Abstract Introduction: Placenta succenturiata (PS) is a rare placenta abnormalities whose prevalence is 1.2%. It appears morphologically with a main lobe and one or several accessory lobes of various sizes joined together by blood vessels. The main risk factors correspond to advanced maternal age and in vitro fertilization. The diagnosis is made clinically after delivery. PS presents important obstetric risks such as postpartum hemorrhage and infection due to retained accessory lobe. Case presentation: It is presented the case of a 35-year-old woman, multiparous of 1, in single pregnancy with a diagnosis of fetal growth restriction and Rh (–) not sensitized. At 26 + 6 weeks a placental malformation is detected with a posterior placental body and accessory lobe on the anterior surface connected to arteriovenous vessels in the uterine fundus, findings compatible with PS. At 37 + 6 weeks the patient had a vaginal delivery; at delivery the diagnosis of PS was confirmed. Conclusion: PS is a rare and underdiagnosed placental malformation that presents significant obstetric risks. Antepartum ultrasound diagnosis of placenta abnormalities is essential to avoid unfavorable maternal and fetal outcomes.

Resumen Introducción: La placenta succenturiata (PS) es una malformación placentaria poco frecuente cuya prevalencia es del 1,2%. Se presenta morfológicamente con un lóbulo principal y uno o varios lóbulos accesorios de diverso tamaño unidos entre sí mediante vasos sanguíneos. Los principales factores de riesgo corresponden a la edad materna avanzada y la fertilización in vitro. El diagnóstico se realiza clínicamente posterior al alumbramiento. La PS presenta riesgos obstétricos importantes como la hemorragia posparto e infección por retención de lóbulo accesorio. Presentación de caso: Se presenta el caso clínico de una mujer de 35 años, multípara de uno, cursando embarazo único con diagnóstico de restricción de crecimiento fetal y Rh (–) no sensibilizada. A las 26 + 6 semanas se detecta una malformación placentaria con un cuerpo placentario posterior y lóbulo accesorio por cara anterior conectado con vasos arteriovenosos en fondo uterino, hallazgos compatibles con PS. A las 37 + 6 semanas la paciente presenta un parto vaginal conducido, en el alumbramiento se corrobora el diagnóstico de PS. Conclusión: La PS es una malformación poco frecuente y subdiagnosticada que presenta riesgos obstétricos importantes. Es fundamental el diagnóstico ecográfico anteparto de las malformaciones placentarias para evitar resultados maternos y fetales desfavorables.

