The effects of COVID-19 lockdown on lifestyle and emotional state in women undergoing assisted reproductive technology: Results of an Italian survey.

PURPOSE: On March 8, 2020, the Italian Government implemented extraordinary measures to limit viral transmission of COV-19/SARS-CoV-2. We evaluated the impact of COVID-19 lockdown on lifestyle and emotional state in women planning infertility treatments. BASIC PROCEDURES: We performed a quantitative research study using a web-based survey, in 140 women referred to Assisted Reproductive Technologies Center. MAIN FINDINGS: We observed changes in body weight during lockdown in 80 % of women, and a significant increase in BMI in comparison to that observed before (p=.001). We observed a high percentage of non-adherence to the Mediterranean pattern during lockdown due to higher frequency of consumption of sweet/pastries, cheese and meat, rather than fruit, vegetables and legumes. Before lockdown 36.4 % women were snack consumers while during lockdown 55 % (p=.002). By considering individuals' attitude to snack consumption, we observed an increase related to boredom (p=<.0001) and anxiety (p=.05) during lockdown. Increased levels of anxiety and sadness were observed in about 30 %, and of boredom in 25 %. The percentage of women worried about their planning infertility treatment was more than 50 %. PRINCIPAL CONCLUSIONS: Quarantine-related restrictions strongly influenced lifestyle psychological behavior leading to an increased burden of cardiovascular disease.

Cardiometabolic and Thrombotic Risk Profile in Women Undergoing Oocyte Donation for Assisted Reproduction.

Background: The last two decades have seen a growing number of pregnancies in women who needed the donation of oocytes. With oocyte donation pregnancies, studies on obstetric outcomes among these women revealed an increased incidence of pre-eclampsia and pregnancy-induced hypertension. Furthermore, several studies have found a higher incidence of low birth weight, preterm birth, and delivery by cesarean section in oocyte donation rather than in women subjected to assisted reproduction techniques (ART) with autologous oocytes. Numerous studies have also shown a deep connection between cardiovascular and thrombotic risk factors and adverse pregnancy outcomes. In this setting, to strictly assess the preconceptional risk for women who undergo egg donation to achieve pregnancy, the aim of our study is to draw a detailed assessment of the vascular risk profile of patients with gamete donation ART indications through the evaluation of comorbidities and cardiometabolic and thrombophilic markers Materials and Methods: Patients undergoing ART with oocyte or sperm donation or double donation of gametes underwent a careful clinical assessment through a detailed personal and family anamnesis and they were evaluated for cardiometabolic and thrombophilic profile. Clinical and demographic characteristics, comorbidities, and biohumoral parameters were collected. The study was approved by the Regional Ethical Committee(Em 2018-017 CINECA 10189). Results: We evaluated 525 women. Around 73.1% were >40 years and 35% of them were older than 45 years. There was a high prevalence of dyslipidemias (58.1%), smoking habit (24.6%), a body mass index >25 in 28.6% of patients, a high abdominal circumference in 58.1% of cases, a prevalence of acquired thrombophilia in about 7% and hereditary of 19.2%. Around 39.2% of patients had total cholesterol >200 mg/dL, 19.5% had high-density lipoprotein <48 mg/dL and 43.6% had low-density lipoprotein >115 mg/dL, and 6.9% had triglyceride values >150 mg/dL. Conclusions: A careful assessment of the preconceptional status of patients undergoing ART programs with oocyte donation can be highly recommended.

'Two countries-two labs': the transnational gamete donation (TGD) programme to support egg donation.

PURPOSE: To evaluate the effectiveness, efficiency, and safety of a transnational gamete donation (TGD) programme based on the shipping of vitrified donor oocytes. METHODS: A retro-prospective observational study was conducted in the Assisted Reproductive Technology Center of the University Hospital of Florence, Italy. The study population included 622 consecutive donor oocyte cycles. A mean number of 6 vitrified oocytes per couple were shipped from two Spanish biobanks. In the receiving centre, gametes were warmed and inseminated and the subsequent embryo transfer (ET) was performed. The main outcome measurement was LBR. Secondary outcomes included oocyte survival rate, ICSI damage rate, normal fertilization, cleavage, and implantation rate (IR) in both 'fresh' and cryotransfer cycles. RESULTS: A total number of 3536 mature oocytes were warmed with 81.4% surviving oocytes. 2PN oocytes were 1941 with an ICSI normal fertilization rate of 70.4% and a cleavage rate of 93.4%; 857 day-3 embryos were transferred in 498 women, 63 blastocysts in 44. Couples with at least one vitrified embryo were 181 (32.3%). IR was 25.1% and 33.1% for day-3 ET and blastocyst stage respectively. Crude pregnancy rate and LBR after the first ET were 35.5% and 27% correspondingly with a conservative cumulative LBR of 34% and an optimal LBR of 51.4%. CONCLUSION: Imported vitrified donor oocytes retain their competence and are capable of resulting in ongoing pregnancies and healthy babies in a proportion comparable to other existing systems as egg donation with vitrification/warming in the same laboratory and transnational fresh oocyte donation.

Bilateral Endometrioma Excision: Surgery-Related Damage to Ovarian Reserve.

