Amniotic Fluid Hormone Research Articles

Application of Ultrasonographic Imaging in Fetal Thyroid Function Measurement of Amniotic Hormone Concentration

Objective: To investigate the fetal thyroid function using ultrasound imaging combined with amniotic fluid hormone concentration. Method: 380 pregnant women who did pregnancy test in our hospital from August 2015 to August 2017 were selected as the subjects. 260 of them had normal fetal growth indicators, all of them were singletons, with a gestational time of 18–41 weeks. 120 pregnant women underwent cesarean section, parturition, and amniocentesis. All pregnant women had no thyroid related diseases, hypertension, or autoimmune diseases. 3DUS (Three-dimensional ultrasound) and 2DUS (two-dimensional ultrasound) were used to measure the size of fetal thyroid gland, and the amniotic hormone content was measured. Result: The repeatability and consistency of the 3DUS-VOCAL (Voual organ computerized analysis) technique were superior to 2DUS in the measurement of fetal thyroid volume in 20 cases. There were 243 fetal thyroid size-related data collected from 260 fetuses at 18 weeks and 41 weeks of pregnancy. Amniotic fluid FT3, T3 had a certain correlation with gestational age, but amniotic fluid FT4, T4, TSH had no significant correlation with gestational age. 4 cases of fetal hypothyroidism were screened by ultrasound combined with amniotic fluid hormone measurement. Conclusion: The volume of fetal thyroid measured by 3DUS-VOCAL is better than that of 2DUS. Ultrasound combined with amniotic fluid thyroid associated hormone detection is helpful to evaluate fetal thyroid function.

Steroid Hormone Levels in Recipient Amniotic Fluid in Twin-Twin Transfusion Syndrome and Their Association with Preterm Delivery.

Preterm delivery following fetoscopic laser surgery (FLS) of twin-twin transfusion syndrome (TTTS) is associated with severe perinatal morbidity and mortality. The role of steroid hormones in amniotic fluid (AF) after FLS remains unknown. A prospective cohort study of consecutive case series of FLS for TTTS was performed from April 2012 to February 2017. Cases were divided into early (≤27 weeks) spontaneous preterm delivery (ED) and late delivery (LD; ≥34 weeks) following FLS and compared. AF supernatants were assessed for protein, estradiol, progesterone and cortisol levels (using the ELISA kit), and normalized to total protein levels to adjust for dilution. A total of 294 consecutive cases of FLS for TTTS in monochorionic-diamniotic twins were performed during the study period. AF was available in 44 ED patients and 50 LD patients. On logistic regression, ED was associated with higher normalized progesterone levels (odds ratio [OR]: 1.25; 95% confidence interval [CI]: 1.12-1.41), lower normalized cortisol (OR: 0.78; 95% CI: 0.64-0.96), and higher estradiol levels (OR: 1.3; 95% CI: 1.03-1.63). Elevated AF normalized progesterone and estradiol, and lower normalized cortisol levels were associated with ED. This novel finding requires further exploration to establish the molecular mechanism operational in pregnancies complicated by TTTS to potentially prevent early preterm birth after fetal surgery.

Quantifying steroid hormones in amniotic fluid by ultra-performance liquid chromatography and tandem mass spectrometry

Background: Simultaneous assessment of steroid hormone concentrations in amniotic fluid is of importance for elucidating long-term consequences of intrauterine processes, and of broad scientific and clinical relevance. The objective of the study was to develop sensitive and specific analytical ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) methods for determination of steroid hormone concentrations in human amniotic fluid, and to provide proof-of-concept of their applicability. Methods: Methods were validated according to linearity, limit-of-detection, limit-of-quantification, recovery, intra- and inter-assay precision, and applied to 275 amniotic fluid samples. Results: Limits-of-quantification (S/N=10:1) were 0.05 ng/mL for cortisol, dehydroepiandrosterone sulfate (DHEAS), estradiol, estriol, and testosterone, and 0.01 ng/mL and 1.0 ng/mL for cortisone and dehydroepiandrosterone (DHEA), respectively. Good inter- and intra-assay precision were observed. Cortisol, cortisone, DHEAS, estradiol and estriol concentration were quantified in all samples. By lack of sensitivity of the analytical method, DHEA was quantified in 11 samples only. Testosterone was quantified in 119 of 275 samples analyzed. 116 of the quantified testosterone samples were from male offspring (out of 138 male offspring). Conclusion: These specific and sensitive methods offer a simple and non-invasive way to measure cortisol, cortisone, DHEAS, estradiol, estriol, and testosterone concentrations in human amniotic fluid.

