ABSTRACT The recognition of oocytes, in their maturational stages, allow estimate the ovarian development and the type of spawning of a species. Although, distinguishing oocytes on histological images requires a visual and subjective interpretation by the specialist. With the development of deep learning techniques, automatic object detection has become an important mechanism for this task. However, studies that use deep learning techniques have not been widely explored for the analysis of fish oocyte samples so far. In this paper, we propose the use of YOLO, a family of convolutional neural networks, for oocyte morphology detection of Centropomus undecimalis fish. The research uses an image database with 5,680 oocytes with different maturation stadiums (PV - pre-vitellogenesis, VI - early vitellogenesis and VF - late vitellogenesis), in histological images, divided into training, testing and validation, and detection performed by YOLOv3, YOLOv4, and YOLOv5 architectures. The results obtained were promising, highlighting that the YOLOv5l model, in the detection of oocytes of the VF class, reached the best values in the metrics precision, recall, mAP@.5 and mAP@.95 with 85.4%, 95.3%, 95.7%, and 75.9%, respectively. When considering all classes, YOLOv5l was the model that obtained the best results in the analyzed metrics.

RESUMO O fluido folicular é um abrigo que proporciona um microambiente para o desenvolvimento e a maturação dos oócitos. É um meio que proporciona um meio de comunicação dentro das células foliculares. A função do fluido folicular no desenvolvimento e na maturação de folículos e oócitos envolve a ação de determinados genes. Aqui, examinamos o impacto da CFF na expressão gênica de oócitos de camelo durante a (IVM). Hipoteticamente, o meio de MIV foi aprimorado com 2,5% de CFF e analisamos a expressão de vários genes, incluindo Bax, Bcl-2, P53, Acta2, TGFβ, Cx43 e Tagln. Consequentemente, o fluido folicular de camelos teve um efeito substancial sobre o nível de transcrição de mRNA do gene BCL2, que está relacionado à apoptose. Os genes Acta2 e Tagln, relacionados à proteína do citoesqueleto “actina” na via do fator de crescimento transformador beta (TGFβ), também foram modulados. No entanto, o fluido folicular aumentou a expressão de TGFβ em oócitos de camelo. Este estudo mostra os efeitos benéficos da CFF na expressão gênica de oócitos de camelo e fornece uma plataforma para o estudo dos muitos mecanismos envolvidos na meiose de oócitos, particularmente aqueles associados à via do TGFβ. Recomendamos o uso de dosagens mais altas e a medição dos níveis de expressão de determinados genes envolvidos na morte celular programada e na inflamação.

ABSTRACT Follicular fluid is a haven that provides a microenvironment for the development and maturation of oocytes. It is a medium that provides a means of communication within the follicular cells. The role of follicular fluid in the development and maturation of follicles and oocytes involves the actions of certain genes. Here, we examined the impact of CFF on the gene expression of camel oocytes during (IVM). Hypothetically, IVM medium was enhanced with 2.5% CFF, and we analyzed the expression of several genes including Bax, Bcl-2, P53, Acta2, TGFβ, Cx43, and Tagln. Consequently, the follicular fluid from camels had a substantial effect on the mRNA transcript level of the BCL2 gene, which is related to apoptosis. Acta2 and Tagln genes that relate to cytoskeletal protein “actin” in transforming growth factor beta (TGFβ) pathway were also modulated. However, the follicular fluid increased the expression of TGFβ in camel oocytes. This study showcases the beneficial effects of CFF on camel oocyte Gene Expression and provides a platform for studying the many mechanisms involved in oocyte meiosis, particularly those associated with the TGFβ pathway. We recommend using higher dosage and measuring the expression levels of certain genes involved in programmed cell death and inflammation.

