Effects of Zishen Yutai Pills on in vitro Fertilization-Embryo Transfer Outcomes in Patients with Diminished Ovarian Reserve: A Prospective, Open-Labeled, Randomized and Controlled Study / 中国结合医学杂志

OBJECTIVE@#To explore the effects of Zishen Yutai Pills (ZYPs) on the quality of oocytes and embryos, as well as pregnancy outcomes in patients with diminished ovarian reserve (DOR) receiving in vitro fertilization-embryo transfer (IVF-ET). The possible mechanisms, involving the regulation of bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9), were also investigated.@*METHODS@#A total of 120 patients with DOR who underwent their IVF-ET cycle were randomly allocated to 2 groups in a 1:1 ratio. The patients in the treatment group (60 cases) received ZYPs from the mid-luteal phase of the former menstrual cycle by using gonadotropin-releasing hormone (GnRH) antagonist protocol. The patients in the control group (60 cases) received the same protocol but without ZYPs. The primary outcomes were the number of oocytes retrieved and high-quality embryos. Secondary outcomes included other oocyte or embryo indices as well as pregnancy outcomes. Adverse events were assessed by comparison of the incidence of ectopic pregnancy, pregnancy complications, pregnancy loss, and preterm birth. Contents of BMP15 and GDF9 in the follicle fluids (FF) were also quantified with enzyme-linked immunosorbent assay.@*RESULTS@#Compared with the control group, the numbers of oocytes retrieved and high-quality embryos were significantly increased in the ZYPs group (both P<0.05). After treatment with ZYPs, a significant regulation of serum sex hormones was observed, including progesterone and estradiol. Both hormones were up-regulated compared with the control group (P=0.014 and 0.008), respectively. No significant differences were observed with regard to pregnancy outcomes including implantation rates, biochemical pregnancy rates, clinical pregnancy rates, live birth rates, and pregnancy loss rates (all P>0.05). The administration of ZYPs did not increase the incidence of adverse events. The expressions of BMP15 and GDF9 in the ZYPs group were significantly up-regulated compared with the control group (both P<0.05).@*CONCLUSIONS@#ZYPs exhibited beneficial effects in DOR patients undergoing IVF-ET, resulting in increments of oocytes and embryos, and up-regulation of BMP15 and GDF9 expressions in the FF. However, the effects of ZYPs on pregnancy outcomes should be assessed in clinical trials with larger sample sizes (Trial reqistration No. ChiCTR2100048441).

Oral dydrogesterone in frozen-thawed embryo transfer cycles

SUMMARY OBJECTIVE: The aim of this study was to compare the use of micronized vaginal progesterone and oral dydrogesterone in the endometrial preparation for frozen-thawed embryo transfer. METHODS: This was a randomized, controlled, open, two-armed clinical trial, with women undergoing frozen-thawed embryo transfer along with hormone replacement therapy for endometrial preparation, between September 2019 and February 2021. A total of 73 patients were randomly selected and orally administered 40 mg/day dydrogesterone (dydrogesterone group, n=36) or 800 mg/day micronized vaginal progesterone (micronized vaginal progesterone group, n=37), after endometrial preparation with transdermal estradiol. The main outcome was a viable ongoing pregnancy with 12 weeks of gestation as evaluated by ultrasound. RESULTS: The reproductive outcomes in frozen-thawed embryo transfer cycles were similar, with pregnancy rates in the dydrogesterone and micronized vaginal progesterone treatment groups being, respectively, 33.3 and 32.4% at 12 weeks pregnancy (confidence interval= -22.4-20.6, p=0.196). CONCLUSIONS: The use of oral dydrogesterone may be a more patient-friendly approach to endometrial preparation in frozen-thawed embryo transfer cycles, avoiding undesirable side effects and discomfort resulting from vaginal administration, while also providing similar reproductive results.

A meta-analysis on uterine transplantation: Redefining the limits of reproductive surgery / Uma metanálise sobre o transplante de útero: redefinindo os limites da cirurgia reprodutiva

SUMMARY In September 2014, the first baby grown in a transplanted uterus was born, which represented an astonishing scientific progress that will mark the history of human reproduction. The recipient was a 32-year-old woman with Rokytanski syndrome who became pregnant after a successful embryo transfer and had an uneventful pregnancy, giving birth to a healthy newborn and marking the beginning of a new era. Patients who do not have a uterus or have a dysfunctional uterus now have the chance of dreaming with pregnancy and motherhood. Combining principles of solid organ transplantation and techniques of human reproduction, uterus transplantation is the first ephemeral transplant performed in order to promote reproductive potential of women and may be removed after successful pregnancy. Worldwide, 11 uterine transplantations were performed in patients. Of these, seven maintained their reproductive potential, with viable transplanted uteri and regular menstrual cycles.

