Determination of prolactin, growth hormone, beta-endorphin, and cortisol in both maternal plasma and amniotic fluid during human gestation

This study focuses on PRL, GH, beta-endorphin and cortisol in maternal blood and amniotic fluid during human pregnancy. Maternal blood and amniotic fluid samples were obtained from 18 normal pregnant women in the second trimester, 12 full-term gravidas having spontaneous delivery, and 10 full-term gravidas having elective cesarean section. Two gravidas bearing anencephalic fetuses in the third trimester were also studied. In the second trimester women, levels of PRL (3215.9 +/- 458.9 micrograms/l), GH (19.1 +/- 1.7 micrograms/l) and beta-endorphin (11.1 +/- 0.9 pmol/l) were significantly higher in the amniotic fluid than in maternal plasma. In addition, PRL was significantly correlated with beta-endorphin (r = 0.670) and with GH (r = 0.547) in the amniotic fluid. However, amniotic fluid cortisol levels (0.27 +/- 0.18 nmol/l) were significantly lower than plasma cortisol levels. The amniotic fluid of the women with anencephalic fetuses had normal levels of PRL, GH and beta-endorphin. In full-term gravidas, plasma PRL levels were significantly lower in women with vaginal delivery than in those with elective cesarean section, and there was a significant negative correlation between plasma PRL and beta-endorphin, and between plasma PRL and cortisol levels. Plasma GH levels in women with vaginal delivery showed no significant difference from those in women with cesarean section. Examination of amniotic fluid yielded no significant differences in the levels of PRL, beta-endorphin and GH between these two groups.(ABSTRACT TRUNCATED AT 250 WORDS)     

Plasma and amniotic fluid immunoreactive neuropeptide-Y level changes during pregnancy, labor, and at parturition

This study was designed to determine the presence of and possible changes in plasma and amniotic fluid immunoreactive neuropeptide-Y (irNPY) levels in pregnant women during gestation and at parturition. We studied 127 healthy pregnant and 12 nonpregnant women. The peptide was extracted from plasma or amniotic fluid with a propanolformic acid mixture and measured by RIA. The mean plasma irNPY concentration in 15 pregnant women during the first trimester of gestation was 129 +/- 12 (+/- SE) pmol/L, compared to 40 +/- 8 pmol/L in nonpregnant women (p less than 0.01). The mean values were 144 +/- 13 and 156 +/- 24 pmol/L, respectively, in 15 pregnant women during the second trimester and 33 women during the third trimester. These values did not differ from that during the first trimester. Amniotic fluid irNPY levels were similar to those in plasma and did not vary among the 3 groups of women studied during the various trimesters of gestation. During labor, plasma irNPY levels progressively increased, reaching the highest levels at the most advanced stages of cervical dilatation (greater than 8 cm, 351 +/- 38 pmol/L) and at the time of vaginal delivery (416 +/- 73 pmol/L). Plasma irNPY levels then decreased significantly 2 h after vaginal delivery. The amniotic fluid irNPY levels in women during the early or late stages of labor were similar. Moreover, plasma and amniotic fluid irNPY levels at the time of elective cesarean section also were similar. These results indicate that pregnant women have high plasma and amniotic fluid irNPY levels and that the stress of labor results in a further increase in plasma levels, suggesting a possible role of NPY in human pregnancy and parturition.     

A radioimmunoassay for measurement of 3,3'-diiodothyronine sulfate: Studies in thyroidal and nonthyroidal diseases, pregnancy, and fetal/neonatal life