Sequential Amniotic Fluid Thyroid Hormone Changes Correlate with Goiter Shrinkage following in utero Thyroxine Therapy

Several isolated reports of fetal goiter treatment have shown limited generalizability of approaches and provide no real guidance for optimal timing, dosages, and treatment strategies. Graves' disease accounts for >60% of these cases. Maternal treatments of hyperthyroidism include antithyroid medications such as methimazole and more commonly propylthiouracil (PTU). Here, our management of a patient with a fetal thyroid goiter from maternal exposure to PTU diagnosed at 23.6 weeks' gestation and the management of other cases allow us propose a general strategy for treatment. Intrauterine therapy with 200 and then 400 μg of levothyroxine (3 weeks apart) showed an 85% reduction in fetal thyroid goiter volume. We collected amniotic fluid samples at the time of treatments and assayed thyroid hormones and associated antibodies which closely reflected the changes in thyroid goiter mass volume. Our observations suggest a weekly or biweekly therapeutic intervention schedule. Utilizing both goiter size as well as a novel approach in using amniotic fluid hormone levels to monitor therapy efficacy might improve the quality of treatments. Only with a standardized approach and collection of amniotic fluid thyroid panels do we have the opportunity to develop the database required to determine the number and timing of treatments needed.

Ambiguous genitalia: What prenatal genetic testing is practical?

Concern for ambiguous genitalia or chromosome-phenotype discordance detected in a prenatal setting has increased over the last two decades. Practitioners faced with this prenatal finding have a variety of genetic tests available to them; however, it is unclear to what extent prenatal testing for disorders of sex development (DSD) is useful or practical. We undertook a retrospective review of the medical records of 140 individuals evaluated through the DSD clinic at Seattle Children's Hospital with birthdates from 01/01/1994 through 08/16/2011 to determine the rate of prenatal detection of ambiguous genitalia in individuals with DSD, what prenatal diagnostic workup was undertaken, and the postnatal outcome, including whether a postnatal genetic diagnosis was confirmed. Of all 140 subjects, 34 (24%) were identified prenatally. The most common postnatal diagnoses were penoscrotal hypospadias with transposition of the scrotum with no known genetic cause (24/140; 17%) and 21-hydroxylase deficiency (20/140; 14%). Apart from these, no single diagnosis comprised more than a few cases. Prenatal diagnostic testing varied widely, from no tests to multiple molecular tests with amniotic fluid hormone concentrations. In the absence of other fetal anomalies or growth retardation on ultrasound, prenatal karyotype with fluorescence in situ hybridization for the SRY gene is the most useful test when ambiguous genitalia is suspected. Further prenatal testing for Smith-Lemli-Opitz syndrome in 46,XY individuals and congenital adrenal hyperplasia in 46,XX individuals may be considered. However, targeted molecular testing for rare DSD conditions in the absence of a family history of DSD has a low yield.