ABSTRACT Introduction Obstructive azoospermia occurs when there is a blockage in the male reproductive tract, leading to a complete absence of sperm in the ejaculate. It constitutes around 40% of all cases of azoospermia (1, 2). Blockages in the male reproductive tract can arise from either congenital or acquired factors, affecting various segments such as the epididymis, vas deferens, and ejaculatory ducts (3). Examples of congenital causes encompass conditions like congenital bilateral absence of the vas deferens and unexplained epididymal blockages (4). Acquired instances of obstructive azoospermia may result from factors like vasectomy, infections, trauma, or unintentional injuries caused by medical procedures (5). This complex condition affecting male fertility, presents two main treatment options: microsurgical reconstruction and surgical extraction of sperm followed by in vitro fertilization (IVF). Microsurgical reconstruction proves to be the most cost-effective option for treating obstructive azoospermia when compared with assisted reproductive techniques (6, 7). However, success rates of reconstruction defined by patency are as high as 99% for vasovasostomy (VV) but decline to around 65% if vasoepididymostomy (VE) is required (8, 9). Thus, continued refinement in technique is necessary in order to attempt to improve patency for patients undergoing VE. In this video, we show a comprehensive demonstration of microsurgical VE, highlighting the innovative epididymal occlusion stitch. The goal of this innovative surgical technique is to improve outcomes for VE. Materials and Methods The patient is a 39-year-old male diagnosed with obstructive azoospermia who presents for surgical reconstruction via VE. His partner is a 37-years-old female with regular menstrual cycles. The comprehensive clinical data encompasses a range of factors, including FSH levels, results from semen analysis, and outcomes from testicular sperm aspiration. This thorough exploration aims to provide a thorough understanding of our innovative surgical technique and its application in addressing complex cases of obstructive azoospermia. Results The procedure was started on the right, the vas deferens was identified and transected. The abdominal side of the vas was intubated and a vasogram performed, there was no obstruction. There was no fluid visible from the testicular side of the vas for analysis, thus we proceeded with VE. Upon inspection of the epididymis dilated tubules were identified. After selecting a tubule for VE, two 10-0 nylon sutures were placed, and it was incised. Upon inspection of the fluid motile sperm was identified. After VE, we performed a novel epididymal occlusion stitch technique. This was completed distal to the anastomosis by placing a 7-0 prolene through the tunica of the epididymis from the medial to lateral side. This stitch was then tightened down with the goal to largely occlude the epididymis so that sperm will preferentially travel through the anastomosis. The steps were then repeated on the left. At 3-month follow up, the patient had no change in testicular size as compared with preoperative size (18cc), he had no testicular or incisional discomfort, and on semen analysis he had presence of motile sperm. After 3 months post-surgery, the patient had motile sperm seen on semen analysis. Discussion The introduction of a novel epididymal occlusion stitch demonstrates a targeted strategy to enhance the success of microscopic VE. Encouragingly, a 3-month post-surgery follow-up reveals the presence of motile sperm, reinforcing the potential efficacy of our approach. This is promising given the historical lower patency, delayed time to patency, and higher delayed failure rates that patients who require VE experience (10). In total, 40% of all azoospermia cases can be attributed to obstruction. The conventional treatments for obstructive azoospermia involve microsurgical reconstruction and surgical sperm retrieval followed by IVF. While microsurgical reconstruction has proven to be economically viable, the quest for enhanced success rates has led to the exploration of innovative techniques. Historically, the evolution of VV and VE procedures, initially performed in the early 20th century, laid the foundation for contemporary microsurgical approaches (11). Notably, the microscopic VV demonstrated significant improvements in patency rates and natural pregnancy likelihood, as evidenced by the seminal Vasovastomy Study Group study in 1991 (8). In contemporary literature, success rates particularly for VE remain unchanged for the past three decades since the original published success rates by the Vasectomy Reversal Study Group (12). VE is associated with a longer time to patency as well with patients taking 2.8 to 6.6 months to have sperm return to ejaculate as compared to 1.7 to 4.3 months for those undergoing VV. Additionally, of those patients who successfully have sperm return to the ejaculate after VE up to 50% will have delayed failure compared to 12% for those undergoing VV who are patent. Finally, of those who experience delayed failure after undergoing VE it usually occurs earlier with studies reporting as early as 6 months post-operatively (20). Given the lack of improvement and significantly worsened outcomes with VE further surgical refinement is a constant goal for surgeons performing this procedure. Conclusion: In conclusion, this video is both a demonstration and a call to action for commitment to surgical innovation. We aim to raise the bar in VE success rates, ultimately bringing tangible benefits to patients and contributing to the ongoing evolution of reproductive medicine. The novel epididymal occlusion stitch emerges as a beacon of progress, promising not only enhanced safety but also potential reductions in patency time. Surgical excellence and methodological refinement, as exemplified in this video, lay the foundation for a future where male reproductive surgery continues to break new ground. 40 1, 1 (1 2. 2 . 2) 3. (3) 4. 4 (4) vasectomy infections trauma 5. 5 (5) fertility options IVF (IVF) costeffective cost effective 6, (6 7. 7 7) However 99 (VV 65 (VE 8, 8 (8 9. 9 9) Thus 39yearold yearold 39 year old 37yearsold yearsold 37 years cycles levels aspiration right transected obstruction 100 10 0 10- placed incised 70 7- left 3month month 18cc, 18cc cc , (18cc) discomfort postsurgery, postsurgery post surgery, Encouragingly followup approach 10. (10) total viable Historically th century 11. 11 (11) Notably likelihood 199 8. (8) literature 12. 12 (12) 28 66 6. 17 1. 43 Additionally 50 patent Finally postoperatively operatively 20. 20 (20) Conclusion conclusion innovation medicine progress ground ( (3 (4 (5 (IVF (18cc (10 (11 19 (12 (20 (2

ABSTRACT: This study determined the zootechnical, metabolic, and health performance of Girolando calves born with high or low birth weight, conceived through in vitro fertilization. The study was carried out on a commercial dairy farm located in Passos, Minas Gerais, Brazil. For this, a hundred Girolando calves were divided into two groups: The Control, which consisted of calves that were born weighing ≤ 35 kg; and high birth weight (HBW) calves, which were born weighing > 35 kg. The calves were monitored for zootechnical parameters; epidemiological indices, for diseases such diarrhea and pneumonia; as well as serum concentrations of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma-glutamyl transferase, cholesterol, triglycerides, paraoxonase1, albumin, urea, and globulin. Calves from the HBW group had a higher general mortality rate, but no effects on the zootechnical performance or metabolism were observed. The results obtained allow us to conclude that Girolando calves generated by IVF and heavier at birth have a greater chance to present diseases such as omphalophlebitis, tympany, and bovine parasitic sadness, and have a higher rate of general mortality. Conversely, birth weight of the calves does not influence diarrhea or pneumonia, the zootechnical performance, or the metabolism. ABSTRACT metabolic fertilization Passos Gerais Brazil this groups Control 3 kg (HBW parameters indices pneumonia aminotransferase phosphatase gammaglutamyl gamma glutamyl transferase cholesterol triglycerides paraoxonase1 paraoxonase albumin urea globulin observed omphalophlebitis tympany sadness Conversely

