Plasma Cortisol and Cortisone in Pregnancies with Normal and Anencephalic Fetuses MONTSERRAT DEM. FENCL, RAPIN OSATHANONDH, AND DAN TULCHINSKY Department of Obstetrics and Gynecology, Harvard Medical School; and the Hormone Laboratory, Boston Hospital for Women, Boston, Massachusetts 02115 those of normal fetuses, suggesting that a main portion of UA cortisol and cortisone originates in the fetal adrenal. The UV and MPV plasma F and E concentrations of patients carrying anencephalic fetuses did not differ, however, from those of normal patients, suggesting that these UV corticoids are derived mainly from maternal sources. The amniotic fluid cortisol levels of the patients carving anencephalic fetuses were lower than those observed in the normal pregnancies, suggesting that amniotic fluid cortisol is derived mainly from fetal sources. (J Clin Endocrinol Metab 43: 80, 1976)
ABSTRACT. Plasma cortisol (F), cortisone (E), and progesterone (P), were measured in the umbilical vein (UV), umiblical artery (UA), and maternal peripheral vein (MPV) of 17 normal patients, and of 8 patients carving anencephalic fetuses. The plasma F in MPV of patients undergoing vaginal delivery after labor of spontaneous onset was significantly higher than that of patients delivered by elective cesarean section, whereas the plasma F concentrations in the UA or UV of the 2 groups were not statistically different from each other. The anencephalic fetuses had UA plasma F and E concentrations which were significantly lower than
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N SEVERAL animal species, elevated cortisol levels in fetal blood are associated with the onset of spontaneous labor (1-4). In humans, the extent of corticosteroid synthesis by the fetal adrenal (5-8) and its possible role in the initiation of labor is still controversial (9-14). In part, this can be attributed to the lack of a suitable model for studying the human. The studies of umbilical plasma corticoid levels have been difficult to interpret, due to the unknown magnitude of the maternal contribution to fetal plasma cortisol level (15,16). Pregnancies associated with anencephalic fetuses provide a unique opportunity for studying corticosteroid dynamics in a state in which the fetal adrenals are hypoplastic while the placenta and maternal adrenals are presumably normal. In this communication, we report on the concentrations of cortisol, cortisone, and progesterone in the maternal and umbilical arterial and venous plasma of patients carrying anencephalic fetuses, and compare them to those of normal term pregnancies. Received November 4, 1975. Reprint requests to: Dr. Montserrat deM. Fencl, Boston Hospital for Women, 221 Longwood Avenue, Boston, Massachusetts 02115.
Materials and Methods Study group Eight patients undergoing vaginal delivery at term after the spontaneous onset of labor, 9 nonnal patients undergoing elective cesarean section at term, and 8 patients who were delivered of anencephalic fetuses at 36-47 weeks of pregnancy (6 vaginally following the spontaneous onset of labor and 2 by cesarean section) were studied. All the patients delivered live-born infants between 8 and 12 AM under epidural anesthesia, and none received oxytocin for the stimulation of labor. The maternal peripheral vein blood (MPV) was obtained with delivery of the fetal head, and the umbilical arterial (UA) and venous (UV) blood was obtained after delivery of the placenta. The plasma was immediately separated and frozen until analyzed. Amniotic fluid samples of 4 patients carrying anencephalics were obtained in 2 cases by amniocenteses before the delivery, and in 2 cases during cesarean sections. None of these patients had evidence of polyhydramnios. Steroid determinations All plasma samples were assayed by radioimmunoassay for their progesterone (P), cortisoi (F), and cortisone (E) concentrations. To each plasma sample (0.1-0.5 ml) were added known amounts (ca. 2500 cpm) of tritiated P, F, and E (SA, 40-50 Ci/mmole), obtained from New
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CORTICOSTEROIDS IN ANENCEPHALY England Nuclear, Boston, Mass., and further purified by paper chromatography. Plasma was extracted with 5 ml methylene chloride (spectroquality, Matheson, Coleman and Bell). After separation, the organic solvent was taken to dryness. The residue was dissolved in 100/^1 of methanol (spectroquality, Matheson, Coleman and Bell) and spotted on paper strips (Whatman No. 1) which had been previously washed three times with methanol. Reference standards (ca. 2 /xg) of F, E, corticosterone (B), and P (Sigma Chemical Co.) were applied as a mixture to the outer strips of the paper. The paper chromatogram was equilibrated overnight in the Bush 5 system (2 benzene: 1 methanol:1 water) at room temperature (23 C), and the reference steroids were then detected with 2,4-dinitrophenylhydrazine reagent (17). The corresponding areas on the sample strips were cut and eluted with 5 ml of methanol. The extracts were taken to dryness and dissolved in 0.4 ml ethanol (USP qualify, Commercial Solvent Corp.). An aliquot of 0.1 ml was counted for radioactivity to determine procedural losses, and another aliquot (10-100 (JL\) was used for analysis of the corresponding steroid by radioimmunoassay (RIA). The technique of RIA was the same as previously described for F (18) and P (19). The antiserum used for E measurement was obtained from a rabbit after three months' immunization with cortisol-21-hemisuccinate (Steraloid, Inc.) coupled to human serum albumin (20). Significant cross reaction occurred only with F and B (133% and 10%, respectively), which were subsequently separated by paper chromatography. The coefficient of variation of plasma P, F, and E detenuinations did not exceed 12%. Recoveries averaged 75 ± 10% (SD), and blank values in the chromatogram were always undetectable (