Increased levels of human placental growth hormone in the amniotic fluid of pregnancies affected by Down syndrome

Objective To evaluate the concentrations of human placental growth hormone (hPGH) in amniotic fluid (AF) at gestational mid-trimester in normal pregnancies and in pregnancies complicated by Down’s syndrome. Design AF samples from 21 women with Down’s syndrome pregnancies were analyzed retrospectively. About 47 AF samples from women with singleton, uncomplicated pregnancies, who gave birth to healthy neonates with birth weight appropriate for gestational age were used as controls. All AF samples were obtained during amniocentesis for fetal karyotyping at 16–23 weeks’ gestation. hPGH levels were measured by a solid phase immunoradiometric assay using two different epitopes. Results The mean hPGH values in the AF of the Down’s syndrome-affected pregnancies were significantly higher ( P < 0.05) compared to those of normal pregnancies, at 16–23 weeks’ gestation: mean-value ± SD in the AF was 1.96 ± 1.35 μg/l vs. 0.82 ± 0.67 μg/l. Conclusions Higher hPGH levels in AF were found in pregnancies affected by Down’s syndrome as compared to normal pregnancies at gestational mid-trimester. hPGH was detected in all AF samples, and it provides evidence that this pregnancy-specific hormone enters the fetal compartment and is not limited to the maternal circulation. The physiological role and effect of hPGH on fetal growth in normal and pathological pregnancies needs further investigation.

The effect of gestational age and labor on placental growth hormone in amniotic fluid

Objective Placental growth hormone (PGH) is produced by trophoblast. This hormone becomes detectable in maternal serum during the first trimester of pregnancy. Its concentration increases as term approaches and becomes undetectable within one hour of delivery. PGH has important biological properties, including somatogenic (growth promotion), lactogenic, and lipolytic activity. Recently, PGH has been detected in amniotic fluid (AF) of midtrimester pregnancies. The purpose of this study was to determine whether PGH concentrations in AF change with advancing gestational age and in labor at term. Design AF was assayed for PGH concentrations in samples obtained from patients undergoing genetic amniocentesis between 14 and 18 weeks of gestation ( n = 67), normal patients at term not in labor ( n = 24), and pregnant women at term in labor ( n = 51). PGH concentrations were determined by ELISA. Non-parametric statistics were used for analysis. Results (1) PGH was detected in all AF samples; (2) patients in the midtrimester had a higher median concentration of PGH in AF than those at term (midtrimester: median: 3140.5 pg/ml; range: 1124.2–13886.5 vs. term: median: 2021.1 pg/ml; range: 181.6–8640.8; p < 0.01); (3) there was no difference in the median concentration of PGH between women at term, not in labor, and those in labor (term not in labor: median: 2113.4 pg/ml; range: 449.3–8640.8 vs. term in labor: median: 2004.1 pg/ml; range: 181.6–8531.5; p = 0.73). Conclusions (1) PGH is detectable in AF at both mid- and third trimesters; (2) the median AF concentration of PGH is significantly lower at term when compared to the second trimester; (3) labor at term is not associated with changes in the AF concentration of PGH. The role of this unique placental hormone now found in the fetal compartment requires further investigation.

Establishment of Reference Intervals for Thyroid-Stimulating Hormone and Free Thyroxine in Amniotic Fluid Using the Bayer ADVIA Centaur

Thyroid hormone concentrations in amniotic fluid (AF) can aid in the diagnosis of fetal hypothyroidism. However, the availability of reference intervals for thyroid hormones in AF using current methods is limited. The purpose of this study was to validate the Bayer ADVIA Centaur (Bayer Healthcare, Tarrytown, NY) third-generation thyroid-stimulating hormone (TSH), total thyroxine (T4), and free thyroxine (FT4) assays for use with AF and establish reference intervals for these thyroid hormones in AF. Third-trimester AF samples were assayed for TSH, T4, and FT4. Reference intervals were calculated using nonparametric analyses. The reference intervals were as follows: TSH (n = 131), 0.04 to 0.51 microIU/mL (0.04-0.51 mIU/L), with a median of 0.10 microIU/mL (0.10 mIU/L); FT4 (n = 133), less than 0.10 to 0.77 ng/dL (1.29-9.93 pmol/L), with a median of 0.26 ng/dL (3.35 pmol/L). T4 in AF was undetectable using the Centaur assay, reinforcing the importance of validating different manufacturers' immunoassays for use with nonserum specimens. These reference intervals represent the first study performed to date using a third-generation TSH assay and a sensitive FT4 assay.

