Abstract Purpose: In this study, the Pre-implantation embryonic sex ratio in 125 couples who had three or more female children and underwent pre-implantation genetic diagnosis (PGD) for non-medical reason was included. Besides, we have aimed to find out whether these couples had more chances of getting a girl child once again. Methods: 125 couples who had three or more female offspring and those who underwent PGD for non-medical sex selection (XY) between 2015 and 2019 were included. Nuclear DNA was analyzed by Fluorescent in situ Hybridization (FISH). 2-chromosome (X, Y), 3-chromosome (21, X, Y), 5-chromosome (13, 18, 21, X, Y) probes were used for FISH. The standard protocol was followed for sperm processing and embryo culture for IVF and PGD. Result: Independent sample t-test showed that there is no significant difference between equal and unequal embryonic groups in patients’ age, husbands’ age, sperm count, sperm motility, total male embryos, total female embryos, normal male embryos, and normal female embryos. For patients with positive pregnancy outcome, 84.6% had unequal embryonic ratio while 15.4% had equal embryonic ratio. Similarly, patients those who were treated by short protocol had 85% of unequal embryonic ratio and 15% had equal ratio. Conclusion: Greater variability in the female to male embryos was found in these couples, confirms the fact that couples previously having girl offspring may predominantly not produce embryo of the same sex every time.

The present study was conducted to compare embryo survival rate and treatment outcomes including pregnan-cy, miscarriage and live birth rates of IVF and ICSI vitrified-thawed embryo transfers. Authors suggested to compare treatment outcomes [clinical pregnancy, miscarriage, ectopic pregnancy, and live birth rates] after transfer of IVF versus ICSI vitrified-thawed embryos. Cohort retrospective study in the IVF unit at a university af-filiated Medical Center which included 845 frozen thawed embryo transfer cycles of vitrified embryos from treatment cycles between the years 2013-2019. Out of 845 FET cycles of vitrified thawed embryos, 223 clinical pregnancies were achieved [26.4%]. Of these 64 [28.4%] following IVF, 131 [25.6%] after ICSI and 28 [25.9%] in the sibling oocytes group [P =0.631]. Live birth rate did not differ between the three groups: 23.6%, 22.3% and 21.5% for groups IVF, ICSI and IVF+ICSI, respectively [p=0.899]. Sub analysis by age below 35 and above 35 years old revealed no statistical difference between IVF and ICSI cycles in terms of clinical pregnancy, miscarriage, ectopic pregnancy, and live birth rates between the groups. The present study suggests that the technique of insemina-tion has no adverse effect related to vitrification.

The ability to predict the likelihood of a live birth after single fresh embryo transfer is important for treatment planning and managing patient expectation, particularly in their first in vitro fertilization (IVF) cycle. Cryopreservation of supernumerary embryos is often regarded as an important prognostic variable and a surrogate marker of success for several reasons. While previous large studies have examined the association between the number of oocytes retrieved and cleavage-stage embryos available, and the odds of a live birth following a fresh embryo transfer, the relationship between the number of supernumerary blastocysts cryopreserved following a fresh embryo transfer has not been rigorously studied. We performed a retrospective analysis of data collected between 2006 and 2018 for all first time IVF patients with a fresh autologous day 5 single blastocyst transfer. The relationship between the likelihood of a live birth and number of supernumerary blastocysts cryopreserved was assessed according to patient age group. In patients aged <35 years and 35-39 years old, the likelihood of a live birth increased linearly from 0.33 (95%CI:0.31–0.34) to 0.80 (95%CI:0.74–0.86; P<.0001) and 0.30 (95%CI:0.28–0.32) to 0.82 (95%CI:0.73–0.91;P<.0001) between 1-6 blastocysts cryopreserved and then non-linearly to 0.95 (95% CI 0.92–0.97; P<.0001) and 0.96 (95%CI:0.93–0.99; P<.0001) if 10 or more blastocysts were cryopreserved, respectively. When aged 40 years and above, the likelihood of a live birth increased linearly from 0.26 (95%CI:0.19–0.32) to 0.83 (95%CI:0.68–0.97; P<.0001) between 1-4 blastocysts cryopreserved and then non-linearly to 0.99 (95%CI:0.98–0.99; P<.0001) if 10 or more blastocysts were cryopreserved. The present study demonstrated a non-linear relationship between the number of supernumerary blastocysts cryopreserved and the likelihood of a live birth after single blastocyst transfer in the first autologous fresh IVF/ICSI cycle across different age groups.

