Amniotic fluid metabolic biomarkers of fetal physiology and pregnancy success.

Need to define and validate an early pregnancy loss screening strategy

Editorial group: Cochrane Gynaecology and Fertility Group. Publication status and date: New search for studies and content updated (no change to conclusions), published in Issue 11, 2015.

PurposeIn some cases of prenatal genetic testing, an ample amount of fetal DNA is needed, to allow for parallel testing (conducting several genetic tests simultaneously). This study investigated the association between amniotic fluid DNA concentration and various factors. We aimed to define the required amount of amniotic fluid to be extracted in amniocentesis, to allow parallel testing throughout gestational weeks.MethodsDNA concentration was analyzed from amniocentesis samples taken during the years 2016–2022. Sex association was also analyzed in postnatal whole blood samples from a separate cohort. Theoretical minimum volume of amniotic fluid needed to ensure enough DNA for chromosomal microarray analysis and exome sequencing was calculated.ResultsWe focused our analysis on 2573 samples, which were taken during weeks 17–23 and 30–35. DNA concentrations increased from weeks 17 to 21, with relatively stable concentrations thereafter. Significantly higher DNA concentrations were seen in pregnancies of female fetuses. DNA concentrations in postnatal whole blood samples did not show this association. Across most weeks, the volume needed to extract 2 µg of DNA from 95% of the samples was about 34 ml.ConclusionDNA concentrations in amniotic fluid vary according to gestational age and are higher in pregnancies of female fetuses. This should be considered when determining the volume of fluid extracted and the timing of amniocentesis, with greater volumes needed in earlier stages of pregnancy.

Consumption of sucrose (table sugar) is high in much of the world. The effects of maternal sucrose intake on the placenta and fetal brain remain unknown. In rats, maternal consumption of sucrose at a human-relevant level alters the mother's physiology and steroids, as well as the adult offspring's brain and behavior. Effects in mothers are impaired glucose tolerance, increased liver lipids, increased adipose inflammation, and decreased corticosterone levels in the blood but not in the brain. In contrast, in adult female offspring, maternal sucrose intake increases corticosterone levels in the blood and the brain. In adult male offspring, maternal sucrose intake increases preference for high-sucrose and high-fat diets as well as motivation for sugar rewards in a progressive ratio task. In this study, we investigate the underlying mechanisms of the observed behavioral and endocrine effects in the adult offspring. We examine anti-inflammatory steroids, steroidogenic enzymes and cytokines in the placenta, amniotic fluid, fetal blood and brain. In our model, we feed rat dams either a high-sucrose diet (26% of kCal) or an isocaloric, matched, control diet (1% sucrose) 10 weeks prior to and during gestation. At embryonic day 19 (E19), we collected maternal serum, placenta, amniotic fluid, fetal blood, and fetal brain. We used Palkovits punch to microdissect the placenta and fetal brain. Next, we used liquid chromatography-tandem mass spectrometry (LC-MS/MS), which is highly sensitive and specific, to measure multiple steroids (e. g. corticosterone, estrone, allopregnanolone). We examined multiple regions of the fetal brain (e. g. prefrontal cortex, nucleus accumbens, hypothalamus, hippocampus). We will also examine neuronal proliferation, microglia and tyrosine hydroxylase immunoreactivity in the fetal brain. Maternal high-sucrose diet increased 11-dehydrocorticosterone (inactive metabolite of corticosterone) and aldosterone in maternal serum and amniotic fluid. Testosterone and androstenedione were significantly higher in the amniotic fluid of male fetuses than female fetuses. Placental steroidogenic enzymes 3β-HSD and CYP19 were not affected by maternal diet or fetal sex; however, 3β-HSD activity was higher in the decidua than in the fetal part of the placenta. Steroid and cytokine data from the placenta, fetal blood and fetal brain are currently being analyzed. Ongoing analyses examine how a maternal high-sucrose diet affects brain development possibly by increasing inflammation.Presentation: No date and time listed

