A number of laboratory parameters were estimated in 54 pregnant women in order to assess whether changes induced by smoking could reflect weight gain in the fetus. There was a significant correlation between maternal weight gain index, oestriol levels, antithrombin III levels and reticulocyte synthesis with infant and placental weight gain. One or more of these tests was found abnormal in 27% of normal pregnant patients, 72% of light smokers, and 89% of heavy smokers. Several other parameters were found to be significantly altered in smoking compared to non-smoking pregnant patients; however, these bore no relationship to infant and placental parameters. These studies indicate that laboratory assessment of a number of maternal parameters could be of some value in determining the likelihood of depressed weight gain by infants of smoking mothers.

We report the preliminary results of a prospective study aimed at evaluating the effectiveness of Down syndrome (DS) screening using second-trimester measurement of maternal serum human chorionic gonadotrophin (hCG) and unconjugated oestriol (uE3) together with maternal age. Reference values for hCG, uE3, and the hCG/uE3 ratio in normal pregnancies were established from more than 3000 normal gestations and found to follow a log-normal statistical distribution. Risk evaluation was made using reference values for affected pregnancies from retrospective studies. Screening of 10,000 women under 38 years resulted in 412 amniocenteses and the prenatal diagnosis of six cases of DS, whereas four cases remained undetected until term. In a parallel study, diagnostic amniocentesis was performed in women over 38 years and in women with a previous affected child, and an evaluation of the risk of fetal DS based on serum hCG and uE3 levels was made in all cases. Fourteen cases of DS were detected. Median values for hCG and uE3 in the 24 affected pregnancies were close to the 90th and tenth centiles of the normal reference values, respectively, and thus are in good agreement with the values reported by others in retrospective studies.

A pilot study of the endogenous steroid concentrations in human breast tumours was performed. The technique of high-resolution molecular-ion monitoring during combined g.l.c.-mass spectrometry was used to determine oestrone, oestradiol-17beta and oestriol in concentrations above 1ng/g wet wt. of tissue, and dehydroepiandrosterone, testosterone, androsterone (3alpha-hydroxy-5alpha-androstan-17-one) and 3beta-hydroxy-5alpha-androstan-17-one in concentrations exceeding 5ng/g, in extracts of five primary breast tumours.

The effects of oestriol sulphate on myometrial activity in a group of young primigravidae, who were at 41 or more weeks' gestation, are compared with those of a placebo in a double-blind trial. Both compounds were administered over a period of six hours directly into the uterine cavity following artificial rupture of membranes. Intra-amniotic pressure recordings demonstrated a peak in the mean intensity in uterine contractions in the oestriol-treated and placebo-treated groups at six and four hours, respectively. The length of labour tended to be shorter in those patients having greater uterine activity.

In a 23-year old primipara who was in her 34th week of pregnancy, serum oestriol could not be identified, with no other abnormal clinical and endocrinological findings, which is why the authors suspected placental sulphatase deficiency. The placental sulphatase activity was tested by the intravenous application of dehydroepiandrosterone sulphate, and compared with the data obtained from a control patient with normal endocrinological findings. There was a slight increase in DHEA from 7.5 ng/ml to 18.0 ng/ml, whereas the serum values of oestrone and oestradiol did not change. In contrast, the control patient showed an excessive increase of DHEA from 5.5 ng/ml to 56.8 ng/ml. Oestrone and oestradiol also reacted with a strong increase of activity from 12.7 ng/ml to 42.7 ng/ml or 25.2 ng/ml to 61.2 ng/ml, respectively. A placental sulphatase deficiency with absence of hydrolysis of the applied DHEAS must be assumed as the reason for the deficient or absent reaction of DHEA and the oestrogens. The patient gave birth to a healthy boy in the 41st week of pregnancy, delivery being performed by caesarian section. Clinically normal course of pregnancy with lowered oestriol values, delivery by caesarian section in the case of primiparae, as well as male sex of the newborn, are criteria of placental sulphatase deficiency. These criteria correspond to the few observations published so far.