SUMMARY: The role of neutrophil extracellular traps (NETs) and monocyte extracellular traps (METs) is well established in fertilization disorders by adverse effects on sperm motility, acrosome integrity, membrane lipoperoxidation, and disturbing oocyte sperm binding. Moreover, NETs/METs remove efficiently excessive spermatozoa within the female reproductive tract (FRT) after natural coitus or artificial insemination (AI). Depending on their location in reproductive organs, NETs/METs may be beneficial to the host by their capability of either limiting the dissemination of infective pathogens, promoting wound healing, and exerting anti-inflammatory properties through the degradation of pro-inflammatory components. Conversely, excessive or imbalanced NETs/METs release within FRT or urogenital tract (UGT) might be associated with worse fertility due to deleterious effects on sperm functions, oocyte fertilization, endometrium microenvironment, and embryo implantation. In the UGT, patients with epididymitis or urogenital bacterial infections and with leukocytospermia spontaneous release of NETs/METs in seminal fluids. Moreover, in infertile male/female patients without infectious etiologies but with autoimmune (i. e. systemic lupus erythematosus, rheumatoid arthritis)-, metabolic (diabetes mellitus, obesity)- and vascular (vasculitis, hypertension, and preeclampsia)- diseases uncontrolled NETs/METs might negatively influence their fertility. Thus, this review aims to provide novel insights into the beneficial as well as adverse effects of NETs/METs on male gametes, oocyte fertilization, and endometrium by highlighting broad implications on early diagnosis of extracellular traps (ETs) in seminal fluid samples of infertility patients with inflammatory etiologies. As well as, for early diagnosis of spontaneous formation of NETs/METs in infertile couples with one or both being affected with chronic inflammatory-, metabolic-, vascular- and/or autoimmune diseases to increase the chance of conception.

RESUMEN: El papel de las trampas extracelulares de neutrófilos (NETs) y las trampas extracelulares de monocitos (METs) está bien establecido en los trastornos de la fertilización por efectos adversos sobre la motilidad de los espermatozoides, la integridad del acrosoma, la lipoperoxidación de la membrana y la alteración de la unión de los espermatozoides al ovocito. Además, las NETs/METs eliminan de manera eficiente el exceso de espermatozoides dentro del tracto reproductivo femenino (FRT) después del coito natural o la inseminación artificial (IA). Dependiendo de su ubicación en los órganos reproductivos, las NET/MET pueden ser beneficiosas para el huésped por su capacidad de limitar la diseminación de patógenos infecciosos, promover la cicatrización de heridas y ejercer propiedades antiinflamatorias a través de la degradación de componentes proinflamatorios. Por el contrario, la liberación excesiva o desequilibrada de NETs/METs dentro del FRT o el tracto urogenital masculino (UGT) podría estar asociada con una baja fertilidad debido a efectos nocivos sobre las funciones de los espermatozoides, la fertilización de los ovocitos, el microambiente del endometrio y la implantación del embrión. En UGT, los pacientes con epididimitis o infecciones bacterianas urogenitales y con leucocitospermia presentan liberación espontánea de NETs/ METs en los fluidos seminales. Además, en pacientes masculinos/ femeninos infértiles sin etiologías infecciosas pero con enfermedades autoinmunes (es decir, lupus eritematoso sistémico, artritis reumatoide), metabólicas (diabetes mellitus, obesidad) y vasculares (vasculitis, hipertensión y preeclampsia), los NETs/ METs no controlados pueden influir negativamente en su fertilidad. Por lo tanto, esta revisión tiene como objetivo proporcionar nuevos conocimientos sobre los efectos beneficiosos y adversos de los NETs/METs en los gametos masculinos, la fertilización de ovocitos y el endometrio, destacando las amplias implicaciones en el diagnóstico temprano de trampas extracelulares (TE) en muestras de fluido seminal de pacientes infértiles con etiologías inflamatorias. Así como para el diagnóstico precoz de la formación espontánea de NETs/METs en parejas infértiles en las que uno o ambos padecen enfermedades inflamatorias, metabólicas, vasculares y/o autoinmunes crónicas, con el fin de aumentar las posibilidades de concepción.