RESUMO: Este estudo determinou o desempenho zootécnico, metabólico e saúde de bezerras Girolando nascidas com alto ou baixo peso, concebidas por fertilização in vitro. O estudo foi realizado em uma fazenda leiteira comercial localizada em Passos, Minas Gerais, Brasil. Cem bezerras Girolando foram divididas em dois grupos: Controle, que consistiu em bezerras que nasceram com peso ≤ 35 kg; e HBW, bezerras que nasceram com peso > 35 kg. As bezerras foram monitoradas quanto aos parâmetros zootécnicos; índices epidemiológicos para doenças como diarreia e pneumonia; bem como as concentrações séricas de aspartato aminotransferase, alanina aminotransferase, fosfatase alcalina, gama-glutamil transferase, colesterol, triglicérides, paraoxonase1, albumina, uréia e globulina. As bezerras do grupo HBW apresentaram maior taxa de mortalidade, mas nenhum efeito no desempenho zootécnico ou metabolismo foram observados. Os resultados permitem concluir que bezerras Girolando provenientes de FIV que nascem mais pesadas têm maior chance de apresentar doenças, como onfaloflebite, timpanismo e tristeza parasitária bovina, tendo uma maior taxa de mortalidade geral. Porém, o peso ao nascimento das bezerras não influenciou a ocorrência de diarreia ou broncopneumonia, o desempenho zootécnico e o perfil metabólico. RESUMO vitro Passos Gerais Brasil grupos Controle 3 kg zootécnicos pneumonia aminotransferase alcalina gamaglutamil gama glutamil transferase colesterol triglicérides paraoxonase1 paraoxonase albumina globulina observados onfaloflebite bovina geral Porém broncopneumonia

RESUMEN Introducción: la infertilidad es una condición de salud con impacto emocional en la pareja acompañada de un aumento de angustia psicológica. La fertilización in vitro es la alternativa más avanzada de reproducción asistida a la que una pareja infértil puede acceder en Cuba. Objetivo: describir el perfil psicológico de parejas infértiles tributarias a tratamientos de fertilización in vitro. Métodos: se realizó una investigación con un enfoque cuanti-cualitativo longitudinal prospectivo en parejas infértiles atendidas en el Centro Provincial de reproducción asistida de baja complejidad del Hospital Ginecoobstétrico Docente “Mariana Grajales” en el período de enero de 2022 a junio de 2023. La muestra se conformó por 21 parejas que fueron seleccionadas mediante un muestreo no probabilístico. Resultados: la muestra se caracterizó por presentar edades entre 20 y 47 años, con una media en la tercera década de vida en ambos sexos. Se determinó la presencia de alteraciones emocionales, con predominio de ansiedad como estado en niveles bajos y medio, el sexo femenino es el más afectado, y en el masculino se determinó un predominio de la incertidumbre como estado emocional predominante. Conclusiones: el perfil psicológico de las parejas en estudio está determinado por alteraciones emocionales, con predominio de la ansiedad como estado en niveles bajos y medio, el sexo femenino fue el más afectado y en el sexo masculino se determinó un predominio de la incertidumbre como estado emocional predominante.

ABSTRACT Introduction: infertility is a health condition with an emotional impact on the couple accompanied by an increase in psychological distress. In vitro fertilization is the most advanced assisted reproduction alternative that an infertile couple can access in Cuba. Objective: to describe the psychological profile of infertile couples undergoing in vitro fertilization treatments. Methods: an investigation was carried out with a prospective longitudinal quantitative-qualitative approach in infertile couples treated at the Provincial Center for low-complexity assisted reproduction of the “Mariana Grajales” Gynecobstetric Teaching Hospital in the period from January 2022 to June 2023. The sample it was made up of 21 couples that were selected through non-probabilistic sampling. Results: the sample was characterized by ages between 20 and 47 years, with an average in the third decade of life in both sexes. The presence of emotional alterations was determined, with a predominance of anxiety as a state at low and medium levels, the female sex is the most affected, and in the male sex a predominance of uncertainty was determined as the predominant emotional state. Conclusions: the psychological profile of the couples in the study is determined by emotional alterations, with a predominance of anxiety as a state at low and medium levels, the female sex was the most affected and in the male sex a predominance of uncertainty as a state was determined predominant emotional.

ABSTRACT The aim of this study was to evaluate the effect of Insulin-Transferrin-Selenium (ITS) supplementation in embryo culture on the blastocyst rate and embryo quality. The bovine zygotes obtained by in vitro maturation (MIV) and fertilization (IVF) were transferred to Evolve medium in 4 groups (T1: Without ITS supplement; T2: ITS the first 48 h of culture (CIV1): T3: ITS after day 2 of culture (CIV2), T4: ITS during all culture time (CIV1+CIV2). All procedures were performed at 38.5 °C,5%CO2and90%relativehumidity. The cleavage and blastocyst rates were determined at 48 h and day 8 of culture. The total cell number and the production of reactive oxygen species (ROS) in the expanded blastocysts were determined by staining the nucleus with Hoechst and the oxidation of the fluorescent probe dichlorodihydrofluorescein diacetate (H2DCFDA). The T4 cultured embryos presented the highest rate of blastocysts (32.6%) and total cell number (145.6) compared to the control group without STI (26.2% and 98.6, respectively; p<0.05). Likewise, the production of ROS in T4 blastocysts was lower (p<0.05) than that obtained in the other study groups. The results show that the addition of the ITS supplement throughout the embryo culture time has beneficial effects on the percentage of blastocysts and embryo quality, assessed by the number of total cells and the production of ROS.