Data suggesting that (1) sulfation of the phenolic hydroxyl of iodothyronines plays an important role in thyroid hormone metabolism and (2) maternal serum 3,3'-diiodothyronone sulfate (3,3'-T(2)S) may reflect on the status of fetal thyroid function stimulated us to develop a radioimmunoassay (RIA) for measurement of T(2)S. Our T(2)S RIA is highly sensitive, practical, and reproducible. T(4)S, T(3)S, and T(1)S crossreacted 3.1%, 0.81%, and 5.3%, respectively; thyroxine (T(4)), triiodothyronine (T(3)), and reverse (r)T(3), 3,3'-T(2) and 3'-T(1) crossreacted <0.1%. Although rT(3) sulfate (rT(3)S) crossreacted 55% in 3,3'-T(2)S RIA, its serum levels are very low and have little influence on serum T(2)S values reported here. T(2)S was measured in ethanol extracts of serum, amniotic fluid, and urine. Recovery of nonradioactive T(2)S added to serum was 96%. The dose-response curves of inhibition of binding of (125)I-T(2)S to anti-T(2)S by serial dilutions of ethanol extracts of serum or urine were essentially parallel to the standard curve. The detection threshold of the RIA varied between 0.17 and 0.50 nmol/L (or 10 and 30 ng/dL). The coefficient of variation (CV) averaged 9% within an assay and 13% between assays. The serum concentration of T(2)S was [mean +/- SE, nmol/L] 0.86 +/- 0.59 in 36 normal subjects, 2.2 +/- 0.06 in 10 hyperthyroid patients (P <.05), 0.73 +/- 0.10 in 11 hypothyroid patients (not significant [NS]), 6.0 +/- 1.5 in 16 patients with systemic nonthyroidal illness (P <.001), 18 +/- 2.5 in 16 newborn cord blood sera (P <.02), 2.7 +/- 0.32 in 25 pregnant women [15 to 40 weeks gestation, P <.001], 0.94 +/- 0.10 in 10 hypothyroid women receiving T(4) replacement therapy (NS), and 2.0 +/- 0.38 in 11 hypothyroid women treated with T(4) replacement and oral contraceptives (P <.02); serum T(2)S levels in the third trimester of pregnancy were similar to those in the second trimester of pregnancy. T(2)S concentration in amniotic fluid was 12.5 +/- 2.7 nmol/L (n = 7) at 15 to 20 weeks gestation, and it decreased markedly to 3.3 +/- 1.3 nmol/L (n = 3) at 35 to 38 weeks gestation. Urinary excretion of T(2)S in random urine samples of 19 normal subjects was 10.9 +/- 1.3 nmol/g creatinine. (1) T(2)S is a normal component of human serum, urine, and amniotic fluid, and serum T(2)S levels change substantially in several physiologic and pathologic conditions; (2) high serum T(2)S in pregnancy may signify increased transfer of T(2)S from fetal to maternal compartment, estrogen-induced increase in T(2)S production, decreased clearance, or a combination of these factors. The data do not support the notion that fetal thyroid function is the only or the predominant factor responsible for high serum T(2)S in pregnant women.     

Amniotic fluid testosterone: relationship with cortisol and gestational age

INTRODUCTION: Foetal exposure to testosterone is increasingly implicated in the programming of future reproductive and non-reproductive behaviour. Some outcomes associated with prenatal exposure to testosterone may be predicted from exposure to prenatal stress, suggesting a link between them. The peak serum levels of testosterone in the foetus are thought to be around 14-18 weeks' gestation, and we explored testosterone levels at different gestations. Although best investigated in foetal plasma, this is now difficult because of the decline in frequency of foetal blood sampling; in this study, we used amniotic fluid as a biomarker to investigate foetal exposure. AIMS: To investigate the relationship between amniotic fluid testosterone, amniotic fluid cortisol, foetal gender, and gestational age. METHODS: Paired amniotic fluid and maternal plasma samples were collected from 264 pregnant women undergoing amniocentesis between 15 and 37 weeks' gestation (median 17 weeks [119 days]). Total testosterone and cortisol in amniotic fluid, and total plasma testosterone (maternal) were measured by radioimmunoassay. RESULTS: Amniotic fluid testosterone levels were higher in male than in female foetuses, with a median (interquartile range) of 0.85 nmol/l (0.60-1.17 nmol/l) and 0.28 nmol/l (0.175-0.45 nmol/l), respectively. No relationship between amniotic fluid testosterone and gestational age was detected in either sex. Amniotic fluid testosterone correlated positively with amniotic fluid cortisol in both sexes (r = 0.30 male foetuses, r = 0.33 female foetuses, P < 0.001 for both), and remained significant in multivariate analysis. CONCLUSION: Testosterone in amniotic fluid did not change with gestation in the second and third trimester, raising questions about the timing of the reported early peak in the male foetus. The positive correlation between cortisol and testosterone in amniotic fluid suggests that increased foetal exposure to cortisol may also be associated with increased exposure to testosterone.     

Growth hormone (GH) replacement therapy in GH-deficient women during pregnancy

OBJECTIVE: The present recommendation is to discontinue GH replacement therapy in hypopituitary women, as they become pregnant. We report our experience of managing GH deficiency with GH replacement therapy in pregnant hypopituitary women. PATIENTS AND MEASUREMENTS: Eight hypopituitary women, median age 30.5 years (range 20-39 years), were followed prospectively during 12 distinct pregnancies. Six women were receiving replacement therapy for other pituitary hormone deficiencies and five pregnancies were achieved with ovulation induction therapy. GH replacement therapy was maintained at the same pregestational dose during the first trimester, with a gradual decrease of the dose during the second trimester and discontinuing the treatment at the beginning of the third trimester. Serum IGF-1 levels were measured in four selected pregnancies at three different points, one in each trimester of gestation. RESULTS: Seven pregnancies resulted in normal vaginal deliveries and five had programmed Caesarian sections. The median gestation age at time of delivery was 40 weeks (range 33-42 weeks). Newborns had a median birthweight SD score of -0.37 (range -1.93 to 1.21) and a median birthlength SD score of 0.07 (range -2.73 to 1.53). No congenital malformations were observed. Three women were able to breastfeed their babies. Before gestation, the median daily dose of GH was 0.5 mg (range 0.3-0.8 mg) and the median serum IGF-1 was 37.5 nmol/l (range 7.6-54.5 nmol/l). IGF-1 levels were fairly constant in the first and second trimesters of gestation, showing an increment during the last trimester, when GH treatment was discontinued. CONCLUSION: The GH replacement regimen presented for pregnant women with GH deficiency was safe. No major side-effects and no negative influences on maternal and fetal outcome were observed.     