Background: Infertility has a tendency to become potential physical and emotional stressors. The current progressive advances in fertility technology have encouraged more infertile couples to seek for the cause of infertility and take measures to improve their fertility. It is not uncommon that these couples experience emotional distress when they are undergoing fertility treatments. Although it is controversial, some experts argue that this emotional distress affects the success of fertility programs, such as intrauterine insemination and In vitro fertilization programs. Objective: The aim of the study was to investigate the relationship between successful pregnancy and anxiety in patients undergoing intrauterine insemination (IUI) and In vitro fertilization (IVF) programs. Material and method: A total of 39 patients undergoing intrauterine insemination and In vitro fertilization program in Aster clinic of Dr. Hasan Sadikin General Hospital during period of September-November 2016 were measured for their anxiety level using Zung Self-Rating Anxiety Scale. The results were then categorized as not anxious (score of 20-44) and anxious (45-80). Data were then compared against successful pregnancy after each program. Results: Of 39 patients, 23 underwent IUI and 16 underwent IVF. In the IUI group, 10 patients were not anxious (43.48%) and 13 people were anxious (56.52%) while in the IVF group 9 were not anxious (56.25%) and 7 were anxious (43.75%). Three patients (18%) who successfully conceived in the insemination program were those who were not anxious. Meanwhile, in the IVF group 3 patients (18.8%) also got pregnant with 2 of them were not anxious and 1 was anxious. A statistical calculation was performed using Fisher test, resulting in p = 0.068 and p = 1.000 for IUI and IVF, respectively. Conclusion: There is no significant correlation between the success of the fertility program and anxiety in patients undergoing IUI and IVF programs.

To study the correlation between fetal sex and human chorionic gonadotropin (hCG) in maternal blood and amniotic fluid. One hundred and thirty uncomplicated pregnancies, 82 of whom were at sixteen and 48 at thirty-five weeks of gestation. The hCG levels were significantly higher in maternal serum than in amniotic fluid. At 16 weeks there were no sex-related differences in the hCG levels, either in maternal blood or in amniotic fluid. At 35 weeks the hCG levels in maternal blood were significantly higher in pregnancies with female fetuses than in those carrying male fetuses (p<0.004), while in amniotic fluid the hCG levels tended to be slightly higher in the female group than in the male. In pregnancies with female fetuses the hCG levels in maternal blood were significantly higher at 35 than at 16 weeks (p<0.02), while in pregnancies with male fetuses the levels were highest at 16 weeks. For both sexes the hCG levels in amniotic fluid were significantly higher at 16 than at 35 weeks of pregnancy (p<0.001). Whereas a significant correlation between hCG levels in maternal blood and amniotic fluid was seen at 16 weeks of gestation for both sexes (p<0.01 and R value 0.45 for males and 0.41 for females), no correlation was observed at 35 weeks. This study shows a significant correlation between hCG and fetal sex at third trimester of gestation only, possibly caused by a gender factor and a shift in synthesis and/or in metabolism of hCG from the second to the third trimester.

In vitro fertilisation for unexplained subfertility.

Risk of miscarriage in women conceiving after medically assisted reproduction with an ultrasound-verified viable pregnancy at 6-8 weeks' gestation.

Amniotic fluid testosterone was measured by radioimmunoassay in 226 samples taken between the 12th week of pregnancy and birth. The effect of fetal sex on the level of this androgen was determined and the usefulness of the parameter for predicting fetal sex assessed. Testosterone was measured using an antibody raised in rabbits against testosterone-3-carboxymethyloxime-BSA. The antiserum cross-reacted strongly with 5alpha-dihydrotestosterone so that the values reported were refered to as measuring "immunoreactive testosterone" levels. Male fetal values were highest between the 12th and 15th week of pregnancy reaching a maximum mean of 501 +/- 185 pg/ml gradually falling during the remainder of pregnancy to 281 +/- 74 pg/ml, the difference in values between the first half and the end of pregnancy being highly significant (p less than 0.0005). Female fetal values did not vary significantly during the course of gestation, the minimum mean value being 179 +/- 50 and maximum 253 +/- 98 pg/ml. The fetal amniotic fluid sex difference was highly significant during the first half of pregnancy (p less than 0.0005) and at 12-15 weeks a successful prediction of fetal sex was possible, using testosterone, in 39 out of 41 cases. At birth on the other hand there was no difference between male and female fetal fluid levels. The method can also be used to detect disturbances of intrauterine sexual development as exemplified by the case of a Klinefelter syndrome where female testosterone values were found at the time of sexual differentiation. Thus the measurement of unconjugated testosterone in amniotic fluid would seem to offer not only an addition to chromosomal analysis for prediction of fetal gender but also be a valuable tool for detection of disturbances of intrauterine sexual development i.e. when genetic sex and sex-specific hormone production are discrepant.