Unconjugated and total oestriol (E3) have been determined in saliva (S) and plasma (P) by radioimmunoassay using a highly specific antiserum against E3. Total oestrogens (E = E1 + E2 + E3) in urine were analysed by the Kober-Ittrich method. No correlation (P greater than 0.05) was found between unconjugated E3 (S) and total E (U) as well as total E3 (S) and total E (U). Total E3 (S) and unconjugated E3 (P), total E3 (S) and total E3 (P) correlated weakly (P less than 0.05 and greater than 0.01). The six remaining correlations were highly significant (P less than 0.001 or less than 0.0001).

There are a great deal of data concerning the role of testosterone in the formation of the sex of the fetus. The synthesis of the testosterone is influenced by the hypophysis and the fetoplacental unit. In this study the authors conferred the progesterone, DHAS, cortisol, oestradiol, oestriol, prolactin and HPL levels of male and female newborns in the maternal vein and in the umbilical vein and artery in the 28th-32th weeks (31 parturients, 15 male and 16 female fetuses), in the 33rd-36th weeks (43 parturients, 20 male and 23 female fetuses) and in the 40th week (34 parturients, 11 male and 23 female fetuses) of pregnancy. The determination of serum hormone concentration was carried out with the RIA method in 2084 samples. There was no significant difference between the serum hormone concentrations of the male and female sex in the 28th-40th weeks of pregnancy. They assume that in the 28th-40th weeks there is no difference in the activity of steroid and peptide hormone secretion caused by the sex of the fetus.

An intra-ovarian role for oestrogens in the control of steroid production was investigated using dispersed thecal cells obtained from porcine follicles. Thecal cells were incubated for 14 h at 37 degrees C and the media subsequently assayed for androstenedione, progesterone and cyclic AMP. LH caused a dose-dependent stimulation of both steroids and the addition of oestradiol at doses of 10 ng-10 micrograms/ml significantly (P less than 0.01) inhibited both basal and LH-stimulated steroid production from doses of 500 ng/ml and upwards. Of other oestrogens investigated, oestrone and oestriol were somewhat less potent than oestradiol in inhibiting steroid synthesis, whereas the synthetic oestrogen diethylstilbestrol (DES) was more potent. The presence of oestradiol at doses of 10 ng-10 micrograms/ml had no significant effect (P less than 0.05) on either basal or LH-stimulated cAMP suggesting that the oestradiol inhibition does not involve inhibition of LH receptor-linked adenylate cyclase. These results demonstrate that physiological doses of oestrogen can act by local negative feedback to control the synthesis of its own precursor and thus regulate intrafollicular steroidogenesis.

Unconjugated oestrone (Oe1), oestradiol-17beta (Oe2), oestriol (Oe3), progesterone (P) and HPL in plasma were determined by radioimmunoassay and the immunological pregnancy-test in urine was carried out in 70 patients with normal pregnancy or imminent abortion from 4th-20th week of gestation. Oe2 and HPL showed the most pronounced rises, Oe3 increased especially after the first trimester. In cases with abortion symptoms and poor prognosis Oe2 and HPL gave the most reliable results concerning the endocrin function of early normal pregnancy. Oe1- and P-values in normal pregnancy did not differ so clearly from concentrations observed during normal menstrual cycles and were thus of less value. The pregnancy-test was positive (greater than 1000 IU/1) even in most cases of dead pregnancy and therefore not reliable. With increasing production of oestrogen precursors in the fetal adrenal cortex after the first trimester determination of Oe3 becomes more important. In cases with abortion symptoms in early pregnancy and subsequent normal development, plasma Oe2- and Oe3- values represented best criteria for a prognosis. -- For the diagnosis and control of the endangered early pregnancy we recommend, as a consequence of this study, determination of Oe2 up to the 13th week of pregnancy and thereafter Oe3 in maternal plasma.