Changing osteocalcin concentrations during pregnancy and lactation: implications for maternal mineral metabolism

We measured serum osteocalcin concentrations in 82 pregnant and 21 nonpregnant women. Osteocalcin values declined in the second trimester, but returned to nonpregnant levels late in the third trimester. The mean serum osteocalcin concentration in 36 women during pregnancy (mean gestation, 26 weeks) of 2.8 ng/mL was significantly lower than that in nonpregnant women (6.4 ng/mL; P less than 0.001) or term pregnant women at delivery (6.1 ng/mL; n = 46). Serum immunoreactive PTH (iPTH) levels were significantly higher during pregnancy than in nonpregnant women [97 +/- 5 vs. 56 +/- 4 ng/L (mean +/- SE); P less than 0.001]. No significant correlations were found between maternal osteocalcin concentrations and serum phosphorus, alkaline phosphatase, or iPTH, but significant negative correlations were found between osteocalcin and total calcium or total protein. Osteocalcin concentrations in midtrimester amniotic fluid were very low (mean, 0.3 +/- 0.1 ng/mL; n = 11). In 29 lactating mothers, the mean serum osteocalcin level was 9.5 +/- 1.5 ng/mL, significantly higher than in any of the other groups (P less than 0.05), but their serum calcium and iPTH levels were normal. There was no correlation between serum osteocalcin and calcium or iPTH concentrations in lactating women. These changes are compatible with a sequence in which bone turnover is reduced during early pregnancy, rebounds in the third trimester, and increases in postpartum lactating women.     

Sublingual administration of testosterone-hydroxypropyl-beta-cyclodextrin inclusion complex simulates episodic androgen release in hypogonadal men

In search of a more physiological testosterone (T) replacement therapy for hypogonadal states, we evaluated an inclusion complex of T with 2-hydroxypropyl-beta-cyclodextrin (HPBCD). HPBCD enhances T solubility and absorption, but HPBCD is not absorbed. Five hypogonadal men (mean age, 32.4 +/- 2.3 yr) with serum T levels below the normal range were treated in two separate experimental phases with either a 2.5- or 5.0-mg tablet of sublingual (SL) T-HPBCD three times daily for 7 days. Acute pharmacodynamic changes were monitored at baseline and 10, 20, and 40 min and 1, 1.5, 2, 3, 4, and 8 h after administration of the first dose. At the 5-mg dose, a maximal concentration (Cmax) of T (85.4 +/- 11.0 nmol/L) was achieved in 20 min (63 +/- 24-fold increase), followed by a rapid decline to below the normal range (less than 12 nmol/L) at 2 h, with an estimated half-life of decline of 1.87 +/- 0.19 h. The dihydrotestosterone (DHT) Cmax (4.1 +/- 0.5 nmol/L) occurred at 32 +/- 5 min (8.9 +/- 1.3-fold increase) and declined to below the normal range (less than 1.2 nmol/L) after 3 h. The integrated 8 h value for the ratio of T/DHT was 10.0 +/- 1.1, which fell within the normal range. The increment in androstenedione paralleled that in T, and the Cmax (6.8 +/- 0.9 nmol/L) was reached in 24 +/- 4 min (2.3 +/- 0.6-fold increase). Compared to baseline, the Cmax was significantly greater for T (P less than 0.005), DHT (P less than 0.0005), and androstenedione (P less than 0.005). Both estradiol (E2) and estrone (E1) remained in the normal range (less than 200 pmol/L), although the Cmax for E1 was significantly greater than baseline (P less than 0.05). Serum LH levels were suppressed (19.0 +/- 2.6%) at 2 h (P less than 0.05), without a significant change in FSH. During 7 days of treatment, there was no cumulative increase in basal T, DHT, and E2 levels or further decline in LH or FSH levels. There was no change in sex hormone-binding globulin levels. Similar results were observed with the 2.5-mg dose, suggesting that the capacity of SL absorption may be limited to a certain dose of T-HPBCD. The fluctuations in T after SL administration of T-HPBCD resemble endogenous episodic secretion. We conclude that T, complexed with HPBCD, is rapidly absorbed by the SL route and quickly metabolized without sustained elevations of DHT or E2.     