The effect of gestational age and labor on placental growth hormone in amniotic fluid

The effect of gestational age and labor on placental growth hormone in amniotic fluid

Profiling Steroid Hormones in Amniotic Fluid of Midpregnancy by Routine Stable Isotope Dilution/Gas Chromatography-Mass Spectrometry: Reference Values and Concentrations in Fetuses at Risk for 21-Hydroxylase Deficiency

Profiling Steroid Hormones in Amniotic Fluid of Midpregnancy by Routine Stable Isotope Dilution/Gas Chromatography-Mass Spectrometry: Reference Values and Concentrations in Fetuses at Risk for 21-Hydroxylase Deficiency

Profiling steroid hormones in amniotic fluid of midpregnancy by routine stable isotope dilution/gas chromatography-mass spectrometry: reference values and concentrations in fetuses at risk for 21-hydroxylase deficiency.

Using routine stable isotope dilution/gas chromatography-mass spectrometry, 17-hydroxyprogesterone, androstenedione, testosterone, dehydroepiandrosterone, androstanediol, and 5alpha-dihydrotestosterone have been profiled in amniotic fluid of midgestation in 77 normal fetuses and 38 untreated or dexamethasone-treated fetuses at risk for 21-hydroxylase deficiency. Dexamethasone was suspended 5-7 days before amniocentesis. In normal fetuses, amniotic fluid concentrations (median, range; nanograms per mL) of 17-hydroxyprogesterone did not reveal a sex difference (1.48, 0.21-4.96), whereas those of androstenedione were lower in females (0.53, 0.00-2.71) than in males (0.93, 0.29-1.98). Testosterone levels were higher in males (0.24, 0.00-0.50) than in females (0.00, 0.00-0.27). No sex difference was found for dehydroepiandrosterone (0.47, 0.19-1.77). Levels of androstanediol and 5alpha-dihydrotestosterone were below the detection limit of our method in most cases. Regarding prenatal diagnosis of 21-hydroxylase deficiency, 17-hydroxyprogesterone and androstenedione presented the diagnostically most valuable steroids and were of equal diagnostic potential. They permitted successful diagnosis in 36 of 37 fetuses at risk: 12 were untreated and unaffected, 13 were treated and unaffected, 4 were untreated and affected (3 salt wasters and 1 simple virilizer), and 8 were treated and affected (5 salt wasters and 3 simple virilizers). In the latter group, one simple virilizer revealed normal steroid concentrations. Isotope dilution/gas chromatography-mass spectrometry, providing the highest specificity in steroid analysis, is proposed for routine use in clinical steroid analysis whenever maximal reliability is requested. Our study provides the first mass spectrometric reference data on amniotic fluid steroid concentrations and underscores the high accuracy of prenatal hormonal diagnosis of 21-hydroxylase deficiency.

Gastrointestinal and pancreatic hormones in the human fetus and mother at 18-21 weeks of gestation.

Several gastrointestinal hormones appear to play an important developmental role in the newborn, particularly in preterm neonates. Although the cells producing these peptides develop towards the end of the first trimester, fetal secretion of these regulatory peptides has not hitherto been demonstrated. Using samples collected by fetoscopy at 19-21 weeks of gestation we have measured concentrations of several gastrointestinal and pancreatic hormones. Maternal venous and amniotic fluid hormone concentrations were measured simultaneously. Concentrations of the pancreatic hormones, insulin, glucagon and pancreatic polypeptide (PP) were similar in fetal and maternal blood. Gastrin and motilin were present in the fetal circulation but at about 30% (p < 0.05) and 60% (p < 0.01) of the maternal levels, respectively. In contrast, enteroglucagon concentrations were more than twofold higher in the fetal circulation compared with maternal levels (p < 0.05). Concentrations of gastric inhibitory polypeptide (GIP) in fetal blood were higher than levels in maternal blood but not significantly. Concentrations of GIP (p < 0.001) were higher in the amniotic fluid than the fetal circulation. Gastrin and glucagon levels were similar in amniotic fluid and fetal blood. In contrast, PP and motilin were present in amniotic fluid, but at lower concentrations than in fetal blood. Enteroglucagon was not detectable in amniotic fluid. In conclusion, several alimentary hormones are secreted in the fetus at midterm. Since these peptides have trophic, secretory and motor effects on the gut, it is likely that these regulatory peptides are involved in the functional development of the fetal intestine.