What serial hCG can tell you, and cannot tell you, about an early pregnancy

Objective: To examine whether prostate-specific antigen (PSA) is present in amniotic fluid, whether the amniotic fluid PSA concentration changes with gestational age, and whether there is an association between amniotic fluid PSA and fetal sex. Methods: The PSA concentration was measured in the amniotic fluid of 48 pregnant women. Thirty-four samples were obtained during routine amniotic fluid analyses performed during gestational weeks 16–18, whereas 14 samples were obtained during cesarean section performed after gestational week 36. Results: PSA was detected in all amniotic fluid samples. The median amniotic fluid PSA was 0.193 ng/ml during gestational weeks 16–18 and 0.39 ng/ml after gestational week 36 (p = 0.1). Furthermore, no significant association was seen between amniotic fluid PSA and fetal sex. The median amniotic fluid PSA level was 0.233 ng/ml for the 21 boys and 0.222 ng/ml for the 27 girls investigated (p = 0.72). Conclusions: These results confirm recent literature reports that PSA may serve as a growth regulator during normal fetal development. However, further studies are necessary to elucidate the exact role of PSA during fetal development.

The cause of infertility is unexplained or poorly explained in 30-40% of couples undergoing standard investigations, and treatment ranges from expectant management to IUI and IVF. The aim of this study was to compare the clinical pregnancy rates and costs of intra-uterine insemination (IUI) and in vitro fertilisation (IVF) in women where the same ovarian stimulation led to the development of two or three mature follicles. A randomised controlled clinical trial compared the efficacy of IUI and IVF in a tertiary fertility centre (ISRCTN28780587). Primary outcome measures were fetal heart positive pregnancy rate and cost per live birth. The selection criteria were age: females 18-42years and males 18-60years, infertility for one year or more, no IVF or IUI for 12months prior to the trial, and no coital, tubal or ovulatory disorders, oligospermia, untreated endometriosis or contraindication for multiple pregnancy. All women (n=102) had the same dose FSH stimulation protocol. Those who developed two or three preovulatory follicles were randomised 3:1 to IUI (n=33) or IVF (n=10). IUI or IVF was performed 36h after hCG administration with single or double embryo transfer on day two. Clinical pregnancy rates (40% vs 12%, P=0.04) and live birth rate (40% vs 6%, P=0.01) were higher for IVF than IUI. The cost per live birth was AU$8735 for IVF compared with $42,487 for IUI. This study provides evidence that IVF is more successful and cost-effective than IUI using the same doses of FSH. Further confirmatory studies are required.

To analyze whether leptin levels of the amniotic fluid elevate during early pregnancy in women destined to develop preeclampsia and to evaluate the relationship between amniotic fluid leptin levels and gestational age, maternal body mass index, and fetal sex. Leptin levels of the amniotic fluid were compared in two groups of women, preeclamptic (n = 20) and normotensive pregnant (n = 40), matched for fetal sex, maternal body mass index at sampling, gravidity and fetal gestational age at sampling. Furthermore, amniotic leptin levels in 400 normotensive pregnant women were analyzed for their correlation with gestational age, maternal body mass index, and fetal sex. Median leptin concentrations were significantly higher (p < 0.001) in the women with preeclampsia (7.3+/-0.7 ng/ml) than in the normotensive pregnant women (4.1 +/- 0.3 ng/ml), independent of fetal sex. The leptin levels in the amniotic fluid decreased with advanced gestational age (r = 0.24, p < 0.001). Amniotic fluid leptin levels in the pregnant women carrying a female fetus (5.6+/-0.3ng/ml) were significantly higher than those carrying a male fetus (4.7+/-0.2 ng/ml) (p = 0.004). Higher amniotic fluid leptin levels were observed in the preeclamptic pregnant women, and they decreased as gestational age advanced. Furthermore, the women with a female fetus were noted to have higher amniotic fluid leptin levels.

Oocyte retrieval versus conversion to intrauterine insemination in patients with poor response to gonadotropin therapy

Artificial intelligence: Friend or foe?