To study whether unoperated ovarian endometrioma(s) or its surgical excision led to a modified pattern of ovarian decay with increasing female age. A sectional analysis of basal follicle stimulating hormone (FSH) and ovarian response to gonadotropins was conducted on women treated with fresh autologous In Vitro Fertilization/Intracytoplasmic sperm injection (IVF/ICSI) cycles. The study group included patients with unoperated ovarian endometrioma(s) (108 cycles); control groups were women with previous surgery for monolateral ovarian endometrioma (101 cycles), surgery for bilateral ovarian endometriomas (39 cycles), and tubal factor infertility (171 cycles). Simple linear regression analyses and the Pearson correlation were used to analyze the correlation between basal FSH, number of dominant follicles, number of retrieved oocytes, and age of patients. The relationship between the variables was significant in case of patients with nonoperated ovarian endometrioma(s) and patients with previous surgery for monolateral endometrioma and tubal factor infertility group. In patients with a history of surgery for bilateral endometriomas, no relationship was found among the variables (basal FSH 95% confidence interval [CI]: -0.475 to 0.319; P = .688; number of dominant follicles 95% CI: -0.484 to 0.382; P = .808; number of retrieved oocytes 95% CI: -0.478 to 0.370, P = .792). In women with unoperated ovarian endometrioma(s) or with a history of surgery for monolateral endometrioma, the remaining ovarian parenchyma maintains the same pattern of ovarian decay as healthy ovaries. Unoperated ovarian endometriotic lesions did not interfere with ovarian reserve and IVF/ICSI cycles' outcomes and were less injurious than surgery. After surgery for bilateral ovarian endometriomas, a decline in ovarian reserve seems independent from the patient's age.

A Biochemical Approach to Detect Oxidative Stress in Infertile Women Undergoing Assisted Reproductive Technology Procedures.

Oxidative stress plays a major role in critical biological processes in human reproduction. However, a reliable and biologically accurate indicator of this condition does not yet exist. On these bases, the aim of this study was to assess and compare the blood and follicular fluid (FF) redox status of 45 infertile subjects (and 45 age-matched controls) undergoing in vitro fertilization (IVF), and explore possible relationships between the assessed redox parameters and IVF outcomes. Reactive Oxygen Species (ROS) production, assessed by flow cytometry analysis in blood leukocytes and granulosa cells, significantly increased (p < 0.05) in infertile patients. Also, oxidative stress markers-ThioBarbituric Acid-Reactive Substances (TBARS) as an index of lipid peroxidation, and Oxygen Radical Absorbance Capacity (ORAC) to account for total antioxidant capacity, both assayed by fluorometric procedures-in blood and FF were significantly (p < 0.001) modified in infertile patients compared to the control group. Moreover, a significant correlation between blood redox markers and FF redox markers was evident. An ORAC/TBARS ratio, defined as the redox index (RI), was obtained in the plasma and FF of the patients and controls. In the patients, the plasma RI was about 3.4-fold (p < 0.0001) lower than the control, and the FF RI was about six-fold (p < 0.0001) lower than the control. Interestingly, both the plasma RI and FF RI results were significantly correlated (p < 0.05) to the considered outcome parameters (metaphase II, fertilization rate, and ongoing pregnancies). Given the reported findings, a strict monitoring of redox parameters in assisted reproductive techniques and infertility management is recommended.

Biological meaning of ubiquitination and DNA fragmentation in human spermatozoa.

The ubiquitin-proteasome is an ubiquitous system mainly devoted to protein degradation. The presence of ubiquitinated proteins in male gametes suggests a role for this system also in reproduction. Available evidence indicate that ubiquitin in spermatozoa may have a role in semen quality control, as ubiquitinated defective spermatozoa in the epididymis are subsequently phagocytosed by epididymal epithelial cells. Moreover, a role both in the regulation of mitochondrial inheritance in mammals (paternal mitochondria are eliminated and their ubiquitination appears to be important for this process) and in sperm-oocyte interaction at fertilization (which is inhibited by an inhibitor of proteasome) have been also suggested. We found that both morphologically normal and abnormal human spermatozoa in semen may be ubiquitinated and that the percentage of ubiquitinated sperm in the ejaculate positively correlates with normal morphology and motility, suggesting that sperm ubiquitination may have a positive role in sperm functions. It remains to be defined if and which patterns of ubiquitination of spermatozoa may distinguish between the different biological functions of this system. In an attempt to answer this question, we set up a method to detect simultaneously ubiquitination and DNA fragmentation by FACScan since the latter parameter is related to a poor quality of semen; in particular, abnormal morphology. We found that DNA fragmented human spermatozoa are also ubiquitinated. Studies are in progress to determine the correlation between the fraction of ubiquitinated-non DNA fragmented spermatozoa and parameters of semen analysis.

Lack of Histocompatibility Leukocyte Antigen-G expression in early embryos is not related to germinal defects or impairment of interleukin-10 production by embryos.

The expression of Histocompatibility Leukocyte Antigen (HLA)-G molecules is a mandatory prerequisite for the development of pregnancy but no hypotheses have yet been advanced regarding the lack of HLA-G modulation expression in a percentage of early embryos obtained by in vitro fertilization (IVF). One possible hypothetical model assumes that the absence of regulation of HLA-G or impaired interleukin (IL)-10 secretion could be related to germinal defects. We investigated the presence of soluble HLA-G antigens in supernatants of single embryo cultures from couples admitted to a second fertilization procedure; these couples showed a complete absence of HLA-G modulation in the first cycle's embryo supernatants (0/31). The results obtained in the second IVF cycle showed embryo supernatants positive for HLA-G (14/40), suggesting that the previous lack of antigen modulation is independent of germinal defects. Furthermore, since it has been reported that oocytes and early embryos can secrete IL-10, an anti-inflammatory cytokine produced by type 2 helper T cells that induces upregulation of HLA-G expression in monocytes and trophoblasts, we investigated the levels of IL-10 and soluble HLA-G in 40 embryo culture supernatants from 21 IVF cycles. No associations were observed between the presence of IL-10 and the production and concentrations of soluble HLA-G, or between IL-10 levels and pregnancy outcome. These results indicate that the lack of HLA-G production in early embryos is not related to germinal defects or to impairment in embryo IL-10 secretion but could be ascribed to possible uncorrected fertilization processes.