Molecular changes, caused by various environmental factors, affect the quality and developmental potential of oocytes. Oxidative stress (OS) is a major factor involved in various gynecologic disorders and/or in aging. Recent studies suggest that elevated reactive oxygen species (ROS) hamper oocyte quality and future embryonic development. Pigment epithelium‐derived factor (PEDF) is a pleiotropic protein, known for its antiangiogenic, anti‐inflammatory, and antioxidative properties. Our previous findings demonstrate the antioxidative role of rPEDF in maintaining granulosa cell viability. In the current study, we examined the ability of PEDF to negate the adverse impact of OS on oocytes. Maturation rate of oocytes exposed to OS was significantly lower than that of control oocytes. The number of mtDNA copies in OS‐exposed oocytes was significantly higher than in control oocytes (>3 times), whereas ATP concentration was significantly lower. Oocytes exposed to OS demonstrated impaired chromosome arrangement at the metaphase plate. PEDF significantly improved maturation rate of untreated OS‐exposed oocytes. Moreover, mtDNA copy number, ATP concentration, and chromosome arrangement at the metaphase plate in rPEDF‐treated OS‐exposed oocytes were restored to the level of control oocytes. Our findings demonstrate that OS hampers the ability of oocytes to undergo proper in vitro maturation. The energetic balance of OS‐exposed oocyte is characterized by excessive mtDNA replication and reduced ATP concentration; it hampers the ability of oocytes to perform high fidelity chromosome segregation. PEDF alleviates this damage, improves the rate of oocyte maturation, and preserves mtDNA level and ATP content, thus enabling oocytes to form proper metaphase plate and improve oocyte competence.

Breast cancer is diagnosed in ~0.3% of pregnant women. Studies that have addressed gestational and neonatal outcomes of chemotherapy during pregnancy have demonstrated increased gestational complications including preeclampsia and intrauterine growth retardation. We hypothesized that anthracycline-induced gestational complications could be derived from direct toxicity on the placenta vasculature. Pregnant ICR mice (day E12.5) were treated with doxorubicin (DXR; 8 mg/kg) or saline, while their umbilical cord blood flow was imaged by pulse-wave (PW) Doppler. Mice were euthanized on day E18.5, and their embryos and placentae were collected for further analysis. Unlike control mice, the DXR-treated mice presented an acute change in the umbilical cord’s blood flow parameters (velocity time integral and heart rate interval), reduced embryos’ weight, reduced placenta efficiency, and modulation in vascular-related pathways of treated placenta proteomics. Apoptosis and proliferation were also enhanced, as demonstrated by TUNEL and proliferating cell nuclear antigen (PCNA) analysis. We further examined the placentae of patients treated with epirubicin (EPI), who had been diagnosed with breast cancer during pregnancy (weeks 27–35). The immunohistochemistry of the EPI-treated human placentae showed enhanced proliferation and apoptosis as compared with matched chemo-naïve placentae, as well as reduced neovascularization (CD34). Our findings suggest that anthracycline-induced vascular insult promotes placental toxicity, and could point to potential agents designated to offset the damage and to reduce gestational complications in pregnant cancer patients.

Polycystic ovary syndrome (PCOS), one of the most common female endocrine disorder, is a prevalent cause of infertility. Hyperandrogenism is a key feature in PCOS and is correlated with increased expression of VEGF and cytokines in the ovaries. We have previously shown that pigment epithelium-derived factor (PEDF), an endogenous protein, presents potent anti-angiogenic and anti-inflammatory activities in the ovary and negates the effects of cytokines and VEGF. Additionally, PEDF plays a role in both pathophysiology and treatment of ovarian-hyperstimulation syndrome (OHSS), frequently seen in PCOS patients. We established hyperandrogenic-PCOS models, both in vivo, using mice exposed prenatally to dihydrotestosterone (DHT) and, in vitro, using human primary granulosa cells (hpGCs) and human granulosa cell line (KGN). In PCOS-induced mice, the mRNA levels of I l-6, V egf and Amh were higher than those of control; yet, treatment with rPEDF decreased these levels. Moreover, treating OHSS-induced PCOS-mice with rPEDF alleviated all OHSS symptoms. Stimulation of hpGCs with DHT resulted in downregulation of PEDF mRNA expression, concomitantly with a significant increase in IL-6 and IL-8 mRNAs expression. However, co-stimulation of DHT with rPEDF attenuated the increase in cytokines expression. The anti-inflammatory effect of PEDF was found to be mediated via PPARγ pathway. Our findings suggest that rPEDF treatment may normalize the ovarian angiogenic-inflammatory imbalance, induced by PCOS-associated hyperandrogenism. Moreover, the therapeutic potency of PEDF in preventing OHSS symptomes offers a rationale for using PEDF as novel physiological treatment for PCOS sequels.

