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.

Effects of music therapy on pain and oxidative stress in oocyte pick-up: A randomized clinical trial / Efeitos da musicoterapia sobre dor e estresse oxidativo na aspiração folicular: estudo clínico randomizado

Abstract Background and objective: The aim was to investigate the effects of Turkish classical music on pain and oxidative stress in patients undergoing oocyte pick-up. Methods: The study was a randomized, controlled trial. The groups included Group NM (Non-Music), control group; Group PM, which comprised patients who listened to music before the operation; and Group CM, which comprised patients who listened to music both before and during the operation. Blood was drawn prior to the operation to measure the oxidative stress values. Pain, hemodynamic parameters, oxidative stress values were assessed postoperatively. Results: The number of patients requiring additional propofol was higher in Group PM than in Groups NM and CM (p = 0.003). The postoperative Visual Analog Scale (VAS) score were lower in Groups PM and CM than in Group NM (p = 0.001, p = 0.007) in the 1st and 60th minutes. The postoperative VAS score was lower in Group CM than in Group NM (p = 0.045) in the 5th minute. The postoperative additional analgesic requirements were lower in Groups PM and CM than in Group NM (p = 0.045). The postoperative blood glutathione peroxidase values were significantly higher in Groups PM and CM than in Group NM (p = 0.001). The postoperative catalase values were significantly higher in Groups PM and CM than in Group NM (p = 0.008 and p < 0.001). The preoperative malondialdehyde values were significantly lower in Groups PM and CM than in Group NM. The preoperative nitric oxide values were higher in Groups PM and CM than in Group NM (p < 0.001), whereas the postoperative nitric oxide values were lower in Groups PM and CM than in Group NM (p < 0.001). Conclusion: Turkish classical music has beneficial effects on pain and oxidative stress in oocyte pick-up patients.

Resumo Justificativa e objetivo: O objetivo deste estudo foi investigar os efeitos da música clássica turca sobre a dor e o estresse oxidativo em pacientes submetidas a aspiração folicular. Método: Estudo randomizado controlado. Os grupos foram: grupo controle NM, sem música; Grupo PM, com pacientes que ouviram música antes da cirurgia; e Grupo CM, com pacientes que ouviram música antes e durante a cirurgia. Foi coletado sangue antes da cirurgia para avaliar os valores de estresse oxidativo. Dor, parâmetros hemodinâmicos e valores de estresse oxidativo foram avaliados após a cirurgia. Resultados: O número de pacientes que necessitaram de propofol adicional foi mais alto no Grupo PM do que nos grupos NM e CM (p = 0,003). A pontuação da Escala Visual Analógica (EVA) pós-operatória foi mais baixa nos Grupos PM e CM do que no Grupo NM (p = 0,001; p = 0,007), no 1° e 60° minutos. A pontuação da EVA pós-operatória foi mais baixa no Grupo CM do que no grupo NM (p = 0,045) no 5° minuto. A necessidade de analgesia pós-operatória adicional foi mais baixa nos Grupos PM e CM do que no Grupo NM (p = 0,045). Os valores pós-operatórios de glutationa peroxidase no sangue foram significantemente mais altos nos Grupos PM e CM do que no Grupo NM (p = 0,001). Os valores pós-operatórios de catalase foram significantemente mais altos nos Grupos PM e CM do que no Grupo NM (p = 0,008 e p≤ 0,001). Os valores pré-operatórios de malondialdeído foram significantemente mais baixos nos grupos PM e CM do que no Grupo NM. Os valores pré-operatórios de óxido nítrico foram mais altos nos grupos PM e CM do que no Grupo NM (p≤ 0,001), ao passo que valores pós-operatórios de óxido nítrico foram mais baixos nos grupos PM e CM do que no Grupo NM (p≤ 0,001). Conclusão: Música clássica turca exerce efeito benéfico sobre a dor e estresse oxidativo em pacientes na aspiração folicular.

Pregnancy outcomes of the first thawing cycle in "freeze-all" strategy of infertility patients with fever during oocyte recruitment: a matched-pair study / 中华医学杂志(英文版)

BACKGROUND@#It is currently unknown whether patients with a fever after controlled ovulation during egg retrieval could increase the risk of pelvic infection or not, and fever itself may affect endometrial receptivity or embryo quality with poor pregnancy outcomes. The aim of this study was to analyze the outcomes of patients with fever during oocyte retrieval after the first frozen-thawed embryo transfer (FET) cycle.@*METHODS@#This was a 1:3 retrospective paired study matched for age. In this study, 58 infertility patients (Group 1) had a fever during the control ovulation, and the time of the oocyte retrieval was within 72 hours, they underwent ovum pick up and whole embryo freezing ("freeze-all" strategy). The control subjects (Group 2) are 174 patients matched for age who underwent whole embryo freezing for other reasons. The baseline characteristics, clinical data of ovarian stimulation, and outcomes, such as the clinical pregnancy rate, ongoing clinical pregnancy rate were compared between the two groups in the subsequent FET cycle.@*RESULTS@#All patients had no pelvic inflammatory disease after oocyte retrieval. Anti-Mullerian hormone (AMH) levels (4.2 vs. 2.2, P <0.001) were higher in group 2, and the number of oocytes retrieved, and fertilization rate were lower in group 1 (P < 0.001), but the endometrial thickness, the number of embryo transfers, and the type of luteal support supplementation were similar between the two groups. Regarding pregnancy outcomes in the subsequent FET cycle, the implantation rate, clinical pregnancy rate, early spontaneous rate, ectopic pregnancy rate, and ongoing pregnancy rate were all not significantly different. Further regression analyses showed that the clinical pregnancy rate and ongoing pregnancy rate were also not significantly different.@*CONCLUSIONS@#Transvaginal ultrasound-guided follicular puncture for oocyte retrieval is a safe and minimally invasive method for patients with fever. Moreover, the fever had almost no effect on embryo quality.

