Embryo development is impaired by sperm mitochondrial-derived ROS

Background Basal energetic metabolism in sperm, particularly oxidative phosphorylation, is known to condition not only their oocyte fertilising ability, but also the subsequent embryo development. While the molecular pathways underlying these events still need to be elucidated, reactive oxygen species (ROS) could have a relevant role. We, therefore, aimed to describe the mechanisms through which mitochondrial activity can influence the first stages of embryo development. Results We first show that embryo development is tightly influenced by both intracellular ROS and mitochondrial activity. In addition, we depict that the inhibition of mitochondrial activity dramatically decreases intracellular ROS levels. Finally, we also demonstrate that the inhibition of mitochondrial respiration positively influences sperm DNA integrity, most likely because of the depletion of intracellular ROS formation. Conclusion Collectively, the data presented in this work reveals that impairment of early embryo development may result from the accumulation of sperm DNA damage caused by mitochondrial-derived ROS.

Fertilización asistida en mujeres de edad reproductiva avanzada, ¿quo vadis?

La donación de ovocitos y embriones se ha convertido en un procedimiento frecuente en mujeres infértiles en edad reproductiva avanzada, con una elevada tasa de embarazo. En estos embarazos aumenta la incidencia de eventos obstétricos adversos, como trastornos hipertensivos, diabetes gestacional, parto operatorio, entre otros, y mortalidad perinatal. Los embarazos múltiples aumentan significativamente los riesgos, por lo que se recomienda la transferencia de un solo embrión. Las pacientes deben someterse inicialmente a un examen médico integral que incluya evaluación cardiovascular y psicológica, evitando el tratamiento en mujeres con enfermedades crónicas que aumentan significativamente los riesgos. En este simposio se revisará el manejo de la infertilidad en mujeres en edad reproductiva avanzada con ovocitos propios y donados, las complicaciones obstétricas y perinatales, así como los aspectos éticos y las consideraciones especiales para tener en cuenta en este tipo de tratamientos.

Oocyte and embryo donation has become a frequent procedure in infertile women of advanced reproductive age, with a high pregnancy rate. In these pregnancies, the incidence of adverse obstetric events (hypertensive disorders, gestational diabetes, operative delivery, among others) and perinatal mortality increases. Multiple pregnancies significantly increase the risks, so single embryo transfer is recommended. Patients should have a previous comprehensive medical examination including cardiovascular and psychological evaluation, avoiding treatment in women with chronic diseases that significantly increase risks. This symposium ill review infertility management in women of advanced reproductive age with own and donated oocytes, obstetric and perinatal complications as well as ethical aspects and special considerations to consider in this type of treatment.

Fertilización asistida con óvulos donados: indicaciones y resultados

Las mujeres retrasan cada vez más la maternidad por diferentes motivos, lo que les ocasiona recurrir a tratamientos de fertilización in vitro (FIV) con óvulos propios u óvulos donados para conseguir embarazo. En los tratamientos de FIV con óvulos donados se realiza una selección estricta de las donantes, quienes son sometidas a estimulación ovárica con posterior aspiración de los folículos. La edad recomendada para donar es entre 21 y 34 años. Se recomienda un máximo de 6 donaciones por donante. La receptora es la persona a quien se le realizará la transferencia del embrión y llevará el embarazo. Las tasas de embarazo con esta técnica de reproducción asistida son altas y las indicaciones más frecuentes son edad materna avanzada y falla ovárica precoz.

Women are increasingly delaying childbearing for different reasons, which causes them to resort to in vitro fertilization (IVF) treatments with their own oocytes or donated oocytes to achieve pregnancy. In IVF treatments with donated oocytes, donors are strictly selected and undergo ovarian stimulation with subsequent follicle aspiration. The recommended age to donate is between 21-34 years old. A maximum of 6 donations per donor is recommended. The recipient is the person to whom the embryo transfer will be performed and who will carry the pregnancy. Pregnancy rates with this assisted reproduction technique are high and the most frequent indications are advanced maternal age and early ovarian failure.

