Amniotic Fluid Steroid Research Articles

Meconium androgens are correlated with ASD-related phenotypic traits in early childhood in a familial enriched risk cohort

BackgroundPrenatal exposure to increased androgens has been suggested as a risk factor for autism spectrum disorder (ASD). This hypothesis has been examined by measurement of steroids in amniotic fluid, cord blood, saliva, and blood with mixed results.MethodsTo provide an orthogonal measure of fetal exposure, this study used meconium, the first stool of a newborn, to measure prenatal androgen exposure from infants in the Early Autism Risk Longitudinal Investigation (EARLI). EARLI is a familial-enriched risk cohort that enrolled pregnant mothers who already had a child with an ASD diagnosis. In the younger child, we investigated the association between meconium unconjugated (u) and total (t) concentrations of major androgens testosterone (T), dehydroepiandrosterone (DHEA), and androstenedione (A4), and ASD-related traits at 12 and 36 months of age. Traits were measured at 12 months with Autism Observation Scale for Infants (AOSI) and at 36 months with total score on the Social Responsiveness Scale (SRS). One hundred and seventy children had meconium and AOSI, 140 had meconium and SRS, and 137 had meconium and both AOSI and SRS.ResultsSeparate robust linear regressions between each of the log-transformed androgens and log-transformed SRS scores revealed three-way interaction between sex of the child, sex of the proband, and testosterone concentration. In the adjusted analyses, t-T, u-A4, and u-DHEA (P ≤ 0.01) were positively associated with AOSI scores, while u-T (P = 0.004) and u-DHEA (P = 0.007) were positively associated with SRS total score among females with female probands (n = 10). Additionally, higher concentrations of u-T (P = 0.01) and t-T (P = 0.01) predicted higher SRS total score in males with male probands (n = 63).LimitationsSince we explored three-way interactions, this resulted in a limited sample size for some analyses. This study was from an enriched-risk cohort which may limit generalizability, and this study used ASD-assessment scales as outcomes instead of diagnostic categories. Additionally, the novel use of meconium in this study limits the ability to compare the results in this cohort to others due to the paucity of research on meconium.ConclusionsThis study supports the utility of meconium for studies of endogenous fetal metabolism and suggests the sex of older siblings with autism should be considered as a biological variable in relevant studies.

A clinical scoring system to predict the need for extensive resuscitation at birth in very low birth weight infants

BackgroundTo analyze the risk factors for extensive cardiopulmonary resuscitation in the delivery room and develop a prediction model for outcomes in very low birth weight (VLBW) infants.MethodsThe sample was 5298 VLBW infants registered in the Korean neonatal network database from 2013 to 2015. Univariate and multivariate analyses were used to analyze the risk factors for extensive resuscitation. In addition, a multivariable model predicting extensive resuscitation in VLBW infants was developed.ResultsUnivariate regression analysis of antenatal factors showed that lower gestational age, lower birth weight, birth weight less than third percentile, male sex, maternal hypertension, abnormal amniotic fluid volume, no antenatal steroid use, outborn, and chorioamnionitis were associated with extensive resuscitation at birth. Lower gestational age (25 to 27 gestational weeks, odds ratio [OR] and 95% confidence interval [CI]: 3.003 [1.977–4.562]; less than 25 gestational weeks, OR and 95% CI: 4.921 [2.926–8.276]), birth weight less than 1000 g (OR and 95% CI: 1.509 [1.013–2.246]), male sex (OR and 95% CI: 1.329 [1.002–1.761]), oligohydramnios (OR and 95% CI: 1.820 [1.286–2.575]), polyhydramnios (OR and 95% CI: 6.203 [3.185–12.081]), and no antenatal steroid use (OR and 95% CI: 2.164 [1.549–3.023]) were associated on multivariate regression analysis. The final prediction model for extensive resuscitation included gestational age, amniotic fluid, and antenatal steroid use. It presented a sensitivity of 0.795 and specificity of 0.575 in predicting extensive resuscitation at birth, corresponding to a score cut-off of 2. The area under the receiver operating characteristic curve was 0.738.ConclusionsLower gestational age, abnormal amniotic fluid volume, and no use of antenatal steroid in VLBW infants are important predictors of extensive resuscitation in the delivery room.

Identification of amniotic fluid metabolomic and placental transcriptomic changes associated with abnormal development of cloned pig fetuses.

