rFSH in medically assisted procreation: Evidence for ovarian follicular hyperplasia and interest of mass spectrometry to measure 17-hydroxyprogesterone and Δ4-androstenedione in serum.

Ovarian monitoring requires the determination of serum estradiol and progesterone levels. We investigated whole follicular steroidogenesis under rFSH in medically assisted procreation (MAP: 26 IVF, 24 ICSI) compared to 11 controls (IUI). Estrone, estradiol, Δ4-androstenedione, testosterone, progesterone and 17-hydroxyprogesterone were measured by immunoassay and mass spectrometry except for estrogens. At the start of a spontaneous or induced cycle, steroids levels fluctuated within normal ranges: estradiol (314-585 pmol/L), estrone (165-379 pmol/L) testosterone (1.3-1.6 nmol/L), Δ4-androstenedione (4.5-5.6 nmol/L), 17-hydroxyprogesterone (2.1-2.2 nmol/L) and progesterone (1.8-1.9 nmol/L). 17-hydroxyprogesterone, Δ 4-androstenedione and estradiol predominated. Then estradiol and oestrone levels rise, but less markedly for oestrone in IUI. In MAP, rFSH injections induce a sharp increase in estrogens associated with a rise in 17-hydroxyprogesterone and Δ4-androstenedione levels, disrupting oestrogen/androgen ratios. rFSH stimulation induces an ovarian hyperplasia and Δ4pathway which could become abnormal. Determining 17-hydroxyprogesterone and Δ4-androstenedione levels with LC-MS/MS may therefore be useful in managing recurrent MAP failures.

hCG in screening for aneuploidy: a possible role for its glycoforms?

Fetal trisomy 21 is associated with elevated maternal serum hCG and its free beta-subunit (hCG-beta) in vivo, and abnormal placental hCG production and glycosylation in vitro. Other maternal serum markers may also be disrupted in major aneuploidies (T21, T18, T13). We evaluated our aneuploidy screening practices, focusing on hCG-beta and hCG glycoforms, and retrospectively analyzed 55 aneuploidy cases diagnosed over a 2 year period, determining maternal serum hCG glycoforms profiles using 2D-electrophoresis. Screening efficiency reached 96.7%. T21 was associated with elevated hCG-beta while T18 presented with diminished serum markers. hCG glycoforms tended to be basic in aneuploidy (mainly T13).

Foetal serum but not urinary ß2-microglobulin correlates with histological injury to the kidney.

In a context of foetal obstructive uropathies, biochemical markers can be helpful to assess the renal function, but most studies to date have focused on their correlation with ultrasound findings and neonatal outcome. Our aim was to evaluate foetal ß2-microglobulin as an index of histological injury to the kidney. ß2-microglobulin was measured in serum and/or urine from 27 foetuses with bilateral obstructive uropathy, and compared to the findings of kidney examination following the termination of pregnancy. In serum, increased ß2-microglobulin levels correlated to a decreased number of glomeruli, a reduction in the blastema and the presence of primitive ducts reflecting renal hypoplasia and dysplasia. However, elevated ß2-microglobulin levels in the urine correlated only to a decreased number of glomeruli.

[ART with sperm donor after intraconjugal ICSI failure]. / Prise en charge en don de sperme après échec(s) d'ICSI intraconjugale.

OBJECTIVES: To evaluate the ART results with sperm donation for couples who do not succeed after IC ICSI treatment for male infertility. PATIENTS AND METHODS: Retrospective study of 71 couples (November 1994-February 2009). Two control populations were determined to analyse this two-steps treatment (IC and sperm donation) based on different criteria such as the date of ICSI, the female age at the time of the first attempt and the male indication. RESULTS: In IC-ICSI, the proportion of azoospermic subjects and/or carrying chromosomal abnormalities was significantly higher in the studied population as compared to control. The fertilization rate (FR) and the embryo quality were significantly lower in the studied population. After donation, 30 couples (42.2%) succeeded in being parents. The donor IA pregnancy rates were similar to those of the control population except for the azoospermic patients with poor results. When IVF/ICSI-D was performed, the pregnancy rates were lower than in the control population. DISCUSSION AND CONCLUSION: Sperm donation constitutes an option for half of the couples initially treated with IC-ICSI for male infertility. For couples who failed in sperm donation, a female implication cannot be excluded, even though not detected.