Effect of Endometriosis on Cumulus ATP, Number of Mitochondria and Oocyte Maturity in Cumulus Oocyte Complex in Mice / Efeito da endometriose no cumulus ATP, número de mitocôndrias e maturidade oocitária no complexo oocitário cumulus em camundongos

Abstract Objective Endometriosis causes a decrease in oocyte quality. However, this mechanism is not fully understood. The present study aimed to analyze the effect of endometriosis on cumulus cell adenosine triphosphate ATP level, the number of mitochondria, and the oocyte maturity level. Methods A true experimental study with a post-test only control group design on experimental animals. Thirty-two mice were divided into control and endometriosis groups. Cumulus oocyte complex (COC) was obtained from all groups. Adenosine triphosphate level on cumulus cells was examined using the Elisa technique, the number of mitochondria was evaluated with a confocal laser scanning microscope and the oocyte maturity level was evaluated with an inverted microscope. Results The ATP level of cumulus cells and the number of mitochondria in the endometriosis group increased significantly (p < 0.05; p < 0.05) while the oocyte maturity level was significantly lower (p < 0.05). There was a significant relationship between ATP level of cumulus cells and the number of mitochondrial oocyte (p < 0.01). There was no significant relationship between cumulus cell ATP level and the number of mitochondrial oocytes with oocyte maturity level (p > 0.01; p > 0.01). The ROC curve showed that the number of mitochondrial oocytes (AUC = 0.672) tended to be more accurate than cumulus cell ATP level (AUC = 0.656) in determining the oocyte maturity level. Conclusion In endometriosis model mice, the ATP level of cumulus cells and the number of mitochondrial oocytes increased while the oocyte maturity level decreased. There was a correlation between the increase in ATP level of cumulus cells and an increase in the number of mitochondrial oocytes.

The role of growth hormone in improving oocyte quality in IVF cycles

Background@#Growth hormone has been used as an adjunct in ovarian stimulation of IVF cycles for the past three decades. However, the exact mechanism of its role in improving oocyte quality has not been thoroughly investigated. Although a clear indication for GH co-treatment is in infertile women with GH deficiency, GH has been given mostly to poor ovarian responders.@*Method@#This paper is a review of the most recent published data on the role of GH supplementation in improving oocyte quality in younger women who are suboptimal or unexpected poor responders to standard ovarian stimulation.@*Results@#Retrospective cohort and randomized trials demonstrated an improvement in oocyte quality through morphological parameters, mitochondrial function, biomarkers, GH receptors, insulin growth factor, markers of oxidative stress, mature oocytes, good quality embryos, implantation rate, clinical pregnancy and live birth.@*Conclusion@#Current data suggest GH supplementation may improve oocyte and embryo qualities, endometrial receptivity, clinical pregnancy and live birth. However, better quality evidence is needed before a recommendation can be made for GH supplementation to be given to infertile women who are suboptimal or poor ovarian responders.

Opinions of female academicians on oocyte freezing: a qualitative study

SUMMARY OBJECTIVE: The aim of this study was to evaluate the opinions of female academicians about oocyte freezing. METHODS: This qualitative study included 12 single academic women who had not yet entered menopause, did not have children, and were continuing their doctoral education in Istanbul, Turkey, between August and September 2022. Data were collected with semi-structured interviews and evaluated by content analysis. RESULTS: Three main themes were "Difficulty of fertility in academics," "Advantages of oocyte freezing," and "Concerns about oocyte freezing." Participants mostly had positive attitudes about the advantages of oocyte cryopreservation, but they had concerns about pregnancies obtained with frozen oocytes. CONCLUSION: The academic women attributed fertility as an obstacle to their career and experienced anxiety about fertility. They were aware of the advantages of oocyte cryopreservation; however, they defined the pregnancy with oocyte freezing as artificial.

Use of Triggers on in vitro Fertilization and Evaluation of Risk Factors for Sub-Optimal Maturation Rate / Uso de gatilhos na fertilização in vitro e avaliação dos fatores de risco para taxa de maturação subótima

Abstract Objective To compare the oocyte maturation rate in the treatment of in vitro fertilization (IVF) in terms of the use of human chorionic gonadotropin (hCG), agonist gonadotropin-releasing hormone (GnRH) and dual trigger and to evaluate the associated risk factors for sub-optimal maturation rates. Methods A retrospective cohort study with 856 women who underwent IVF. They performed oocyte retrieval and were classified into 3 groups (1 - hCG, 2 - GnRHagonist, 3 - dual trigger). The primary outcome was maturation rate per trigger, and the secondary outcomes were the pregnancy rate per oocyte retrieval and the correlations between low maturation rate as well as the clinical and treatment characteristics of women. Results The maturation rate was 77% in group 1; 76% in group 2, and 83% in group 3 (p=0.003). Group 2 showed women with better ovarian reserve, greater number of oocytes collected, and more mature oocytes and embryos compared with the other groups (p<0.001). The cumulative clinical pregnancy rate was no different between the groups (p=0.755). Low ovarian reserve and low doses of follicle-stimulating hormone (FSH) administered during the stimulus were associated with a higher chance of null maturation rate. Conclusion The oocyte maturation rates and IVF results were similar in all groups. Low ovarian reserve is associated with the worst treatment results.