Efeito da técnica de coleta e do estágio do ciclo estral na recuperação de oócitos de boa qualidade em felinos e caninos domésticos / The effect of recovery technique and stage of estrous cycle on the recoveryof good quality oocytes in domestic felines and canines

A obtenção de oócitos de boa qualidade é essencial para o sucesso de diversas biotécnicas reprodutivas. Objetivou-se determinar o efeito de duas técnicas na recuperação de oócitos de boa qualidade em gatas e cadelas em diferentes estágios reprodutivos. Foram utilizados 43 pares de ovários de gata e 35 de cadela após realização da ovariosalpingohisterectomia eletiva. A fase do ciclo estral foi classificada em inativa, folicular ou luteal. Os ovários da fase folicular foram divididos em três grupos: PUN) punção dos folículos com agulha; PUN+FAT) fatiamento do mesmo ovário já puncionado; e FAT) fatiamento do segundo ovário. Os ovários das fêmeas em fase luteal e inativa foram submetidos ao FAT. Foram obtidos no total 974 oócitos (~23/animal) nas fêmeas felinas e 940 (~27/animal) nas caninas. O fatiamento recuperou número superior (P0,05) entre as técnicas de coleta na qualidade de estruturas recuperadas. A quantidade de oócitos recuperados em cada fase foi similar (P>0,05). Contudo, a fase inativa foi superior à luteal (P<0,05) e semelhante à folicular na quantidade de oócitos de boa qualidade em gatas e não houve diferença em cadelas. Conclui-se que o fatiamento recupera maior quantidade de oócitos, não influenciando em sua qualidade. As fases inativa e folicular recuperam maior quantidade de oócitos de boa qualidade em gatas e não afetam a recuperação em cadelas. Portanto, para otimizar o uso das biotecnologias, deve-se levar em consideração o estágio do ciclo estral em fêmeas felinas e a técnica de coleta utilizada na recuperação de oócitos.

The recovery of good quality oocytes is essential for the success of various reproductive biotechniques. The aim of this study was to determine the effect of two techniques on the recovery of good quality oocytes in queens and bitches at different reproductive stages. A total of 43 pairs of ovaries of queens and 35 of bitches after elective ovariosalpingohisterectomy were performed. The estrous cycle phase was classified as inactive, follicular or luteal. The ovaries of the follicular phase were allocated into three groups: PUN) puncture of the follicles with a needle; PUN + SLI) slicing of the same ovary already punctured; and SLI) slicing of the second ovary. The ovaries of luteal and inactive females were submitted to SLI. A total of 974 oocytes (~23/animal) were obtained in feline females and 940 (~27/animal) in canines females. The SLI technique recovered superior number (P0.05) between the collection techniques in the quality of recovered structures. The number of oocytes recovered in each phase was similar (P>0.05). However, the inactive phase was higher than luteal (P<0.05) and similar to the follicular phase in the quantity of good-quality oocytes in queens and there was no difference in bitches. In conclusion, it is preferable to perform the slicing technique to recover more oocytes in both species. Moreover, in queens it is possible to obtain good quality oocytes in the inactive phase and in bitches the estrous cycle phase does not influence the quality.

Efeito da técnica de coleta e do estágio do ciclo estral na recuperação de oócitos de boa qualidade em felinos e caninos domésticos / The effect of recovery technique and stage of estrous cycle on the recovery of good quality oocytes in domestic felines and canines

A obtenção de oócitos de boa qualidade é essencial para o sucesso de diversas biotécnicas reprodutivas. Objetivou-se determinar o efeito de duas técnicas na recuperação de oócitos de boa qualidade em gatas e cadelas em diferentes estágios reprodutivos. Foram utilizados 43 pares de ovários de gata e 35 de cadela após realização da ovariosalpingohisterectomia eletiva. A fase do ciclo estral foi classificada em inativa, folicular ou luteal. Os ovários da fase folicular foram divididos em três grupos: PUN) punção dos folículos com agulha; PUN+FAT) fatiamento do mesmo ovário já puncionado; e FAT) fatiamento do segundo ovário. Os ovários das fêmeas em fase luteal e inativa foram submetidos ao FAT. Foram obtidos no total 974 oócitos (~23/animal) nas fêmeas felinas e 940 (~27/animal) nas caninas. O fatiamento recuperou número superior (P<0,05) de oócitos. Não houve diferença (P>0,05) entre as técnicas de coleta na qualidade de estruturas recuperadas. A quantidade de oócitos recuperados em cada fase foi similar (P>0,05). Contudo, a fase inativa foi superior à luteal (P<0,05) e semelhante à folicular na quantidade de oócitos de boa qualidade em gatas e não houve diferença em cadelas. Conclui-se que o fatiamento recupera maior quantidade de oócitos, não influenciando em sua qualidade. As fases inativa e folicular recuperam maior quantidade de oócitos de boa qualidade em gatas e não afetam a recuperação em cadelas. Portanto, para otimizar o uso das biotecnologias, deve-se levar em consideração o estágio do ciclo estral em fêmeas felinas e a técnica de coleta utilizada na recuperação de oócitos.