A longitudinal analysis of maternal serum insulin-like growth factor I (IGF-I) and total and nonphosphorylated IGF-binding protein-1 in human pregnancies complicated by intrauterine growth restriction

In cord blood and late gestation maternal serum, IGF-I is positively correlated with birth weight, whereas IGF-binding protein-1 (IGFBP-1) is inversely correlated with birth weight. Our goal was to determine whether maternal serum or amniotic fluid concentrations of IGF-I, IGFBP-1, or nonphosphorylated IGFBP-1 (npIGFBP-1) in early gestation predict later fetal growth abnormalities. Maternal serum was collected prospectively across gestation (5-40 wk) from 749 pregnant subjects. Amniotic fluid was collected after amniocentesis during wk 15-26 from 207 subjects. We compared median serum concentrations of IGF-I, IGFBP-1, and npIGFBP-1 in 38 subjects who delivered growth-restricted infants with the control group of 236 subjects with normal weight infants for each gestational age grouping, wk 5-12, 13-23, and 24-34. In the control group median IGF-I concentrations were 14.8, 11, and 15.6 nmol/liter for wk 5-12, 13-23, and 24-34, respectively, compared with 13.7, 14.3, and 10.6 nmol/liter in the intrauterine growth restriction (IUGR) group. Median IGFBP-1 concentrations were 8.5, 30.4, and 24.4 nmol/liter, respectively, in controls, compared with 11.4, 28.6, and 25.5 nmol/liter in the IUGR group. Median npIGFBP-1 concentrations were 6.9, 22, and 17.4 nmol/liter, respectively, in controls, compared with 5.0, 32.1, and 24.2 nmol/liter in the IUGR group. In the control group the median amniotic fluid IGFBP-1 level was 13,160 nmol/liter, and the median npIGFBP-1 level was 15,970 nmol/liter; in the IUGR group these levels were 13,440 and 18,440 nmol/liter, respectively. No clinically useful differences were found between the IUGR and control groups. Our results do not support the use of maternal serum IGF-I or IGFBP-1 or amniotic fluid IGFBP-1 or npIGFBP-1 early in gestation to predict later fetal growth restriction.     

Amniotic fluid levels of dimeric inhibins, pro-αC inhibin, activin A and follistatin in Down''s syndrome

OBJECTIVE: In the second trimester of pregnancy, inhibin A is significantly increased in maternal serum and decreased in amniotic fluid in Down's syndrome pregnancies compared to normal. We wished to further evaluate the levels of inhibin A, inhibin B, pro-alpha C inhibin, activin A and the binding protein follistatin in amniotic fluid in Down's syndrome and control pregnancies. DESIGN: Case-matched control study. PATIENTS: 29 Down's syndrome and 290 chromosomally normal control pregnancies were identified from records and amniotic fluid, collected at second trimester amniocentesis, retrieved from routine storage for analysis. MEASUREMENTS: Inhibin A, inhibin B, pro-alpha C inhibin, total activin A and follistatin were measured using sensitive and specific enzyme linked immunosorbent assays. RESULTS: The median (10th-90th percentiles) amniotic fluid inhibin A level in the control pregnancies increased from 334 (122-553) ng/l at 14 weeks' to 695 (316-1475) ng/l at 19 weeks' gestation. The corresponding figures for inhibin B and the alpha-subunit precursor inhibin pro-alpha C were 632 (185-1354) and 2062 (1237-3381) ng/l, respectively at 14 weeks' and 2439 (748-5307) and 3115 (2021-6567) ng/l, respectively at 19 weeks' gestation. Total activin A increased from 3795 (1554-5296) at 14 weeks' to 5086 (3059-8224) at 18 weeks' gestation. Expressed as multiples of the median (MoM) the median (95% CI) amniotic fluid levels of inhibin A, inhibin B, pro-alpha C inhibin and acitivin A in the Down's syndrome samples were 0.77 (0.59-0.85), 0.94 (0.63-1.23), 0.77 (0.49-0.84) and 0.77 (0.53-0.87), respectively. Compared to controls the levels of inhibin A, pro-alpha C inhibin and activin A were significantly lower in Down's syndrome pregnancies (P < 0.01, Mann-Whitney U test). Follistatin levels in the controls declined slightly from 2106 (1421-3538) ng/l at 14 weeks' to 1600 (1281-2543) ng/l at 18 weeks' gestation. Levels in the Downs' syndrome pregnancies were similar to controls. CONCLUSIONS: The data suggest that the production, secretion or metabolism of the inhibin alpha- and beta A-subunits is altered in Down's syndrome pregnancies in the second trimester.     