Hyperglycosylated hCG is a marker of early human trophoblast invasion.

CONTEXT: Human chorionic gonadotropin (hCG) is the major pregnancy glycoprotein hormone whose maternal concentration and glycan structure change all along pregnancy. hCG is mainly secreted by the syncytiotrophoblast covering the chorionic villi, but little is known about the source of hyperglycosylated hCG (hCG-H) production. OBJECTIVE: The objective of the study was to analyze expression and secretion of hCG and hCG-H in vitro during human trophoblastic cell differentiation, in situ in first-trimester placentas, and in maternal sera during early pregnancy. DESIGN: hCG and hCG-H were measured in cell supernatants from primary cultures of first-trimester placenta trophoblastic cells, which differentiate in vitro into syncytiotrophoblast or invasive extravillous cytotrophoblasts (evct). hCG-H immunodetection were performed on 9 wk gestation (WG) placental tissue sections. Total hCG and hCG-H were quantified by chemiluminometric assay in 539 maternal sera collected between 9 and 19 WG during normal pregnancies. RESULTS: In vitro, hCG secretion reached 37 ng/ml per µg DNA during syncytiotrophoblast formation but contained few hCG-H (2-5% of total hCG). In contrast, hCG secretion (20 ng/ml per µg DNA) in evct supernatants contained 10-20% hCG-H. In situ, hCG-H immunostaining was strong in invasive and endovascular evct, weaker in mononucleated villous cytotrophoblasts, but negative in the syncytiotrophoblast. In maternal sera, hCG-H concentrations continuously decreased during pregnancy from 406 ± 222 ng/ml at 9 WG to 8 ± 6 ng/ml at 19 WG, whereas total hCG picked up at 11 WG and then decreased. CONCLUSIONS: This study suggests that the high levels of hCG-H observed in first-trimester maternal sera are mainly from invasive evct origin, reflecting the early trophoblast invasion process.

[Assessment of thyroid hormones and androgens in amniotic fluid: clinical interest]. / Dosage des hormones thyroïdiennes et surrénaliennes dans le liquide amniotique: aide au diagnostic.

Ultrasound scanning is able to detect foetal goiter due either to an hypothyroidy either to an hyperthyroidy, or clitoris hypertrophia resulting from adrenal hyperplasia in female, during the second half of pregnancy. The diagnosis of these rare diseases is of interest because the treatment can be started during pregnancy. An amniotic fluid punction can be discussed and its biochemical analysis may be of interest even though very few commercial assays have been tested on amniotic fluid. Our aim was two investigate the practicability and the value of free thyroxin (FT4), thyrotropin (TSH), 17alpha hydroxyprogesterone (17-OHP) and delta 4 androstenedione (Delta4A) measurement on amniotic fluid using commercially available assays for serum. FT4 and TSH are detectable at low levels in amniotic fluid. FT4 significantly increases from 2.1 pmol/L to 4.2 pmol/L while TSH significantly decreases from 0.27 mU/L to 0.12 mU/L during the second half of pregnancy. An increase in amniotic fluid TSH concentration contributes to the diagnosis of foetal hypothyroidy while the measurement of amniotic fluid FT4 is not informative in case of foetal goiter. 17-OHP and Delta4A are present in amniotic fluid at the same level as in serum. 17-OHP significantly decreases from 1.9 ng/mL to 1 ng/mL during the second half of pregnancy while Delta4A significantly increases from 0.5 ng/mL to 0.8 ng/mL. Absence of increase in their concentrations excludes any severe adrenal hyperplasia.