RESUMEN El estudio tuvo como objetivo evaluar el efecto del suplemento Insulina-Transferrina-Selenio (ITS) en el cultivo embrionario sobre la tasa de blastocistos y calidad embrionaria. Los cigotos de bovino obtenidos por maduración (MIV) y fertilización in vitro (FIV) fueron transferidos a medio Evolve en 4 grupos (T1: Sin suplemento ITS; T2: ITS las primeras 48 h de cultivo (CIV1): T3: ITS después del día 2 de cultivo (CIV2), T4: ITS todo el tiempo de cultivo (CIV1 + CIV2). Todos los procedimientos se realizaron a 38.5 °C, 5% de CO2 y 90% de humedad relativa. Las tasas de clivaje y de blastocistos se determinaron a las 48 h y día 8 de cultivo, respectivamente. El número de células totales y la producción de especies reactivas de oxígeno (EROs) en los blastocistos expandidos fueron determinados por tinción del núcleo con Hoechst y la oxidación de la sonda fluorescente diacetato diclorodihidrofluoresceína (H2DCFDA). Los embriones cultivados de T4 presentaron la mayor tasa de blastocistos (32.6%) y número de células totales (145.6) comparada al grupo control sin ITS (26.2% y 98.6, respectivamente; p<0.05). Asimismo, la producción de EROs en los blastocistos de T4 fue menor (p<0.05) que la obtenida en los demás grupos de estudio. Los resultados demuestran que la adición del suplemento ITS durante todo el tiempo de cultivo embrionario tiene efectos benéficos sobre el porcentaje de blastocistos y calidad embrionaria, valorada por el número de células totales y la producción de EROs.

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.

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 Introduction: Infertility is a disease present in the 17.5% of the adult population. COVID-19 caused overload in health centers, producing an unclear impact in the reproductive health units that perform assisted reproduction therapies. Objectives: To compare clinical characteristics and the attention times of patients subjected to in vitro fertilization in the Reproductive Medicine Unit of the Gustavo Fricke Hospital between the 2017-2019 and 2020-2022 periods. Method: Descriptive observational study in which an anonymized database of 82 patients subjected to in vitro fertilization in the Reproductive Medicine Unit of the Gustavo Fricke Hospital was used. Results: The average age was 34.2 ± 4.2 years old. The infertility time was 5.1 ± 4.3 years. The most frequent infertility factors were tuboperitoneal (59.8%) and male factors (46.3%). During the first pre-pandemic 20 months, there was a higher tendency towards early referral to the Center for Human Reproduction of the Valparaiso University (odds ratio: 3.84). Conclusions: In the course of the pandemic, there was an increase of the infertility time, the pregnancies median, prior abortions, and male factor frequency. On the other hand, there were not considerable changes when it comes to referral times to in vitro fertilization, however, early referrals decreased during the pandemic.

Resumen Introducción: La infertilidad es una enfermedad presente en el 17.5% de la población adulta. La COVID-19 provocó sobrecargas en atención de salud, generando un impacto incierto en unidades de salud reproductiva que realizan terapias de reproducción asistida. Objetivos: Comparar características clínicas y tiempos de atención de pacientes sometidas a fertilización in vitro de la Unidad de Medicina Reproductiva del Hospital Gustavo Fricke entre los periodos 2017-2019 y 2020-2022. Método: Estudio observacional descriptivo, utilizando base de datos anonimizada de la Unidad de Medicina Reproductiva del Hospital Gustavo Fricke con 82 pacientes sometidas a fertilización in vitro. Resultados: La media de edad fue de 34.2 ± 4.2 años. El tiempo de infertilidad fue de 5.1 ± 4.3 años. Los factores de infertilidad más frecuentes son el tuboperitoneal (59,8%) y masculino (46,3%). Durante los primeros 20 meses en prepandemia hubo mayor tendencia a derivación precoz al Centro de Reproducción Humana de la Universidad de Valparaíso (odds ratio: 3.84). Conclusiones: En pandemia se evidenció un aumento en el tiempo de infertilidad, mediana de gestaciones, abortos previos y frecuencia de factor masculino. No hubo cambios significativos en tiempos de derivación a fertilización in vitro, no obstante, durante la pandemia disminuyeron las derivaciones precoces.

Abstract 17. The study aimed to evaluate the Fertilization Rate of four independent in vitro treatments of bovine oocytes with Sexed (SX) and Conventional (Conv) semen, capacitated (B.O and Percoll) using the Swim-Up technique. Cleavage was assessed at 48 hours post-fertilization and on the seventh day of embryonic development. The results showed a cleavage percentage of 59.4% in T1 (Conv. Percoll), 60% in T2 (Conv. B.O), 75% in T3 (SX. Percoll), and 46.6% in T4 (SX. B.O), with a statistically significant difference (p>0.05) observed specifically between T3 and T4. The embryo development rate on the seventh day was 34.4% in T1, 20% in T2, 15% in T3, and 10.51% in T4. In this case, a statistically significant difference was found in the use of conventional medium with Percoll compared to SX B.O medium (p<0.05) (T1 and T4, respectively). The results also showed similar production of morula and early blastocysts and protruded embryos among the four treatments, demonstrating the feasibility of in vitro fertilization (IVF) technique for producing bovine embryos. In conclusion, the preliminary evaluation highlights the significant influence of the sexing method and treatment on semen quality and embryonic development in animal reproduction programs. Although Sexed semen with Percoll shows potential to improve cleavage rates, conventional semen, especially in T1 treatment (Conv. Percoll), exhibits superiority in individual motility and concentration. These findings provide crucial insights for optimizing reproductive programs.