Piglets cloned by somatic cell nuclear transfer (SCNT) show a high incidence of malformations and a high death rate during the perinatal period. To investigate the underlying mechanisms for abnormal development of cloned pig fetuses, we compared body weight, amniotic fluid (AF) metabolome, and placental transcriptome between SCNT- and artificial insemination (AI)-derived pig fetuses. Results showed that the body weight of SCNT pig fetuses was significantly lower than that of AI pig fetuses. The identified differential metabolites between the two groups of AF were mainly involved in bile acids and steroid hormones. The levels of all detected bile acids in SCNT AF were significantly higher than those in AI AF. The increase in the AF bile acid levels in SCNT fetuses was linked with the downregulation of placental bile acid transporter expression and the abnormal development of placental folds (PFs), both of which negatively affected the transfer of bile acids from AF across the placenta into the mother's circulation. Alteration in the AF steroid hormone levels in cloned fetuses was associated with decreased expression of enzymes responsible for steroid hormone biosynthesis in the placenta. In conclusion, cloned pig fetuses undergo abnormal intrauterine development associated with alteration of bile acid and steroid hormone levels in AF, which may be due to the poor development of PFs and the erroneous expression of bile acid transporters and enzymes responsible for steroid hormone biosynthesis in the placentas.

Characterizing the steroidal milieu in amniotic fluid of mid-gestation: A LC–MS/MS study

Growth and development of an embryo or fetus during human pregnancy mainly depend on intact hormone biosynthesis and metabolism in maternal amniotic fluid (AF). We investigated the hormonal milieu in AF and developed a liquid chromatography-tandem mass spectrometry (LC–MS/MS) method for the determination of 14 sulfated and 6 unconjugated steroids in AF. 65 A F samples (male: female = 35: 30) of mid-gestation ranging from 16th week of gestation to 25th week of gestation were analyzed. Reference data of 20 steroid levels in AF of healthy women were provided. 13 sulfated and 3 unconjugated steroids were for the first time quantified in AF by LC–MS/MS. Highest concentrations were found for pregnenolone sulfate (PregS: mean ± SD, 8.6 ± 3.7 ng/mL), 17α-hydroxypregnenolone sulfate (17OHPregS: 4.9 ± 2.0 ng/mL), epitestosterone sulfate (eTS: 7.3 ± 3.6 ng/mL), 16α-hydroxydehydroepiandrosterone sulfate (16OH-DHEAS: 21.5 ± 10.7 ng/mL), androsterone sulfate (AnS: 9.2 ± 7.4 ng/mL), estrone sulfate (E1S: 3.0 ± 3.0 ng/mL), estriol 3-sulfate (E3S: 8.1 ± 4.0 ng/mL) and estriol (E3: 1.2 ± 0.4 ng/mL). Only testosterone (T) showed a significant sex difference (p < 0.0001). Correlations between AF steroids mirrored the steroid metabolism of the feto-placental unit, and not only confirmed the classical steroid pathway, but also pointed to a sulfated steroid pathway.

Development of a new panel of 9 steroids in amniotic fluid in LC–MSMS

Background Since 1980 (Forest et al. [1] ) steroid analysis in amniotic fluid (AF) by radioimmunoassay were used in our laboratory as a tool for the prenatal diagnosis of 21 hydroxylase deficiency or other disorder of sex development (DSD). LC–MSMS became reference method for several reasons: analytical performance, possibility of studying a panel of several steroids. Objective Establishment of reference values for 21-deoxycortisol (21DF), deoxycorticosterone (DOC), corticosterone (Cortico), 17-hydroxyprogesterone (17OHP), 11-deoxycortisol (CS), dehydroepiandrosterone (DHEA), delta4-androstenedione (D4), testosterone (T) and dihydrotestosterone (DHT) in AF by LC–MS/MS method according to sex and term of pregnancy. Method Liquid chromatography was performed on an Agilent Technologies UPLC 1290 Infinity and selected steroids were analyzed with an Agilent Technologies 6460 triple quadrupole. Internal standard were added in 1 mL of the sample before supported liquid extraction. Materials After informed consent, we included 193 pregnant women AF performed from 14 to 36 weeks of amenorrhea, for increased maternal serum markers. All the samples associated with fetus ultrasound morphological, karyotypic or neonatal abnormalities were excluded. Results We validate the method allowing quantification of the 9 steroids in amniotic fluid with variability coefficient less than 10% intra- and inter-assay. The average concentration of these steroids was similar in both sex for 21DF( Conclusion LC–MS/MS is a rapid, sensitive method for steroid analysis in AF. References values will be very useful for the management of a mismatch between external genitalia observed by ultrasound and karyotype.

