miR-532-3p: a possible altered miRNA in cumulus cells of infertile women with advanced endometriosis.

RESEARCH QUESTION: Is the profile of microRNA (miRNA) altered in cumulus cells of infertile women with early (EI/II) and advanced (EIII/IV) endometriosis? DESIGN: In this prospective case-control study, a miRNA profile including 754 targets was evaluated in samples of cumulus cells from infertile women with endometriosis (5 EI/II, 5 EIII/IV) and infertile controls (5, male and/or tubal factor) undergoing ovarian stimulation for intracytoplasmic sperm injection, using TaqMan® Array Human MicroRNA Cards A and B. The groups were compared with Kruskal-Wallis test, followed by Benjamini-Hochberg correction and Dunn's post hoc test. An in silico enrichment analysis was performed to list the possibly altered pathways in which the altered miRNA target genes are involved. RESULTS: Only the miRNA miR-532-3p showed significant differences among the analysed groups, being down-regulated in the EIII/IV group compared with the infertile control group, as well as compared with the EI/II group. The enrichment analysis showed that some genes regulated by this miRNA are involved in important pathways for the acquisition of oocyte competence, such as the oxytocin, calcium, Wnt, FoxO, ErbB and Ras signalling pathways, as well as the oocyte meiosis pathway. CONCLUSION: The present findings bring new perspectives to understanding the follicular microenvironment of infertile women with different stages of endometriosis. It is suggested that the dysregulation of miR-532-3p may be a potential mechanism involved in the aetiopathogenesis of endometriosis-related infertility. Further studies are needed to evaluate these pathways in cumulus cells of infertile women with the disease, as well as their impact on the acquisition of oocyte competence.

Inhibition of LINE-1 retrotransposition represses telomere reprogramming during mouse 2-cell embryo development.

PURPOSE: To investigate whether inhibition of LINE-1 affects telomere reprogramming during 2-cell embryo development. METHODS: Mouse zygotes were cultured with or without 1 µM azidothymidine (AZT) for up to 15 h (early 2-cell, G1/S) or 24 h (late 2-cell, S/G2). Gene expression and DNA copy number were determined by RT-qPCR and qPCR respectively. Immunostaining and telomeric PNA-FISH were performed for co-localization between telomeres and ZSCAN4 or LINE-1-Orf1p. RESULTS: LINE-1 copy number was remarkably reduced in later 2-cell embryos by exposure to 1 µM AZT, and telomere lengths in late 2-cell embryos with AZT were significantly shorter compared to control embryos (P = 0.0002). Additionally, in the absence of LINE-1 inhibition, Dux, Zscan4, and LINE-1 were highly transcribed in early 2-cell embryos, as compared to late 2-cell embryos (P < 0.0001), suggesting that these 2-cell genes are activated at the early 2-cell stage. However, in early 2-cell embryos with AZT treatment, mRNA levels of Dux, Zscan4, and LINE-1 were significantly decreased. Furthermore, both Zscan4 and LINE-1 encoded proteins localized to telomere regions in 2-cell embryos, but this co-localization was dramatically reduced after AZT treatment (P < 0.001). CONCLUSIONS: Upon inhibition of LINE-1 retrotransposition in mouse 2-cell embryos, Dux, Zscan4, and LINE-1 were significantly downregulated, and telomere elongation was blocked. ZSCAN4 foci and their co-localization with telomeres were also significantly decreased, indicating that ZSCAN4 is an essential component of the telomere reprogramming that occurs in mice at the 2-cell stage. Our findings also suggest that LINE-1 may directly contribute to telomere reprogramming in addition to regulating gene expression.

Systemic oxidative stress as a possible mechanism underlying the pathogenesis of mild endometriosis-related infertility.