Resumen 13. El objetivo del presente estudio fue evaluar la Tasa de Fecundación in vitro de ovocitos bovinos con semen sexado (SX) y convencional (Conv.) capacitados (B.O y Percoll) con la técnica Swim-Up. Se evaluó el clivaje a las 48 horas post fecundación y al séptimo día del desarrollo embrionario. Los resultados mostraron un porcentaje de clivaje del 59,4% en el T1 (Conv. Percoll), 60% en el T2 (Conv. B.O), 75% en el T3 (SX. Percoll) y 46,6% en el T4 (SX. B.O), mostrando diferencia estadística entre T3 y T4 (p>0,05). La tasa de desarrollo del embrión al séptimo día fue 34,4% T1, 20% en T2, 15% en T3 y 10,51% en T4; en este caso, se encontró diferencia estadísticamente significativa en el uso del medio convencional más Percoll en comparación con el medio de SX B.O (p<0,05) (T1 y T4, respectivamente). Los resultados también mostraron una producción similar de mórulas y blastocitos tempranos y protruidos entre los cuatro tratamientos, y demuestran la viabilidad del desarrollo e implementación de la técnica de fertilización in vitro (FIV) para producir embriones bovinos. Concluyendo la evaluación preliminar, destaca la significativa influencia del método de sexado y del tratamiento en la calidad del semen y desarrollo embrionario en programas de reproducción animal. Aunque el semen sexado con Percoll muestra potencial para mejorar tasas de clivaje, el semen convencional, especialmente en el tratamiento T1 (Conv. Percoll), exhibe una superioridad en motilidad individual y concentración. Estos resultados informan decisiones cruciales para optimizar programas reproductivos.

ABSTRACT: Cadmium (Cd) is a heavy metal that is extremely dangerous to human health and can be found naturally in soils, deposited through industrial waste or phosphate fertilization. In this study, we evaluated the morphological responses of in vitro grown plants of potato in the presence of Cd, and define a procedure for assessing Cd sensitivity of different clones. The potato clone SMIJ461-1 was cultured in the presence of Cd at concentrations of 0, 20, 40, 60, 80, 90, 110, 120, 140, and 160 µM. Survival, rooting, number and height of shoots, leaf and root counts were evaluated at 7, 14, 21, and 28 days of cultivation. Adverse effects of Cd on the growth of the potato plants, exerted in a concentration-dependent manner, were observed for shoot height, leaf and root counts. Results of this study indicated that Cd concentrations of 0, 45, 90, and 135 µM can be used for future in vitro evaluation of potato clone sensitivity to Cd exposure. ABSTRACT (Cd soils fertilization clones SMIJ4611 SMIJ SMIJ461 1 SMIJ461- 0 20 40 60 80 90 110 120 140 16 Survival rooting shoots 7 14 21 2 cultivation concentrationdependent concentration dependent manner 45 13 exposure SMIJ46 4 6 8 9 11 12 SMIJ4

RESUMO: O cádmio (Cd) é um metal pesado extremamente perigoso à saúde humana, podendo ser encontrado naturalmente nos solos, depositado através de resíduos industriais ou da adubação fosfatada. Este trabalho teve por objetivo avaliar as respostas morfológicas de plantas de batata cultivadas in vitro na presença de Cd e definir uma metodologia para inferir sobre a sensibilidade de diferentes clones. Foi estudado o clone SMIJ461-1 de batata, nas concentrações de 0, 20, 40, 60, 80, 90, 110, 120, 140, 160 µM de Cd adicionadas ao meio de cultura. A sobrevivência, o enraizamento, o número de brotações, a altura da parte aérea e o número de folhas e raízes foram avaliados aos 7, 14, 21 e 28 dias de cultivo. Foram observados efeitos adversos de concentrações de Cd sobre o crescimento in vitro das plantas de batata do clone SMIJ461-1 para a altura da parte aérea e o número de folhas e raízes, sendo estes efeitos dependentes das concentrações de Cd adicionadas ao meio de cultura. Os resultados deste estudo indicam que as concentrações de 0, 45, 90 e 135 µM de Cd podem ser utilizadas para a avaliação in vitro da sensibilidade de clones de batata a presença de Cd. RESUMO (Cd humana solos fosfatada SMIJ4611 SMIJ SMIJ461 1 SMIJ461- 0 20 40 60 80 110 120 140 16 cultura sobrevivência enraizamento brotações 7 14 2 cultivo 45 9 13 SMIJ46 4 6 8 11 12 SMIJ4