Early development in mammals is characterized by the ability of each cell to produce a complete organism plus the extraembryonic, or placental, cells, defined as pluripotency. During subsequent development, pluripotency is lost, and cells begin to differentiate to a particular cell fate. This review summarizes the current knowledge of pluripotency features of bovine embryos cultured in vitro, focusing on the core of pluripotency genes (OCT4, NANOG, SOX2, and CDX2), and main chemical strategies for controlling pluripotent networks during early development. Finally, we will discuss the applicability of manipulating pluripotency during morula to blastocyst transition in cattle species.

No major breakthroughs have entered mainstream clinical fertility practice since egg donation and intracytoplasmic sperm injection decades ago, and oocyte deficits secondary to advanced age continue as the main manifestation of diminished ovarian reserve. In the meantime, several unproven IVF ‘accessories’ have emerged including so-called ovarian rejuvenation which entails placing fresh autologous platelet-rich plasma (PRP) directly into ovarian tissue. Among cellular responses attributed to this intervention are reduced oxidative stress, slowed apoptosis, and improved metabolism. Besides impacting the existing follicle pool, platelet growth factors might also facilitate de novo oocyte recruitment by specified gene upregulation targeting uncommitted ovarian stem cells. Because disordered activity at mechanistic target of rapamycin (mTOR) has been shown to exacerbate or accelerate ovarian aging, PRP-discharged plasma cytokines combined with mTOR suppression by pulsed/cyclic rapamycin represents a novel fusion technique to enhance ovarian function. While beneficial effects have already been observed experimentally in oocytes and embryos with mTOR inhibition alone, this is the first discussion of intraovarian platelet cytokines followed by low-dose, phased rapamycin. For refractory cases, this investigational, tailored approach could amplify or sustain ovarian capacity sufficient to permit retrieval of competent oocytes via distinct but complementary pathways—thus reducing dependency on oocyte donation.

The use of probiotics in reproductive-related dysbiosis is an area of continuous progress due to the growing interest from clinicians and patients suffering recurrent reproductive microbiota disorders. An imbalance in the natural colonization sites related to reproductive health: vaginal, cervicovaginal, endometrial and also pregnancy-related altered microbiota could play decisive role in reproductive outcomes. Oral and vaginal administrations are in continuous discussion regarding the clinical effect pursued, but probiotics as oral supplement therapy is the route administration better studied. To complement and summarise with qualitative and quantitative information of vaginal probiotics clinical studies, the main objective of this work was to retrieve the standardised protocols commonly used and their microbiota modulation capacities. The studies selected were related to treat bacterial vaginosis (BV) as the most commonly disorder, few studies on vulvovaginal candidiasis (VVC) and also administration to stabilise microbiota before in vitro fertilisation (IVF). Probiotic doses administered were similar to oral probiotics protocols, ranging from ≥ 107 CFU/day to 2.5 x 1010 CFU/day, and variable regarding posology duration from 1 day to 12 weeks, being 1 week commonly applied. Moderate modulation was achieved regarding the relative abundance decrease of abnormal microbiota, coinciding with parallel increase in Lactobacillus species.

In vitro fertilization (IVF) has revolutionized the field of assisted reproductive technologies (ART) by offering hope to individuals and couples struggling with infertility. Nevertheless, traditional IVF methods are not exempt from drawbacks, such as suboptimal success rates, intrusive procedures, and ethical considerations. The introduction of micro/nanorobotics in IVF offers a revolutionary chance to tackle these difficulties. This paper presents a short review of the utilization of micro/nanorobotics in IVF, investigating progress in the manipulation of sperm, retrieval of oocytes, culture of embryos, and potential future developments in this fast progressing area. In this discussion, we explore the fundamental concepts, advantages, difficulties, and possible influence of micro/nanorobotics in transforming IVF processes and enhancing patient outcomes.