Corticotrophin-Releasing Hormone Levels in Human Plasma and Amniotic Fluid During Gestation

Summary OBJECTIVE Corticotrophin releasing hormone, a hypothalamic neuropeptide also made in placenta, may regulate fetal maturation in a stress-responsive manner. The objectives of this study were: (1) to determine if levels of corticotrophin releasing hormone in the amniotic fluid correlate with fetal lung maturation; (2) to confirm that third trimester plasma levels of corticotrophin releasing hormone are increased in patients with pregnancy-induced hypertension compared to normotensives, and (3) to increase the recovery of extracted corticotrophin releasing hormone from plasma and amniotic fluid. DESIGN (1) Levels of corticotrophin releasing hormone in amniotic fluid during the third trimester were compared with those of saturated phosphatidyl choline. (2) Corticotrophin releasing hormone levels were measured in a group of normotensive pregnant women during the entire gestation period. Corticotrophin releasing hormone levels during the third trimester were compared in normotensives and patients with pregnancy-induced hypertension. PATIENTS Twenty-one non-pregnant normal volunteers and 63 pregnant women. MEASUREMENTS Blood pressure, corticotrophin releasing hormone in plasma and amniotic fluid, and saturated phosphatidyl choline in amniotic fluid. RESULTS Corticotrophin releasing hormone levels in amniotic fluid samples during the third trimester ranged from 12 to 98 pmol/l and positively correlated with the saturated phosphatidyl choline levels, but not with gestational age. A significant difference existed in plasma corticotrophin releasing hormone concentration between gestational age-matched third trimester normotensive and hypertensive gravids: corticotrophin releasing hormone levels were significantly lower in normotensives (223 ± 65 pmol/l) than in patients with pregnancy-induced hypertension (544 ± 106 pmol/l, P= 0.001). Plasma corticotrophin releasing hormone increased with gestational age from 51 pmol/l (range 8.4–85) at 25–32 weeks to 375 pmol/l (range 35–1386) at 33–40 weeks. During the third trimester the rise in plasma corticotrophin releasing hormone conformed to an exponential mathematical model of a positive feedback loop between placental corticotrophin releasing hormone and fetal adrenal Cortisol. CONCLUSIONS During the third trimester of pregnancy there is a positive correlation between the level of amniotic fluid corticotrophin releasing hormone and that of saturated phosphatidyl choline. The positive correlation between amniotic fluid corticotrophin releasing hormone and saturated phosphatidyl choline, but not between amniotic fluid corticotrophin releasing hormone and gestational age, suggests that a factor(s), such as stress, may affect both amniotic fluid corticotrophin releasing hormone and saturated phosphatidyl choline in parallel. Furthermore, our data are consistent with the hypothesis that the rise in placental corticotrophin releasing hormone is coupled to an increase in fetal glucocorticoid and lung maturation, and that stresses such as pregnancy-induced hypertension may accelerate this process.

Corticotrophin releasing hormone levels in human plasma and amniotic fluid during gestation