Response to corticotropin-releasing hormone during pregnancy in the baboon

CRH is secreted by the placenta into human maternal and fetal plasma during gestation. In the present study plasma CRH was measured in the plasma of five pregnant baboons and their fetuses to ascertain whether the baboon is a suitable model for study of placental CRH. Studies were performed in chronically catheterized animals that exhibited no behavioral or endocrinological signs of stress; maternal animals moved freely about the cage. Mean maternal plasma CRH was 620 +/- 110 pmol/L (2970 pg/mL) at 146 +/- 11 days gestation, and mean fetal plasma CRH was 133 +/- 29 pmol/L (640 pg/mL) at delivery in four animals. Plasma CRH was undetectable (less than 8.5 pmol/L; less than 41 pg/mL) in nonpregnant animals and in animals 8 h after delivery. Maternal and fetal plasma CRH levels in the chronically catheterized baboon were very similar to human maternal and umbilical cord CRH levels at comparable gestational ages. In addition, the majority of maternal plasma CRH eluted in the same position as synthetic human CRH by gel filtration. CRH stimulation tests were performed in the chronically catheterized maternal baboon to investigate whether pituitary-adrenal function during pregnancy is similar to that observed after chronic CRH infusion; blunted ACTH and cortisol responses to acute injections of CRH are observed after chronic CRH infusion. The administration of 0.5 micrograms/kg ovine CRH (oCRH) failed to result in an ACTH or cortisol rise in four pregnant baboons. Baseline ACTH levels were 5.2 +/- 0.4 pmol/L (23.5 pg/mL), and baseline cortisol levels were 800 +/- 55 nmol/L (29.1 micrograms/dL); neither rose after CRH administration. In contrast, 0.5 micrograms/kg oCRH did result in significant ACTH and cortisol elevations in five nonpregnant baboons [ACTH: baseline, 5.9 +/- 1.4; peak, 16 +/- 4.8 pmol/L (P less than 0.05); cortisol: baseline, 430 +/- 55 nmol/L; peak, 960 +/- 200 nmol/L (P less than 0.05)]. In contrast, the administration of a larger dose of oCRH (5.0 micrograms/kg) led to stimulation of ACTH release in five pregnant baboons (baseline, 6.6 +/- 1.3 pmol/L; peak, 34.1 +/- 6.4; P less than 0.001). After this dose cortisol levels also rose in the pregnant animals (baseline = 1040 +/- 30 nmol/L; peak, 1620 +/- 130); however, this response was blunted compared to that in the nonpregnant animals (P less than 0.05). CRH (5.0 micrograms/kg) significantly stimulated both ACTH and cortisol in the nonpregnant animals.(ABSTRACT TRUNCATED AT 400 WORDS)     

Increased human placental growth hormone at midtrimester pregnancies may be an index of intrauterine growth retardation related to preeclampsia

OBJECTIVE: To evaluate the relationship between maternal serum and amniotic fluid levels of human Placental Growth Hormone (hPGH) with the fetal intrauterine growth retardation (IUGR) related to preeclampsia. DESIGN: We analyzed samples in pairs of serum and amniotic fluid retrospectively from 25 women, who manifested preeclampsia and IUGR in the late second or the third trimester of gestation. The samples were obtained at 16-22 weeks' gestation during amniocentesis for fetal karyotyping. At this time, there was no clinical or sonographic evidence of preeclampsia or IUGR, respectively. Sixty-two serum samples were used as controls which were obtained at 16-22 weeks' gestation from women with singleton, uncomplicated pregnancies, with normal outcome, and appropriate for gestational age neonatal birth weight. Forty-seven amniotic fluid samples were also used as controls which were obtained at 16-22 weeks' gestation from the women that were included in the control group who underwent an amniocentesis. hPGH levels were measured by a solid phase immunoradiometric assay. RESULTS: The mean hPGH values in the serum and the amniotic fluid of the IUGR related to preeclampsia affected pregnancies were significantly higher (P<0.05) than those of the normal pregnancies at 16-22 weeks' gestation: mean+/-SD in the serum was 13.16+/-10.52 ng/ml vs. 4.39+/-2.23 ng/ml; mean+/-SD in the amniotic fluid 2.49+/-1.6 ng/ml vs. 0.82+/-0.67 ng/ml. CONCLUSION: hPGH levels in maternal serum and amniotic fluid were found to be higher at 16-22 weeks' gestation in pregnancies that will be complicated subsequently by IUGR related to preeclampsia. Our findings suggest that the evaluation of the changes of hPGH levels at midtrimester should be further investigated for the possibility to provide a potential predictive index of IUGR and preeclampsia.     

Highest concentrations of prorenin and human chorionic gonadotropin in gestational sacs during early human pregnancy.

Prorenin is not only the biosynthetic precursor of renin; under certain circumstances in vitro prorenin exhibits reversible intrinsic renin activity and can form angiotensin from renin substrate with or without cleavage of the prosequence. Prorenin is the predominant form of renin synthesized by reproductive organs (ovary, chorion laeve of the placenta, uterine decidua). Its plasma concentrations increases 10-fold throughout pregnancy to 10-100 times that of renin; amniotic fluid prorenin concentration is even higher. No data are available of gestational fluid prorenin concentrations during early pregnancy. For the first 10 weeks there are two gestational cavities; the chorionic cavity then disappears and the smaller amniotic cavity becomes predominant. In this study we measured prorenin, renin, renin substrate and hCG in fluid aspirated from gestational sacs during the first trimester of gestation (predominantly chorionic) and during the second and third trimesters (amniotic). Seventeen patients had amniocentesis during the second or third trimester. Nine patients underwent selective abortion of multiple pregnancy at 7-12 weeks gestation. One patient underwent surgery at 5 5/7 weeks (26 days after conception) for a tubal pregnancy. Second and third trimester amniotic fluid prorenin maximum velocity (Vmax) (16 and 3 sacs, respectively) averaged 6,100 +/- 1,700 (SD) and 1,930 +/- 760 ng/mL.h, respectively (i.e. 1,700 and 540 ng/L.s). In gestational fluid collected before 8 weeks, prorenin Vmax was 10-fold higher, averaging 62,500 +/- 40,000 ng/mL.h (17,000 ng/L.s). The concentration was 140,000 ng/mL.h (39,000 ng/L.s) in the 5 5/7 week tubal pregnancy. In sharp contrast, at 10-12 weeks gestation (n = 3) prorenin Vmax was only 260 +/- 114 ng/mL.h (72 ng/L.s); human CG was also highest before 8 weeks (276,500 +/- 110,900 IU/L) and lowest at 10-12 weeks (1210 +/- 540 IU/L) with intermediate levels occurring later in pregnancy. This study shows that the highest biological levels of prorenin yet detected (close to 1 micrograms protein/mL) occur in gestational sacs in early pregnancy, consistent with a role for the renin-angiotensin system in embryonic development or placentation.     