ABSTRACT. The spotted rose snapper (Lutjanus guttatus) is a commercially important species that provides economic income to small-scale fisheries in the Pacific coastal regions of Guatemala; however, there are no studies on this species that establish a baseline of knowledge on their reproductive aspects and potential management measures. The present study was carried out to obtain the first knowledge on the reproductive behavior of this resource, which will contribute to the establishment of future management strategies for the fisheries. Aspects of gonadal-maturity phases and reproductive indicators such as the condition factor, gonadosomatic index, and temporal oocyte diameter were analyzed to estimate reproductive development and the size at first maturity. The results suggest that the spotted rose snapper has two well-defined reproductive periods - from October to December and March to May. These periods were closely related to temperatures that ranged from 29 to 31°C. In addition, according to the monthly monitoring of the oocyte diameter density distribution, it is suggested that spotted rose snapper in the region could participate in up to three spawning events during the reproductive season, with gamete development occurring throughout most of the entire study period. The first size at maturity was a total length (TL) of 33.49 cm (confidence interval = 30.95-36.14 cm). This information is important for sustainable fishery management and provides a baseline for future research.

Resumo Introdução e Objetivos: A maternidade tardia é hoje uma realidade, principalmente para mulheres com níveis de escolaridade superiores. A criopreservação eletiva de ovócitos permite diminuir o risco de infertilidade futura. A maioria das mulheres não está adequadamente informada sobre o declínio da fertilidade associado à idade nem sobre os fatores de risco associados. O objetivo do estudo foi avaliar o conhecimento e atitudes em relação à maternidade, fertilidade e à criopreservação eletiva de ovócitos das jovens universitárias em Portugal. Tipo de estudo: Estudo observacional e descritivo. População: Estudantes universitárias do sexo feminino. Metodologia: Dados recolhidos através de inquérito online Google Forms® realizado durante os meses de agosto e setembro de 2023. Resultados: Foram incluídas no estudo 266 estudantes do sexo feminino, com uma média de idade de 22,46 anos (± 4,2). Quando inquiridas quanto à vontade de terem filhos no futuro, 65% (n=173) das participantes respondeu “sim”, 24,8% (n=66) “talvez” e 10,2% (n=27) “não”. O principal motivo apontado para as respostas “não” e “talvez” foi a priorização da carreira (68,8%). Cerca de 53,6% (n=121) das participantes apenas desejam ter filhos a partir dos 30 anos. Para 69,2% (n=184) a fertilidade futura é uma preocupação, 72,9% (n=194) está familiarizada com o conceito de reserva ovárica e 82,7% (n=220) com a existência de métodos de preservação da fertilidade. Quando questionadas quanto à possibilidade de recorrer à criopreservação eletiva de ovócitos, apenas 14,3% (n=38) exclui essa possibilidade. A maioria das jovens (87,2%, n=232) gostaria de estar mais informada sobre a temática e 87,6% (n=233) gostaria de ver discutido o tema com o seu médico. Conclusões: A maioria das estudantes equaciona a maternidade, contudo a carreira profissional é destacada como principal prioridade para os próximos 10 anos. O conhecimento sobre declínio da reserva ovárica e eficácia da criopreservação de ovócitos está ainda aquém do desejado.

Abstract Overview and Aims: Delayed motherhood is a reality today, especially for women with higher levels of education. Elective oocyte cryopreservation reduces the risk of future infertility. Despite increasing discussions in the media and social networks, most women are not adequately informed about the fertility decline associated with age and the associated risk factors. The aim of our study was to access the knowledge and attitudes regarding motherhood, fertility, and elective oocyte cryopreservation among young female university students in Portugal. Study Design: Observational and descriptive study. Population: Female college students. Methods: Data were collected through an online Google Forms® survey during August and September 2023. Results: A total of 266 female students were included in the study, with an average age of 22.46 years (±4.2). When asked about their willingness to have children in the future, 65% (n=173) of participants responded “yes”, 24.8% (n=66) “maybe”, and 10.2% (n=27) “no”. The main reasons cited for “no” and “maybe” responses were career prioritization (68.8%), fear of childbirth (45.2%), fear of motherhood (43%) or socioeconomic issues (40%). Approximately 53.6% (n=121) of participants wished to have children after the age of 30 years. Concerns about future fertility were present in 69.2% (n=184) of participants, and 72.9% (n=194) were familiar with the concept of ovarian reserve, while 82.7% (n=220) were aware of fertility preservation methods. When asked about the possibility of opting for elective oocyte cryopreservation, only 14.3% (n=38) ruled out this possibility. Most young women (87,2%, n=232) wished to be more informed about the topic, and 87.6% (n=233) would like to discuss the issue with their gynaecologist. Conclusions: The majority of students considered motherhood, however their professional career was highlighted as the main priority for the next 10 years. Knowledge about the ovarian reserve decline and the effectiveness of oocyte cryopreservation was not satisfactory.