Abstract: Important variables that affect the quality and functionality of sperm and may affect male reproductive health and fertility include telomere length, mitochondrial content, and the ratio of mitochondrial DNA (mtDNA) to nuclear DNA (nDNA). The purpose of this study was to look at the connections between these three sperm characteristics and how they might affect a woman's ability to conceive. Methods: Data were collected from 82 men who underwent IVF/ICSI at the University Hospital of Ioannina's IVF Unit in the Obstetrics and Gynecology Department. Evaluations included sperm morphology, sperm count, sperm motility, and participant history. To address this, males of reproductive age who were categorized into three body mass index (ΒΜΙ) groups—normal, overweight, and obese—had their sperm samples tested. Results: Our findings indicate a negative correlation between relative telomere length and ΒΜΙ for both the normal and overweight groups. Conversely, a positive correlation was observed for the obese group, indicating a possible relationship between mitochondrial health and telomere maintenance. The results were statistically significant only for the obese group. Furthermore, a negative correlation was observed between telomere length and mitochondrial content in both the normal and overweight groups, indicating that a lower mitochondrial content may be linked to longer telomeres. Nonetheless, there was a positive correlation for the group that was obese. For all three groups, the data did not reach statistical significance. Given that mitochondrial con-centration and telomere length are both markers of cellular integrity and health, these correla-tions may have an impact on the quality of sperm. Furthermore, the relative telomere length of the normal and overweight groups had a negative correlation with the ratio of mtDNA to nDNA, though the obese group showed a positive correlation. The findings were not statistical-ly significant in any of the groups under investigation. According to this, male fertility may be negatively impacted by an imbalance in the copy number of the mitochondrial genome com-pared to the nuclear DNA in sperm. Conclusions: Our research, in essence, emphasizes the interaction among human sperm telo-mere length, mitochondrial concentration, and the mtDNA to nDNA ratio. Gaining knowledge of these correlations could help explain the reasons behind male infertility and open up new treatment options for reproductive health problems. The functional importance of these correla-tions and possible uses in therapeutic contexts require more investigation.

Study question: What are the current trends of research in Human Assisted Reproduction around the world? Summary answer: USA is the leading country, followed by the UK, China, France and Italy. The largest research area is “laboratory techniques”, although other areas such as “public health”, “quality, ethics and law” and “female factor” are gaining ground worldwide. What is known already: Scientific research, especially in health and medical sciences, aims at addressing specific needs that society (and, especially, patients) perceives as pressing. One of the main challenges for policymakers and research funders alike is therefore to align research priorities to societal needs. We can thus think of research agendas in terms of a demand side (societal needs) and a supply side (research outputs). Research output in Human Assisted Reproduction has expanded in the past years, as indicated by the increasing number of scientific publications in indexed journals in this area. Nevertheless, no map of research related to assisted reproduction has been produced so far, hindering the identification of potential areas of improvement and need. Study design, size, duration: 26,000+ scientific publications (articles, letters, and reviews) on Human Assisted Reproduction produced worldwide between 2005 and 2016 were analyzed. These publications were indexed in PubMed or obtained from reference list of indexed publications included in the analysis.Participants/materials, setting, methods: The corpus of publications was obtained by combining the MeSH terms: “Reproductive techniques”, “Reproductive medicine”, “Reproductive health”, “Fertility”, “Infertility”, and “Germ cells”. Then it was analyzed by means of text mining algorithms (Topic Modeling (TM) based on Latent Dirichlet Allocation (LDA)), in order to obtain the main topics of interest. Finally, these categories were analyzed across world regions and time. Main results and the role of chance: We identified 44 main topics, which were further grouped in 11 macro categories, form larger to smaller: “laboratory techniques”, “male factor”, “quality, ethics and law”, “female factor”, “public health and infectious diseases”, “basic research and genetics”, “pregnancy complications and risks”, “general infertility and ART”, “psychosocial aspects”, “cancer”, and “research methodology”. The USA was the leading country in number of publications, followed by the UK, China, France and Italy. Interestingly, research contents in high income countries is fairly homogeneous across macro-categories, and it is dominated by “laboratory techniques” in Western and Southern Europe, and by “quality, ethics and law” in North America, Australia and New Zealand. In middle income countries we observe that research is mainly performed on “male factor”, and noticeably less on “female factor”. Finally, research on “public health and infectious diseases” predominates in low-income countries. Regarding temporal evolution of research, “laboratory techniques” is the most abundant topic on a yearly basis, and relatively constant over time. However, since production in most of the other categories is increasing, the relative contribution of this research category is actually decreasing. Publication is especially increasing in “public health and infectious diseases” (in all world regions, but especially in low income countries), “quality, ethics and law” (high income countries), and “female factor” (middle income countries). Limitations, reasons for caution: Three main factors might limit the robustness of our work: the textual corpus analyzed is based on abstract and titles, the reproducibility of the stochastic algorithms applied, which may produce slightly differing results at each run, and the interpretation of the topics obtained. Wider implications of the findings: This study should prove beneficial in the design of research strategies and policies that foster the alignment between supply (assisted reproduction research) and demand (society). Study funding/competing interest(s): PTQ-14-06718 of the Spanish MINECO Torres Quevedo programme (FAM).