SUMMARY OBJECTIVE: The aim of this study was to determine the effect of anxiety and depression on pregnancy outcome in couples receiving in vitro fertilization treatment. METHODS: A total of 102 couples (102 females and 102 males) with unexplained infertility were included in the study. Personal Information Form was used to collect data, Case Follow-up Form to record the treatment process, and Spielberger’s State-Trait Anxiety Inventory and Beck Depression Inventory to measure the anxiety and depression levels of couples. Couples were measured twice: before treatment and on oocyte pickup day. RESULTS: There was no statistically significant difference between the anxiety and depression levels and oocyte count of women (p>0.05). There were statistically significant differences between State-Trait Anxiety Inventory level and sperm count and between BID level and sperm motility (p<0.05). There was no statistically significant difference between the anxiety and depression levels and pregnancy outcomes of women (p>0.05). CONCLUSION: Anxiety and depression had no effect on pregnancy outcome. More studies are needed to investigate the effect of anxiety and depression on pregnancy outcome in unexplained infertility. OBJECTIVE METHODS 10 (10 males data Followup Follow up process Spielbergers Spielberger s StateTrait State Trait twice day RESULTS p>0.05. p005 p p>0.05 . 0 05 (p>0.05) p<0.05. p<0.05 (p<0.05) CONCLUSION 1 (1 p00 p>0.0 (p>0.05 p<0.0 (p<0.05 ( p0 p>0. (p>0.0 p<0. (p<0.0 p>0 (p>0. p<0 (p<0. p> (p>0 p< (p<0 (p> (p< (p

Urea is the most widely used nitrogen fertilizer worldwide. However, ammonia volatilization, resulting from applying urea to the soil surface, causes economic and environmental losses; thus, urease inhibitors have been developed to mitigate these losses. In this work, the anti-ureolytic activity of Schiff’s base 4-(3-hydroxybenzylideneamino) phenol (3B4) and its amine-derived (3B4a) was evaluated. The most promising urease inhibitor in soil was 3B4 (55.0 ± 3.9% inhibition), with comparable results to N-(butyl) thiophosphoric triamide (p = 0.659). In the in vitro analysis (Canavalia ensiformis), the results of anti-ureolytic activity were similar, 22.6 ± 6.9% for 3B4 and 24.2 ± 9.6% for 3B4a. Biophysical interaction studies were also carried out through molecular docking studies and molecular fluorescence spectroscopy. These studies showed that both substances are preferentially competitive inhibitors, with the interaction between 3B4a and urease forming a more stable complex. In the analysis by Fourier transform infrared spectroscopy, no interaction was observed when 3B4 or 3B4a was mixed with urea (1:1) for 48 h, providing evidence of compatibility. Thus, the Schiff base 3B4 and its corresponding amine 3B4a may represent potential additives for urea fertilization aiming to assist in the urease inhibition process. worldwide However volatilization surface losses thus work antiureolytic anti ureolytic Schiffs s 43hydroxybenzylideneamino hydroxybenzylideneamino 4 3 4-(3-hydroxybenzylideneamino B (3B4 aminederived derived Ba (3B4a evaluated 3B 55.0 550 55 0 (55. 39 9 3.9 inhibition, , inhibition) Nbutyl N butyl N-(butyl p 0.659. 0659 0.659 . 659 0.659) Canavalia ensiformis, ensiformis ensiformis) similar 226 22 6 22. 69 6.9 242 24 2 24. 96 9.6 spectroscopy complex 11 1 (1:1 h compatibility Thus process (3B 55. 5 (55 3. 065 0.65 65 6. 9. (1: (5 06 0.6 (1 ( 0.

ABSTRACT This study aimed to evaluate the in vitro gas production from three pearl millet cultivars submitted to four nitrogen levels. The treatments consisted of three pearl millet cultivars (ADR 500, ADR 7020, and LABH 70732) and four nitrogen levels (0, 50, 100, and 200 kg/ha), using urea as the nitrogen source. A 3 x 4 factorial experimental design, employing a randomized block setup, was used to assess both gas production and digestibility. No interaction was observed between nitrogen doses and fermentative parameters on in vitro gas production from the dry matter of pearl millet cultivars. Nitrogen had no impact on the dry matter content or chemical composition of the cultivars. Consistency prevailed in the chemical composition across the various cultivars, a trend that extended to the gas production rates for both soluble and insoluble contents. Notably, nitrogen fertilizer exerted an influence on the fermentation pattern of organic matter, gas production, and lag time for the cultivars. Nitrogen fertilization is advised solely for enhancing fermentative attributes in the case of the cultivar ADR 7020. 500 7020 70732 0, 0 (0 50 100 20 kg/ha, kgha kg/ha , kg ha kg/ha) source design setup digestibility contents Notably 702 7073 ( 5 10 2 70 707 1 7

RESUMO Objetivou-se avaliar a produção “in vitro” de gás de três cultivares de milheto forrageiro sob adubação nitrogenada. Os tratamentos foram constituídos por três cultivares de milheto: (ADR 500, ADR 7020 e LABH 70732) e quatro doses de N(ureia) (0; 50; 100 e 200 kg/ha). Para as avaliações da produção de gases e digestibilidade o delineamento experimental foi blocos casualizados em esquema fatorial 3 x 4, com quatro repetições. Não houve interação entre as doses de adubação nitrogenada e os parâmetros fermentativos da produção de gases “in vitro” da matéria seca das cultivares de milheto. A adubação nitrogenada não influenciou os teores de matéria seca ou a composição bromatológica das cultivares. Não houve diferenças na composição bromatológica entre as cultivares. O mesmo foi observado para as taxas de produção de gás a partir do conteúdo solúvel e insolúvel. A adubação nitrogenada influenciou o padrão de fermentação, a produção de gases e o tempo de colonização por bactérias da matéria orgânica das três cultivares. A adubação nitrogenada é recomendada para incrementar as características fermentativas apenas da cultivar ADR 7020. Objetivouse Objetivou se in vitro 500 702 70732 Nureia N ureia N(ureia 0 (0 50 10 20 kg/ha. kgha kg/ha . kg ha kg/ha) 4 repetições insolúvel fermentação 70 7073 ( 5 1 2 7 707