Measurement of amniotic fluid steroids of midgestation via LC–MS/MS

IntroductionAnalysis of steroids by mass spectrometry (MS) has evolved into a reliable tool for the simultaneous detection of multiple steroids. As amniotic fluid (AF) and fetal serum composition of early pregnancy are closely related, the analysis of AF can yield information on the physiological status of the developing fetus. We evaluated the use of liquid-chromatography tandem mass spectrometry (LC–MS/MS) for AF steroid analysis, including the analysis of its sensitivity and accuracy for gender verification in healthy subjects. Materials and methodsAF of 78 male and 94 female healthy newborns was analyzed by LC–MS/MS at 16 weeks of gestation. The levels of androstenedione, corticosterone, cortisol, cortisone, deoxycorticosterone, 11-deoxycortisol, dehydroepiandrosterone (DHEA), dehydroepiandrosterone-sulfate (DHEA-S), 17-hydroxyprogesterone, progesterone (17-OHP) and testosterone were measured. Steroid levels were compared to RIA and GC–MS levels of midgestation from the literature. Cross-validated logistic regression was used to obtain statistical predictions of gender at birth from testosterone and the above steroids. ResultsLC–MS/MS analysis of AF steroids yielded comparable results with published GC–MS data. Gender specific differences were found for androstenedione and testosterone concentrations with higher levels in the male fetus. In contrast to published RIA data no gender specific differences were observed for 17-hydroxyprogesterone and dehydroepiandrosterone AF concentrations. Testosterone concentrations yielded highly accurate predictions for male gender at birth. Additional analysis of further steroids did neither increase the accuracy, sensitivity nor specificity of this prediction. The estimated optimal cut-off value for amniotic testosterone level was 0.074μg/L for healthy male newborns. ConclusionsLC–MS/MS is a reliable method for the determination of steroids in amniotic fluid. The determination of testosterone in amniotic fluid by LC–MS/MS in early pregnancy of healthy subjects can be used to offer a reliable prediction of fetal gender at birth.

Quantitative high-performance liquid chromatographic determination of delta 4-3 ketonic free steroids in human plasma and amniotic fluid.

Summary A high-performance liquid chromatographic method is described for the measurement and identification of eight steroids possessing the Δ4-3 ketonic compounds in human specimens. The eight steroids (Cortisone, Cortisol, Corticosterone, DOC, Δ4-Androstendione, Testosterone, 17-Hydroxyprogesterone and Progesterone) were separated with a methanol-THF-water system on an IRICA RP-18 column. Effluents were detected by UV absorption at 245 nm. Quantitative analysis was performed by measurement of peak heights, which were proportional to the amount of each substance. This method is rapid, highly sensitive, easy to perform and reproducible.

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.

Prenatal diagnosis of congenital lipoid adrenal hyperplasia.

Congenital lipoid adrenal hyperplasia (lipoid CAH) is a rare genetic disorder of adrenal and gonadal steroidogenesis of unknown cause in which cholesterol cannot be converted to pregnenolone. As a result, affected individuals can make no steroid hormones, so that all affected newborns are phenotypic females, irrespective of karyotype. We studied two pregnancies in a family with two previously affected children by examining fetal karyotype, genital ultrasonography, and amniotic fluid steroid concentrations and by performing ACTH tests on family members. Prenatal diagnosis correctly identified both an unaffected XX fetus and an affected XY fetus. In the affected pregnancy, amniotic fluid concentrations of progesterone and pregnenolone were 30% and 50% of normal, respectively, but concentrations of 17 alpha-hydroxypregnenolone, 17 alpha-hydroxyprogesterone, cortisol, dehydroepiandrosterone, androstenedione, and estriol were either extremely low or undetectable, suggesting that these detected steroids were donated by maternal steroidogenesis. Fetal cord blood obtained at the termination of pregnancy showed very low concentrations of estrogens donated by the mother's circulation. Absent fetal steroidogenesis was confirmed by gas chromatography and mass spectrometry of both fetal and maternal serum. The responses of 10 different steroids to adrenal stimulation with ACTH in the obligately heterozygous parents were normal. Thus, unlike the case with other forms of CAH, heterozygosity cannot be determined by hormonal responses to provocative testing with ACTH. Immunocytochemistry and Western blotting showed that the affected placental tissue contained P450scc protein, confirming that P450scc is intact in these patients.