BACKGROUND. One of the biggest issues in reproductive medicine is treating women with low ovarian reserves or a poor ovarian response (POR) to stimulation. Oxidative stress was suggested as one of the major contributors to POR. However, the pathophysiology remains unknown. Focusing on certain subpopulations within the diverse group of poor responders may help determine the best therapy for these patients. OBJECTIVE. To investigate the effect of adding coenzyme A Q10 to dehydroepiandrosterone (DHEA) in young, poor responder infertile females undergoing in vitro fertilisation (IVF)/intracytoplasmic sperm injection (ICSI) cycles. METHODS. One hundred and sixty-eight females with poor ovarian response (Poseidon Group 3) from multiple centres were randomly divided into two groups. The first group received DHEA orally along with coenzyme 10, while the second group received only DHEA. Both groups underwent controlled ovarian stimulation. RESULTS. There was a significant improvement in the quality of oocytes and the number and quality of embryos transferred in the coenzyme A Q10 group. However, no significant differences were observed in other parameters between the groups. CONCLUSION. In young, poor-responder women with decreased ovarian reserve, pretreatment with CoQ10 enhances oocyte and embryo quality and boosts ovarian response to stimulation. An improvement in clinical pregnancy and live birth rates may be possible.

RESUMEN El estudio tuvo como objetivo evaluar el efecto del suero fetal bovino (FCS) agregado al medio de cultivo los días 5 a 7 después de la fertilización in vitro sobre la producción y posterior criotolerancia de blastocistos de vaquillonas criollas ecuatorianas. Se recolectaron complejos cúmulo-ovocitos (COCs) inmaduros mediante aspiración intravaginal guiada por ultrasonido (OPU) de 10 vaquillonas criollas y de ovarios de matadero en ocho sesiones. Los COCs se sometieron a maduración in vitro (MIV), fecundación (FIV) y cultivo (IVC). El día 1 después de la FIV, los presuntos embriones de cada fuente de ovocitos (OPU [O] y Matadero [A]) se asignaron aleatoriamente en dos grupos según si se añadió FCS al 2.5% (v/v) al medio de cultivo el día 5 después de la FIV: 1) O-FCS+, 2) O-FCS-, 3) A-FCS+ y 4) A-FCS-. El día 7 después de la FIV se vitrificaron embriones de alta calidad. La criotolerancia de los embriones vitrificados-calentados se evaluó según la reexpansión del blastocele a las 2 h y la posterior reexpansión y eclosión a las 24 y 48 h de incubación. La tasa de blastocistos el día 7 no difirió entre la fuente de ovocitos y el grupo FCS. Después del proceso de vitrificación/ calentamiento, la adición de FCS solo afectó la tasa de reexpansión a las 2 h, independientemente de la fuente de ovocitos (p<0.05). Asimismo, la tasa de eclosión de los blastocistos a las 48 h de incubación se vio drásticamente afectada únicamente en los ovocitos derivados de OPU (p<0.01). En conclusión, la suplementación de FCS el día 5 después de la FIV no mejoró la producción de blastocistos y afectó negativamente la criotolerancia de embriones bovinos in vitro derivados de vaquillonas criollas criadas en los Andes ecuatorianos.