ABSTRACT. Plasma cortisol (F), cortisone (E), and progesterone (P), were measured in the umbilical vein (UV), umiblical artery (UA), and maternal peripheral vein (MPV) of 17 normal patients, and of 8 patients carving anencephalic fetuses. The plasma F in MPV of patients undergoing vaginal delivery after labor of spontaneous onset was significantly higher than that of patients delivered by elective cesarean section, whereas the plasma F concentrations in the UA or UV of the 2 groups were not statistically different from each other. The anencephalic fetuses had UA plasma F and E concentrations which were significantly lower than
I
N SEVERAL animal species, elevated cortisol levels in fetal blood are associated with the onset of spontaneous labor (1-4). In humans, the extent of corticosteroid synthesis by the fetal adrenal (5-8) and its possible role in the initiation of labor is still controversial (9-14). In part, this can be attributed to the lack of a suitable model for studying the human. The studies of umbilical plasma corticoid levels have been difficult to interpret, due to the unknown magnitude of the maternal contribution to fetal plasma cortisol level (15,16). Pregnancies associated with anencephalic fetuses provide a unique opportunity for studying corticosteroid dynamics in a state in which the fetal adrenals are hypoplastic while the placenta and maternal adrenals are presumably normal. In this communication, we report on the concentrations of cortisol, cortisone, and progesterone in the maternal and umbilical arterial and venous plasma of patients carrying anencephalic fetuses, and compare them to those of normal term pregnancies. Received November 4, 1975. Reprint requests to: Dr. Montserrat deM. Fencl, Boston Hospital for Women, 221 Longwood Avenue, Boston, Massachusetts 02115.
Materials and Methods Study group Eight patients undergoing vaginal delivery at term after the spontaneous onset of labor, 9 nonnal patients undergoing elective cesarean section at term, and 8 patients who were delivered of anencephalic fetuses at 36-47 weeks of pregnancy (6 vaginally following the spontaneous onset of labor and 2 by cesarean section) were studied. All the patients delivered live-born infants between 8 and 12 AM under epidural anesthesia, and none received oxytocin for the stimulation of labor. The maternal peripheral vein blood (MPV) was obtained with delivery of the fetal head, and the umbilical arterial (UA) and venous (UV) blood was obtained after delivery of the placenta. The plasma was immediately separated and frozen until analyzed. Amniotic fluid samples of 4 patients carrying anencephalics were obtained in 2 cases by amniocenteses before the delivery, and in 2 cases during cesarean sections. None of these patients had evidence of polyhydramnios. Steroid determinations All plasma samples were assayed by radioimmunoassay for their progesterone (P), cortisoi (F), and cortisone (E) concentrations. To each plasma sample (0.1-0.5 ml) were added known amounts (ca. 2500 cpm) of tritiated P, F, and E (SA, 40-50 Ci/mmole), obtained from New
80
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 14 November 2015. at 18:09 For personal use only. No other uses without permission. . All rights reserved.
81
CORTICOSTEROIDS IN ANENCEPHALY England Nuclear, Boston, Mass., and further purified by paper chromatography. Plasma was extracted with 5 ml methylene chloride (spectroquality, Matheson, Coleman and Bell). After separation, the organic solvent was taken to dryness. The residue was dissolved in 100/^1 of methanol (spectroquality, Matheson, Coleman and Bell) and spotted on paper strips (Whatman No. 1) which had been previously washed three times with methanol. Reference standards (ca. 2 /xg) of F, E, corticosterone (B), and P (Sigma Chemical Co.) were applied as a mixture to the outer strips of the paper. The paper chromatogram was equilibrated overnight in the Bush 5 system (2 benzene: 1 methanol:1 water) at room temperature (23 C), and the reference steroids were then detected with 2,4-dinitrophenylhydrazine reagent (17). The corresponding areas on the sample strips were cut and eluted with 5 ml of methanol. The extracts were taken to dryness and dissolved in 0.4 ml ethanol (USP qualify, Commercial Solvent Corp.). An aliquot of 0.1 ml was counted for radioactivity to determine procedural losses, and another aliquot (10-100 (JL\) was used for analysis of the corresponding steroid by radioimmunoassay (RIA). The technique of RIA was the same as previously described for F (18) and P (19). The antiserum used for E measurement was obtained from a rabbit after three months' immunization with cortisol-21-hemisuccinate (Steraloid, Inc.) coupled to human serum albumin (20). Significant cross reaction occurred only with F and B (133% and 10%, respectively), which were subsequently separated by paper chromatography. The coefficient of variation of plasma P, F, and E detenuinations did not exceed 12%. Recoveries averaged 75 ± 10% (SD), and blank values in the chromatogram were always undetectable (