PRENATAL DIAGNOSIS OF CONGENITAL LIPOID ADRENAL HYPERPLASIA (LIPOID CAH)

Inability to convert cholesterol to pregnenolone causes lipoid CAH, the most severe form of CAH. We now report the prenatal diagnosis of lipoid CAH in a previously evaluated Korean family (JCI 88:1955, 1991). Steroidal responses to ACTH stimulation in both obligatory heterozygous parents were indistinguishable from normal data (JCEM 73:679, 1991). In two pregnancies after diagnosis of the affected proband one male fetus was diagnosed ultrasonographically as unaffected and confirmed postnatally by endocrine testing, and a second XY male fetus lacked sonographically detectable genitalia. Amniotic fluid steroids were low but not absent (Table). The fetus was aborted at 18 weeks. The fetal adrenals were of normal size but contained lipoid droplets; the testes were histologically normal.In summary: these data indicate that prenatal diagnosis of lipoid CAH can be made readily, and that maternally derived steroids (particularly Prog) play an essential role in carrying such pregnancies to term.

HORMONAL MONITORING AND SIDE EFFECTS OF PRENATAL DEXAMETHASONE (DEX) TREATMENT FOR 21-HYDROXYIASE DEFICIENCY CONGENITAL ADRENAL HYPERPIASIA (CAH)

Maternal Dex therapy for fetal CAH at 1-1.5 mg daily initiated between 5-8 weeks gestation and continued to birth, resulted in normal genital outcome, mild virilization, and significant genital virilization in the three CAH newborns treated, and significant side effects in the three mothers, including excess weight gain (9-26 Ibs greater at 35-37 weeks gestation compared to previous full-term pregnancies), glucose intolerance, facial hirsutism, and Cushingoid features including moon-face, plethora, markedly discolored striae on arms, abdomen, breast, and thighs, which resulted in permanent scarring. In addition, hypertension, epigastric pain, and emotional lability were noted, but not consistently. Amniotic fluid 17-OH Progesterone, androstenedione, and testosterone, were elevated in all three Dex treated cases. Maternal EI level was consistently suppressed in the case with the normal genital outcome, and were initially unsuppressed in the cases with mild and severe genital virilization. Conclusion: Dex therapy for prenatal treatment of CAH at a dose of 1-1.5 mg daily was associated with significant maternal side effects. Amniotic fluid steroid levels may not be useful, while maternal serum E1 level may be useful for therapeutic monitoring of fetal CAH, this however requires further investigation.

Prenatal dexamethasone treatment in pregnancies at risk for congenital adrenal hyperplasia due to 21-hydroxylase deficiency: effect on midgestational amniotic fluid steroid levels.

Prenatal diagnosis of congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency by amniotic fluid (AF) steroid analysis is not possible in those cases in which prenatal dexamethasone (DEX) therapy is initiated to prevent virilization of female CAH fetuses because AF steroid levels are suppressed if DEX therapy is continued beyond amniocentesis (AC). In order to use AF steroids for prenatal diagnosis, it is necessary to discontinue DEX therapy for 5 days before AC. To study the effects of this interruption on AF steroid levels, we measured levels of aldosterone, corticosterone, 11-deoxycorticosterone, progesterone, 17-hydroxyprogesterone (170HP), 11-deoxycortisol, cortisol, and cortisone as well as androstenedione (delta 4-A) in AF samples (16-18 weeks) obtained from 25 pregnancies at risk for CAH treated with Dex (daily dosage: 1.0-1.5 mg). The prenatal diagnosis of 14 normal fetuses and 11 affected CAH fetuses was postnatally confirmed in all cases. Additionally, steroid levels were measured in AF samples (16-18 weeks) from 8 untreated CAH fetuses and in 19 AF samples (weeks 16-20) obtained in normal pregnancies. In 17/19 prenatally diagnosed CAH fetuses, the affected sibs had the salt wasting (SW)-form, in 2 cases the simple virilizing (SV)-form. All steroids were measured by RIA after extraction and Sephadex LH-20 chromatography. AF levels of aldosterone, corticosterone, deoxycorticosterone, progesterone, cortisol, cortisone, and 11-deoxycortisol were not different between CAH fetuses, prenatally DEX-treated normal fetuses and untreated controls. The 170HP-levels of the CAH-SW-fetuses (range: 19.9-59.8 mmol/L) were clearly above the normal range (3.74-11.6), but normal in the SV-fetuses (10.9, 11.5), whereas delta-4 A-levels (normal range: 0.87-5.13 mmol/L) were elevated both in the SW-(range: 6.53-37.6) and the SV-form (9.37,6.25) of CAH. 170HP and delta-4 A levels of prenatally DEX-treated pregnancies with normal fetuses were not different from levels found in normal pregnancies. Mean 170HP and delta-4 A AF steroid levels of prenatally DEX-treated CAH-pregnancies were slightly lower (NS) than levels of untreated CAH-pregnancies (170HP: 30.5 vs. 40.7; delta-4 A: 15.8 vs. 21.1). 170HP levels are elevated in the SW-form of CAH, but not in the SV-form. However, with the combination of 170HP and delta-4 A levels it is possible to diagnose prenatally both forms. There is no rebound phenomenon of AF steroid levels if DEX therapy is interrupted 5 days before amniocentesis.

