Abstract Intracytoplasmic sperm injection (ICSI) requires selecting a suitable spermatozoon from an adequate pool of candidates. The order of density gradient centrifugation (DGC) and magnetic-activated cell sorting (MACS) may influence the characteristics of the recovered sperm population, but its clinical significance remains uncertain. This commentary hypothesizes that DGC followed by MACS (DGC-MACS) may offer practical advantages over the reverse sequence (MACS-DGC) when sperm availability is sufficient. DGC enriches motile, apparently morphologically normal spermatozoa while reducing debris and other cells; subsequent MACS enriches the population for spermatozoa without detectable externalized phosphatidylserine (PS). However, PS negativity does not establish complete functional competence, and DGC has not been validated as a measure of intrinsic sperm stress resistance. We therefore discuss membrane sialidase activity and mitochondrial membrane potential (MMP) as illustrative, exploratory biomarkers of complementary functional domains. Their assessment is proposed in parallel research aliquots, not for real-time selection of individual spermatozoa for ICSI. We also introduce a provisional PS-negative Enrichment Ratio (PSNER) based on the relative proportions of PS-negative spermatozoa before and after DGC-MACS, while acknowledging its dependence on processing conditions, baseline semen quality, and selective recovery. Direct randomized comparisons of DGC-MACS and MACS-DGC, with live birth as the primary outcome, alongside validation of the proposed biomarkers and index, are needed before clinical implementation. Lay Summary In some fertility treatments, a specialist helps fertilization by injecting a single sperm directly into an egg. Before this step, laboratories use different methods to prepare sperm samples and increase the chances of selecting a suitable sperm. Two methods can be used together. One separates sperm from unwanted cells and other material, while the other helps remove sperm showing certain changes on their surface. However, it is not yet clear whether the order in which these methods are performed affects the chances of having a baby. In this paper, we suggest that using the first method before the second may offer practical advantages when enough sperm are available. We also propose studying additional signs of sperm health and developing a new way to describe changes in sperm samples during preparation. These ideas have not yet been proven to improve fertility treatment outcomes. Further studies are needed to compare the two methods in different orders and determine whether either approach increases the chances of having a baby.
Abstract Current treatments of endometriosis, a complex and polygenic disease characterized by the growth of endometrial-like tissue outside the uterus, are ineffective in many patients and prone of side effects. Systems pharmacology development offered new avenues to study complex diseases at a systematic level for prioritizing more effective therapies. Here, we apply this strategy for gene disease and drug prioritization by using six gene expression datasets from Gene Expression Omnibus of healthy, eutopic and ectopic endometrium. A similar molecular basis of endometriosis disease and infertility-associated genes was highlighted since they share a gene overlap higher than randomly expected (210 genes through direct protein-protein interactions from 553 infertility-related and 2,931 endometriosis disease-related genes, p-value=1.9 × 10-11) and these genes were closer physically interacting in the network than random expectation (z-score=-1.75, p-value=0.03). Sixteen potential candidate drugs were identified, which had superior molecular efficacy treating the disease as they target more endometriosis-related genes (p-value=0.01) than randomly expected compared with current treatments (p-value=0.97). Zinc, copper and resveratrol, which were found among the clinically safest candidate compounds, suppressed endometriosis-related phenotypes in endometriosis cells (cell proliferation, migration, and overexpression of endometriosis-related biomarkers). This systems pharmacology approach, together with the in vitro validation in the endometriosis cell model, allowed to identify zinc and copper, nutraceutical compounds, as potential endometriosis treatments addressing the causes of the disease rather than symptoms like current therapies. Additionally, since both compounds are already approved for other indications, they can be proposed for repurposing for endometriosis therapies, reducing time and costs of clinical trials. Lay Summary Current treatments for endometriosis address symptoms rather than causes, are not always effective, and produce side effects. We used a computer-based approach to compare gene expression in healthy endometrial tissue and in endometrial tissue affected by endometriosis. This helped us identify which genes may be involved in the disease and which existing drugs might be worth testing as possible treatments. We found that endometriosis disease and infertility-associated genes share similarities, suggesting that both conditions may have similar underlying causes. We looked for existing drugs and compounds that might target those underlying causes more effectively than current treatments. We found that zinc and copper might be promising treatments for endometriosis because they may target the disease’s underlying causes. Since these drugs are already used for other purposes, repurposing them may reduce the time and cost of clinical trials.
LAY SUMMARY When embryos are frozen and thawed for fertility treatment, they sometimes collapse, but does this mean they are no longer viable? This study followed 438 frozen embryo transfer cycles to understand whether collapsed embryos can recover, and whether giving them more time in the incubator before transfer makes a difference. The key finding is that what matters most is how an embryo looks at the time of transfer, not immediately after thawing. Embryos that re-expanded before being transferred had similar success rates to those that never collapsed. Only embryos that remained collapsed at transfer had significantly poorer outcomes. Importantly, for every additional hour spent in the incubator, a collapsed embryo's odds of recovering improved substantially. This means that rather than discarding a collapsed embryo, clinicians may simply need to wait, giving it time to recover could meaningfully improve the chances of a successful pregnancy. Collapsed embryos that are given longer to recover in the incubator before transfer may have the same chance of achieving a pregnancy as embryos that never collapsed.