Resumo Objetivo Comparar a taxa de maturação oocitária no tratamento de fertilização in vitro (FIV) emrelação so o uso de gonadotrofina coriônica humana (hCG), agonista de hormônio liberador de gonadotrofina (GnRH), e gatilho duplo e avaliar os fatores de risco associados a taxas de maturação subótimas. Métodos Estudo de coorte retrospectivo com 856 mulheres submetidas à FIV. Elas foram classificadas em 3 grupos (1 - hCG, 2 - GnRH agonista, 3 - gatilho duplo). O desfecho primário foi a taxa de maturação por gatilho, e os desfechos secundários foram a taxa de gravidez por recuperação de oócitos e as correlações entre a baixa taxa de maturação bem como as características clínicas e do tratamento das mulheres. Resultados A taxa de maturação foi de 77% no grupo 1; 76% no grupo 2, e 83% no grupo 3 (p=0,003). O grupo 2 apresentou mulheres com melhor reserva ovariana, maior número de oócitos coletados, oócitosmaduros, e embriões, emcomparação aos demais grupos (p<0,001). A taxa cumulativa de gravidez clínica não foi diferente entre os grupos (p=0,755). Baixa reserva ovariana e baixas doses de hormônio folículoestimulante (FSH) administradas durante o estímulo foram associadas a uma maior chance de taxa de maturação nula. Conclusão As taxas de maturação oocitárias e os resultados de FIV foram semelhantes em todos os grupos. A baixa reserva ovariana está associada aos piores resultados do tratamento.

Prevention of Premature Ovulation by Administration of Gonadotropin Releasing Hormone Antagonist the day After Ovulation Triggering in Diminished Ovarian Reserve Patients / Prevenção da ovulação prematura por meio da administração de antagonismo de liberação de gonadotropismo no dia seguinte à ovulação

Abstract Objective The aim of the present retrospective study was to investigate the effectiveness of single-dose gonadotropin releasing hormone (GnRH) antagonist administration, the day after human chorionic gonadotropin (hCG) triggering for final oocyte maturation, on the prevention of premature luteinization in patients with diminished ovarian reserve in in-vitro fertilization (IVF) cycles. The secondary objective of the study was to search the effect of this protocol on pregnancy outcomes. Methods This is a retrospective study including 267 infertile patients who have single antral follicle seen with ultrasonography on the 2nd or 3rd day of the menstrual cycle before starting IVF treatment. We randomized patients into two groups. The case group comprised patients who had single-dose GnRH antagonist injection the day after hCG triggering formed, and the patients who had the standard treatment regime formed the control group. In both groups, the oocytes were collected 36 hours after hCG injection. Results The premature ovulation rate was significantly low in the case group compared with the control group (6.86 versus 20.6% per scheduled cycle) (p=0.022). Also, the oocyte retrieval rate (93.14 versus 67.87% per scheduled cycle) (p=0.013), the oocyte maturity rate (79.42 versus 47.87%) (p=0.041), the fertilization rate (65.68 versus 34.54%) (p=0.018), and the embryo transfer rate per scheduled cycle (44.11 versus 18.78%) (p=0.003) were higher in the GnRH antagonist group than in the control group. Conclusion The administration of GnRH antagonist the day after hCG trigger in IVF treatments of patients with diminished ovarian reserve enabled a significant decrease in the rate of premature ovulation but had no effect on live birth rate.