RESUMO Em 2014, nasce o primeiro bebê oriundo de um útero transplantado, um avanço científico que ficará marcado na história da reprodução humana. A paciente, portadora da síndrome de Rokytanski, engravidou após transplante de útero e transferência de embrião. Teve uma gestação sem complicações e deu à luz um recém-nascido saudável, marcando o início de uma nova realidade. Pacientes que não possuem útero ou possuem útero disfuncional agora podem sonhar em ser mães e gestar um filho. Combinando princípios de transplante de órgão sólido e técnicas de reprodução humana, o transplante de útero consiste no primeiro transplante efêmero realizado com o intuito de promover potencial reprodutivo à mulher, podendo ser removido após gestação bem-sucedida. Mundialmente, 11 transplantes de útero foram realizados em humanos. Dentre os casos, sete mantêm potencial reprodutivo, com úteros viáveis e ciclos menstruais regulares.

Follicle and corpus luteum size and vascularity as predictors of fertility at the time of artificial insemination and embryo transfer in beef cattle / Tamanho e vascularização do folículo e corpo lúteo como preditores de fertilidade após inseminação artificial e transferência de embriões em gado de corte

Two ultrasound based fertility prediction methods were tested prior to embryo transfer (ET) and artificial insemination (AI) in cattle. Female bovines were submitted to estrous synchronization prior to ET and AI. Animals were scanned immediately before ET and AI procedure to target follicle and corpus luteum (CL) size and vascularity. In addition, inseminated animals were also scanned eleven days after insemination to target CL size and vascularity. All data was compared with fertility by using gestational diagnosis 35 days after ovulation. Prior to ET, CL vascularity showed a positive correlation with fertility, and no pregnancy occurred in animals with less than 40% of CL vascularity. Prior to AI and also eleven days after AI, no relationship with fertility was seen in all parameters analyzed (follicle and CL size and vascularity), and contrary, cows with CL vascularity greater than 70% exhibit lower fertility. In inseminated animals, follicle size and vascularity was positive related with CL size and vascularity, as shown by the presence of greater CL size and vascularity originated from follicle with also greater size and vascularity. This is the first time that ultrasound based fertility prediction methods were tested prior to ET and AI and showed an application in ET, but not in AI programs. Further studies are needed including hormone profile evaluation to improve conclusion.

Duas técnicas de predição de fertilidade, baseadas em ultrassonografia, foram testadas no momento da transferência de embriões (TE) e inseminação artificial (IA) em bovinos. Fêmeas bovinas foram submetidas a protocolos de sincronização de estro para TE e IA. Os animais foram escaneados por ultrassonografia imediatamente antes do procedimento de TE e IA para identificar o tamanho e vascularização do folículo e corpo lúteo (CL). Além disso, os animais inseminados foram escaneados onze dias após a inseminação para identificar o tamanho e vascularização do CL. Todos os dados foram comparados com a fertilidade utilizando-se do diagnóstico gestacional 35 dias após a ovulação. No momento da TE, a vascularização do CL apresentou-se positivamente relacionada com a fertilidade, sendo que animais com menos de 40% de vascularização do CL não ficaram gestantes. No momento da IA, assim como onze dias após a IA, nenhuma relação foi encontrada entre fertilidade e os parâmetros analizados (tamanho e vascularização do folículo e CL), enquanto que contrariamente, houve uma queda na fertilidade em vacas com vascularização do CL acima de 70%. Nos animais inseminados, o tamanho e vascularização do folículo foi positivamente relacionado ao tamanho e vascularização do CL, demonstrado pela presença de um CL maior e mais vascularizado proveniente de um folículo maior e mais vascularizado. O presente estudo é o primeiro a mostrar métodos de predição de fertilidade baseados em ultrassonografia no momento da IA e TE, demonstrando uma aplicabilidade prática no momento da TE. Novos estudos são necessários para suportar os resultados mostrados, incluindo análises hormonais.

Viable offspring after successful non-surgical embryo transfer in goats / Nascimento de fetos viáveis após transferência de embriões não-cirúrgica em caprinos

O objetivo do presente estudo foi avaliar a viabilidade da técnica de transferência não cirúrgica em cabras. Quatro cabras não-lactantes pluríparas da raça Toggenburg foram utilizadas como receptoras de embriões, sendo que duas receberam um embriões e duas receberam dois embriões coletados não cirurgicamente cabras doadoras. Os corpos lúteos das receptoras foram detectados um dia antes da transferência de embriões por ultrassonografia transretal. Uma seringa de 5mL contendo 2mL de meio holding foi acoplada em um cateter tomcat, no qual os embriões foram aspirados em uma coluna central a duas outras colunas. Um espéculo Colin número 2 foi inserido na vulva e na vagina, e com o uso de uma fonte de luz, a cerviz foi localizada e imobilizada com uma pinça de Allis. Um cateter uretral número seis acoplado a um mandril e lubrificado com meio PBS foi inserido na cérvix, e assim os aneis cervicais foram gradualmente transpostos. Após perder a resistência, o cateter uretral foi movido lateralmente para o corno uterino desejado. O mandril e a pinça de Allis foram retirados e o conjunto seringa e tomcat foi acoplado ao cateter uretral e o conteúdo injetado no corno uterino ipsilateral ao corpo lúteo com posterior retirada do cateter. Cabras que ovularam em apenas um ovário foram usadas para testar a eficiência da deposição do embrião. O tempo gasto entre a inserção do espéculo e a sua remoção foi inferior a três minutos. O tempo para transpor a cérvix foi inferior a um minuto. A ultrassonografia revelou a deposição de líquido no corno desejado. Receptoras que receberam dois embriões tornaram-se gestantes e pariram três crias. Estes primeiros resultados encorajam a técnica e demonstram que a transferência de embriões em caprinos pode ser feita totalmente por procedimentos não cirúrgicos...