Abstract In reproductive technologies, uncovering the molecular aspects of oocyte and embryo competence under different conditions is crucial for refining protocols and enhancing efficiency. RNA-seq generates high-throughput data and provides transcriptomes that can undergo additional computational analyses. This study presented the transcriptomic profiles of in vitro matured oocytes and blastocysts produced in vitro from buffalo crossbred (Bubalus bubalis), coupled with gene co-expression and module preservation analysis. Cumulus Oophorus Complexes, obtained from slaughterhouse-derived ovaries, were subjected to in vitro maturation to yield metaphase II oocytes (616) or followed in vitro fertilization and culture to yield blastocysts for sequencing (526). Oocyte maturation (72%, ±3.34 sd) and embryo development (21.3%, ±4.18 sd) rates were obtained from three in vitro embryo production routines following standard protocols. Sequencing of 410 metaphase II oocytes and 70 hatched blastocysts (grade 1 and 2) identified a total of 13,976 genes, with 62% being ubiquitously expressed (8,649). Among them, the differentially expressed genes (4,153) and the strongly variable genes with the higher expression (fold-change above 11) were highlighted in oocytes (BMP15, UCHL1, WEE1, NLRPs, KPNA7, ZP2, and ZP4) and blastocysts (APOA1, KRT18, ANXA2, S100A14, SLC34A2, PRSS8 and ANXA2) as representative indicators of molecular quality. Additionally, genes exclusively found in oocytes (224) and blastocysts (2,200) with specific biological functions were identified. Gene co-expression network and module preservation analysis revealed strong preservation of functional modules related to exosome components, steroid metabolism, cell proliferation, and morphogenesis. However, cell cycle and amino acid transport modules exhibited weak preservation, which may reflect differences in embryo development kinetics and the activation of cell signaling pathways between buffalo and bovine. This comprehensive transcriptomic profile serves as a valuable resource for assessing the molecular quality of buffalo oocytes and embryos in future in vitro embryo production assays. technologies efficiency RNAseq RNA seq highthroughput high throughput analyses Bubalus bubalis, bubalis , bubalis) coexpression co Complexes slaughterhousederived slaughterhouse derived ovaries 616 (616 526. 526 . (526) 72%, 72 (72% 334 3 34 ±3.3 sd 21.3%, 213 21 (21.3% 418 4 18 ±4.1 41 7 grade 2 13976 13 976 13,97 62 8,649. 8649 8,649 8 649 (8,649) them 4,153 4153 153 (4,153 foldchange fold change 11 BMP15, BMP15 BMP (BMP15 UCHL1 UCHL WEE1 WEE NLRPs KPNA7 KPNA ZP2 ZP ZP4 APOA1, APOA1 APOA (APOA1 KRT18 KRT ANXA2 ANXA S100A14 SA S A SLC34A2 SLCA SLC PRSS Additionally 224 (224 2,200 2200 200 (2,200 components metabolism proliferation morphogenesis However bovine assays 61 (61 52 (526 72% (72 33 ±3. 21.3% (21.3 ±4. 1397 97 13,9 6 864 8,64 64 (8,649 4,15 415 15 (4,15 BMP1 (BMP1 (APOA KRT1 S100A1 SLC34A 22 (22 2,20 220 20 (2,20 (6 5 (52 (7 ±3 21.3 (21. ±4 139 9 13, 86 8,6 (8,64 4,1 (4,1 (BMP S100A (2 2,2 (2,2 ( (5 ± 21. (21 8, (8,6 4, (4, 2, (2, (8, (4 (8

