Proteomic profile of maternal-aged blastocoel fluid suggests a novel role for ubiquitin system in blastocyst quality

Purpose: The etiology of maternal aging, a common cause of female factor infertility and a rate-limiting step in vitro fertilization (IVF) success, remains still unclear. Proteomic changes responsible for the impaired successful pregnancy outcome after IVF with aged blastocysts have not been yet evaluated. The objective of this prospective study was to employ proteomic techniques and bioinformatic tools to enlight differences at the protein level in blastocoel fluid of aged and younger woman. Methods: Protein composition of human blastocoel fluid isolated by micromanipulation from 46 blastocysts of women aged <37 years (group A) and 29 of women aged 6537 years (group B) have been identified by a shotgun proteomic approach based on high-resolution nano-liquid chromatography electrospray-ionization-tandem mass spectrometry (nLC-ESI-MS/MS) using label free for the relative quantification of their expression levels. Results: The proteomic analysis leads to the identification and quantification of 148 proteins; 132 and 116 proteins were identified in groups A and B, respectively. Interestingly, the identified proteins are mainly involved in processes aimed at fine tuning embryo implantation and development. Among the 100 proteins commonly expressed in both groups, 17 proteins are upregulated and 44 downregulated in group B compared to group A. Overall, the analysis identified 33 proteins, which were increased or present only in B while 76 were decreased in B or present only in A. Conclusions: Data revealed that maternal aging mainly affects blastocyst survival and implantation through unbalancing the equilibrium of the ubiquitin system known to play a crucial role in fine-tuning several aspects required to ensure successful pregnancy outcome

Implantation failure in assisted reproduction technology and a critical approach to treatment

In this article, we review the literature and our personal experience regarding the many factors that appear to influence implantation rate. Oocyte quality, as determined by patient age and aneuploidies, probably plays a major role in RIF. However, a panoply of other factors have been brought under investigation, quite often with contradictory results and additional intriguing questions to be studied. Infections of the vagina, cervix, and endometrium, the role of mucus aspiration and washing of the cervix on transfer, the role of catheter guidance for a correct transfer and potion of embryos, the effect of mock transfer, and the role of hysteroscopy and its timing before transfer procedures are analyzed both as a review of the literature and as opinions and data from our experience. Many of these factors are interlaced and from the apparently simple issue of trauma, to infections and immune modulation of hatching and implantation, a biological continuum can easily be identified. The impact of abnormalities of the immune system and of homeostasis abnormalities is also covered in a brief overview of reported works and our experience. These latter areas probably constitute the common biological background of all other external factors that, however, the skilled must equip themselves for improving implantation success

Oocyte slow freezing and oocyte vitrification: data from the italian ART register 2007-2011

analyze slow freezing/rapid thawing or vitrification in one of the largest worldwide experience of oocyte cryopreservation

Five years (2004-2009) of a restrictive law-regulating ART in Italy significantly reduced delivery rate: analysis of 10 706 cycles

Study question: Law 40/2004 enacted by the Italian Parliament established a long list of restrictions for ART procedures. We investigated the differences in the delivery rate of all ART cycles performed before and after the introduction of the law. Summary answer: This large and extensive comparative analysis of ART outcomes prior and after the introduction of the Law 40 revealed a significant reduction in pregnancy and delivery rates per cycle, independent of age or other clinical variables, once the law went into effect. What is known and what this paper adds: Several studies have been published on the effect of Law 40/2004 on ART outcomes, some authors demonstrating a negative impact of the law also in relation to specific aetiologies of infertility, other authors showing opposite conclusions. Our study, the first based on the analysis of a so large number of cycles, clearly demonstrated a negative effect of Law 40 restrictions on ART procedures, as an independent factor responsible for the reduced delivery rate of IVF cycles. Design: Retrospective clinical study Partecipants and setting: 10,706 cycles were analysed, 3,808 performed before and 6,898 cycles after the application of the Law. An intention to treat statistical analysis was performed to detect pregnancy and delivery rates (pregnancies ≥ 24 weeks) per started cycle. A p value <0.05 was considered statistically significant. We analysed different outcomes: differences in fertilization, pregnancy, and delivery rate, multiple pregnancies and miscarriage rates between the two time periods. Main results and the role of chance: The delivery rate for started cycle was 20% before and 16.0% after the introduction of the Law representing a 25% reduction (p<0.001). The multivariate analysis, corrected by age, basal FSH level, number of retrieved oocytes and total progressive motile count, showed a 16% lower delivery rate (OR 0.84 CI 0.75-0.94) and confirmed the effect of the Law as an independent factor responsible for the reduced chances of a successful outcome