The recovery of good quality oocytes is essential for the success of various reproductive biotechniques. The aim of this study was to determine the effect of two techniques on the recovery of good quality oocytes in queens and bitches at different reproductive stages. A total of 43 pairs of ovaries of queens and 35 of bitches after elective ovariosalpingohisterectomy were performed. The estrous cycle phase was classified as inactive, follicular or luteal. The ovaries of the follicular phase were allocated into three groups: PUN) puncture of the follicles with a needle; PUN + SLI) slicing of the same ovary already punctured; and SLI) slicing of the second ovary. The ovaries of luteal and inactive females were submitted to SLI. A total of 974 oocytes (~23/animal) were obtained in feline females and 940 (~27/animal) in canines females. The SLI technique recovered superior number (P<0.05) of oocytes. There was no difference (P>0.05) between the collection techniques in the quality of recovered structures. The number of oocytes recovered in each phase was similar (P>0.05). However, the inactive phase was higher than luteal (P<0.05) and similar to the follicular phase in the quantity of good-quality oocytes in queens and there was no difference in bitches. In conclusion, it is preferable to perform the slicing technique to recover more oocytes in both species. Moreover, in queens it is possible to obtain good quality oocytes in the inactive phase and in bitches the estrous cycle phase does not influence the quality.

Comparison of two surgical approaches for Laparoscopic Ovum Pick Up in Ewes / Comparação de duas abordagens cirúrgicas para aspiração folicular laparoscópica em ovelhas

The present study had as an aim to evaluate a right lateral access as an alternative method to laparoscopic ovum pick-up (LOPU) in sheep. Twenty-four Santa Ines ewes were randomly assigned in two groups with twelve animals each: RLD - positioned in right lateral decubitus, with 10º head-down tilt; and DD - positioned in dorsal decubitus with 35º head-down tilt. The following parameters were evaluated every 10 minutes during the procedure: total surgical time (ST), visualized follicles (VF), aspirated follicles (AF), recovered oocytes (RO), mean arterial pressure (MAP), heart rate (HR), respiratory rate (fR) and end tidal CO2 pressure (EtCO2). Pre and postoperative arterial hemogasometry parameters (PaO2, PaCO2, pH, CHCO3 and BE) were also evaluated; and serum fibrinogen levels (SFL) on postoperative period. The values of VF, AF, RO, fR, PaO2, pH, CHCO3, BE and SFL were similar between groups, although ST, HR, MAP, EtCO2 and PaCO2 were higher in LG. Regarding operative periods, PaO2 and pH were lower after surgery (PaO2: 79.1±16.4; 79.2±11.7mmHg; pH: 7.30±0.09; 7.32±0.08) in both groups when compared to preoperative (PaO2: 80.1±14.3; 83.4±10.5 mmHg; pH: 7.38±0.05; 7.39±0.05) while PaCO2 (43.6±4.6; 41.9±5.4mmHg) and CHCO3 (22.8±1.5; 22.7±3.0mmol/L) increased (PaCO2: 54.3±10.9; 46.9±6.3mmHg; CHCO3: 24.8±3.4; 24.4±2.7mmol/L) postoperative. This alternative decubitus presented is a viable procedure and did not differ in oocyte recovery rates in ewes. However, entails cardiorespiratory major alterations compared to conventional procedure, making its practical applicability limited.(AU)