Prenatal dexamethasone treatment in pregnancies at risk for congenital adrenal hyperplasia due to 21-hydroxylase deficiency: effect on midgestational amniotic fluid steroid levels

Prenatal dexamethasone treatment in pregnancies at risk for congenital adrenal hyperplasia due to 21-hydroxylase deficiency: effect on midgestational amniotic fluid steroid levels

EVIDENCE THAT GONADAL HORMONE LEVELS IN AMNIOTIC FLUDI ARE DECREASED IN MALES BORN TO ALCOHOL USERS IN HUMANS

Biochemical evaluation of amniotic fluid content is often used to monitor fetal secretory and excretory functions. It is known that alcohol freely passes through the placenta and enters the fetal tissues. The study was designed to investigate whether alcohol use during pregnancy affects amniotic fluid steroid and peptide hormone levels. Five alcohol users who drank between 40 and 45 drinks per month and consumed approximately 1 oz. of alcohol per day from beer drinking participated in the study. Thirteen controls with no history of alcohol use including essential and non-essential drugs were selected for the study. The final selection of participants was made among those who delivered male offsprings. Amniotic fluid was aspirated between 16 and 20 weeks of gestation by amniocentesis and an aliquot was used to assay steroid hormones (4-androstenedione [-4-AD], testosterone [T], estradiol [E2], dehydroepiandrosterone sulfate [DHEA-S]), and peptide hormones (FSH and LH). Birth weights, placental weights and gestation age were recorded at birth. The results showed that -4-AD, T and E2 levels were significantly decreased (P less than 0.01) in alcohol users; DHEA-S, FSH and LH levels were unaffected. Birth weights were significantly decreased (P less than 0.05) in alcohol users, the placental weights and gestation age were not affected. We conclude that alcohol passes through the placenta and affects fetal testes, capacity to synthesize steroids and the effect appears to be direct and not via the hypothalamic-hypophyseal axis.

Congenital Adrenal Hyperplasia

The incidence of congenital adrenal hyperplasia in Saudi Arabia and the frequency of the carrier rate are not well known. Both figures should be high, considering the commonness of first-degree consanguinity in this part of the world. We present 25 cases of congenital adrenal hyperplasia, most of them due to a defect of the 21 hydroxylase enzyme, diagnosed and followed over three years at Suleimania Children's Hospital in Riyadh. There were ten boys and 15 girls exhibiting all degrees of masculinization, mostly Grade III (53%) and Grade IV and V (13.3% each). The degree of masculinization of the external genitalia in the girls (46XX) was sufficiently pronounced in seven patients to lead to sex identity errors at birth. This aggravated the psychological reactions in the families, particularly before the surgical correction. The frequency of neonatal deaths in the patients' families related to a similar disorder was high (12%). The mean age at onset of the salt-losing crises in affected infants was 9 days in females and 25 days in males and this was the most common clinical presentation of the disorder (80%). Systemic neonatal screening and intrauterine diagnosis in at risk families must be done to provide hormonal intervention and thus prevent the formation of ambiguous genitalia and avoid the resulting plastic surgery that must be performed in female patients.

