P.E. Levi Setti

Power spectral analysis of the heart rate of the human fetus at 26 and 36 weeks of gestation

The fetal electrocardiogram (ECG) was recorded from the maternal abdomen together with sonographic detection of fetal states in four pregnant patients at 26 and 36 weeks of gestation. Computerised algorithms obtained a good recognition of maternal ECG and fetal ECG. The corresponding power spectral density (PSD) was estimated during epochs of quiet or breathing movements for 256 consecutive heart beats. At 36 weeks of gestation the PSD analysis showed heart rate variability components similar to an adult heart. A low frequency component was observed together with a high frequency component in the range of fetal breathing movements. This high frequency component (0.6 - 0.9 Hz) disappeared from the PSD analysis during epochs of apnea. To our knowledge this is the first demonstration of such a mechanism in the human fetus in utero. At 26 weeks no respiratory component was observed in the PSD analysis. The maturation of control mechanisms could be responsible for this difference.

Prenatal Diagnosis after ART Success: The Role of Early Combined Screening Tests in Counselling Pregnant Patients

First-trimester Down syndrome screening may cause a higher false positive rate in pregnant patients who have undergone ART (assisted reproductive technologies). The aim of this paper is to contribute to this analysis with the second largest series of combined biophysical and biochemical tests in the first trimester of pregnancy after ART. One hundred and forty-two singleton successful ART pregnancies were selected for this study: 50 pregnancies induced by using in-vitro fertilization (IVF), and 92 using intracytoplasmic sperm injection (ICSI). Each patient was matched with three naturally conceived pregnancies based on maternal age and gestational age. Free beta-HCG and PAPP-A were measured on dried blood spots and converted to MoMs. Nuchal translucency (NT) was measured by certified operators. Mean maternal age was 33 +/- 4. NT, free beta-HCG and PAPP-A values of the control cases were not significantly different from local standards evaluated on 3043 cases. NT between ART pregnancies and matched controls was not significantly different. PAPP-A was reduced but not significantly lower in ART pregnancies. Free beta-HCG was the only analyte that resulted in significantly higher values in ART pregnancies (1.12 MoM) versus controls (0.99 MoM). No significant differences were found for biochemical values observed between ICSI and IVF patients. The screen positive rates observed in ART and control pregnancies were 5.5 per cent and 4.6 per cent respectively. NT measurements were not affected by ART pregnancies. Our results (non-significant lower values of PAPP-A and significantly higher free beta-HCG values) were consistent with other reported series. The increase in the screen positive rate determined by these biological variations was not greater than 0.9 per cent. This higher false positive rate has a negligible impact on counselling ART patients. The algorithm used to calculate the relative risk after the combined tests should not be changed until the detection rate of trisomies in ART pregnancies is not fully disclosed by larger series.

The Impact of Embryo Transfer on Implantation—A Review

Embryo transfer has received little clinical attention and has been, until recently, the most inefficient step in in-vitro fertilization (IVF). In this article, the authors review the literature and their personal experience regarding the process of intrauterine transfer of embryos, which remains the object of much discussion. Factors which appear to influence implantation rates are: contamination of the catheter tip with cervical bacteria, stimulation of uterine contractions during the procedure, the type of catheter, ultrasound guidance during the transfer, and the position of the embryos in the uterine cavity. Easy and atraumatic transfer is essential for successful implantation and the embryos need to be placed in the middle of the cavity, away from the fundus. Knowing, beforehand, the position and length of the uterus can provide better results and may reduce the rate of ectopic pregnancies. Evidence from randomized studies has supported this claim. Despite the number of available studies controlling certain variables, most authors, even using the same catheter, ultrasound guidance and/or a trial transfer use different protocols or similar instruments in different ways. Standardization of the transcervical intrauterine transfer of embryos in a large randomized study is needed before definitive conclusions can be drawn. The goal of improved implantation and pregnancy rates deserve these efforts.