O presente estudo teve como objetivo avaliar o acesso lateral direito como um método alternativo para a recuperação de oócitos por laparoscopia (LOPU) em ovelha. Vinte e quatro ovelhas Santa Inês foram distribuídas aleatoriamente em dois grupos com 12 animais: grupo RLD - posicionado em decúbito lateral direito, cefalodeclive com 10º de inclinação; grupo DD - posicionado em decúbito dorsal em cefalodeclive, inclinação de 35º. Foram avaliados, a cada 10 minutos, durante o procedimento cirúrgico: tempo total da cirurgia (ST), folículos visualizados (VF), folículos aspirados (AF), oócitos recuperados (RO), pressão arterial média (MAP), frequência cardíaca (FC), frequência respiratória (fR) e pressão final de CO2 (EtCO2). Também foram avaliados os parâmetros de hemogasometria arterial pré-operatória e pós-operatória (PaO2, PaCO2, pH, CHCO3 e BE), bem como os níveis séricos de fibrinogênio (SFL) no período pós-operatório. Os valores de VF, AF, RO, fR, PaO2, pH, CHCO3, BE e SFL foram semelhantes entre os grupos, embora ST, FC, MAP, EtCO2 e PaCO2 tenham sido maiores em RLD. Os parâmetros PaO2 e pH foram menores após a cirurgia (PaO2: 79,1±16,4; 79,2±11,7mmHg; pH: 7,30±0,09; 7,32±0,08) em ambos os grupos em relação ao momento pré-cirúrgico (PaO2: 80,1±14,3; 83,4±10,5mmHg; pH: 7,38±0,05; 7,39±0,05), enquanto PaCO2 (43,6±4.6; 41,9±5,4mmHg) e CHCO3 (22,8±1,5; 22,7±3.0mmol/L) aumentaram (PaCO2: 54,3±10,9; 46,9±6,3mmHg; CHCO3: 24,8±3,4; 24,4±2,7mmol/L) após a cirurgia. O decúbito lateral é uma alternativa viável para LOPU e não apresenta diferença para a taxa de recuperação oocitária em ovelhas. No entanto, promove alterações cardiorrespiratórias em comparação com o decúbito dorsal, tornando a sua aplicabilidade prática limitada.(AU)

Iatrogenic ureteral obstruction during transvaginal ocyte retrieval

ABSTRACT Transvaginal oocyte retrieval is a crucial step in assisted reproductive technology. Various complications may arise during this procedure. Ureteral injury is a rare, but a serious complication in gynecological practice. During oocyte retrieval, ureteral injuries, detachment and obstruction can be seen, though rare. In this study, we will present ureteral obstruction that develops secondary to small hematoma, which mimics ovarian cyst torsion or ruptured ovarian cyst.

Could surgical management improve the IVF outcomes in infertile women with endometrioma?: a review

Endometriosis is a chronic inflammatory condition that affects fertility and could be toxic to the ovary. Endometrioma per se and surgical interventions for endometrioma significantly reduce the ovarian reserve. Therefore, to prepare for surgical intervention for endometrioma, the high-risk group with decreased ovarian reserve must be considered. There is no evidence to support the use of surgical intervention before in vitro fertilization (IVF) to improve the reproductive outcomes of subsequent IVF in infertile women with advanced-stage endometriosis or endometrioma. As surgical treatment has few benefits, IVF could be recommended immediately for aiding conception in these women. However, the reproductive prognosis of IVF may be worse in the more advanced stages of endometriosis. When dysmenorrhea is severe or when cancer is suspected, surgery prior to IVF may be necessary and justified. When the size of the endometrioma is very large, surgery could be required prior to IVF to facilitate access to follicles during oocyte retrieval or to improve the ovarian response to controlled ovarian stimulation. Prolonged pituitary downregulation in women with surgically diagnosed endometriosis may be helpful to increase the clinical pregnancy rate in subsequent IVF cycles. The purpose of this paper was to review the efficiency and clinical application of the surgical intervention and IVF for infertile women with advanced-stage endometriosis or endometrioma.

Effect of Sperm DNA Fragmentation on Embryo Quality in Normal Responder Women in In Vitro Fertilization and Intracytoplasmic Sperm Injection

PURPOSE: To investigate the associations between sperm DNA fragmentation (SDF) and embryo formation rate in normal responder women to in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI). MATERIALS AND METHODS: Fifty-three consecutive, fresh IVF/ICSI cycles performed from 2014 to 2017 were selected. All women were normal responders (4 to 14 mature oocytes were retrieved) and at least one normally fertilized oocyte with two pronuclei was obtained in all cycles. Semen was collected on the day of oocyte retrieval, and SDF levels were measured by sperm chromatin dispersion test (Halosperm assay). At day 3 after insemination, embryo quality was evaluated by morphologic criteria and categorized as A/B/C/D. Top quality embryo were defined as grade A embryos with seven cells or more. RESULTS: SDF levels showed a positive linear correlation with the male's age (r=0.307, p=0.025) and a negative linear correlation with sperm motility (r=−0.491, p70%, the cut-off value SDF was <30.7% for each. Among individuals with SDF <30.7%, the median top-quality or grade A embryo formation rate was significantly higher than that among individuals with SDF ≥30.7% (38.1% vs. 20.0%, p=0.038; 50% vs. 25.0%, p=0.017). CONCLUSION: In normal responder women, high SDF level resulted in low day 3 embryo formation rates. Our results suggest a paternal effect on embryo quality in IVF/ICSI cycles.