BackgroundTo test whether poor quality day-3 embryos can undergo successful blastulation and implantation.MethodsA prospective cohort study was conducted. Whether or not a good quality embryo was transferred on day-3, poor quality (rejected) embryos were further cultured and followed. The clinical outcome of each embryo was assessed.ResultsA total of 694 rejected embryos (from 205 patients) were included, with a blastulation rate of 21.2% (147 embryos) compared to 64.2% general blastulation rate reported by our laboratory (P < 0.01). In a multivariate logistic regression model, only their grade on day-3 significantly affected blastulation (P = 0.01). A total of 97 embryos attained eligibility for fresh transfer or cryopreservation, only 6 of which resulted from a day-3 embryo scored < 2. Of these, 52 were transferred, resulting in 21 pregnancies (16 clinical and 5 chemical). In summary, 694 cultured embryos yielded 16 clinical pregnancies; a 2.3% clinical pregnancy rate.ConclusionsLow score day-3 embryos can result in successful blastulation and clinical pregnancies. However, the normal blastulation rate is poor.

No abstract

Study question What is the clinical outcome of instant irregular cleavage (IDC) from zygote to three cells? Summary answer IDC embryos, did not develop to blastocysts and did not acieve pregnancy; FC embryos that reached blastocyst stage exerted similar pregnancy rates as control embryos What is known already Evaluation of the zygote morphology is one of the earliest embryonic stages investigated. Several zygote grading systems were proposed based on pronuclei size and NPB (nucleolus precursor bodies) distribution, thus is an effective indicator of the embryo’s potential to result in a life birth. The first cleavage of a zygote is a highly coordinated event. Disruptive timing of the first cleavage, named direct cleavage (DC, less than 5 hours), is associated with formation of poor embryo quality. Early identification of pathological embryos is challenging and impacts the timing of embryo fate decision in ART. Study design, size, duration A total of 1978 fresh IVF cycles from single unit were analyzed and a total of 4012 embryos were studied. Only 2pn embryos, cultured exclusively in time-lapse system for 5 days were included. The morphokinetics of embryos and their outcome were recorded, from z-score analysis through day 5. We discriminated between IDC, FC and normal cleavage pattern of embryos and compared clinical outcome of various morphokinetic parameters. We also evaluated possible early predictors for IDC. Participants/materials, setting, methods We generated three study groups according to embryo cleavage pattern: (I) Control, normal cleavage (n = 551); (II) FC, zygote to three cells within 5 hours (n = 1587); (III) IDC, instant cleavage from zygote to three cells (n = 922). The association between z-score of 138 IDC embryos and their sibling random controls was thoroughly investigated. All time lapse annotations were performed by senior embryologists. Statistical analysis was performed by SPSS software. Main results and the role of chance IDC embryos were mainly arrested at day 3 and the number of usable embryos (reached blastocyst stage and were suitable for embryo transfer or cryopreservation) was negligible; 4/922 (0.4%). In comparison, the amount of usable FC embryos was 108/1587 (6.6%) and control embryos usage reached 180/551 (32.7%). While the pregnancy rate of control and FC embryos, which reached embryo transfer were similar (40.35% and 42.55%, respectively); transfer of IDC embryos did not result in pregnancy. Additionally, the timetable as measured by time of PN fading and time from fading to first cleavage, differed significantly between the three groups. Therefore, we performed a thorough analysis of zygote morphology data of IDC embryos compared to control sibling embryos in search of early possible markers for these findings. We have not detected significant differences in z-score analysis, reflected by number and size of nucleoli, as well as pronuclear symmetry. Limitations, reasons for caution IDC embryos number used for z-score analysis was limited since many didn’t meet the inclusion criteria (lack of overlapping of the pronuclei, presence of random of control embryo and good quality image). Wider implications of the findings The decision regarding the fate of IDC and FC embryos should include the pattern of first cycle cleavage. Culture IDC and FC embryos for 5 days up to the blastocyst will spare transfer of embryos that are fated to arrest even when their morphological grade on day 3 is acceptable. Trial registration number 0043-22-MMC

Reproductive aging is characterized by a decline in ovarian function and in oocytes’ quantity and quality. Pigment epithelium-derived factor (PEDF), a pivotal player in ovarian angiogenic and oxidative balance, was evaluated for its involvement in reproductive aging. Our work examines the initial stage of reproductive aging in women and mice, and the involvement of PEDF in the process. Granulosa cells from reproductively-aged (RA) women and mice (36–44 years old and 9–10 months old, respectively) indicated an increase in the level of PEDF mRNA (qPCR), with yet unchanged levels of AMH and FSHR mRNAs. However, the PEDF protein level in individual women showed an intra-cellular decrease (ELISA), along with a decrease in the corresponding follicular fluid, which reflects the secreted fraction of the protein. The in vitro maturation (IVM) rate in the oocytes of RA mice was lower compared with the oocytes of young mice, demonstrated by a reduced polar body extrusion (PBE) rate. The supplementation of PEDF improved the hampered PBE rate, manifested by a higher number of energetically-competent oocytes (ATP concentration and mtDNA copy number of individual oocytes). Our findings propose PEDF as an early marker of reproductive aging, and a possible therapeutic in vitro agent that could enhance the number of good-quality oocytes in older IVF patients.