Corticotrophin releasing hormone, a hypothalamic neuropeptide also made in placenta, may regulate fetal maturation in a stress-responsive manner. The objectives of this study were: (1) to determine if levels of corticotrophin releasing hormone in the amniotic fluid correlate with fetal lung maturation; (2) to confirm that third trimester plasma levels of corticotrophin releasing hormone are increased in patients with pregnancy-induced hypertension compared to normotensives, and (3) to increase the recovery of extracted corticotrophin releasing hormone from plasma and amniotic fluid. (1) Levels of corticotrophin releasing hormone in amniotic fluid during the third trimester were compared with those of saturated phosphatidyl choline. (2) Corticotrophin releasing hormone levels were measured in a group of normotensive pregnant women during the entire gestation period. Corticotrophin releasing hormone levels during the third trimester were compared in normotensives and patients with pregnancy-induced hypertension. Twenty-one non-pregnant normal volunteers and 63 pregnant women. Blood pressure, corticotrophin releasing hormone in plasma and amniotic fluid, and saturated phosphatidyl choline in amniotic fluid. Corticotrophin releasing hormone levels in amniotic fluid samples during the third trimester ranged from 12 to 98 pmol/l and positively correlated with the saturated phosphatidyl choline levels, but not with gestational age. A significant difference existed in plasma corticotrophin releasing hormone concentration between gestational age-matched third trimester normotensive and hypertensive gravids: corticotrophin releasing hormone levels were significantly lower in normotensives (223 +/- 65 pmol/l) than in patients with pregnancy-induced hypertension (544 +/- 106 pmol/l, P = 0.001). Plasma corticotrophin releasing hormone increased with gestational age from 51 pmol/l (range 8.4-85) at 25-32 weeks to 375 pmol/l (range 35-1386) at 33-40 weeks. During the third trimester the rise in plasma corticotrophin releasing hormone conformed to an exponential mathematical model of a positive feedback loop between placental corticotrophin releasing hormone and fetal adrenal cortisol. During the third trimester of pregnancy there is a positive correlation between the level of amniotic fluid corticotrophin releasing hormone and that of saturated phosphatidyl choline. The positive correlation between amniotic fluid corticotrophin releasing hormone and saturated phosphatidyl choline, but not between amniotic fluid corticotrophin releasing hormone and gestational age, suggests that a factor(s), such as stress, may affect both amniotic fluid corticotrophin releasing hormone and saturated phosphatidyl choline in parallel. Furthermore, our data are consistent with the hypothesis that the rise in placental corticotrophin releasing hormone is coupled to an increase in fetal glucocorticoid and lung maturation, and that stresses such as pregnancy-induced hypertension may accelerate this process.

Immunoreactive corticotropin-releasing hormone in amniotic fluid

Immunoreactive corticotropin-releasing hormone in the amniotic fluid of both human beings and rats was measured by a specific radioimmunoassay. In human subjects the hormone was detectable in all amniotic fluid samples (obtained during the sixteenth and eighteenth weeks of gestation) (2.5 +/- 1.7 fmol/ml, mean +/- SD, n = 17) and the thirty-eighth to fortieth weeks (9.3 +/- 5.4 fmol/ml, n = 24). The levels of concentration of this hormone in this amniotic fluid correlated significantly with the levels in both maternal plasma and placenta for each patient. Gel filtration of amniotic fluid extracts revealed two major peaks of immunoreactive corticotropin-releasing hormone, one at the elution position of the rat hormone and the other at a small-molecular-weight region. Immunoreactive corticotropin-releasing hormone was not detectable in rat amniotic fluid or placenta. We concluded that immunoreactive corticotropin-releasing hormone, which may be derived from the placenta, is present in human amniotic fluid and that its detection in the human placenta but not in rat placentas suggests that the mechanism of corticotropin-releasing hormone gene expression in the placenta is species specific.

Corticotropin-releasing hormone in amniotic fluid during gestation and labor and in relation to fetal lung maturation

Corticotropin-releasing hormone was discovered in the placenta, and its concentration in the maternal plasma was found to increase greatly during the latter half of pregnancy. We studied the concentration of immunoreactive corticotropin-releasing hormone in amniotic fluid in 59 uncomplicated and in 73 complicated pregnancies. The mean (+/- SE) value of corticotropin-releasing hormone in amniotic fluid in uncomplicated pregnancies was significantly higher in the third (24.1 +/- 3.3 pmol/L) than in the second (9.1 +/- 0.7 pmol/L) trimester, but no change was found during labor. In groups matched by gestational age, larger mean values of corticotropin-releasing hormone and cortisol were observed in the group in which the lecithin/sphingomyelin ratio was greater than 2 or the phosphatidylglycerol test was positive than in the group with a lecithin/sphingomyelin ratio less than 2 or a negative phosphatidylglycerol test result. In samples taken at an interval of 1 to 3 weeks, concomitant increases in corticotropin-releasing hormone and cortisol levels were found with the appearance of phosphatidylglycerol. Concentrations of corticotropin-releasing hormone in amniotic fluid were elevated in patients with diabetes and in women with preeclampsia and intrauterine growth retardation. We conclude that the intrauterine release of corticotropin-releasing hormone increases during the last trimester. This may stimulate the fetal pituitary-adrenal axis and promote fetal maturation.