ABSTRACT This experiment aimed to assess the effect of fetal calf serum (FCS) added to culture medium on day 5 to 7 after in vitro fertilization on production and subsequent cryotolerance of blastocysts from Ecuadorian Creole heifers. Immature cumulus-oocyte complexes (COCs) were collected by ovum pick-up (OPU) from 10 Creole heifers and from ovaries collected at abattoir in eight collection sessions. COCs were subjected to in vitro maturation (IVM), fertilization (IVF), and culture (IVC). On day 1 after IVF, presumptive embryos from each oocyte source (OPU [O] and Abattoir [A]) were allocated randomly in two groups according to whether 2.5% (v/v) FCS was added to culture medium on day 5 after IVF: 1) O-FCS+, 2) O-FCS-, 3) A-FCS+, and 4) A-FCS-. On day 7 after IVF, high quality embryos were vitrified. Cryotolerance of vitrified-warmed embryos was assessed according to blastocele re-expansion at 2 h and subsequent re-expansion and hatching at 24 and 48 h of re-incubation. Blastocysts rate on day 7 did not differ between oocyte source and FCS group. After vitrification/warming process, addition of FCS affected the re-expansion rate only at 2 h, irrespective of the oocyte source (p<0.05). Likewise, blastocysts hatching rate at 48 h of incubation was drastically affected only in OPU derived oocyte (p<0.01). Supplementation of FCS on day 5 after IVF did not improve blastocyst production and adversely affected the cryotolerance of in vitro bovine embryos derived from creole heifers raised on Ecuadorian Andean highlands.

BACKGROUND: Rhinoceros are currently one of the most threatened mammal species globally. Slow population growth, increased poaching and habitat destruction have led to increased conservation efforts for each species. Assisted reproductive technologies (ART) have been implemented in an attempt to aid reproductive outputs for the conservation of these endangered species. Developing species-specific ART programmes for wildlife have been challenging. Temperature control during oocyte recovery is essential for ensuring in vitro success OBJECTIVE: This study is the first to investigate the effect of enema warming prior to trans-rectal ovum pick-up (OPU) on in vitro oocyte maturation in Southern white rhinoceros (Ceratotherium simum simum). METHODS: OPUs were performed on 20 rhinoceros cows from three different game farms in South Africa; oocytes were transported to one of two in vitro fertilisation laboratories for culture. The enema fluid was either warmed to 32 °C or not warmed prior to the OPU. Location of the farm, the different laboratories, ambient temperature, season, aspiration probe temperature, media type and enema temperature were investigated as predictor variables for oocyte maturation success RESULTS: After considering all other potential covariates, warming of the enema fluid was the only independent predictor of in vitro oocyte maturation success during this study CONCLUSION: Oocytes retrieved from rhinoceros cows that received an enema warmed to 32 °C were 2.3 times more likely to mature in vitro compared to oocytes from cows that received an unwarmed enema; the findings can be implemented in other rhinoceros ART programmes and in conservation efforts of other endangered mammalian species

BACKGROUND: The implementation of assisted reproductive techniques and use of in vitro embryo production has highlighted their potential in attempts to prevent the extinction of the northern white rhinoceros (Ceratotherium simum cottoni). However, in vitro embryo production has proved challenging. Transrectal ovum pick-up (OPU), while done successfully with custom-made aspiration probes (AP), may expose oocytes to detrimental temperature shock OBJECTIVE: This study compares the temperature regulation properties between two AP, made from different materials and used during OPU in southern white rhinoceros (C. s. simum METHODS: The initial PVC plastic AP, produced using a computer numerical control machine, was compared to a modified aluminium AP. A rhinoceros model was constructed using a 210 L water container fitted with heating elements and a water pump. Water was warmed to 37.5-38.5 °C to simulate rhinoceros body temperature. The AP was equilibrated to room temperature (22 simulating summer) or in a walk-in fridge (4 simulating winter) overnight. Temperature loggers were placed inside and on the outside of the AP RESULTS: The aluminium AP equilibrated to the model water temperature within 15 minutes of submersion in the model, while the temperature of the PVC AP remained significantly lower during the three-hour period. The inside of the PVC AP remained significantly colder compared to outside during both winter and summer (33.23 ± 4.28°C vs. 36.51 ± 2.13 °C and 36.04 ± 1.69 °C vs. 37.22 ± 0.92 °C, respectively). There was no significant difference between the inside temperature of the aluminium AP compared to the outside temperature during both the winter and summer simulations (36.75 ± 0.49 °C vs. 37.83 ± 0.40 °C and 37.86 ± 0.16 °C vs. 38.04 ± 0.11 °C, respectively CONCLUSION: This research highlights the importance of using material with good thermal properties during assisted reproductive procedures. It also contributes to further advancement during the establishment of a successful in vitro embryo production protocol for rhinoceros in South Africa

