fluid (AFPRL) was measured in 75 samples obtained in the third trimester of 14 isoimmunized women. There was a uniform decline in prolactin levels with advancing gestation in each pregnancy (r = -0.89 to -0.99). The decline in AFPRL was similar in uncomplicated pregnancies. AFPRL levels were not predictive of umbilical cord hemoglobin or bilirubin levels and amniotic fluid lecithin/sphingomyelin ratio.

Pulmonary phospholipids in amniotic fluid of pathologic pregnancies: relationship with clinical status of the newborn.

We evaluated phospholipids, C-peptide and cortisol levels in amniotic fluid of 203 pathologic pregnancies (63, class A, B and C diabetics; 11 class D, F and H diabetics; 44 preclampsia and 85 Rh-isoimmunization); the control group was 82 normal pregnant women. There was an acceleration of fetal pulmonary maturation in women with preclampsia and severe Rh-isoimmunization in class D, F and H diabetics (at 34 weeks gestation the incidence of mature surfactant (lecithin/sphingomyelin greater than or equal to 2.7 and presence of phosphatidyl-glycerol) in these groups was 30%, 50% and 100%, respectively, while it was zero in the control group). At 37 and 38 weeks only 44.4% of the class A, B and C diabetics had mature surfactant and there was a significant difference with respect to the control group (x2 = 4.9; p less than 0.05); C-peptide levels in these diabetics (class A, B and C) were higher than in controls (p less than 0.001); in pregnant women with accelerated fetal lung maturation they were lower. We demonstrated a close relationship between fetal pulmonary maturity and the type of surfactant in amniotic fluid, which was independent of gestational age.

Amniotic fluid erythropoietin predicts fetal distress in Rh-immunized pregnancies.

Repeated amniotic fluid erythropoietin measurements in 23 Rh-immunized pregnancies were done to evaluate erythropoietin levels of amniotic fluid as an indicator of fetal distress (umbilical artery, pH 7.14 or less, or 1-minute Apgar score of 4 or less). Amniotic fluid erythropoietin levels did not vary significantly between 168 and 273 gestational days in the pregnancies without fetal distress. Increasing levels of amniotic fluid erythropoietin predicted highly reliably severe fetal distress at birth. Whether erythropoietin levels of amniotic fluid can also predict fetal distress in other pathologic pregnancies needs further study.

Hormone release by primary amniotic fluid cell cultures.

Amniotic fluid cells have been widely used in prenatal diagnosis; however, there is great heterogeneity of the cells and their origin. In this study we analyze the karyotype and release of human chorionic gonadotropin (hCG), human chorionic somatomammotropin (hCS), free estriol (E 3), prolactin (PRL) and progesterone (P) of amniotic fluid cells from primary cultures of six normal and two anencephalic fetuses. In all the amniotic fluid samples there was release of hCG; in one amniotic fluid, in which several tetraploid colonies were found. PRL and P were also released. The heterogeneity of amniotic fluid cell morphology and their hormone release in culture was confirmed. The presence of hormones like hCG supports the trophoblastic origin of some amniotic fluid cells from normal and anencephalic fetuses. Other hormones, such as PRL and P could be used in the differential diagnosis between the karyotype of fetal membranes and the true fetal karyotype. Amniotic fluid cell cultures used in prenatal diagnosis yielded second trimester placental cells without any elaborate methods that could be used as cell models for hormone studies.

Measurement of fetal surfactant production by fluorescence polarization of amniotic fluid in complicated pregnancies.

The results are presented of fluorescence polarization as a method for measurement of surfactant production in 159 specimens of amniotic fluid collected from pregnant women with diabetes, hypertension, Rh immunization, premature rupture of membranes (for more than 48 h prior to delivery) and intrauterine growth retardation (IUGR). The predictability of the development of respiratory distress syndrome has been assessed by this assay. Its specificity, sensitivity and overall accuracy were similar to the lecithin/sphingomyelin (L/S) ratio. The influence of the conditions detailed on fetal lung maturation was determined, lung development being enhanced until near term by Rh immunization, rupture of membranes and hypertension with IUGR.

There is evidence that amniotic fluid arginine vasopressin is a marker for foetal stress in rhesus erythroblastosis.

In response to different stress stimuli the foetal neurohypophysis releases arginine vasopressin (AVP). Part of the AVP is cleared from circulation by urinary excretion into the amniotic fluid (AF). Increased AF AVP levels may therefore indicate foetal stress, all the more because AF AVP solely is of foetal origin. We therefore studied AF AVP levels in 13 patients with rhesus erythroblastosis from 22 to 34 weeks of gestation. Twenty-eight patients from 14 to 34 weeks of gestation served as controls. The AVP levels were measured by RIA. Spectral absorption curves were performed and delta/E values determined at 450 nm. Mean AF AVP levels in controls were 2.39 pg/ml and were not normally distributed. There was no significant change in AF AVP levels with different gestational age. If in rhesus erythroblastosis patients the delta/E value was low (n = 7; x = 0.048 +/- 0.007 SE), the AF AVP values were not increased. If the delta/E values were within zone III (n = 6; x = 0.22 +/- 0.035 SE), indicating severe haemolysis, the AF AVP levels were significantly elevated (4.7 pg/ml +/- 0.51 SE; P = 0.001). Linear regression analysis showed a significant correlation between delta/E and AF AVP values (P = 0.05; y = 1.94 +/- 10.88 x). We conclude that there is evidence for the role of AF AVP as a marker for foetal stress in rhesus erythroblastosis.