Midgestational amniotic fluid (AF) steroid levels in CAH-pregnancies: effect of prenatal dexamethasone (DEX) treatment

Midgestational amniotic fluid (AF) steroid levels in CAH-pregnancies: effect of prenatal dexamethasone (DEX) treatment

Prenatal diagnosis of congenital adrenal hyperplasia: reliability of amniotic fluid steroid analysis.

The concentration of 170H-progesterone was measured in amniotic fluid samples collected from 55 mothers who had previously had a child with congenital adrenal hyperplasia due to 21-hydroxylase deficiency. In eight pregnancies the levels of 170H-progesterone were raised; the parents elected to terminate in four and examinations of the fetus confirmed the diagnosis of congenital adrenal hyperplasia. In each case, the affected sib was a salt loser. The remaining four affected pregnancies proceeded to term and each infant had salt losing 21-hydroxylase deficiency. All 47 infants predicted to be unaffected were normal at birth. However, an increased plasma concentration of 170H-progesterone was documented in a male non-salt loser at three months of age. Prenatal diagnosis of congenital adrenal hyperplasia by amniotic fluid steroid analysis is reliable only for the salt losing variant of 21-hydroxylase deficiency. Of the affected sibs in this study, 20% died during infancy in a salt losing crisis. This simple and rapid prenatal test is sufficiently reliable to predict the group of infants most at risk in early infancy.

Amniotic fluid steroid levels and fetal adrenal weight in congenital adrenal hyperplasia.

The concentration of 17-OH-progesterone was determined in second trimester amniotic fluid collected from 58 pregnancies at risk for fetal 21-hydroxylase deficiency. The prediction was incorrect in 1 male nonsalt-loser who had an increased plasma 17-OH-progesterone concentration at 3 months of age. All 11 infants predicted to be affected were salt-losers. The adrenals from 2 affected fetuses available for study were significantly enlarged in comparison with adrenal size in 84 normal fetuses from 15 to 26 weeks' gestation. Amniotic fluid steroid analysis reliably predicts the fetus with 21-hydroxylase deficiency most at risk in early infancy. There is no evidence from this study to indicate that ACTH is not the main trophic factor for fetal adrenal growth and steroidogenesis.

Pitfalls in prenatal diagnosis of 21-hydroxylase deficiency by amniotic fluid steroid analysis? A six years experience in 102 pregnancies at risk.

Pitfalls in prenatal diagnosis of 21-hydroxylase deficiency by amniotic fluid steroid analysis? A six years experience in 102 pregnancies at risk.

124 PRENATAL DIAGNOSIS OF 17-KETOSTEROID REDUCTASE DEFICIENCY

In the index case, first child of the family, diagnosis of 17-ketosteroid reductase deficiency(17-KSRD) was made at 3 months of age on the response of testosterone(T) and Δ4-androstenedone(Δ4) after an hCG test : low levels (ng/dl) of T(101) but Δ4 values (394) 10 times greater than the normal mean. At that time, the mother being pregnant again, requested a prenatal diagnosis. The proposed protocol included fetal karyotype, amniotic fluid (AF) steroid analysis at mid-pregnancy and ultrasound follow up of the genitalia. Prior to this study, AF levels of T,Δ4 and individual Δ4/T ratio were determined in 150 controls. In normal pregnancies AF Δ4/T ratios are clearly (p < 0.01) lower in male(4±1.6) than in female(12.8±4.5)fetuses. Based on the results (46,XY, AF levels low for T(5.9), high for Δ4 (217) and extremely high Δ4/T ratio (36.8), no phallic growth at ultrasound) the fetus was predicted affected. 17-KSRD was confirmed after birth : female phenotype, testes palpable in labia majora, typical response to hCG(T : 16→140; Δ4: 48→601). At the mother's request the same protocol was used for prenatal diagnosis during the third pregnancy : karyotype was 46,XY but AF levels of T(14), Δ4 (57) and Δ4/T ratio (4.2) were normal for a male fetus ; apparently normal genitalia were observed at ultrasound. The prediction was confirmed, since the boy was perfectly normal at birth.In conclusion. This study demonstrates for the first time the feasability of a prenatal diagnosis of 17-KSRD, as reported for an other testicular enzyme defect (17, 20-desmolase) (JCEM 1980, 50, 826).