Abstract 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.

Abstract Background Helicase for meiosis 1 (HFM1), a putative DNA helicase expressed in germ-line cells, has been reported to be closely associated with premature ovarian insufficiency (POI). However, the underlying molecular mechanism has not been clearly elucidated. The aim of this study was to investigate the function of HFM1 in the first meiotic prophase of mouse oocytes. Results The results suggested that the deficiency of HFM1 resulting in increased apoptosis and depletion of oocytes in mice, while the oocytes were arrested in the pachytene stage of the first meiotic prophase. In addition, impaired DNA double-strand break repair and disrupted synapsis were observed in the absence of HFM1. Further investigation revealed that knockout of HFM1 promoted ubiquitination and degradation of FUS protein mediated by FBXW11. Additionally, the depletion of HFM1 altered the intranuclear localization of FUS and regulated meiotic- and oocyte development-related genes in oocytes by modulating the expression of BRCA1. Conclusions These findings elaborated that the critical role of HFM1 in orchestrating the regulation of DNA double-strand break repair and synapsis to ensure meiosis procession and primordial follicle formation. This study provided insights into the pathogenesis of POI and highlighted the importance of HFM1 in maintaining proper meiotic function in mouse oocytes.

Abstract Follicular dynamics in dairy sheep is poorly understood. This study assessed the follicular dynamics of dairy sheep within and out of the breeding season by observing the effect of two progesterone concentrations (60 and 120 mg) used in vaginal pessaries in pre-synchronization. The experiment was conducted in April/May and October/November using 40 healthy multiparous dairy ewes predominantly of the Lacaune breed, which underwent daily transrectal ultrasound examinations. The number of ovulations (1.35 vs 1.05), ovulatory follicle diameter (mm) (5.97 vs 5.05), length of the luteal phase (11 vs 9.14 days), and length of the estrous cycle (16.83 vs 16.6 days) were assessed within and out of the breeding season, respectively. The sheep out of the breeding season did not present a corpus luteum at the time of insertion of the vaginal pessary, characterizing anestrus. The ovulatory follicle had a larger diameter in the breeding season even with the shorter length of the follicular phase, suggesting higher viability and quality of the ovulated oocyte. The application of exogenous progesterone extends the luteal phase in the breeding season (11 vs 9.14 days). The anestrus observed out of season suggests the need for a higher progesterone concentration in reproductive protocols. Therefore, breeding protocols can be adapted for different seasons, suggesting the use of vaginal pessaries with a higher progesterone concentration (120 mg) out of the breeding season. The data allowed the characterization of follicular dynamics in dairy sheep, demonstrating their differences in each season based on the use of different progesterone doses in pre-synchronization. understood 60 (6 12 mg presynchronization. presynchronization pre synchronization. synchronization pre-synchronization AprilMay April May OctoberNovember October November 4 breed examinations 1.35 135 1 35 (1.3 1.05, 105 1.05 , 05 1.05) mm (mm 5.97 597 5 97 (5.9 5.05, 505 5.05 5.05) 11 (1 914 9 14 9.1 days, days 16.83 1683 16 83 (16.8 166 6 16. respectively pessary oocyte days. . Therefore seasons (12 ( 1.3 13 3 (1. 10 1.0 0 5.9 59 (5. 50 5.0 91 9. 16.8 168 8 (16. 1. 5. (5 (16