Resumo Objetivo O objetivo do presente estudo retrospectivo foi investigar a eficácia da administração do antagonista do hormônio liberador da gonadotrofina (GnRH) em dose única no dia seguinte ao desencadeamento da gonadotrofina coriônica humana (hCG) para a maturação final do oócito, na prevenção da luteinização prematura em pacientes com diminuição do ovário reserva em ciclos de fertilização in vitro (FIV). O objetivo secundário do estudo foi pesquisar o efeito deste protocolo nos resultados da gravidez. Métodos Trata-se de um estudo retrospectivo incluindo 267 pacientes inférteis que apresentam um único folículo antral visto por ultrassonografia no 2° ou 3° dia do ciclo menstrual antes de iniciar o tratamento de FIV. Nós randomizamos os pacientes em dois grupos. Os pacientes que receberam injeção de antagonista de GnRH em dose única no dia seguinte ao desencadeamento do hCG formaram o grupo caso, e os pacientes que receberam o regime de tratamento padrão formaram o grupo controle. Em ambos os grupos, os oócitos foram coletados 36 horas após a injeção de hCG. Resultados A taxa de ovulação prematura foi significativamente baixa no grupo caso em comparação com o grupo controle (6,86 versus 20,6% por ciclo programado) (p=0,022). Além disso, a taxa de recuperação de oócitos (93,14 versus 67,87% por ciclo programado) (p=0,013), a taxa de maturidade do oócito (79,42 versus 47,87%) (p=0,041), a taxa de fertilização (65,68 versus 34,54%) (p=0,018) e a taxa de transferência de embriões por ciclo programado (44,11 versus 18,78%) (p=0,003) foram maiores no grupo antagonista de GnRH do que no grupo controle. Conclusão A administração de antagonista de GnRH, no dia seguinte ao desencadeamento de hCG em tratamentos de FIV de pacientes com reserva ovariana diminuída permitiu uma redução significativa na taxa de ovulação precoce,mas não teve efeito na taxa de nascidos vivos.

Application of transabdominal wall press and vaginal B-ultrasound probe to adjust ovarian position in oocyte retrieval / 中国医师杂志

Objective:The aim of this study was to investigate the efficacy and safety of rtansabdominal wall press to adjust ovarian position in patients with difficulty in oocyte retrieval.Methods:A retrospective case-control study was conducted to collect 177 patients with difficulty in oocyte retrieval from January 2020 to December 2021 in the Reproductive Medical Center of Peking University Shenzhen Hospital. Among the 177 patients with difficulty in oocyte retrieval, 76 patients used transabdominal wall press to adjust the position of the ovary (pushing group), and the other 101 patients were treated with routine oocyte retrieval (control group). The oocyte retrieval rate, the number of oocyte and mature oocyte, the number of transferable embryos and high-quality embryos were compared between the two groups, and the incidence of complications was recorded.Results:There was no significant difference between the two groups in the ovarian stimulation program, the dosage of ovulation promoting drugs and the number of follicles ≥14 mm on human chorionic gonadotropin (HCG) day (all P>0.05). The days of promoting ovulation in the pushing group were longer than those in the control group, and the level of estradiol on HCG day was significantly higher than that in the control group (all P<0.05). The rate of oocyte retrieval, the number of oocytes retrieved, the number of transferable embryos and high-quality embryos in the pushing group were significantly higher than those in the control group (all P<0.05). The incidence of complications during oocyte retrieval in the pushing group was significantly lower than that in the control group ( P=0.003). Conclusions:Transabdominal wall press to adjust ovarian position can improve oocyte retrieval rate and reduce the incidence of complications in patients with difficulty in oocyte retrieval.

The effect of artificial oocyte activation on the short- and long-term development of offspring / 中华实用儿科临床杂志

Intracytoplasmic sperm injection (ICSI) is an important treatment option for male infertility at pre-sent.However, a few patients still suffer from repeated ICSI fertilization failure because their sperm is unable to activate oocytes.Artificial oocyte activation (AOA) technology can improve the fertilization rate, pregnancy rate, live birth rate, etc., but it remains unknown whether AOA has short- and long-term effect on offspring.In this article, recent literature about the effect of AOA technology on perinatal outcomes, genetics, physical development and neurological development of offspring was summarized.This paper aims to provide reference for reproductive medicine workers and pediatricians in clinical practice.