RESEARCH QUESTION: Does systemic oxidative stress occur during the early follicular phase of the menstrual cycle in infertile women with minimal (stage I) or mild (stage II) endometriosis? Are serum oxidative stress markers during the early follicular phase of the menstrual cycle good predictors of successful gestation in these women who undergo ovarian stimulation for intracytoplasmic sperm injection (ICSI)? MATERIALS AND METHODS: A pilot study (prospective case-control study) was conducted in a University Hospital. Serum samples were obtained during the early follicular phase of the natural cycle preceding ovarian stimulation for ICSI of infertile women (with and without stage I and II endometriosis, the latter having male factor infertility). Total hydroperoxides (FOX1), malondialdehyde, advanced oxidation protein products, reduced glutathione, superoxide dismutase, total antioxidant capacity (TAC), 8-hydroxy-2'-deoxyguanosine (8OHdG) and vitamin E were analysed in serum from 35 women with stage I or II endometriosis and 60 control women. The accuracy of oxidative stress markers for predicting clinical pregnancy and live births was determined by receiver operator characteristic curves. RESULTS: Women with stage I and II endometriosis showed lower serum 8OHdG concentrations (16.02 ng/ml) compared with the control group (22.08 ng/ml). The best predictor for clinical pregnancy and live births was TAC, whereas FOX1 was the best predictor of clinical pregnancy in the control group. CONCLUSIONS: Infertile women with stage I and II endometriosis present systemic oxidative stress during the early follicular phase of the menstrual cycle. Some oxidative stress markers were good predictors of clinical pregnancy and live births after ICSI. Serum TAC was predictive of clinical pregnancy and live births after ICSI in women with stage I or II endometriosis.

Peritoneal fluid of women with endometriosis reduces SOD1 in bovine oocytes in vitro maturation.

Studies have demonstrated oxidative stress in peritoneal fluid (PF) from women with endometriosis and the importance of enzymatic antioxidant machinery to avoid oocyte oxidative damage. Considering that PF constantly surrounds the ovaries and has direct contact with the oocyte at ovulation, we wonder if PF from women with endometriosis may affect antioxidant enzyme gene expression. Thus, the present study aims to evaluate the PF impact from infertile women with minimal and mild endometriosis and from fertile control women without endometriosis on SOD1, CAT, GSR gene's expression in experimental bovine oocytes matured in vitro. Samples of PF were obtained from women who underwent videolaparoscopy-7 infertile with EI/II and 7 fertile without endometriosis. Immature bovine oocytes underwent in vitro maturation in the absence of PF and in the presence of three concentrations (1, 5 and 10%) of PF from fertile and from infertile women with EI/II. After 22 to 24 h of IVM, oocytes were denuded and stored for analysis of SOD1, CAT and GSR by real-time polymerase chain reaction. Oocyte SOD1 expression was significantly lower in the 10% endometriosis group (0.67 ± 0.32) when compared with no-peritoneal fluid (1.05 ± 0.24, p < 0.008) and 10% control groups (1.06 ± 0.22, p < 0.006). These findings raise the possibility of a deleterious influence of PF from women with EI/II on the oocyte, not only after ovulation but also during the maturation process, which could contribute to worsening oocyte quality, being one of the mechanisms related to infertility in patients with endometriosis.

Oxidative stress and oocyte quality: ethiopathogenic mechanisms of minimal/mild endometriosis-related infertility.

Endometriosis, a highly prevalent gynecological disease, is often associated with infertility, even in its milder forms (minimal and mild endometriosis). However, no consensus has been established with regard to this relationship and the possible mechanisms involved have not been completely elucidated. The oocyte is believed to have an important role in the infertility presented by these patients. Hence, oxidative stress events associated with alterations in the peritoneal, serum and/or follicular microenvironments might result in poor oocyte quality and compromise the reproductive potential of these women. Here, we review possible mechanisms involved in oocyte quality impairment that might lead to infertility in patients with early endometriosis.

Oxidative stress and polycystic ovary syndrome: evaluation during ovarian stimulation for ICSI.