Maternal serum alpha 1-antitrypsin concentrations were measured serially in pregnant women who were normotensive and those with mild, moderate, and severe hypertension of pregnancy from 27 weeks' gestation to term. alpha 1-antitrypsin concentrations increased with advancing gestation in all four groups. In addition, the hypertensive pregnancies showed higher than normal concentrations at each stage of pregnancy, with values in the severe hypertension group being higher than values in the other two hypertensive groups. At 35-36 weeks' gestation to term the increase in alpha 1-antitrypsin in the severe hypertension group was significant (p less than 0.05) when compared with the normotensive group. Although plasma oestriol and progesterone concentrations increased with advancing gestation in all groups, there was no direct relation between their concentrations and the increase in alpha 1-antitrypsin concentration in the hypertensive groups.

Dehydroepiandrosterone sulphate (DHEA-S), estradiol (E2), estriol (E3), progesterone (P4) and cortisol (F) maternal plasma concentrations were determined using enzyme or fluoroimmunoassays in 28 healthy pregnant women prior to the induction of labour, performed for various reasons, in order to evaluate their impact on cervical ripening. Mean maternal and gestational ages were 24.7 +/- 3.6 years and 39.1 +/- 1.9 weeks, respectively, and the gravidity ranged from 0-4. The subjects were divided according to cervical maturity into Group A (n = 13, Bishop scores>> 4) and Group B (n = 15, Bishop scores < 4). There were no statistically significant differences observed between the two groups as to concentrations of oestradiol, oestriol, progesterone and cortisol. However, significantly higher concentrations of DHEA-S were found in group A compared to group B (1335 +/- 885 ng/ml and 558.2 +/- 191.4 ng/ml, respectively), (p < 0.001). In conclusion, the high DHEA-S concentrations in maternal plasma may play an important role in pregnancy by producing favourable cervical conditions for delivery or by triggering the labour itself.

Umbilical cord arterial and venous, and maternal peripheral vein blood samples were collected during 20 deliveries after uncomplicated pregnancies. Using a gas chromatographic method, oestriol, oestriol sulphate, progesterone and a number of neutral steroid mono- and disulphates were quantitated in the cord plasma samples and a number of neutral steroid mono- and disulphates in the maternal plasma samples.

Daily injections of oestriol, spanning a wide dose range, produced mitotic responses in the uterine luminal epithelium of mice. At the higher doses the response was comparable to that obtained with oestradiol. The maximum response occurred on day 1, after which mitosis was significantly reduced on days 2 and 3. This reduction in mitosis following an initial peak was seen at all doses and was not therefore consequent on a critical cell number being reached. Proliferation was also induced in the basal epithelium of the vagina, though to a lesser extent. Oestriol and progesterone together stimulated DNA synthesis and mitosis in the epithelium of the mammary gland.

The structure-activity relationships of various natural and synthetic oestrogens, anti-oestrogens and some other steroid hormones with respect to early uterine albumin accumulation (EAV) was studied in sprayed mice. The results were compared with the activity of these agents to produce a late anabolic response (LAR). The potency ranking order among the oestrogens with respect to EAV was found to be oestradiol-17 beta, greater than oestriol, greater than meso-hexoestrol, greater than erythro-deoxyhexoestrol, greater than ent-oestradiol-17 beta, greater than (-)methallenestril phenol. Generally the oestrogens were much more potent with respect to EAV than LAR. No correlation was found when the ability to promote EAV and LAR was compared. Among other agents tested for effect on the EAV, progesterone, the synthetic progestogen, DU-41165 and the anti-hormones MER-25 and U-11100A (nafoxidine) showed no effect. However, hydrocortisone diminished EAV, while dihydrotestosterone increased EAV substantially.