One of the main factors that influence the fertility of cattle in grazing systems in hot tropical climates is heat stress. The objective of this study was to evaluate the effect of season, breed, hormonal and physiological condition on the quantity and quality of cumulus-oocyte complexes (COCs) and embryos produced in vitro, from Romosinuano (RM) and Tropical Milking (TM) donors. Three ovum pick-up and in vitro fertilization (OPU-IVF) were performed, one per season; hot dry (HD; 10, 10), hot humid (HH; 9, 9) and fresh dry (FD; 7, 10) in RM and LT donors. Serum levels of cortisol, insulin and glucose were measured, in addition to heart rate (HR), respiratory rate (RR) and rectal temperature (RT). Effect of season x genotype interaction (p≤0.05) was observed in all COCs variables and only in cleavage embryos (CLI) (p≤0.05). Body weight (BW) affected all COCs variables (p≤0.01), except unviable (UNV) although affected degenerated embryos (DEG) (p≤0.01) and total blastocysts (BLAST) (p≤0.01). Cow age only affected viable COCs (VIAB) (p ≤ 0.05), code one blastocysts (BC1) and BLAST (p≤0.01). Cortisol affected total COCs (COCsT), VIAB and total matured in vitro (TMIV) (p≤0.01), as well as CLI, BC1 (p≤0.01) and BLAST (p≤0.05). Insulin affected COCsT (p≤0.01), UNV (p≤0.05), denuded oocytes (DE) (p≤0.01), BC1 and code two blastocysts (BC2) (p≤0.01). Glucose affected all COCs variables (p≤0.01), except UNV and all embryo variables except BC2. HR affected COCsT, DE, TMIV (p≤0.01), CLI, BLAST and DEG (p≤0.05). RR affected COCsT, UNV, VIAB, CLI (p≤0.05), BC1, BLAST and DEG (p≤0.01). RT only affected DE, VIAB (p≤0.01) and BLAST (p≤0.05). The seasonal climatic year variation of Veracruz and changes in physiological and hormonal variables have diverse effects on the cumulus-oocyte complexes and embryos produced by RM and TM donors.

Oxidative stress negatively affects folliculogenesis and embryo development. However, a reliable and biologically accurate indicator of oxidative stress does not yet exist. On these bases, the aim of this study was to assess -and compare- blood and follicular fluid (FF) redox status in 45 infertile subjects (and 45 age-matched controls) undergoing in vitro fertilization (IVF) and to establish its connection with the outcome of IVF. Blood and FF were obtained at the time of egg retrieval and immediately analyzed. Firstly, ROS production in blood leukocytes and in granulosa cells was assessed. Oxidative stress markers in blood and in granulosa cells resulted significantly (p<0.001) increased in infertile patients compared to controls. Then, a redox index was obtained in plasma and in FF of patients and controls. The main findings emerging from our study in infertile women are: i) blood oxidative stress reflects FF oxidative stress as demonstrated by the significant correlation between blood redox markers and FF redox markers; ii) a significant correlation between oxidative stress parameters and the considered IVF outcomes is present. We suggest the strict monitoring of the redox parameters for the improvement of assisted reproductive techniques success rate and infertility management.