11 Beta-hydroxylase deficiency congenital adrenal hyperplasia: update of prenatal diagnosis

Hormonal measurements in maternal urine and amniotic fluid (AF) during pregnancy and/or at delivery correctly predicted the postnatal diagnosis of 11 beta-hydroxylase deficiency congenital adrenal hyperplasia (11 beta-OH deficiency CAH) in 7 fetuses at risk. In the 4 affected ones, maternal urinary tetrahydro-11-deoxycortisol (THS) excretion was high during the first trimester [0.3-2.2 mg/day (1.1-7.7 mumol/day)] and rose further during the third trimester [0.5-3.5 mg/day (1.8-12.3 mumol/day)] compared to urinary THS excretion in 20 normal pregnancies of the same gestational age (P less than 0.01). In 1 mother, dexamethasone administration (2 mg/day for 72 h) greatly reduced urinary THS excretion (and plasma steroid levels). Urinary THS excretion was low after delivery in these mothers, in normal pregnancies, and in parents of affected individuals [less than 0.05 mg/day (less than 0.08 mumol/day); P = NS]. However, 2 of the 3 heterozygous mothers who carried nonaffected fetuses excreted moderately increased amounts of THS during pregnancy, ranging from 0.15-0.26 mg/day (0.53-0.91 mumol/day), significantly higher than normal (P less than 0.01). Although urinary THS excretion in these mothers was similar to that in 2 mothers with affected fetuses early in pregnancy, urinary THS excretion was higher in mothers with affected compared to those with nonaffected fetuses after the first trimester (P less than 0.01). AF THS and 11-deoxycortisol concentrations were markedly elevated in pregnancies with affected fetuses (P less than 0.01), but normal in nonaffected ones. AF delta 4-androstenedione levels were high in 2 pregnancies and borderline elevated in a third. Although the AF tetrahydrocortisol and tetrahydrocortisone levels were always within the normal range, the AF THS to tetrahydrocortisol plus tetrahydrocortisone ratio was significantly elevated in all pregnancies with affected fetuses (2.8-5.5; P less than 0.01) and normal in nonaffected ones (0.48-1.2; P = NS) compared to that in 160 normal pregnancies [0.64 +/- 0.34 (+/- SD)]. AF 17-hydroxyprogesterone, testosterone, and 11-deoxycorticosterone levels were normal in all pregnancies. Maternal plasma 11-deoxycortisol and delta 4-androstenedione concentrations, determined sequentially throughout gestation, were variable and did not contribute to prenatal diagnosis. All affected infants were born hyperpigmented, 2 were large for gestational age, and the female was severely virilized. In the first week of life 2 males developed severe hypertension with seizures and adrenal insufficiency, respectively.(ABSTRACT TRUNCATED AT 400 WORDS)     

AMNIOTIC FLUID CONCENTRATIONS OF 3,3'',5''-TRI-IODOTHYRONINE (REVERSE T3), 3,3''-DI-IODOTHYRONINE, 3,5,3''-TRI-IODOTHYRONINE (T3) AND THYROXINE (T4) IN NORMAL AND COMPLICATED PREGNANCY

Amniotic fluid concentrations of 3,3',5'-tri-iodothyronine (rT3), 3,3'-Di-iodothyronine (3,3'-T2), 3,5,3'-tri-iodothyronine (T3) and T4 were studied in 384 women during normal and complicated pregnancy. An inverse correlation was observed between decreasing rT3 and increasing 3,3'-T2 concentrations in amniotic fluid with gestational age. The mean rT3 level in normal pregnancy was 2.81 nmol/1 at 12-20 weeks and decreased significantly to 1.06 nmol/1 at 36-42 weeks of gestation. The mean 3,3'-T2 concentration was 49.1 pmol/1 at12-20 weeks increasing to 119 pmol/1 at 36-42 weeks. The mean T4 value of 3.83 nmol/1 at 12-20 weeks was about half that of later periods. The T3 concentration in a random sample of 45 amniotic fluids ranged from less than 28 to 370 pmol/1 (mean 102 pmol/1). The mean rT3, 3,3'-T2 and T4 values measured in patients with intra-uterine malnutrition, gestation diabetes, tocolysis, placental insufficiency and rhesus incompatibility at 31-40 weeks of gestation were not significantly different from those in uncomplicated pregnancy. Significantly decreased rT3 and T4 concentrations were found in toxaemia. From the results obtained in complicated pregnancy it may be concluded that measurements of iodothyronines, especially rT3, in amniotic fluid have insignificant diagnostic value in the recognition of intra-uterine lesions with the probable exception of fetal hypothyroidism. The analysis of the dependence of iodothyronine concentrations on the gestational age showed a maximum of rT3 and T4 levels between 20 and 30 weeks of pregnancy. This marked rise of iodothyronine concentrations in amniotic fluid at mid-gestation may be due to the onsetting maturation of the hypothalamic-pituitary-thyroid control system of the fetus.     