Correlation between Ovarian Reserve and Incidence of Ectopic Pregnancy after In Vitro Fertilization and Embryo Transfer

PURPOSE: To elucidate the correlation between ovarian reserve and the incidence of ectopic pregnancy (EP) following in vitro fertilization and embryo transfer (IVF/ET) cycles. MATERIALS AND METHODS: In this observational study, 430 fresh IVF/ET cycles were examined from patient data of two university hospital infertility clinics. All included patients were positive for β-human chorionic gonadotropin (hCG) at 2 weeks after oocyte retrieval via controlled ovarian stimulation. For each cycle, information on age, duration of infertility, basal follicle stimulating hormone (FSH), anti-Müllerian hormone (AMH), days of ovarian stimulation, numbers of retrieved oocytes and transferred embryos, and pregnancy outcomes was collected. Patients with AMH lower than 1.0 ng/dL or basal FSH higher than 10 mIU/mL were classified into the decreased ovarian reserve (DOR) group, and the remaining patients were classified into the normal ovarian reserve (NOR) group. RESULTS: In total, 355 cycles showed NOR, and 75 cycles DOR. There were no significant differences between the DOR and NOR groups regarding intrauterine (74.7% vs. 83.4%, respectively) or chemical (14.7% vs. 14.1%, respectively) pregnancies. The DOR group had a higher EP than that of NOR group [10.7% (8/75) vs. 2.5% (9/355), p=0.004]. In both univariate [odds ratio (OR) 5.6, 95% confidence interval (CI) 1.4–9.6, p=0.011] and multivariate (adjusted OR 5.1, 95 % CI 1.1–18.7, p=0.012) analysis, DOR was associated with a higher risk of EP. CONCLUSION: DOR may be associated with a higher risk of EP in IVF/ET cycles with controlled ovarian stimulation. More careful monitoring may be necessary for pregnant women with DOR.

Oxidative stress in granulosa cells contributes to poor oocyte quality and IVF-ET outcomes in women with polycystic ovary syndrome / 医学前沿

The increased levels of intracellular reactive oxygen species (ROS) in granulosa cells (GCs) may affect the pregnancy results in women with polycystic ovary syndrome (PCOS). In this study, we compared the in vitro fertilization and embryo transfer (IVF-ET) results of 22 patients with PCOS and 25 patients with tubal factor infertility and detected the ROS levels in the GCs of these two groups. Results showed that the PCOS group had significantly larger follicles on the administration day for human chorionic gonadotropin than the tubal factor group (P 0.05). PCOS group had slightly lower fertilization, cleavage, grade I/II embryo, clinical pregnancy, and implantation rates and higher miscarriage rate than the tubal factor group (P > 0.05). We further found a significantly higher ROS level of GCs in the PCOS group than in the tubal factor group (P < 0.05). The increased ROS levels in GCs caused GC apoptosis, whereas NADPH oxidase 2 (NOX2) specific inhibitors (diphenyleneiodonium and apocynin) significantly reduced the ROS production in the PCOS group. In conclusion, the increased ROS expression levels in PCOS GCs greatly induced cell apoptosis, which further affected the oocyte quality and reduced the positive IVF-ET pregnancy results of women with PCOS. NADPH oxidase pathway may be involved in the mechanism of ROS production in GCs of women with PCOS.

Use of different pressures for transvaginal follicular aspiration in mares / Uso de diferentes pressões para aspiração folicular transvaginal em éguas

The success of transvaginal follicular aspiration in mares can be influenced by several factors, such as vacuum pump pressure levels. The present study aimed to investigate the effect of different negative pressures (150, 280 and 400mmHg) of the vacuum pump on the oocyte recovery in the mares. The mares (n=10) were undergoing follicular aspiration using three different negative pressures for three consecutive estrous cycles as follows: G150 = 150mmHg (n = 10); G280 = 280mmHg (n = 10); G400 = 400mmHg (n = 10). Every estrous cycle, the group that the mare would participate was drawn, and each animal participated once in each group. Only preovulatory follicle was used, about 30 to 36 hours after application of hCG. To compare the results, the chi-square test was used (5% significance) and Fisher exact test, when recommended. Thirty preovulatory follicles (diameter 36.1±1.80mm) were aspirated and ten oocytes were recovered (33.3%). There was no statistical difference between the experimental groups (p=0.59). Thus, accord to the results observed in this study, we could conclude that the negative pressure of the vacuum pump used was not efficient to increase oocyte recovery.(AU)

O sucesso da técnica de aspiração folicular transvaginal em éguas pode ser influenciado de maneira determinante por diversos fatores, tais como níveis de pressão da bomba de vácuo. Diante disso, o presente experimento visou investigar o efeito de diferentes pressões negativas (150, 280 e 400mmHg) da bomba de vácuo sobre a taxa de recuperação de oócitos em éguas. As éguas (n=10) foram submetidas à aspiração folicular utilizando-se três diferentes pressões negativas por três ciclos estrais consecutivos, da seguinte maneira: G150= 150mmHg (n=10); G280= 280mmHg (n=10); G400= 400mmHg (n=10). A cada ciclo estral, sorteava-se o grupo do qual a égua participaria, sendo que cada animal integrou um grupo somente uma vez. Foi puncionado somente folículo pré-ovulatório, em torno de 30 a 36 horas após a aplicação do hCG. Os resultados foram comparados utilizando-se o teste qui-quadrado (a 5% de significância) e o Fisher Exato, quando recomendados. Foram aspirados 30 folículos pré-ovulatórios (diâmetro 36,1±1,80mm) e recuperados 10 oócitos (33,3%). Não houve diferença estatística entre os grupos experimentais (P=0,59). Dessa forma, mediante os resultados obtidos no presente estudo, foi possível concluir que a pressão negativa da bomba de vácuo utilizada não se mostrou determinante para elevar a recuperação oocitária.(AU)