Is letrozole needed for controlled ovarian stimulation in patients with estrogen receptor-positive breast cancer?

Abstract Objective: To assess the advantages and disadvantages of using Letrozole for controlled ovarian stimulation (COH) in young patients with estrogen receptor-positive (ER+) breast cancer, wishing to cryopreserve oocytes. Design: Retrospective cohort analysis. Setting: Sixteen Italian units for reproductive medicine and in vitro fertilization. Methods: Data of 50 ER+ breast cancer patients undergoing COH to cryopreserve oocytes before gonadotoxic chemotherapy with a Letrozole plus gonadotropins (Le+Gn) protocol were compared with those of 25 young women with ER– breast cancer, submitted to COH using a protocol with gonadotropins alone (Gn-only). Results: The Le+Gn protocol implied a significantly lower total Gn consumption and allowed to maintain significantly lower circulating E2 levels at all checkpoints throughout stimulation (peak E2 value 446 ± 357 versus 1553 ± 908 pg/ml, respectively; p = 0.001). On the other side, the Le+Gn protocol allowed a significantly lower yield of oocytes available for cryostorage (6.6 ± 3.5 versus 8 ± 5, respectively; p = 0.038). Conclusions: In breast cancer patients, the association of Letrozole to Gn significantly reduces the number of oocytes available for cryostorage in comparison with the use of Gn alone. On the other side, it is associated with significantly lower E2 levels during the whole stimulation cycle, a safety issue that has been traditionally considered advantageous in case of ER+ cancers.

Cancer Risk in Male Factor-infertility

Severe forms of male-factor infertility are associated with an increased risk of testicular cancer and scrotal ultrasonography is widely used for diagnosis. In this study, 2172 male members of infertile couples referred to our Reproductive Medicine Unit were submitted to scrotal ultrasonography and 835 selected patients had been followed during a 2-year period. Eight out of nine neoplastic nodules found at the initial examination were unpalpable and discovered by ultrasonography. Ten tumoral lesions were found in 370 testicular biopsies performed for diagnostic purposes or to extract spermatozoa; and eight additional neoplastic lesions were discovered during the 2-year follow-up of 835 patients. The cumulative rate of neoplastic disease was 3.2%. Thirteen cases (1.5%) were malignant (12 germ cell tumours and one non-Hodgkin lymphoma of testicular origin); the remaining 14 were benign forms (Leydig cell tumours and hyperplasias, Sertoli cell nodules, adenomatoid tumours). Testicular volume (cut-off: 12ml) resulted weakly correlated with germ cell cancer (p=n.s., odds ratio 2.01) while low total sperm count (<40x10(6)) (p=0.002, odds ratio 8.4), previous cryptorchidism (p=0.04, odds ratio 7.5) and hypergonadotrophic hypogonadism (p=0.04, odds ratio 7.9) were associated with an increased risk. But a stronger correlation with germ cell cancer was found in the patients with some utrasonographic anomalies, i.e. testicular microlithiasis (p=0.0015, odds ratio 37.1) or larger calcifications not fitting the description of testicular microlithiasis (p<0.0001, odds ratio 69.5). Our findings indicate that scrotum ultrasonography should always be advised in subfertile men with <40x10(6) spermatozoa/ejaculate or hypergonadotrophic hypogonadism or previous cryptorchidism, and that particular care should be taken in the presence of testicular microlithiasis or testicular calcifications. These men should be aware of the existence of higher risk of testicular cancer and trained in testicular self-examination.