Two immunoreactive binding proteins for insulin-like growth factors in human amniotic fluid: relationship to fetal maturity

A binding protein for insulin-like growth factors (IGFs) has been purified from human amniotic fluid by IGF-I affinity chromatography and high performance reverse phase chromatography. This protein, with an apparent molecular mass of 28K nonreduced and 34K reduced, had an identical amino-terminus to a previously purified binding protein from amniotic fluid and to placental protein 12. The purified preparation (BP-28) bound both IGFs with high affinity [Ka, 6.55 +/- 2.24 (+/- SD) L/nmol for IGF-I and 3.23 +/- 1.05 L/nmol for IGF-II], with approximately 0.5 mol binding sites/mol BP-28 for either ligand. A 53K IGF-binding protein purified from human plasma (BP-53) did not cross-react in a RIA for BP-28, and BP-28 had less than 0.1% molar cross-reactivity in a RIA for BP-53. Human amniotic fluid reacted strongly in both assays. Fractionation of amniotic fluid samples by reverse phase chromatography showed that BP-28 and BP-53 immunoreactivities were present on separate proteins. In 40 third trimester amniotic fluid samples selected to cover a wide range of lecithin to sphingomyelin ratios, the mean concentrations of BP-28 and BP-53 were 37.6 +/- 17.6 (+/- SD) and 4.6 +/- 1.6 mg/L, respectively. Significant negative correlations were found between the levels of both BP-28 and BP-53 and the lecithin to sphingomyelin ratio, suggesting an association between the levels of both proteins and the degree of fetal maturity. A significant positive association was also found between the levels of BP-28 and BP-53. We conclude that the 28K IGF-binding protein from amniotic fluid, like the previously purified 53K binding protein, has high affinity for both IGF-I and IGF-II, that it coexists in amniotic fluid with BP-53 or a related protein, and that the levels of both proteins decline with increasing fetal maturity.     

Elevated first trimester maternal levels of soluble fibrin polymer are associated with lower birthweight in twin gestation.

We assessed soluble fibrin polymer longitudinally in normal pregnancy, thrombophilic pregnancy and twin gestation Thirty-three thrombophilic pregnancies, 34 uncomplicated multiple gestations and 23 singleton normal pregnancies were studied. Maternal plasma samples were collected in the first (6-12 weeks), 2nd (13-25 weeks) and 3rd trimesters of pregnancy (26-40 weeks) and were stored at -70 degrees C until assay. Soluble fibrin polymer was assayed by enzyme-linked immunosorbent assay (ABS, Copiague, New York, USA). Statistical analysis were made by Spearman test and Levine's test for equality of variance (P < 0.05). First soluble fibrin polymer maternal levels in twin gestation were significantly higher than in normal pregnancy and thrombophilic pregnancy, (23.8 +/- 4.5 mug/ml versus 9.2 +/- 3.1 and 10.0 +/- 2.0 mug/ml respectively, P < 0.005). Second and third trimester maternal levels of SFP in twins were also significantly higher than in normal pregnancy. First trimester soluble fibrin polymer was highly correlated with birthweight in twin gestation (r = -1, P < 0.01). In the third trimester, maternal soluble fibrin polymer correlated with placental weight in twin gestation (r = 0.639, P < 0.01). Overall, soluble fibrin polymer was correlated with placental weight and birthweight in the three groups but this did not reach statistical significance. Elevated maternal plasma levels of soluble fibrin polymer in twin gestation may derive from an accelerated coagulation process due to extensive vascular remodelling. Current studies are underway to determine the utility of soluble fibrin polymer in assessing fetal growth abnormalities.     