The effect of timing of oocyte denudation from oocyte retrieval in the total fertilization failure among in vitro fertilization - Intracytoplasmic sperm injection cycles

@#<p>Objective: This study aimed to evaluate the effect of oocyte incubation after retrieval in TFF among IVF-ICSI and identify factors affecting total fertilization failure (TFF).</p><p>Methods: This is a retrospective cohort study, involving 995 IVF cycles using the antagonist protocol that were clustered into three timings of oocyte denudation from retrieval : Group 1: <1 hour, Group 2: ?1 hour to <2hours and Group 3: ?2hours. Other variables considered were etiology of infertility, female age, days of stimulation and total number of oocytes retrieved.</p><p>Results: Overall TFF was 4.5%. TFF among groups were 4.8%, 5.8% and 3.2%, respectively. Multiple logistic regression analysis showed that oocyte incubation prior to denudation for ? 2 hours tend to decrease TFF incidence. Among factors studied, male factor infertility and a low number of oocytes adversely affect TFF.</p><p>Conclusion: Timing of incubation of oocyte did not significantly affect the occurrence of TFF. Among factors studied, male factor infertility and a low number of oocytes adversely affect TFF.</p>

Oocyte quality assessment in marine invertebrates: a novel approach by fluorescence spectroscopy

BACKGROUND: The assessment of oocyte quality is, nowadays, a major challenge in aquaculture, oocyte cryopreservation, and environmental science. Oocyte quality is a determining factor in fertilization and embryo development; however, there is still a lack of rapid and sensitive cellular markers for its assessment. Currently, its estimation is pre-dominantly based on morphological analysis, which is subjective and does not consistently reflect the developmental competence of the oocytes. Despite several recent studies investigating molecular markers related to oocyte quality, methods currently available for their determination pose various technical challenges and limitations. In this study, we developed a novel approach based on fluorescence spectroscopy to assess different intrinsic physiological parameters that can be employed to evaluate egg quality in marine invertebrates that are widely used as animal models such as sea urchins and mussels. RESULTS: Different physiological parameters, such as viability, mitochondrial activity, intracellular ROS levels, plasma membrane lipid peroxidation, and intracellular pH, for egg quality evaluation have been successfully assessed in sea urchins and mussels by using specific fluorescent dyes and detecting the fluorescent signals in eggs through fluorescence spectroscopy. CONCLUSIONS: Based on our findings, we propose these physiological markers as useful predictors of egg quality in marine invertebrates; they can be estimated rapidly, selectively, and sensitively by employing this novel approach, which, due to the speed of analysis, the low cost, and easy use can be considered a powerful analytical tool for the egg quality assessment.

Sperm DNA damage compromises embryo development, but not oocyte fertilisation in pigs

BACKGROUND: The assessment of sperm DNA integrity has been proposed as a complementary test to conventional mammalian semen analysis. In this sense, single-strand (SSB) and double-strand (DSB) DNA breaks, the two types of sperm DNA fragmentation (SDF), have been reported to have different aetiologies and to be associated to different fertility outcomes in bovine and humans. Considering that no studies in porcine have addressed how SDF may affect sperm quality and fertility outcomes, the present work aimed to determine the impact of global DNA damage, SSB and DSB on sperm quality and in vitro fertilising ability. To this end, 24 ejaculates (one per boar) were split into three aliquots: the first was used to assess sperm quality parameters through a computer-assisted sperm analysis (CASA) system and flow cytometry; the second was used to perform in vitro fertilisation, and the third, to evaluate sperm DNA integrity using alkaline and neutral Comet assays. RESULTS: The results showed that global DNA damage negatively correlates (P 0.05). CONCLUSION: Considering all these findings, this work sets a useful model to study how SDF negatively influences fertility.

Sudden death of an egg donor during oocyte retrieval due to ovarian hyperstimulation syndrome

ABSTRACT Ovarian Hyperstimulation Syndrome (OHSS) is uncommon among oocyte donors during in vitro fertilization (IVF) procedure and is rarely associated with death. We report a case of a 23-year-old oocyte donor who suddenly died on the operation table during oocyte retrieval. She had no risk factors in her menstrual history, laboratory, or clinical parameters. The antagonist cycle, triggered with the GnRH agonist protocol, was carried out. The cause of death at autopsy was attributed to respiratory failure due to acute massive pulmonary edema, which developed due to the complication of OHSS. Only a few autopsy cases associated with OHSS have been published, but, as far as we know, no clinical or autopsy cases of sudden death caused by OHSS have been reported.