Oxidative stress (OS) may affect natural fertility and the results of assisted reproduction techniques (ARTs). Subfertility associated with polycystic ovary syndrome (PCOS) may be related to OS. This process may intensify during controlled ovarian stimulation (COS) for ARTs because of increased ovarian metabolic activity and hypoestrogenism with the use of gonadotropin-releasing hormone agonists (GnRHas). The objective of this study was to investigate the presence of systemic OS in non-stimulated cycles and to determine OS markers (malondialdehyde [MDA], advanced oxidation protein products [AOPP], hydroperoxides [FOX], glutathione [GSH], and vitamin E) during COS in non-obese infertile women with and without PCOS who were subjected to ARTs. A prospective cohort study was conducted on non-obese women (16 with PCOS, and 60 ovulatory patients with infertility due to male and/or tubal factors). The OS markers were determined during the following time-points: the follicular phase of the natural cycle (D1), after pituitary downregulation with GnRHa and before the use of gonadotropins (D2), on the day of administration of human chorionic gonadotropin (D3), and at oocyte retrieval (D4). Intergroup analysis showed that serum MDA concentrations were higher in the PCOS group at D3 (P=0.048) and D4 (P=0.002). On an intragroup analysis, the control group had higher MDA concentrations at D2 than at D1 (P=0.01) or D4 (P=0.004). The AOPP concentrations were higher at D2 (P<0.0001), D3 (P<0.001) and D4 (P<0.0001) compared to D1. The FOX concentrations were lower at D2 (P<0.0001), D3 (P<0.0001), and D4 (P<0.001) than at D1. Serum GSH concentrations were significantly higher at D4 than at D1 (P=0.02). An intragroup analysis of the PCOS subjects showed that the five OS markers did not differ significantly among the four time-points when they were analyzed (D1, D2, D3 and D4). In conclusion, non-obese infertile women with PCOS showed evidence of systemic OS after COS with gonadotropins for ICSI. On the other hand, non-obese ovulatory infertile women, and women with infertility due to male and/or tubal factors showed a possible systemic oxidative balance until the final of COS.

Assessment of Sexual Function in Infertile Women in a Gynecological Care Setting.

INTRODUCTION: Infertility has a high prevalence worldwide. There is also a high prevalence of sexual problems, mainly in gynecological care settings, but many women are unlikely to discuss sexual problems with their physicians. AIMS: To verify how second-year gynecology residents (SGRs) assess the sexual function of infertile women who are undergoing assisted reproductive techniques (ART) at a single infertility tertiary care center in Brazil. MAIN OUTCOME MEASURES: Medical records of patients. METHODS: This retrospective cohort study evaluated all medical records of women who underwent in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) between January 2011 and December 2012 at a fertility clinic of the Hospital das Clinicas of Ribeirão Preto Medical School, University of São Paulo. RESULTS: A total of 616 women underwent ART during the study period. The mean patient age was 34.5 ± 4.4 years, mean weight was 65.6 ± 12.4 kg, mean height was 163 ± 0.6 cm, and mean body mass index (BMI) was 24.8 ± 4.3 kg/m(2). We classified the methods that medical residents used to assess the sexual frequency of these women as a numerical method, by categorization, or none (no assessment). A total of 26.7% (n = 166) of the SGRs did not assess female sexual function and 26.2% (n = 163) made assessments using categorization. SGRs who used a numerical method rather than categorization to classify the sexual frequency of their female patients were more likely to record answers to other questions on sexual desire, arousal, and orgasm. CONCLUSION: SGRs typically do not assess female sexual function in infertile couples. There was considerable heterogeneity among SGRs in their assessment of coital frequency and female sexual function.

Factors Related to Coital Frequency of Women in Their Thirties.

This cross-sectional study aimed to identify factors related to coital frequency (CF) among 254 women in their 30s using a semistructured interview to collect sociodemographic, anthropometric, reproductive, clinical, and relationship data. CF was characterized as (a) never, (b) rarely (≤1 times/month), (c) occasionally (≤1 times /week), (d) regularly (2-3 times/week), or (e) frequently (>3 times/week). The mean age was 34.38 ± 0.43 years, mean body mass index (BMI) was 27.86 ± 6.52 kg, mean family income was US$1,044.18 ± 796.19, mean number of children was 1.71 ± 0.89, and mean relationship duration was 8.87 ± 5.11 years. Eighty-seven women (35.2%) were taking hormonal contraceptives, 143 (98.0%) were employed, 239 (96.48%) had a secondary or higher education, and 9 (3.62%) had primary schooling. CF was classified as >3 times/week in 22 (8.66%), 2-3 times/week for 98 (38.58%), ≤1 times/week in 40 (15.75%), ≤1 times/month in 14 (5.51%), and never in 5 (1.97%). Women who reported having coitus >3 times/week a week had significantly higher body mass index (BMI; 32.72 ± 7.42 kg/m(2)) than those who had coitus 2-3 times/week (28.45 ± 6.76 kg/m(2)) and ≤1 times /week (26.81 ± 5.39 kg/m(2)) (p < 0.01 for both comparisons). Thus, coital frequency varies in women in their 30s. Obese women had a higher CF than normal-weight and overweight women.