Periovulatory follicular fluid hormone levels in spontaneous human cycles

We measured follicular fluid hormone levels in 48 normally cycling infertile women who underwent follicle puncture and oocyte retrieval during diagnostic laparoscopy at time-bracketed intervals after an endogenous LH surge. Follicular fluid LH, FSH, PRL, estrone (E1), estradiol (E2), progesterone (P), androstenedione (A), and testosterone (T) concentrations and P/E2 and A/E2 ratios were determined. Oocytes were classified as germinal vesicle (gv), metaphase I (mI), metaphase II (mII), or degenerating (dg). Follicular fluid (ff) hormone levels then were correlated with the stage of oocyte maturation. There were no differences in ff E1 or E2 levels at any stage of oocyte maturation, except that the mean ff E2 concentration was significantly (P less than 0.05) lower in ff containing dg oocytes [2,474 +/- 1,435 (+/- SE) nmol/L] than in those containing the other oocyte stages. The mean P levels were significantly (P less than 0.0001) higher in ff containing mI (48,781 +/- 10,240 nmol/L) and mII (41,801 +/- 11,098 nmol/L) oocytes than in ff containing gv oocytes (1371 +/- 696 nmol/L). The mean A level was highest (P less than 0.01) in dg-associated ff. Similarly, T was highest (P less than 0.05) in ff containing dg (52 +/- 14 nmol/L) oocytes than in ff containing mI (10.7 +/- 10.1 nmol/L) or mII (10.1 +/- 4 nmol/L) oocytes, and it was also elevated (P less than 0.05) in gv ff (72 +/- 33 nmol/L) compared to mII ff. The above differences also were reflected in the P/E2 ratio, which was significantly higher (P less than 0.05) in mI and mII ff, as well as in the A/E2 ratio, which was higher (P less than 0.05) in ff containing mI and mII oocytes compared to ff containing gv or dg oocytes. These data define the evolving changes in the microenvironment of the follicular fluid of preovulatory follicles of normally cycling women. They also provide reference points for analysis of ff obtained from women during stimulated cycles intended for in vitro fertilization.     

Circulating placental lactogen levels in dairy and beef cattle.

Levels of bovine placental lactogen (bPL) have been measured in the serum of dairy and beef cattle and in the milk and amniotic fluid of pregnant animals with a highly specific radioimmunoassay. In both dairy and beef cows, serum bPL levels remain low (less than 50 ng/ml) during the first two trimesters and then rise rapidly between 160 and 200 days of gestation to a plateau. The bPL levels do not decline prior to parturition. During the last trimester, serum levels in dairy cows, 1103+/-342 ng/ml, are significantly higher than those in beef cattle, 650+/-37 ng/ml (P less than 0.01); furthermore, dairy cows having a high milk production also tend to have high bPL levels. Serum levels are almost twice as high in twin pregnancies and are not correlated with fetal sex or birth weight. bPL levels in milk and amniotic fluid from dairy cattle during the last trimester are approximately 86% and 25% of the serum values, respectively, suggesting that bPL enters these fluids by passive diffusion.     

Total homocyst(e)ine in plasma and amniotic fluid of pregnant women

Total homocyst(e)ine was determined by the quantitation of protein-bound homocyst(e)ine in the stored plasma and amniotic fluid from 25 pregnant women and in the stored plasma from 17 nonpregnant women. The mean +/- SE of plasma total homocyst(e)ine was 29.8 +/- 2.4 nmol/g protein in pregnant women and 52.4 +/- 3.8 nmol/g protein in nonpregnant women. In contrast, the mean +/- SE of total homocyst(e)ine in amniotic fluid obtained at 16 weeks of gestation was 36.3 +/- 2.9 nmol/g protein. There was a statistically significant difference in the plasma total homocyst(e)ine concentrations from pregnant and nonpregnant women (P less than 0.01). Similarly, there was also a statistically significant difference between plasma total homocyst(e)ine from nonpregnant women and amniotic fluid total homocyst(e)ine (P less than 0.01). These observations suggested that the metabolism of homocysteine to cysteine was more efficient in pregnant women. In addition, the concentrations of total homocyst(e)ine in amniotic fluids were within narrow limits in normal pregnancies. Hence, total homocyst(e)ine concentration might be very valuable as a rapid assessment of fetuses for congenital defects of homocysteine metabolism.     

Determination of insulin-like growth factor-2 in feto-maternal circulation during human pregnancy.

In order to clarify the roles of insulin-like growth factors (IGFs) on the human maternal-fetal environment, IGF-2 and IGF-1 levels were investigated in human plasma and amniotic fluid during pregnancy. Initially, new radio-immunoassay (RIA) systems for human IGF-2 could be developed. The sensitivity of this assay was 17.5 pg/tube and the cross-reactivity with IGF-1 was 0.64%. The pattern of change of maternal plasma IGF-2 in early pregnancy differed from that of IGF-1, but both IGF levels increased progressively in the second half of gestation, and decreased to non-pregnancy levels in the puerperium. Maternal levels of IGF-2 were approximately seven times greater than those of IGF-1. The ratio of IGF-2 to IGF-1 was 3.2 in amniotic fluid. The IGF concentrations in amniotic fluid obtained in the second trimester were significantly greater than those of term specimens, and closely related to those of prolactin (PRL) in amniotic fluid. The highest IGF-2 to IGF-1 ratio (15.9) was found in umbilical vein plasma. On Sephadex G-150 gel-chromatography of maternal and fetal plasma at term, two apparent peaks of unsaturated IGF-2 binding protein (BP) could be detected in both 150 and 40 kilo dalton (kD) regions. One main peak of unsaturated IGF-2 BP could be determined in the 40 kD region in the amniotic fluid at term. High concentration of IGF-2 could be detected in feto-maternal circulation during human pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)