Resumo A dinâmica folicular em ovelhas leiteiras é pouco compreendida. Neste trabalho foi avaliado a dinâmica folicular de ovelhas leiteiras, dentro e fora da estação reprodutiva observando o efeito de duas concentrações de progesterona (60 e 120mg) utilizadas em pessários vaginais na pré-sincronização. O experimento foi realizado nos meses de abril/maio e outubro/novembro, utilizando 40 ovelhas multíparas leiteiras hígidas, predominantemente da raça Lacaune, que foram submetidas a exames ultrassonográficos diários por via transretal. Na estação reprodutiva foram avaliados o número de ovulações (1,35 vs 1,05), o diâmetro(mm) do folículo ovulatório (5,97 vs 5,05), a duração da fase lútea (11 vs 9,14 dias) e a duração do ciclo estral (16,83 vs 16,6 dias) dentro da estação em comparação com fora da estação reprodutiva respectivamente. Fora da estação as ovelhas não apresentaram corpo lúteo no momento da inserção do pessário vaginal, caracterizando anestro. Mesmo com menor duração da fase folicular, o folículo ovulatório apresentou maior diâmetro na estação reprodutiva, sugerindo maior viabilidade e qualidade do oócito ovulado. A aplicação de progesterona exógena amplia a fase lútea na estação reprodutiva (11 vs 9,14 dias). O anestro observado fora de época sugere a necessidade de maior concentração de progesterona nos protocolos reprodutivos. Desta forma, é possível adaptar protocolos reprodutivos para diferentes épocas, sugerindo o uso de pessários vaginais com maior concentração de progesterona (120 mg) fora da estação reprodutiva. Os dados obtidos permitiram caracterizar a dinâmica folicular em ovelhas leiteiras, demonstrando suas diferenças em cada estação a partir do uso de diferentes doses de progesterona na pré-sincronização compreendida 60 (6 120mg mg présincronização. présincronização pré sincronização. sincronização abrilmaio abril maio outubronovembro outubro novembro outubro/novembro 4 hígidas Lacaune transretal 1,35 135 1 35 (1,3 1,05, 105 1,05 , 05 1,05) diâmetromm mm diâmetro(mm 5,97 597 5 97 (5,9 5,05, 505 5,05 5,05) 11 (1 914 9 14 9,1 dias 16,83 1683 16 83 (16,8 166 6 16, respectivamente vaginal ovulado dias. . forma épocas 120 (12 ( 1,3 13 3 (1, 10 1,0 0 5,9 59 (5, 50 5,0 91 9, 16,8 168 8 (16, 12 1, 5, (5 (16

Abstract Lipid metabolism is essential for ensuring oocyte maturation and embryo development. β-Oxidized fatty acids (FA) are a potent source of energy for cells, particularly for bovine somatic follicular cells. Superstimulatory protocols using follicle stimulating hormone (FSH) or FSH combined with equine chorionic gonadotropin (eCG) are capable of stimulating the follicular microenvironment and drive the expression of biomarker genes associated with lipid metabolism in the cumulus-oocyte complex (COC) for better embryo development. In this study, we assesed the effects of FSH and FSH/eCG protocols on the expression of genes related to lipid metabolism in bovine granulosa cells (GCs). Further, we measured triglyceride levels in follicular fluid (FF) obtained from both superstimulatd and non-superstimulated cows (synchronized cows). In summary, superstimulation with gonadotropins maintained the TG levels in bovine FF and ensured GCs mRNA abundance of ACSL1, ACSL3, ACSL6, SCD, ELOVL5, ELOVL6, FASN, FADS2, and SREBP1. We, however, found the abundance of CPTIB mRNA to be lower in GCs obtained from cows subjected to FSH/eCG protocols than synchronized cows. In conclusion, the findings of this study showed that ovarian superstimulation around the preovulatory phase has a mild impact on the lipid metabolism in GCs. development βOxidized β Oxidized FA (FA (FSH eCG (eCG cumulusoocyte cumulus COC (COC FSHeCG . (GCs) Further (FF nonsuperstimulated non superstimulated cows) summary ACSL1 ACSL ACSL3 ACSL6 SCD ELOVL5 ELOVL ELOVL6 FASN FADS2 FADS SREBP1 SREBP We however conclusion (GCs

