Does advancing male age influence the expression levels and localisation patterns of phospholipase C zeta (PLCζ) in human sperm?

Socio-economic factors have led to an increasing trend for couples to delay parenthood. However, advancing age exerts detrimental effects upon gametes which can have serious consequences upon embryo viability. While such effects are well documented for the oocyte, relatively little is known with regard to the sperm. One fundamental role of sperm is to activate the oocyte at fertilisation, a process initiated by phospholipase C zeta (PLCζ), a sperm-specific protein. While PLCζ deficiency can lead to oocyte activation deficiency and infertility, it is currently unknown whether the expression or function of PLCζ is compromised by advancing male age. Here, we evaluate sperm motility and the proportion of sperm expressing PLCζ in 71 males (22–54 years; 44 fertile controls and 27 infertile patients), along with total levels and localisation patterns of PLCζ within the sperm head. Three different statistical approaches were deployed with male age considered both as a categorical and a continuous factor. While progressive motility was negatively correlated with male age, all three statistical models concurred that no PLCζ–related parameter was associated with male age, suggesting that advancing male age is unlikely to cause problems in terms of the sperm’s fundamental ability to activate an oocyt

Beyond deep learning: automatic morpho-kinetic parameter extraction using spatial attention

International audienceAbstract Study question Can interpretable deep learning Artificial Intelligence automatically learn to extract morphokinetic parameters from time-lapse embryo videos? Summary answer We trained several popular interpretable AI architectures that were successfully able to recognize most embryo developmental stages from day 1 to 5. What is known already Several deep learning AI approaches have been applied to IVF, leading to promising but heterogeneous and perfectible results. However, none of these « black box » models are interpretable, preventing users from having access to the features included in the model and thus from critically analyzing its results. Some interpretable AIs, have been recently developed and could help users visualize the regions of interest in a picture. These interpretable AI have not been adapted and tested to embryo IVF images/videos but could be extremely relevant in this field, providing more insight into AI functioning, and allowing critical analysis. Study design, size, duration In this monocentric study we used 756 randomly selected time lapse videos recorded between 2011 and 2016 (approximately 10% of all the videos generated over this period) and used half of the dataset for the training phase of the AI and the other half for the final evaluation. Participants/materials, setting, methods We used popular deep learning architectures augmented with spatial attention modules that allow the AI to focus on specific areas of the image. We used ResNet-50 as the principal model and combined it with B-CNN, BR-NPA, ProtoPNet, and ProtoTree attention modules. We also applied state-of-the-art generic explanation methods like Grad-CAM, Grad-CAM ++, or Score-CAM. We used standard training procedures and visualized the explanation map produced with a heatmap. Main results and the role of chance We trained several popular interpretable AI architectures to distinguish the 16 development phases of embryos from day 1 to 5 and obtained overall 70 % accuracy, as compared to manual annotation by experts. The inspection of the explanation maps highlighted various patterns of focus on the embryo of one particular model, BR-NPA. At the fertilization stage, this interpretable AI focused on the pro-nuclei, whereas from phase t5 to t9+ the model focused on the separate cells. At the blastocyst stage, the model was able to separate the ICM from the rest of the embryo (trophectoderm). This shows that interpretable AI can indeed be applied to time-lapse videos and focus on relevant regions of the embryo at all developmental stages, producing interpretable results. The role of chance was minimal in this study as we obtained consistent results across various AI architectures, the dataset was selected randomly among all available videos and the dataset was large enough to ensure minimal selection bias. Limitations, reasons for caution Our study suffers from the inherent limitations of monocentric studies. Indeed, lab procedures and characteristics of the population can vary from one setting to another and question the generalizability of our findings. However, deep learning is versatile and similar results can probably be obtained on data from other centers. Wider implications of the findings We postulate that this original approach will help improve the acceptability and the trust of embryologists in AI-based software, which will improve data handling and consistency, ultimately benefiting infertile patients with improved clinical success rates. Trial registration number Not applicabl