Immunochemical characterization and quantification of the sex steroid-binding protein (SBP) in human amniotic fluid

The sex steroid-binding protein (SBP) is a plasma protein whose concentration in the maternal circulation increases during pregnancy. Using monospecific antibodies raised against human SBP, we could demonstrate the antigenic identity of the protein in human amniotic fluid. In this fluid, we found that the SBP concentration was correlated with the total protein concentration throughout gestation. The concentration gradient of SBP between maternal serum and amniotic fluid was compared to that of other serum proteins, in relation to their relative molecular mass, and it was concluded that SBP enters amniotic fluid in a non-specific manner similar to that of other serum proteins. It is suggested that SBP could act to sequester the sex steroid hormones in amniotic fluid.

Study of the profiles of prolactin and other hormones in both maternal plasma and amniotic fluid during labor

The effects of a dopamine antagonist and stress of labor on prolactin (PRL) concentrations were studied in maternal plasma and amniotic fluid during term delivery. In procedure 1, maternal blood and amniotic fluid samples were obtained from 5 normal full-term deliveries at 20-minute intervals for 180 minutes after the intravenous bolus of 10 mg metoclopramide (MCP) during labor just before delivery. Amniotic fluid samples were drawn through a transcervical intrauterine pressure catheter to avoid contamination with maternal blood. PRL, 17 beta-estradiol (E2), progesterone (P) and cortisol were measured by means of radioimmunoassay (RIA), and free MCP concentrations were measured by high pressure liquid chromatography. In procedure 2, maternal blood and amniotic fluid samples were obtained from 12 full-term pregnancies with spontaneous delivery and 10 cases of elective cesarean section when birth was imminent. The amniotic fluid was obtained by direct sampling from the forewaters in spontaneous delivery and by means of a syringe inserted through amniotic membrane after the uterine wall had been incised in cesarean section. PRL, beta-endorphin (beta-EP) and cortisol were measured by RIA. The PRL levels in maternal plasma increased significantly (p less than 0.01) after the MCP injection. The peak value of PRL net increase (delta PRL) was 676.5 +/- 189.6 ng/ml. However, the PRL levels in amniotic fluid did not change significantly after the administration of this drug. Although the delta PRL levels in maternal plasma were significantly (p less than 0.001) correlated with MCP concentrations (r = 0.812) after the MCP injection, there was no correlation between delta PRL and MCP concentrations in amniotic fluid. No significant changes in E2, P and cortisol levels in both samples were observed after the MCP injection. The plasma PRL levels in vaginal delivery cases were significantly (p less than 0.05) lower than those in elective cesarean section cases (123.3 +/- 15.8 ng/ml vs. 181.0 +/- 22.2 ng/ml), and the plasma beta-EP and cortisol levels in vaginal delivery cases were significantly (p less than 0.01, p less than 0.001) higher than those in elective cesarean section cases respectively (beta-EP: 134.8 +/- 25.6 pg/ml vs. 49.7 +/- 12.3 pg/ml, cortisol: 79.4 +/- 5.8 micrograms/dl vs. 30.2 +/- 3.3 micrograms/dl). Therefore, a significant reduction in plasma PRL levels was accompanied by a marked rise in plasma beta-EP and cortisol levels during labor. In amniotic fluid, however, there were no significant differences of PRL, beta-EP and cortisol levels in vaginal delivery cases and elective cesarean cases.(ABSTRACT TRUNCATED AT 400 WORDS)

Gastro-entero-pancreatic hormones in amniotic fluid from normal and diabetic pregnant women.