Chemotherapy-induced damage to ovary: mechanisms and clinical impact.

Cancer is a major public health problem around the world. Currently, about 5% of women diagnosed with cancer are of reproductive age. These young survivors may face compromised fertility. The effects of chemotherapeutic agents on ovarian reserve and its clinical consequences are generally inferred from a variety of surrogate markers of ovarian reserve, all aiming to provide prognostic information on fertility or the likelihood of success of infertility treatment. Until recently, the mechanisms that are responsible for chemotherapy-induced ovarian damage were not fully elucidated. The understanding of these mechanisms may lead to targeted treatments to preserve fertility. In this manuscript, we will review the current knowledge on the mechanism of ovarian damage and clinical impact of chemotherapy agents on fertility.

Sexual function of women practicing sex in nonconventional settings.

The quality of sexual intercourse in the context of conjugal visits by women to their jailed partners is unknown. This study aimed to assess the quality of the sex lives and psychological conditions of women attending conjugal visits with their jailed inmate partners. This controlled study involved 124 women between the ages of 18 to 40 years who engaged in sexual relations with their inmate partners (conjugal visit group) or with their partners at home (control group). Sexual function was assessed using a semi-structured questionnaire and the Female Sexual Function Index, and psychological parameters were evaluated using the Hospital Anxiety and Depression scale. The total Female Sexual Function Index scores was similar in the 2 groups. The percentage of women reporting good quality of the relationship was significantly higher in the conjugal visit group. Also, the Hospital Anxiety and Depression scale scores were higher in the conjugal visit group. Depression was a risk factor for sexual dysfunction and had a negative effect on scores in the desire, excitement, lubrication, and sexual satisfaction domains, whereas anxiety was associated with lower sexual desire scores. A regular + poor quality of the relationship and being religious were factors associated with sexual dysfunction. Sexual practices in jail were not a risk for sexual dysfunction in this sample.

Importance of lipid accumulation product index as a marker of CVD risk in PCOS women.

BACKGROUND: The polycystic ovary syndrome (PCOS) is considered the most common endocrine disease during the woman's reproductive life, with prevalence ranging from 5 to 10% of women of reproductive age. There is a paucity of studies regarding the use of the lipid accumulation product (LAP) as a risk marker for the development of cardiovascular disease (CVD). METHODS: A cross-sectional study was conducted on 78 women aged 18 to 42 years seen at University Hospital of Maranhão, with a diagnosis of polycystic ovary syndrome according to the Rotterdam criteria. The following variables of interest were recorded on a protocol form: sociodemographic and behavioral data, body mass index, waist circumference, fasting glucose, total cholesterol, triglycerides, low density lipoprotein cholesterol, high-density lipoprotein cholesterol, and systolic and diastolic blood pressure. RESULTS: Logistic regression showed that, except for HDL, all cardiovascular risk markers presented a higher chance of being altered when the lipid accumulation product was above the cut off value of 37.9 cm.mmol/L. CONCLUSION: The lipid accumulation product seems to be sufficient to indicate a risk of cardiovascular diseases in women with polycystic ovary syndrome.

Fertility Preservation and Ovarian Hyperstimulation Syndrome Management in Cancer Care: A Pathophysiological Perspective on Gonadotropin-Releasing Hormone Agonists and Antagonists.