Abstract Objective To compare outcomes in patients with repeated implantation failure undergoing Intracytoplasmic Sperm Injection/In vitro fertilization (IVF/ICSI) plus immunosuppressants such as prednisolone, prednisone, or cyclosporine A versus the use of IVF/ICSI alone. Data source Databases were systematically searched in PubMed, Cochrane, and Embase databases in September 2023. Study Selection Randomized clinical trials and observational studies with the outcomes of interest were included. Data collect We computed odds ratios (ORs) for binary endpoints, with 95% confidence intervals (CIs). Heterogeneity was assessed using I2 statistics. Data were analyzed using Review Manager 5.4.The main outcomes were live birth, miscarriage, implantation rate, clinical pregnancy, and biochemical pregnancy. Data synthesis Seven studies with 2,829 patients were included. Immunosuppressive treatments were used in 1,312 (46.37%). Cyclosporine A improved implantation rate (OR 1.48; 95% CI 1.01-2.18) and clinical pregnancy (1.89, 95% CI 1.14-3.14). Compared to non-immunosuppressive treatment, prednisolone and prednisone did not improve live birth (OR 1.13, 95% CI 0.88-1.46) and miscarriage (OR 1.49, 95% CI 1.07-2.09). Prednisolone showed no significant effect in patients undergoing IVF/ICSI, clinical pregnancy (OR 1.34; 95% CI 0.76-2.36), or implantation rate (OR 1.36; 95% CI 0.76-2.42). Conclusion Cyclosporine A may promote implantation and clinical pregnancy rates. However, given the limited sample size, it is important to approach these findings with caution. Our results indicate that prednisolone and prednisone do not have any beneficial effects on clinical outcomes of IVF/ICSI patients with repeated implantation failure. PROSPERO CRD42023449655 InjectionIn Injection In IVFICSI IVF ICSI (IVF/ICSI alone PubMed Cochrane 2023 included ORs (ORs endpoints 95 CIs. CIs . (CIs) I statistics 54The The 5 4 2829 2 829 2,82 1312 1 312 1,31 46.37%. 4637 46.37% 46 37 (46.37%) OR 1.48 148 48 1.012.18 101218 1.01 2.18 01 18 1.01-2.18 1.89, 189 89 (1.89 1.143.14. 114314 1.14 3.14 14 3 1.14-3.14) nonimmunosuppressive non immunosuppressive treatment 113 13 1.13 0.881.46 088146 0.88 1.46 0 88 0.88-1.46 149 49 1.49 1.072.09. 107209 1.07 2.09 07 09 1.07-2.09) 1.34 134 34 0.762.36, 076236 0.76 2.36 , 76 36 0.76-2.36) 1.36 136 0.762.42. 076242 2.42 42 0.76-2.42) rates However size caution CRD CRD4202344965 202 9 (CIs 282 82 2,8 131 31 1,3 463 46.37 (46.37% 1.4 012 1.012.1 10121 101 1.0 218 2.1 1.01-2.1 1.89 8 (1.8 143 1.143.14 11431 114 1.1 314 3.1 1.14-3.14 11 881 0.881.4 08814 088 0.8 146 0.88-1.4 072 1.072.09 10720 107 209 2.0 1.07-2.09 1.3 762 0.762.36 07623 076 0.7 236 2.3 7 0.76-2.36 0.762.42 07624 242 2.4 0.76-2.42 CRD420234496 20 28 2, 1, 46.3 (46.37 1. 1.012. 1012 10 21 2. 1.01-2. 1.8 (1. 1.143.1 1143 3. 1.14-3.1 0.881. 0881 08 0. 0.88-1. 1.072.0 1072 1.07-2.0 0.762.3 0762 23 0.76-2.3 0.762.4 24 0.76-2.4 CRD42023449 46. (46.3 1.012 1.01-2 (1 1.143. 1.14-3. 0.881 0.88-1 1.072. 1.07-2. 0.762. 0.76-2. CRD4202344 (46. 1.01- ( 1.143 1.14-3 0.88- 1.072 1.07-2 0.762 0.76-2 CRD420234 (46 1.14- 1.07- 0.76- CRD42023 (4 CRD4202 CRD420 CRD42 CRD4

SUMMARY OBJECTIVE: The aim of this study was to determine the effects of seasons (winter vs. summer) on oocyte quality in infertile women undergoing ovulation induction for in vitro fertilization. METHODS: This retrospective cross-sectional study assessed 155 cycles of in vitro fertilization-induced ovulation in women, with 71 and 84 cycles occurring in the summer and winter, respectively. Oocytes were evaluated for quality, with 788 and 713 assessed during summer and winter, and classified according to Nikiforov's categories: (a) category I, good quality; (b) category 2, medium quality; and (c) category 3, low quality. RESULTS: Thickened zona pellucida (p<0.001), increased perivitelline space (p<0.001), oocyte shape abnormalities (p=0.01), and the presence of refractile bodies (p<0.0001) were more frequent in the summer cycles, whereas cytoplasmic granularity (p<0.001) was more frequent in the winter cycles. In winter, we observed a higher frequency of category 3 (p<0.001) and category 2 (p<0.001) oocytes and a lower frequency of category 1 (p<0.001) oocytes. CONCLUSION: Oocyte dysmorphisms were found in 70–80% of cases and were more common in winter. The main features include a thickened zona pellucida, enlarged perivitelline space, irregular shape, and cytoplasmic granularity. This implies better-quality oocytes in the summer than in the winter. However, retrospective studies have limitations due to data collection biases and potential confounding variables such as diet and exercise. Future research is needed to confirm these findings and explore the underlying mechanisms. OBJECTIVE vs fertilization METHODS crosssectional cross sectional 15 fertilizationinduced induced 7 8 respectively 78 Nikiforovs Nikiforov s categories (a I b (b c (c RESULTS p<0.001, p0001 p p<0.001 , 0 001 p=0.01, p001 p=0.01 01 (p=0.01) p<0.0001 p00001 0001 (p<0.0001 (p<0.001 CONCLUSION 7080 70 80 70–80 betterquality better However exercise mechanisms p000 p<0.00 00 p00 p=0.0 (p=0.01 p<0.000 p0000 000 (p<0.000 (p<0.00 708 70–8 p<0.0 p0 p=0. (p=0.0 (p<0.0 70– p<0. p=0 (p=0. (p<0. p<0 p= (p=0 (p<0 p< (p= (p< (p