Effects of mono-[2-ethylhexyl] phthalate and DI-[2-ethylhexyl] phthalate administrations on oocyte meiotic maturation, apoptosis and gene quantification in mouse model

Objective: Phthalates, which are commonly used to render plastics into soft and flexible materials, have also been determined as developmental and reproductive toxicants in human and animals. The purpose of this study was to evaluate the effect of mono-[2-ethylhexyl] phthalate [MEHP] and di-[2-ethylhexyl] phthalate [DEHP] oral administrations on maturation of mouse oocytes, apoptosis and gene transcription levels

Materials and Methods: In this experimental study, immature oocytes recovered from Naval Medical Research Institute [NMRI] mouse strain [6-8 weeks], were divided into seven different experimental and control groups. Control group oocytes were retrieved from mice that received only normal saline. The experimental groups I, II or III oocytes were retrieved from mice treated with 50, 100 or 200 micro l DEHP [2.56 micro M] solution, respectively. The experimental groups IV, V or VI oocytes were retrieved from mouse exposed to 50, 100 or 200 micro l MEHP [2.56 micro M] solution, respectively. Fertilization and embryonic development were carried out in OMM and T6 medium. Apoptosis was assessed by annexin V-FITC/Dead Cell Apoptosis Kit, with PI staining. In addition, the mRNA levels of Pou5f1, Ccna1 and Asah1 were examined in oocytes. Finally, mouse embryo at early blastocyst stage was stained with acridine-orange [AO] and ethidium-bromide [EB], in order to access their viability

Results: The proportion of oocytes that progressed up to metaphase II [MII] and 2-cells embryo formation stage was significantly decreased by exposure to MEHP or DEHP, in a dose-dependent manner. Annexin V and PI positive oocytes showed greater quantity in the treated mice than control. Quantitative reverse transcriptase-polymerase chain reaction [qRT-PCR] revealed that expression levels of Pou5f1, Asah1 and Ccna1 were significantly lower in the treated mouse oocytes than control. The total cell count for blastocyst developed from the treated mouse oocytes was lower than the controls

Conclusion: These results indicate that oral administration of MEHP and DEHP could negatively affect mouse oocyte meiotic maturation and development in vivo, suggesting that phthalates could be risk factors for mammalians' reproductive health. Additionally, phthalate-induced changes in Pou5f1, Asah1 and Ccna1 transcription level could explain in part, the reduced developmental ability of mouse-treated oocytes

Impact of sperm DNA fragmentation on clinical in vitro fertilization outcomes / 대한생식의학회지

OBJECTIVE: We studied the association between sperm DNA fragmentation (SDF) and several clinical in vitro fertilization outcomes. METHODS: We retrospectively analyzed 169 consecutive fresh IVF cycles. Semen was collected on the day of oocyte retrieval, and we assessed standard semen parameters and the SDF level (by terminal deoxynucleotidyl transferase dUTP nick-end labeling). Poor ovarian response (POR) was defined as the collection of three or fewer mature oocytes. Oocytes were inseminated by the conventional method or intracytoplasmic sperm injection. RESULTS: SDF did not affect the fertilization or pregnancy rate, but did have a significant effect on the miscarriage rate. In the miscarriage group (n=10), the SDF level was significantly higher (23.9% vs. 14.1%) and number of mature oocytes was significantly lower (4.3 vs. 7.6) than in the live birth group (n=45). Multiple regression analysis showed that SDF was an independent predictor of miscarriage (odds ratio, 1.051; 95% confidence interval, 1.001–1.104). The cutoffs for the SDF level and number of mature oocytes that could predict miscarriage were >13% and ≤3, respectively. In the low-SDF group (≤13%), the miscarriage rate was similar in POR patients and those with a normal ovarian response (NOR; 14.2% vs. 4.3%). In the high-SDF group (>13%), the miscarriage rate was significantly higher in the POR group than in the NOR group (60.0% vs. 13.3%, p=0.045). CONCLUSION: Our study demonstrated that a high SDF level (>13%) was associated with a high miscarriage rate, and that it mainly contributed to miscarriage in the POR group. The results suggest that SDF measurements should be considered in couples with POR in order to predict the prognosis of the pregnancy.

The association of follicular fluid volume with human oolemma stretchability during intracytoplasmic sperm injection / 대한생식의학회지