Semen cultures analysis: retrospective study during a 6-year period and interest in the management of infertility

International audienceIn the past decade, the number of couples consulting an assisted reproductive techniques (ART) center for infertility has increased in most European countries. In France, sperm bacterial examination must be performed every 6 months in couples undergoing in vitro fertilization (IVF) cycles, according to 2010 French ART Guidelines. The aim of this study was to analyze the results from such samples at Nantes University Hospital and to assess their potential value for infertile couples undergoing ART cycles. Between 2003 and 2008, semen cultures performed were analyzed according to bacterial enumeration, type and number of bacterial species, and their antibiotic resistance profile. A total of 14,119 semen cultures were performed, showing an annual increase of 45% from the start to the end of the study. The proportion of positive semen cultures was stable throughout the study period (40 to 45%). Many bacterial species were considered as contaminants (coagulase-negative staphylococci, alpha-hemolytic streptococci). For pathogen agents (in most cases, Enterobacteriaceae), the antibiotic resistance profile revealed mostly a susceptible phenotype. Finally, every positive bacterial result had direct consequences on the IVF cycle management, with subsequent reinforced advice on the hygiene procedure before sample collection and/or antibiotics prescription

Prospective evaluation of serum anti-Müllerian hormone dynamics in 250 women of reproductive age treated with chemotherapy for breast cancer

International audienceAim: Women of reproductive age with breast cancer generally receive gonadotoxic chemotherapy. Fertility issues are of great concern for them. However, little is known on ovarian damage during chemotherapy and its evolution during long-term follow-up. The Anti-Mü llerian hormone; Ovarian reserve; Amenorrhea aim of this study was to provide a detailed description of serum anti-Mü llerian hormone (AMH) evolution during chemotherapy and 24-month follow-up.Methods: This prospective cohort study was conducted in 250 patients, aged 18e39 years, diagnosed with breast cancer and treated with adjuvant/neoadjuvant chemotherapy. Each patient underwent blood AMH measurement at each chemotherapy cycle, and at 6, 12 and 24 months after chemotherapy. Menses occurrence was also recorded.Results: Mean basal AMH level was 4.19 AE 4.84 ng/mL, and was negatively correlated with age. Serum AMH level rapidly decreased in all patients after each chemotherapy cycle to undetectable levels in most of them, and slowly increased in 45% of the patients during the 24-month follow-up. AMH decrease was significantly associated with age and basal AMH level, but not with cyclophosphamide dose and tamoxifen use. The prevalence of chemotherapy-related amenorrhoea was 92.4% at the end of chemotherapy; women with amenorrhoea being significantly older and having lower basal AMH than women who resumed menses.Conclusions: Our study confirms rapid and deep ovarian reserve alteration in young women receiving chemotherapy for breast cancer, and shows moderate AMH recovery in some patients. Although AMH cannot alone predict fertility potential, these new data emphasise the need for post-treatment ovarian insufficiency follow-up, strongly support the use of fertility preservation strategies and may provide new tools for improved counselling

Parallel derivation of isogenic human primed and naive induced pluripotent stem cells

Induced pluripotent stem cells (iPSCs) have considerably impacted human developmental biology and regenerative medicine, notably because they circumvent the use of cells from embryonic origin and offer the potential to generate patient-specific pluripotent stem cells. However, conventional reprogramming protocols produce developmentally advanced, or primed, human iPSCs (hiPSCs), restricting their use to postimplantation human development modeling. Hence, there is a need for hiPSCs resembling preimplantation naive epiblast. Here, we develop a method to generate naive hiPSCs directly from somatic cells, using OKMS overexpression and specific culture conditions, further enabling parallel generation of their isogenic primed counterparts. We benchmark naive hiPSCs against human preimplantation epiblast and reveal a remarkable concordance in their transcriptome, dependency on mitochondrial respiration and X chromosome status. Collectively, our results are essential for the understanding of pluripotency regulation throughout preimplantation development and generate new opportunities for disease modeling and regenerative medicine.status: publishe