This narrative review delves into the evolving landscape of fertility preservation techniques, with a particular focus on their use in patients undergoing oncology treatment that carries a risk of ovarian insufficiency. Advances in established methods such as cryopreservation of oocytes and embryos are highlighted, and the increasing use of gonadotropin-releasing hormone (GnRH) agonists is discussed. The review also addresses the complexities and controversies associated with these approaches, such as the 'flare-up' effect associated with GnRH agonists and the potential of GnRH antagonists to reduce the risk of ovarian hyperstimulation syndrome. Despite advances in fertility preservation, the report highlights the challenges we face, including the need for personalized treatment protocols and the management of associated risks. It calls for continued research and collaboration between healthcare professionals to refine these techniques and ultimately improve reproductive outcomes for patients facing the prospect of fertility-impairing treatment.

Intrauterine insemination: prognostic factors.

OBJECTIVE: To evaluate the impact of possible maternal and paternal prognostic factors and ovarian stimulation protocols on clinical pregnancy and live birth rates in intrauterine insemination (IUI) cycles. METHODS: Retrospective observational study of 341 IUI cycles performed from January 2016 to November 2020 at the Assisted Reproduction Service of the Clinics Hospital of the Ribeirão Preto Medical School, University of São Paulo. Clinical pregnancy and live birth rates and their potential prognostic factors were evaluated. Wilcoxon's non-parametric test was used to compare quantitative variables, and the chi-square test to compare qualitative variables, adopting a significance level of p<0.05. A logistic regression model was performed to verify which exploratory variables are predictive factors for pregnancy outcome. RESULTS: The ovulation induction protocol using gonadotropins plus letrozole (p=0.0097; OR 4.3286, CI 1.3040 - 14.3684) and post-capacitation progressive sperm ≥ 5million/mL (p=0.0253) showed a statistically significant correlation with the live birth rate. Female and male age, etiology of infertility, obesity, multifollicular growth, endometrial thickness ≥ 7 mm, and time between human chorionic gonadotropin administration and IUI performance were not associated with the primary outcomes. In the group of patients with ideal characteristics (women aged< 40 years, BMI < 30 kg/m2, antral follicle count ≥ 5, partner aged< 45 years, and post-capacitation semen with progressive spermatozoa ≥ 5 million/mL), the rate of clinical pregnancy was 14.8%, while that of live birth, 9.9%. CONCLUSIONS: In this study, the ovulation induction protocol with gonadotropins plus letrozole and post-capacitation progressive sperm ≥ 5 million/mL were the only variables that significantly correlated with intrauterine insemination success.

Omega-3 fatty acids and L-carnitine prevent meiotic oocyte damage induced by follicular fluid from infertile women with endometriosis: an experimental study.

OBJECTIVE: To assess whether follicular fluid (FF) from infertile women with endometriosis in advanced stages [moderate/severe (EIII/IV) without or with endometrioma (Endometrioma)] induce more oocyte damages than in early stages (minimal/mild: EI/II); and whether supplementation with L-carnitine (LC) and omega 3 (n3) can prevent these oocyte damages. METHODS: Experimental study using bovine oocytes (obtained of ovaries from slaughterhouse), and human FF (samples were obtained during oocyte recovery for ICSI). Bovine oocytes were submitted to in vitro maturation (IVM) divided into 9 groups: no FF(No-FF), with 1% FF from infertile women without endometriosis (FFC), with EI/II, EIII/IV and Endometrioma, and with (or not) LC+n3 addition. After IVM, oocytes were fluorescently labelled and visualized by confocal microscopy to analyze chromosomes and spindle. RESULTS: FF from endometriosis decreased rate of normal MII (spindle assembly and chromosome alignment) compared to No-FF (87.2%) and FFC (87.2%). FFEIII/IV (80.7%) and FFEndometrioma (69.3%) decreased total MII rate compared to No-FF (91.9%) and FFC (89.2%), and FFEndometrioma had lower total MII rate compared to other groups. LC+n3 increased MII rate in the FFEIII/IV (80.7% vs. 90.8%) and the Endometrioma (69.3% vs. 86.4%), and it prevented damages in spindle and chromosomes in MII oocytes in the FFEI/II group (62.2% vs. 84.5%) and the FFEIII/IV group (70.2% vs. 84.1%). CONCLUSIONS: FF of endometriosis damaged the meiotic spindle of bovine MII oocytes. EIII/IV led to impaired nuclear maturation; FF from women with endometrioma had further negative impact in oocyte maturation. LC+n3 completely prevented the effects of FF from women with endometriosis on oocyte.