OBJECTIVE: Oocyte degeneration often occurs after intracytoplasmic sperm injection (ICSI), and the risk factor is low-quality oocytes. The follicular fluid (FF) provides a crucial microenvironment for oocyte development. We investigated the relationships between the FF volume aspirated from individual follicles and oocyte retrieval, oocyte maturity, oolemma stretchability, fertilization, and development. METHODS: This retrospective study included data obtained from 229 ICSI cycles. Ovarian stimulation was performed according to a gonadotropin-releasing hormone antagonist protocol. Each follicle was individually aspirated and divided into six groups according to FF volume (<1.0, 1.0 to <2.0, 2.0 to <3.0, 3.0 to <4.0, 4.0 to <5.0, and ≥5.0 mL). Oolemma stretchability during ICSI was evaluated using a mechanical stimulus for oolemma penetration, that is, the stretchability was assessed by oolemma penetration with aspiration (high stretchability) or without aspiration (low stretchability). RESULTS: Oocyte retrieval rates were significantly lower in the <1.0 mL group than in the ≥1.0 mL groups (46.0% [86/187] vs. 67.5%–74.3% [172/255 to 124/167], respectively; p<0.01). Low oolemma stretchability was significantly more common in the <1.0 mL group than in the ≥1.0 mL groups during ICSI (22.0% [13/59] vs. 5.8%–9.4% [6/104 to 13/139], respectively; p=0.018). There was a relationship between FF volume and oolemma stretchability. However, there were no significant differences in the rates of fertilization, cleavage, ≥7 cells at day 3, and blastocyst development among all groups. CONCLUSION: FF volume is potentially associated with the stretchability of metaphase II oolemma during ICSI. Regarding oolemma stretchability, ensuring a uniform follicular size during ovarian stimulation is crucial to obtain good-quality oocytes.

Expression of the genes for peroxisome proliferator-activated receptor-γ, cyclooxygenase-2, and proinflammatory cytokines in granulosa cells from women with polycystic ovary syndrome / 대한생식의학회지

OBJECTIVE: To identify differences in the expression of the genes for peroxisome proliferator-activated receptor (PPAR)-γ, cyclooxygenase (COX)-2, and the proinflammatory cytokines interleukin (IL)-6 and tumor necrosis factor (TNF)-α in granulosa cells (GCs) from polycystic ovary syndrome (PCOS) patients and controls undergoing controlled ovarian stimulation. METHODS: Nine patients with PCOS and six controls were enrolled in this study. On the day of oocyte retrieval, GCs were collected from pooled follicular fluid. Total mRNA was extracted from GCs. Reverse transcription was performed and gene expression levels were quantified by realtime quantitative polymerase chain reaction. RESULTS: There were no significant differences in age, body mass index, and total gonadotropin dose, except for the ratio of luteinizing hormone to follicle-stimulating hormone between the PCOS and control groups. PPAR-γ and COX-2 mRNA was significantly downregulated in the GCs of PCOS women compared with controls (p=0.034 and p=0.018, respectively), but the expression of IL-6 and TNF-α mRNA did not show significant differences. No significant correlation was detected between the expression of these mRNA sequences and clinical characteristics, including the number of retrieved oocytes, oocyte maturity, cleavage, or the good embryo rate. Positive correlations were found among the PPAR-γ, COX-2, IL-6, and TNF-α mRNA levels. CONCLUSION: Our data may provide novel clues regarding ovarian GC dysfunction in PCOS, and indirectly provide evidence that the effect of PPAR-γ agonists in PCOS might result from alterations in the ovarian follicular environment. Further studies with a larger sample size are required to confirm these proposals.

Randomized double-blind clinical trial comparing two anesthetic techniques for ultrasound-guided transvaginal follicular puncture / Estudo clínico duplo-cego randomizado comparando duas técnicas anestésicas para punção folicular guiada por ultrassom transvaginal

ABSTRACT Objective: To compare the anesthetic techniques using propofol and fentanyl versus midazolam and remifentanil associated with a paracervical block with lidocaine in performing ultrasound-guided transvaginal oocyte aspiration. Methods: A randomized double-blind clinical trial (#RBR-8kqqxh) performed in 61 women submitted to assisted reproductive treatment. The patients were divided into two groups: anesthetic induction with 1mcg/kg of fentanyl associated with 1.5mg/kg of propofol (FP Group, n=32), in comparison with anesthetic induction using 0.075mg/kg of midazolam associated with 0.25mcg/kg/min of remifentanil, and paracervical block with 3mL of 2% lidocaine (MRPB Group, n=29). Main outcome measures: human reproduction outcomes, modified Aldrete-Kroulik index, hemodynamic parameters, and salivary cortisol. Results: The results revealed a higher number of embryos formed in the FP Group (p50=2 versus 1; p=0.025), gestation rate two times higher in the FP Group (44.4% versus 22.2%; p=0.127), less time to reach AK=10 in the MRPB Group (p50=10 versus 2; p<0.001), and lower mean of hemodynamic parameters in the MRPB Group (p<0.05). Conclusion: Anesthesia with fentanyl and propofol as well as with midazolam, remifentanil, and paracervical block offered satisfactory anesthetic conditions when performing assisted reproduction procedures, providing comfort for the patient and physician.