In vitro maturation medium supplementation: utilization of repaglinide, L-carnitine, and mesenchymal stem cell-conditioned medium to improve developmental competence of oocytes derived from endometriosis mouse models

Endometriosis (EMS) is one of the most prevalent causes for female infertility. Herein, we investigated the effect of the repaglinide (RG), L-carnitine (LC), and bone marrow mesenchymal stem cell-conditioned medium (BMSC-CM) supplementation during in vitro maturation (IVM) on the quality, maturation, and fertilization rates, as well as embryonic quality and development of oocytes derived from normal and EMS mouse model. Immature oocytes were collected from two groups of normal and EMS-induced female NMRI mice at 6-8 weeks of age. Oocytes were cultured in IVM medium unsupplemented (control group), or supplemented with 1 M RG, 0.3 and 0.6 mg/mL LC, and 25 and 50% BMSC-CM. After 24 h of oocyte incubation, IVM rate and antioxidant status were assessed. Subsequently, the rates of fertilization, cleavage, blastulation, and embryonic development were assessed. Our results demonstrated that supplementation of IVM medium with LC and BMSC-CM, especially 50% BMSC-CM, significantly enhanced IVM and fertilization rates, and markedly improved blastocyst development and total blastocyst cell numbers in EMS-induced mice compared to the control group (53.28±0.24 vs 18.09±0.10%). Additionally, LC and BMSC-CM were able to significantly modulate EMS-induced nitro-oxidative stress by boosting total antioxidant capacity (TAC) and mitigating nitric oxide (NO) levels. Collectively, LC and BMSC-CM supplementation improved oocyte quality and IVM rates, pre-implantation developmental competence of oocytes after in vitro fertilization, and enhanced total blastocyst cell numbers probably by attenuating nitro-oxidative stress and accelerating nuclear maturation of oocytes. These outcomes may provide novel approaches to refining the IVM conditions that can advance the efficiency of assisted reproductive technologies in infertile couples.

Molecular Mechanism of Aurora Kinase A Regulating the Meiosis of Oocyte / 中国医学科学院学报

Aurora kinase A (AURKA),a family member of aurora kinases,is involved in mitotic entry,maturation and separation of centrosome,assembly and stabilization of bipolar spindle,and condensation and separation of chromosome.Studies have demonstrated that AURKA plays a similar role in meiosis,while the specific mechanism and the similarities and differences in its role between meiosis and mitosis remain unclear.Therefore,we reviewed the studies about the localization and activation of AURKA in oocyte meiosis,and compared the role of AURKA in regulating spindle formation,activating spindle assembly checkpoint,and correcting the kinetochore-microtubule attachment between the meiosis of oocytes and the mitosis of somatic cells.This review will lay a theoretical foundation for revealing the mechanism of AURKA in the regulation of cell division and for the clinical research related to cancer and reproduction.

El consumo semicrónico moderado de alcohol altera la foliculogénesis y la calidad núcleo-citoplasmática oocitaria, en el ratón / Moderate semi-chronic alcohol consumption alters folliculogenesis and oocyte nuclear-cytoplasmic quality in mice

Resumen El consumo crónico de alcohol es un problema de salud mundial que afecta particularmente a la población femenina. Sin embargo, los efectos de la ingesta semicrónica en cantidades moderadas a bajas en el ovario y el oocito son poco conocidos. En un modelo murino, se administró etanol al 10% en agua de bebida (hembras tratadas) o agua (hembras control) por 15 días, y luego de la superovulación o no (ovulación espontánea), se analizó el ciclo estral y la calidad ovárico-gamética. En las hembras tratadas, la frecuencia y duración del diestro aumentó, y las frecuencias de folículos y cuerpos lúteos disminuyeron vs hembras controles, valores que se restauraron luego de la superovulación. Sin embargo, en las hembras tratadas, la tasa de proliferación celular folicular y el desbalance de la expresión ovárica de VEGF (factor de crecimiento endotelial) persistieron luego de la superovulación. El número de ovocitos ovulados con metafase II anormal, fragmentados y activados partenogenéticamente fue mayor en las hembras tratadas respecto las controles. En conclusión, el consumo semicrónico moderado de alcohol produce anestro, ciclo estral irregular, foliculogénesis deficiente y anomalías núcleo-citoplasmáticas en los oocitos ovulados. Estas alteraciones podrían constituirse en un factor etiológico de pérdida gestacional temprana y desarrollo embrionario anormal luego del consumo de alcohol.