Risk of ovarian hyperstimulation syndrome in women with malignancies undergoing treatment with long-acting gonadotropin-releasing hormone agonist after controlled ovarian hyperstimulation for fertility preservation: a systematic review.

Background: Fertility preservation is an important quality of life issue for women of reproductive age undergoing gonadotoxic treatment. The possibility of administering an adjuvant long-acting gonadotropin-releasing hormone agonist (GnRHa) with the aim of reducing the number of follicles susceptible to the effects of chemotherapy and thus reducing the risk of ovarian damage is considered in some international society guidelines, particularly in certain cancers such as breast cancer. Nowadays, the administration of long-acting GnRHa after controlled ovarian hyperstimulation (COH) for fertility preservation by cryopreservation of oocytes or embryos is increasingly used. However, cases of ovarian hyperstimulation syndrome (OHSS) have been reported following the use of long-acting GnRHa after COH for fertility preservation, indicating that the potential adverse effects of this treatment need to be further investigated. Objectives: The aim of this systematic review was to comprehensively characterize patients who developed OHSS after treatment with long-acting GnRHa following COH for fertility preservation. Methods: A comprehensive search of major electronic databases through January 2023 was performed. Studies reporting the use of long-acting GnRHa after COH for fertility preservation and the development of OHSS were included. Risk of bias was assessed using a modified version of the Newcastle-Ottawa scale. Results were synthesized qualitatively. Results: Three studies with five patients met the eligibility criteria. The majority of patients were diagnosed with breast cancer and all patients underwent COH for oocyte cryopreservation. OHSS occurred in all patients after administration of long-acting GnRHa. The interval between ovulation induction and administration of long-acting GnRHa thereafter ranged from 3 to 5 days. All patients were treated conservatively and recovered without complications. Conclusion: Current evidence suggests that the use of long-acting GnRHa after COH for fertility preservation may be associated with OHSS. Healthcare providers should thoroughly discuss the benefits and risks of this intervention with their patients before making a decision. Further studies are needed to fully elucidate the causal relationship between long-acting GnRHa and OHSS in this population.

Feasibility and safety of combined laparoscopic and transvaginal oocyte retrieval in a woman with vaginal recurrence of cervical adenocarcinoma: a case report.

Introduction: Oocyte cryopreservation is an established technique for fertility preservation in women diagnosed with cancer. However, some clinical scenarios may preclude the commonly used transvaginal approach to oocyte retrieval. In such cases, a laparoscopic approach may be required. Here, we report the feasibility and safety of a combined laparoscopic and transvaginal approach for oocyte retrieval in a woman with vaginal recurrence of cervical adenocarcinoma. This approach allowed for oocyte cryopreservation prior to cancer treatment, representing a novel application in this clinical context. Methods: A 31-year-old woman with endocervical adenocarcinoma underwent laparoscopic radical hysterectomy and pelvic lymph node dissection. She presented with vaginal recurrence and was referred for fertility preservation by oocyte cryopreservation before chemotherapy and radiotherapy/brachytherapy. Ovarian stimulation was initiated with a gonadotropin antagonist protocol combined with aromatase inhibitors, and oocyte retrieval was performed with a combined laparoscopic and transvaginal approach. Results: A total of 18 oocytes were retrieved and 10 mature oocytes were cryopreserved. Peritoneal fluid cytology was negative for malignancy. The patient underwent chemotherapy and radiotherapy/brachytherapy and was disease-free after oocyte retrieval. Conclusion: The combined laparoscopic and transvaginal approach for oocyte retrieval emerges as a practical and efficacious method for fertility preservation in cases of cervical adenocarcinoma with vaginal recurrence. Further comprehensive studies are warranted to establish the reproducibility, safety, and long-term outcomes associated with this innovative approach.