Clinical applications of analyses of hormones in amniotic fluid (AF) have recently been increased. In diabetic pregnancy, determinations of insulin and C-peptide in AF have been suggested as good indicators of the status of the foetus. We have investigated the pancreatic alpha and beta cell function by measuring insulin (IRI), C-peptide (CPR), glucagon (IRG), somatostatin (SLI), and gastric inhibitory polypeptide (GIP) in amniotic fluid collected during basal conditions or 2 h after an arginine test in 92 diabetic and 32 non-diabetic pregnant women. During basal conditions, in diabetic pregnant women, IRI, CPR and the insulin: glucagon molar ratio (I/G) were all significantly higher while amniotic fluid-IRG was significantly lower than in the controls. After arginine stimulation, IRI increased in AF of the diabetic pregnant women but not in AF of the controls while no differences were observed in AF-GIP and AF-SLI concentrations. Higher IRI and CPR, as well as lower IRG values were significantly related to poor maternal metabolic control. The occurrence of neonatal morbidity including macrosomia was significantly associated with increased AF, IRI and CPR concentrations after an arginine challenge and these factors were the most sensitive predictors of neonatal morbidity in infants of diabetic mothers. Increased AF glucose concentrations and I/G ratios were related to neonatal hypoglycaemia; jaundice and respiratory distress syndrome were associated to low concentrations of SF-IRG.

Pitfalls of Prenatal Diagnosis of 21-Hydroxylase Deficiency Congenital Adrenal Hyperplasia*

Hormonal measurements and HLA genotyping of amniotic fluid at midgestation correctly predicted the postnatal diagnosis of congenital adrenal hyperplasia (CAH) in 29 of 32 fetuses at risk. Of these 29, 7 were predicted to have prenatal-onset classical 21-hydroxylase deficiency (21-OH def) based on elevated amniotic fluid 17-hydroxyprogesterone (17-OHP) and delta 4-androstenedione (delta 4-A) levels. These 7 fetuses and their index cases were ultimately proven to have salt-wasting classical 21-OH def. Of 5 who were HLA typed, the genotype was identical to the index case in 4; in one, HLA prediction was not possible, because the parents shared identical HLA antigens. Normal amniotic fluid 17-OHP and delta 4-A levels in the remaining 22 fetuses predicted that they were not affected with classical CAH. These children have been clinically asymptomatic to date or proven biochemically not to have classical or nonclassical CAH. Of the 22 fetuses, 11 were predicted by HLA genotyping to be homozygous normal or heterozygous for 21-OH def. In 3 of the 32 fetuses, prenatal diagnosis was incorrect. In one, the fetus was predicted to have CAH based on HLA identity to the index case. However, amniotic fluid 17-OHP and delta 4-A were normal, and the fetus was normal. The index case of this family did not have CAH, but was a normal child. Thus, amniotic fluid hormone levels accurately predicted a normal fetus, while HLA genotyping was not relevant in prenatal diagnosis because the index case was unaffected. The second fetus was predicted to be affected on the basis of HLA genotyping and to be unaffected based on normal amniotic fluid 17-OHP and delta 4-A. During infancy, this female infant had postnatal-onset nonclassical CAH. The index case in this family, presumed to have classical simple virilizing CAH, was later diagnosed to have nonclassical CAH. Thus, in nonclassical CAH, hormonal measurement of 17-OHP and delta 4-A is not useful in prenatal diagnosis; only HLA genotyping of the fetus is valuable. In the third case, the fetus was predicted to be a heterozygote by HLA genotyping and to be unaffected by hormonal measurement. Postnatally, at age 2 7/12 yr, the male child was found to have classical simple virilizing CAH and to be HLA-B-DR identical to his brother (index case) who also has classical simple virilizing CAH.(ABSTRACT TRUNCATED AT 400 WORDS)