The COVID-19 pandemic had profound negative effects on millions of couples affected by infertility and in need to resort to assisted reproductive technologies. There is no consensus over the optimal way and moment of screening triage-negative asymptomatic patients and staff. We present SARS-CoV-2 antibodies’ (IgM, IgG) seroprevalence in 516 triage-negative patients and 30 fertility care providers. The sampling for SARS-CoV-2 serological assays took place from the lockdown release throughout the second half of 2020 (17.05 - 01.12.2020). It revealed an increased seroprevalence of antibodies that closely followed the local epidemiology of COVID-19, with the highest rate of seropositivity coincident with the peak of the second wave. From 546 triage-negative individuals whose blood samples were assessed for SARS-CoV-2 antibodies, 6% yielded positive results. The overall seroconversion rate was 2.8% for IgG and 5.1% for IgM. In the group with positive IgM, we observed a negative predictive value for IgM of 98.36% (95% CI: 88.79 – 99.78%), which is clinically meaningful. Serological testing of triage-negative patients up to seven days prior to the actual fertility procedure might avoid the more expensive and not more sensitive molecular testing currently being used for patient screening in most fertility units.

Age-related mitochondrial markers may facilitate the prognosis of artificial reproductive technology outcomes. In this report, we present our study concerning the ratio of cf-mtDNA/cf-nDNA, namely the amount of cell-free mitochondrial DNA relative to cell-free nuclear DNA, in the follicular fluid (FF) of women undergoing IVF, aiming to generate a molecular fingerprint of oocyte quality. The values of this ratio were measured and compared among three groups of women (101 in total): (A)31 women with polycystic ovary syndrome (PCOS), (B) 34 women younger than 36 years, and (C) 36 women older than 35 years of age. Real-time quantitative PCR (qPCR) was performed to quantify the ratio by using nuclear- and mitochondrial-specific primers and analyzed for potential correlation with age and pregnancy rate. Our analysis showed that the level of FF-cf-mtDNA was lower in the group of advanced-age women than in the groups of PCOS and non-PCOS women. Moreover, a significant positive correlation between FF-cf-mtDNA and the number of mature (MII) oocytes was observed. Collectively, the data show that the relative ratio of cf- mtDNA to cf-nDNA content in human FF can be an effective predictor for assessing the corresponding oocyte’s age-related performance in IVF.

Vascular endothelial growth factors (VEGFs) are regarded as the key vasculogenic mediators in the human body, of which VEGF-A is considered as one of the important types. It is the major growth factor possessing increasing vascular permeability as well as angiogenesis properties. This article provides intuition into the different effects of VEGF-A on vessels in ovarian hyper stimulation syndrome (OHSS) which is defined as a complication of in-vitro fertilization for infertility treatment, and obesity, a universally health problem. These are two dissimilar pathways outcomes for VEGF-A effects, thereby This article reviews updated information about the mentioned disorders and reveals that how this kind of growth factor yields different results on vasculature by activating distinct intracellular signaling in endothelial cells.

To investigate factors associated with the success of switching to faricimab for type 1 macular neovascularization (MNV) refractory to intravitreal aflibercept (IVA). This retrospective cohort study included patients with type 1 MNV who were switched to faricimab because they were refractory to IVA at two centers. The main outcome was an extension of the treatment interval after 6 months, and factors related to the success or failure of the extension and visual and anatomical outcomes were assessed. The analysis included 43 eyes of 43 patients. Extended dosing intervals of more than 2 weeks were identified in 14 eyes (32.6%). A short dosing interval before the switch, the absence of polypoidal lesions, and a thin central choroidal thickness before the switch were identified as factors involved in successful extension. For patients with refractory type 1 MNV, switching to faricimab is a safe and potential option to extend from existing dosing intervals.