Abstract During oocyte meiosis resumption, a coordinated program of transcript translation and decay machinery promotes a remodeling of mRNA stores, which determines the success of the acquisition of competence and early embryo development. We investigated levels of two genes related to mRNA translation (CPEB1 and CPEB4) and two related to mRNA degradation (CNOT7 and ZFP36L2) machinery and found ZFP36L2 downregulated in in vitro-matured bovine oocytes compared to in vivo counterparts. Thereafter, we tested the effects of a pre-IVM step with NPPC and a modified IVM with AREG on the modulation of members of mRNA translation and degradation pathways in cumulus cells and oocytes. Our data showed a massive upregulation of genes associated with translational and decay processes in cumulus cells, promoted by NPPC and AREG supplementation, up to 9h of IVM. The oocytes were less affected by NPPC and AREG, and even though ZFP36L2 transcript and protein levels were downregulated at 9 and 19h of IVM, only one (KDM4C) from the ten target genes evaluated was differently expressed in these treatments. These data suggest that cumulus cells are more prone to respond to NPPC and AREG supplementation in vitro, regarding translational and mRNA decay programs. Given the important nursing role of these cells, further studies could contribute to a better understanding of the impact of these modulators in maternal mRNA modulation and improve IVM outcomes. resumption stores development CPEB1 CPEB (CPEB CPEB4 CNOT7 CNOT (CNOT ZFPL ZFP L ZFP36L vitromatured vitro matured counterparts Thereafter preIVM pre h KDM4C KDMC KDM C (KDM4C treatments programs outcomes

Objective: Fertility preservation is a priority in oncology for female cancer patients. However, there is a lack of communication between infertility specialists and oncologists. This study aimed to evaluate infertility specialists’ perceptions and experiences regarding fertility preservation. Methods: Conduct an online survey to profile infertility specialists. Participants were infertility affiliated with the Brazilian Federation of Gynecology and Obstetrics Associations (FEBRASGO). The specialists received an online survey, which response rate were 30.9%, most of whom were in southern and southeastern. The survey consisted on 14 questions about the infertility specialists’ location, techniques in clinical practice, treatment successful rate, patients idea, etc. Results: The average experience in human reproduction were 15.5 ± 10.2 years (mean ± standard deviation, range 1-40). Among reproductive-aged female cancer patients recommended for fertility preservation, 60.3 ± 28.8% (range 10-100%) underwent preservation procedures. Main barriers were cost (41%), oncologists’ knowledge or acceptance (35%) and accessibility (9%). Most infertility specialists (58%) considered 40 years the limit for fertility preservation. Leukemia, lymphoma, breast and ovarian cancers were prioritized for fertility preservation, while lung, thyroid, gastric, and brain cancers were less relevant. Conclusion: This is the first Brazilian study about infertility specialists’ perceptions on oncology patients access to fertility preservation. These patients primarily receive treatment in the public health system, while infertility specialists mainly work in the private healthcare. This healthcare mode is currently fragmented, but integrating these experts is enhancing patient access to fertility preservation. Studies on this topic are still warranted. Objective However oncologists Methods FEBRASGO. FEBRASGO . (FEBRASGO) 309 30 9 30.9% southeastern 1 location practice idea etc Results 155 15 5 15. 102 10 2 10. mean deviation 140. 140 1-40) reproductiveaged reproductive aged 603 60 3 60. 288 28 8 28.8 10100% 10100 100% 100 10-100% procedures 41%, 41 41% , (41%) 35% 35 (35% 9%. 9% (9%) 58% 58 (58% 4 Leukemia lymphoma lung thyroid gastric relevant Conclusion system fragmented warranted (FEBRASGO 30.9 1-40 6 28. 1010 10-100 (41% (35 (9% (58 30. 1-4 101 10-10 (41 (3 (9 (5 1- 10-1 (4 ( 10-