Preliminary evidence for high anti-PLAC1 antibody levels in infertile patients with repeated unexplained implantation failure

OBJECTIVE Placenta-specific1 (PLAC1) is a trophoblast-specific gene encoding for a protein that is highly expressed in human placenta, on the surface of the syncytiotrophoblast. PLAC1 was found to elicit spontaneous antibody responses in cancer patients. We aimed to determine the levels of anti-PLAC1 antibodies in infertile women with a history of unexplained repeated implantation failure after IVF cycles as compared to fertile women. STUDY DESIGN An observational case-control clinical study. MAIN OUTCOME MEASURE(S) Two groups of patients were analysed in two different experimental settings: 21 infertile women and 81 control patients were enrolled in the first group, 16 infertile women and 67 fertile controls in the second group. Anti-PLAC1 antibody levels and ranking were analysed by ELISA test. RESULTS In both groups of infertile patients enrolled, optical densities (OD) from ELISA test ranked significantly higher than those of controls (0.27 ± 0.2 vs. 0.13 ± 0.1 respectively; p = 0.0009 in the first group), (0.62 ± 0.38 vs. 0.39 ± 0.35 respectively; p = 0.0044 in the second experiment). In the first group about one case in four (29%) had OD levels above the 95th percentile (0.337) for healthy controls (p = 0.005). In the second experiment 4 out of 16 cases (25%) had OD levels above the 95th percentile (0.878) for healthy controls (p = 0.023). CONCLUSIONS Anti-PLAC1 antibodies could represent a biomarker associated with infertility and with high probability of repeated implantation failure after ovarian stimulation and IVF-ET, greatly improving the diagnostic work up of infertile couples.

Gynecological fertility-sparing surgery.

The implementation of early detection protocols and advanced treatment strategies has significantly improved survival outcomes for gynecologic cancer patients. The improvement of oncological outcomes has led to an increased attention toward Quality of Life issues, including the childbearing potential for young women. Traditionally the surgical treatment of cervical, endometrial and ovarian cancers involves the removal of the uterus and adnexa, irrespective of the impact on fertility and parenthood and regardless of patient desires. For young women affected by gynecological malignancies at an apparently early stage, fertility-sparing procedures could be offered. The aim of our review is to going through the available evidence in the Literature and to evaluate the current state of art regarding fertility-sparing procedures for women with gynecological malignancies in terms of oncological and fertility outcomes.

Preface: A window into the reproductive era research.

I am proud to welcome you to this special edition of Placenta which contains the proceedings of the third symposium on reproductive medicine to be held in our Institution, organized by Serono Symposia International Foundation, with myself as scientific organizer. In the field of reproductive health, two needs are emerging that apparently conflict: research is providing continuous innovations in daily procedures, but the demand for infertility treatment is increasing all over the world. The key point of conflict between these two aspects is cost, because innovation requires investment, at least in the early phases, and thewidening of ART to geographical areas with lower average incomes requires careful evaluation of the cost/benefit balance. The aim of this symposium, which follows the previous two successfully organized in 2003 and 2008 together with Serono Symposia International Foundation, is to provide participants with overviews of the most innovative procedures that will be introduced into ART in the near future, and with the background needed to offer the most up-to-date infertility treatment, according to different local conditions. The conference is organized into four sessionswith two key note lectures. The first session ‘Innovative surgical procedures in ART’ is scheduled to show how innovation in surgical techniques and better survival rates can impact the patient’s quality of life and provide new opportunities to preserve ovarian function and enhance the future utilization rate of frozen tissue, even in oncological patients who previously had a poor prognosis. Cryopreservation and transplantation techniques of both full ovary and ovarian fragments will be discussed in the context of growing experience in endoscopy and robotics. The first key note lecture ’Benefit and risk of application of European tissue management regulation in ART’ will fully discuss the effects of tissue management regulation in the context of safety and a cost-benefit analysis. In 2004 the European Community included ART in cell factory and banking regulation and these new rules for approval and application in all countries created a revolution in infertility treatment management. The second session ‘Innovation in ART laboratories’will focus on innovative air and environment quality control, sequential media selection, incubation techniques, and sperm selection that will impact on embryo quality and implantation potential. The third session ‘Explore the chaos: the early stages of embryo development’ will discuss recent developments in