Live birth after single euploid frozen embryo transfer in a 39-year-old woman with high-grade mosaic Turner syndrome.

OBJECTIVE: To describe the reproductive and obstetric outcomes of an intracytoplasmic sperm injection cycle with preimplantation genetic testing for aneuploidy in an advanced reproductive-age woman with high-grade mosaic Turner syndrome. METHODS: Case report of a 39-year-old woman diagnosed with mosaic Turner Syndrome 45,X[90]/46,XX[10] karyotype who underwent in vitro fertilization treatment with blastocyst trophectoderm biopsy for preimplantation genetic testing using next-generation sequencing. RESULT(S): Two of the four blastocysts biopsied were euploid. The patient achieved ongoing pregnancy after the first single euploid frozen embryo transfer, followed by the birth of a healthy child. CONCLUSION: Autologous intracytoplasmic sperm injection cycles can be considered in a select group of advanced reproductive-age women diagnosed with high-grade mosaic Turner syndrome.

Comparing the effects of a commercial and a prototype vitrification medium on meiotic spindle morphology and survival rate of mouse oocytes.

OBJECTIVE: To compare oocyte survival and meiotic spindle normality between vitrified-warmed oocytes in a mouse embryo assay using Tvitri-4 or Ingámed vitrification media. METHODS: C57BL/6 female mice aged 8-12 weeks were submitted to superovulation with pregnant mare's serum gonadotropin and human chorionic gonadotropin (hCG) for obtaining of in vivo matured oocytes. The oocytes were randomly distributed into one of the following three groups: CTR - control (fresh oocytes); ING - oocytes vitrified-warmed in a standard commercial kit supplied by Ingámed, and T4 - oocytes vitrified-warmed in the novel prototype Tvitri-4 medium. After warming and recovery culture, oocytes were assessed with respect to survival rate (SR) and both meiotic spindle morphology and chromosome alignment of each oocyte fixed in the sagittal position after immunostaining and analysis by confocal microscopy. RESULTS: A total of 354 mature oocytes were vitrified in ING (n=178) and T4 (n=176), out of which 299 (85%) survived after warming. Oocyte survival rates were not statistically different (p=0.08) between ING (145/178=81.5%) and T4 (154/176=87.5%). Regarding meiotic normality, there were no significant changes in the proportion of oocytes with normal meiotic spindle morphology and chromosome structure between ING (52,2%) and T4 (63.4%) after warming (RR: 0.95, 95% CI: 0.92-1.607). When the meiotic normality was assessed using the CTR group as a reference in the analysis of relative risk, no significant differences were observed between T4 (63.4%) and CTR (70.5%) (RR: 0.95, 95% CI: 0.72-1.12). On the other hand, the percentage of oocytes retaining normal meiotic spindle morphology and chromosome configuration in ING (52.2%) was lower than in the CTR group (RR: 0.95, 95% CI: 0.57-0.97). CONCLUSIONS: The novel prototype Tvitri-4 medium was efficient in preserve survival rate and meiotic spindle normality of oocytes and, with further verification, may be able to replace commercially available media in future clinical applications.

The Impact of Controlled Ovarian Stimulation on Serum Oxidative Stress Markers in Infertile Women with Endometriosis Undergoing ICSI.

Endometriosis-related infertility is associated with oxidative stress (OS). The present study aims to compare serum OS markers of infertile women with endometriosis and controls during the follicular phase of the natural cycle (D1), after pituitary downregulation using a GnRH agonist (D2), after controlled ovarian stimulation (COS) on the day of human chorionic gonadotropin administration (D3), and on the day of oocyte retrieval (D4). One hundred and eight serum samples (58 controls and 35 early and 18 advanced endometriosis cases) were collected at these four timepoints. OS markers were compared among the groups and timepoints using a linear regression model with mixed effects and a post-test using orthogonal contrasts. The significance was set at 5%. We observed altered OS markers in the endometriosis patients during the D1, D2, D3, and D4 timepoints compared to the controls. The evidence of systemic OS in infertile patients with endometriosis during COS suggests the mobilization of potent antioxidants in an attempt to protect the oocyte from oxidative damage, especially on the day of oocyte retrieval.