RESUMO Objetivo: Comparar as técnicas anestésicas utilizando propofol e fentanil contra midazolam e remifentanil associados a um bloqueio paracervical com lidocaína na aspiração de oócitos transvaginal guiada por ultrassom. Métodos: Ensaio clínico randomizado duplocego (#RBR-8kqqxh) realizado em 61 mulheres submetidas ao tratamento de reprodução assistida. As pacientes foram divididas em dois grupos: um grupo foi submetido à indução da anestesia com 1mcg/kg de fentanil associada com 1,5mg/kg de propofol (Grupo FP, n=32), em comparação com ao grupo submetido à indução da anestesia utilizando 0,075mg/kg de midazolam associada com 0,25mcg/kg/min de remifentanil, e bloqueio paracervical com 3mL de lidocaína a 2% (Grupo MRBP, n=29). Foram avaliados os resultados reprodutivos, índice modificado de Aldrete e Kroulik, parâmetros hemodinâmicos e cortisol salivar. Resultados: Foi encontrado um número mais elevado de embriões formados no Grupo FP (p50=2 versus 1; p=0,025), taxa de gestação duas vezes mais elevada no Grupo FP (44,4% versus 22,2%; p=0,127), menos tempo para alcançar AK=10 no Grupo MRBP (p50=10 versus 2; p<0,001) e média mais baixa de parâmetros hemodinâmicos do Grupo MRBP (p<0,05). Conclusão: Ambas as anestesias (com fentanil e propofol, e com midazolam, remifentanil e bloqueio paracervical) ofereceram condições anestésicas satisfatórias na realização de procedimentos realizados em reprodução assistida, proporcionando conforto para o paciente e médico.

Random-start controlled ovarian stimulation for emergency fertility preservation in a patient with myelodysplastic syndrome: a case report

This study reports a case of a gonadotropin-releasing hormone agonist trigger in a young female with myelodysplastic syndrome (MDS) who underwent fertility preservation using random-start controlled ovarian stimulation. This method involves the stimulation of the ovary regardless of a patient's menstrual-cycle phase. A review of the related literature is also provided. A 17-year-old patient was diagnosed with MDS and required initiation of peripheral blood stem cell transplantation within a maximum of 3 weeks and was in the luteal phase of the menstrual cycle when the possibility of attempting preservation of fertility was presented to her. She opted for a random-start controlled ovarian stimulation with gonadotropins. With successful hemorrhagic prophylaxis, 17 oocytes were retrieved including 10 mature and 7 immature oocytes. Of the immature oocytes, 3 were successfully matured in vitro and a vitrification protocol was used to freeze the 13 mature oocytes.

Effects of Cangfu Congxian Decoction on Oxidative Stress in Polycystic Ovary Syndrome Patients / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Cangfu Congxian Decoction (CCD) on oxidative stress in granulosa cells of polycystic ovary syndrome (PCOS) patients.</p><p><b>METHODS</b>Forty PCOS patients underwent in vitro fertilization-embryo transfer (IVF-ET) were assigned to the treatment group and the control group 1 according to random digit table, 20 in each group. Patients in the treatment group took CCD (200 mL, once in the morning and once in the afternoon) 2 months before IVF-ET, while those in the control group 1 took no Chinese medical decoction. Recruited were another 20 patients undergoing IVF-ET for tubal factors (as the control group 2). The clinical effect of IVF-ET were observed, including oocyte retrieval number, 2 pronuclear (2PN) fertilization rate, good quality embryo rate, clinical pregnancy rate, and ovarian hyperstimulation syndrome (OHSS) induced transplantation cancel rate. The expression of relative oxygen species (ROS) in granulosa cells was detected using cell immunofluorescence combined with confocal microscopy and FCM.</p><p><b>RESULTS</b>Compared with the control group 1, occyte retrieval number, 2PN fertilization rate, and good quality embryo rate increased in the control group 2 and the treatment group (P <0. 05). OHSS induced transplantation cancel rate decreased in the control group 2 (P < 0.05). Fluorescence intensity of ROS decreased in the treatment group and the control group 2, as compared with the control group 1 (P < 0.01).</p><p><b>CONCLUSION</b>CCD increased good quality embryo rate by down-regulating the expression of ROS protein in ovarian granulosa cells, and correcting in vivo oxidative stress.</p>

The rate of morphologically normal sperm does not affect the clinical outcomes of conventional IVF in patients with one retrieved oocyte / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the influence of the rate of morphologically normal sperm (MNS) on the clinical outcomes of conventional in vitro fertilization (IVF) in patients with one retrieved oocyte.</p><p><b>METHODS</b>From January 2013 to January 2015, a total of 256 couples with one retrieved oocyte underwent conventional IVF in our center. According to the rate of MNS, the patients were divided into two groups: MNS < 4% (134 cycles) and MNS ≥ 4% (122 cycles). We compared the rates of no transferrable embryo cycles, fertilization, cleavage, normal fertilization, abnormal fertilization, high-quality embryo and transferrable embryo between the two groups. A total of 75 fresh embryo transfer cycles were performed, 43 in the MNS < 4% group and the other 32 in the MNS ≥ 4% group. We also compared the rates of implantation, clinical pregnancy and abortion between the two groups.</p><p><b>RESULTS</b>There were no statistically significant differences between the two groups in the rates of no transferrable embryo cycles, fertilization, cleavage, normal fertilization, abnormal fertilization, high-quality embryo and transferrable embryo (P > 0.05). The rates of implantation, clinical pregnancy and abortion exhibited no remarkable differences either in the fresh embryo transfer cycles between the two groups (P > 0.05).</p><p><b>CONCLUSION</b>The rate of MNS does not affect the clinical outcomes of conventional IVF in patients with one retrieved oocyte.</p>