Abstract Chronic alcohol consumption is a global health problem that particularly affects the female population. However, the ef-fects of semi-chronic ethanol intake in low-moderate amounts on the ovary and oocyte are poorly understood. In a mouse model, 10% ethanol was administered in drinking water (treated females) or water (control females) for 15 days, and after superovulation or not (spontaneous ovulation), the estrous cycle and ovarian-gametic quality were analyzed. In treated females, the frequency and duration of the diestrus increased, and the frequencies of follicles and corpus luteum decreased vs control females, values that restored after superovulation. However, in treated females, the follicular cell proliferation rate and the imbalance in ovarian expression of VEGF (endothelial growth factor) persisted after superovulation. The number of ovulated oocytes with abnormal metaphase II, fragmented and parthenogenetically activated was higher in treated females than in control ones. In conclusion, moderate semi-chronic alcohol consumption produces anestrum, irregular estrous cycle, poor folliculogenesis, and nuclear-cytoplasmic abnormalities in ovulated oocytes. These alterations could constitute an etiological factor of early gestational loss and abnormal embryonic development after alcohol consumption.

Ex vivo retrieval of mature oocytes for fertility preservation in a patient with bilateral borderline ovarian tumor / Recuperação ex vivo de oócitos maduros para preservação da fertilidade em paciente com tumor ovariano borderline bilateral

Abstract We report a case of ultrasound-guided ex vivo oocyte retrieval for fertility preservation in a woman with bilateral borderline ovarian tumor, for whom conventional transvaginal oocyte retrieval was deemed unsafe because of the increased risk of malignant cell spillage. Ovarian stimulation with gonadotropins was performed. Surgery was scheduled according to the ovarian response to exogenous gonadotropic stimulation; oophorectomized specimens were obtained by laparoscopy, and oocyte retrieval was performed ~ 37 hours after the ovulatory trigger. The sum of 20 ovarian follicles were aspirated, and 16 oocytes were obtained.We performed vitrification of 12 metaphase II oocytes and 3 oocytes matured in vitro. Our result emphasizes the viability of ex vivo mature oocyte retrieval after controlled ovarian stimulation for those with high risk of malignant dissemination by conventional approach.

Resumo Relatamos um caso de obtenção ex vivo de óvulos, guiada por ultrassonografia, para preservação da fertilidade em uma mulher com tumor ovariano borderline bilateral, para quem a recuperação transvaginal convencional foi considerada insegura, devido ao aumento do risco de disseminação de célulasmalignas. Foi realizada estimulação ovariana com gonadotrofinas. A cirurgia foi agendada de acordo com a resposta ovariana à estimulação gonadotrófica exógena; após ooforectomia por laparoscopia, ~ 37 horas após a maturação folicular, procedeu-se à recuperação extracorpórea de oócitos. Umtotal de 20 folículos ovarianos foi aspirado e 16 complexos cumulus foramobtidos, resultando na vitrificação de 12 oócitos maduros e de 3 oócitos imaturos amadurecidos in vitro. Nosso resultado enfatiza a viabilidade da recuperação ex vivo de oócitos maduros após estimulação ovariana controlada para mulheres com alto risco de disseminação maligna pela captação oocitária realizada convencionalmente pela via transvaginal.

Follicular fluid from infertile women with mild endometriosis impairs in vitro bovine embryo development: potential role of oxidative stress / Fluido folicular de mulheres inférteis com endometriose leve prejudica o desenvolvimento in vitro de embriões bovinos: Potencial papel do estresse oxidativo