Oestradiol (E2), oestriol (E3) and oestrone (E1) levels were measured in plasma and endometrium, myometrium and vagina of 29 postmenopausal women who underwent hysterectomy. The influence of vaginal E3, compared to vaginal E2 therapy at a dose one-tenth that of E3 on the basal steroid levels was examined. We found (1) no correlation between basal tissue and plasma concentrations of the oestrogens in untreated postmenopausal women, however, after vaginal E3 therapy we did find a positive correlation between them, (2) E2 to be the oestrogen in the highest basal concentration in endometrium and myometrium as well as in the vagina, (3) higher basal concentrations of all three oestrogens in endometrium compared to myometrium and vagina, (4) a long term (at least 12 h) elevation of the plasma and tissue E3 concentrations after vaginal E3 therapy (0.5 mg per day), (5) no significant changes of the plasma and tissue E2 concentrations after 0.05 mg per day vaginal E2 therapy, measured 12 h after the last application and (6) no signs of a difference between vagina and uterus in uptake and retention of E3 or E2. In conclusion, there was no difference on this level of mechanism of action in vagina and uterus which can account for the supposed vaginotrophicity and non-uterotrophicity observed with E3 but not E2.

Data in the Caucasian population suggest that maternal serum alpha-fetoprotein (AFP) and unconjugated oestriol concentrations are reduced and human chorionic gonadotrophin (HCG) concentrations are elevated in pregnancies conceived after in-vitro fertilization (IVF), leading to a higher than expected Down's syndrome screen-positive rate. There are no previous reports on the serum marker values in pregnancies conceived after intracytoplasmic injection (ICSI). Between 1996 and 1998, we measured maternal serum total HCG and AFP concentrations between 15 and 20 weeks gestation in 42 in-vitro fertilization (IVF) pregnancies and 23 ICSI pregnancies with known normal outcome. The results were compared with that of 2799 naturally occurring singleton pregnancies who were known to have a normal outcome. Median AFP multiple of the median (MOM) in ICSI pregnancies was significantly reduced to 0.76 compared with both that of the controls and that of the IVF pregnancies. For the IVF pregnancies, median HCG MOM was elevated to 1.15, and median AFP MOM was reduced to 0.88 compared with the controls, but these differences were not statistically significant. In both the IVF and ICSI pregnancies the changes might result in a falsely high Down's syndrome risk. In particular, the reduced AFP concentration in ICSI pregnancies was substantial. If this preliminary finding is substantiated by other series, the appropriate adjustment needs to be made to allow for valid interpretation of the screen result and to avoid an unnecessarily high false positive rate.

BACKGROUND

The recommendations of the UK National Screening Committee (NSC) are based on the findings of the Serum, Urine and Ultrasound Screening Study (SURUSS). Our study sought to establish if the SURUSS findings, in terms of detection rates (DR) and false-positive rates (FPR) for various second-trimester marker combinations, could be replicated in a local setting.

METHODS

We investigated the effects of adding inhibin-A and unconjugated oestriol to our existing double test protocol. This retrospective study examined 1000 control pregnancies and 128 affected pregnancies.

RESULTS

The inhibin-A method was associated with considerable assay drift and very marked within-batch imprecision (intrabatch percent coefficient of variation [CV] = 17%). At a cut-off of 1 in 250, the quadruple test showed a DR of 72%, the triple test 70% and the double test 63%. There were no significant differences between the FPRs for any of the combinations, which were all between 6.6% and 7.0% for a 1 in 250 cut-off.

CONCLUSIONS

In our view, the current inhibin-A assay is unacceptable as a screening marker due to poor assay performance. We have reviewed the NSC benchmark programme outcomes, and would suggest that the 2005 target of a DR of at least 60% with an FPR of less than 5% is achievable using triple testing in the second trimester in conjunction with universal scan dating. Our results suggest that the April 2007 target of a DR of 75% with an FPR of less than 3% is unachievable using current second-trimester maternal serum screening.