Abstract Objective To investigate whether follicular fluid (FF) from infertile women with mild endometriosis (ME) alters in vitro bovine embryo development, and whether the antioxidants N-acetyl-cysteine (NAC) and/or L-carnitine (LC) could prevent such damages. Methods Follicular fluid was obtained from infertile women (11 with ME and 11 control). Bovine oocytes were matured in vitro divided in: No-FF, with 1% of FF from control women (CFF) or ME women (MEFF); with 1.5mM NAC (CFF + NAC, MEFF + NAC), with 0.6mg/mL LC (CFF + LC, MEFF + LC), or both antioxidants (CFF + NAC + LC, MEFF + NAC + LC). After in vitro fertilization, in vitro embryo culture was performed for 9 days. Results A total of 883 presumptive zygotes were cultured in vitro. No differences were observed in cleavage rate (p = 0.5376) and blastocyst formation rate (p = 0.4249). However, the MEFF group (12.5%) had lower hatching rate than the No-FF (42.1%, p = 0.029) and CFF (42.9%, p = 0.036) groups. Addition of antioxidants in the group with CFF did not alter hatching rate (p ≥ 0.56), and in groups with MEFF, just NAC increased the hatching rate [(MEFF: 12.5% versus MEFF + NAC: 44.4% (p = 0.02); vs MEFF + LC: 18.8% (p = 0.79); versus MEFF + NAC + LC: 30.8% (p = 0.22)]. Conclusion Therefore, FF from infertile women with ME added to medium of in vitro maturation of bovine oocytes impairs hatching rate, and NAC prevented these damages, suggesting involvement of oxidative stress in worst of oocyte and embryo quality of women with ME.

Resumo Objetivo Investigar se o fluido folicular (FF) de mulheres inférteis com endometriose leve (ME, na sigla eminglês) altera o desenvolvimento in vitro de embriões bovinos, e se os antioxidantes N-acetil-cisteína (NAC) e/ou L-carnitina (LC) poderiam prevenir possíveis danos. Métodos O FF foi obtido de mulheres inférteis (11 com ME e 11 controles). Oócitos bovinos foram maturados in vitro divididos em: sem FF (No-FF), com 1% de FF de mulheres controle (CFF) ou mulheres comME (MEFF); com 1,5mMde NAC (CFF + NAC, MEFF + NAC), com 0,6mg/mL de LC (CFF + LC, MEFF + LC), ou ambos antioxidantes (CFF + NAC + LC, MEFF + NAC + LC). Depois da fertilização in vitro, o cultivo in vitro de embriões foi realizado por 9 dias. Resultados Um total de 883 zigotos presumidos foram cultivados in vitro. Nenhuma diferença foi observada na taxa de clivagem (p = 0,5376) e na taxa de formação de blastocistos (p = 0,4249). Entretanto, o grupo MEFF (12.5%) teve menor taxa de eclosão de blastocistos do que os grupos No-FF (42,1%, p = 0,029) e CFF (42,9%, p = 0,036). Adição de antioxidantes no grupo comCFF não alterou a taxa de eclosão (p ≥ 0.56), e nos grupos com MEFF, somente a NAC aumentou a taxa de eclosão [(MEFF: 12.5% versus MEFF + NAC: 44.4% (p = 0.02); versus MEFF + LC: 18.8% (p = 0.79); versus MEFF + NAC + LC: 30.8% (p = 0.22)]. Conclusão Portanto, o FF de mulheres inférteis com ME adicionado ao meio de maturação in vitro de oócitos bovinos prejudica a taxa de closão embrionária, e a NAC preveniu esses danos, sugerindo o envolvimento do estresse oxidativo na piora da qualidade oocitária e embrionária de mulheres com ME.

ADAMTS proteoglycanases downregulation with impaired oocyte quality in PCOS

ABSTRACT Objective: A disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS-4) and ADAMTS-5 normal expression levels are essential for ovulation and subsequent fertilization. The objective of the present study was to assess expression pattern of these genes in cumulus cells (CCs) taken from patients with polycystic ovary syndrome (PCOS) and to investigate any possible relationship with the oocyte quality. Subjects and methods: ADAMTS-4 and -5 expression levels within CCs containing oocytes at the metaphase II (MII) and germinal vesicle (GV) stages, taken from 35 patients with PCOS and 35 women with normal ovarian function, were investigated using RT-qPCR. Moreover, possible correlations between ADAMTS-4, ADAMTS-5, and progesterone receptors (PRs) expression as well as oocyte quality were evaluated. Results: ADAMTS-4 and -5 expression levels were dramatically diminished in the CCs of the PCOS patients when compared to the controls. ADAMTS-4 and -5 expression levels were correlated with each other and with the oocyte quality. Furthermore, lower expression levels of ADAMTS-4 and -5 in the PCOS patients were strongly correlated with the diminished PRs expression levels. Conclusions: Downregulation of ADAMTS-4 and -5 in the human CCs of the PCOS patients correlated with the decline in the PRs expression, and impaired oocyte quality may cause lower oocyte recovery, maturation, and fertilization rate.