La criopreservación embrionaria en estadios más avanzados de desarrollo disminuye la frecuencia de pacientes con embriones criopreservados y el número de embriones criopreservados / Cryopreservation in blastocyst stage effectively reduce the number of embryos cryopreserved

Background: Multiple pregnancies are the main complication associated to assisted reproduction, due to the transfer of more than one embryo. Embryo cryopre-servation allows the sequential transfer of all generated embryos, thus diminishing the risk of multiple pregnancies. However, it leads to accumulation of cryopreserved embryos. To reduce their accumulation in our unit, we started to preserve embryos as blastocysts, that have a rate of successful pregnancies of approximately 40%. Aim: To perform a sensitivity analysis of this change of policy on the accumulation of embryos. Material and Methods: Records of 571 cycles of in vitro fertilization since 2007 were reviewed, assuming a transference rate of two embryos in women aged less than 35 years and three embryos in older women. The number of embryos that would be preserved as zygotes, eight cell stage or blastocysts, was analyzed. Results: Multiple component logistic regression analysis showed a 20% reduction in the odds ratio of cryopreservation per year of age. There was a 95% reduction in the ratio, when comparing the preservation of third and first day embryos and a 99% reduction when comparing preservation of embryos in blastocyst stage and first day embryos. Conclusions: Cryopreservation at blastocyst stage effectively decreased the frequency of embryo cryopreservation.

Influencia de la subespecie de la donadora en la cantidad deembriones totales obtenidos in vitro: estudio retrospectivo / Influence of the subespecie of the donator cow in the quantity oftotal embryos obtained in vitro: a retrospective study / Influência da subespécie da vaca doadora na quantidade de embriões totais obtidos in vitro: estudo retrospectivo

A partir de los registros de transferencia de embriones producidos in vitro de tantos años de la central genética en el municipio de San Pedro de los Milagros Antioquia – Colombia, se buscó retrospectivamente si existía unarelación sobre la cantidad de ovocitos y embriones viables producidos por las donadoras. Dependiendo de la raza,se buscó determinar si existe una influencia de la donante. Las variables analizadas fueron: el toro, la raza, el lote, la historia reproductiva de las donadoras, las cuales se encontraban en iguales condiciones de manejo, tanto sanitarias y nutricionales. La evaluación de los resultados se realizó mediante el modelo estadístico de Chi-cuadrado.Para la comparación entre el porcentaje de ovocitos viables y el porcentaje de embriones producidos se realizó unaprueba de análisis de varianza (ANOVA) y una prueba de Fisher con un intervalo de confianza del 95% y un valorde p>0.05. Se encontraron diferencias estadísticamente significativas en la producción de embriones (40% vs 29%), taurus e indicus, respectivamente. Se concluye, que las donadoras taurus producen mayor número de embrionesque las indicus.

From the many years of transfer records of embryos produced in vitro at the genetics center in the municipality ofSan Pedro de los Milagros, Antioquia, Colombia, we examined retrospectively if there was a relationship betweenthe number of viable oocytes and embryos produced by the donors. Depending on the race, we sought to determine whether there is an influence by the donor. The variables analyzed were: the bull, race, batch, and reproductive history of the donors, all of which were in similar health and nutritional conditions. The evaluation of the results was carried out with the Chi- Square test statistical model. In order to compare between the percentage of viable oocytes and the percentage of embryos, an analysis of variance (ANOVA) was performed as well as a Fischer test, with a confidence interval of 95% and a p-value of p>0.05. Statistically significant differences were found in the production of embryos (40% vs 29%) taurus and indicus, respectively. The conclusion is that taurus donors produce greater numbers of embryos than indicus donors.

A partir dos registros de transferência de embriões produzidos in vitro nos muitos anos da central genética nomunicípio de San Pedro de los Milagros, Antioquia- Colômbia procuramos retrospectivamente se há uma relaçãoentre a quantidade do número de oócitos viáveis e embriões produzidos por doadores. Dependendo da raça,buscou-se determinar se há uma influência do doador. As variáveis analisadas foram: o touro, a raça, o lote, a história reprodutiva dos doadores, os quais encontravam-se nas mesmas condições de manejo, saúde e nutrição. A avaliação dos resultados foi realizada utilizando o modelo de teste estatístico Chi-quadrado. Para a comparação entrea porcentagem de oócitos viáveis e a porcentagem de embriões produzidos foram realizados um teste pela análisede variância (ANOVA) e um teste de Fisher, com intervalo de confiança de 95% e um valor de p>0.05. Foramencontradas diferenças estatisticamente significativas na produção de embriões (40% vs 29%), taurus e indicus,respectivamente. A conclusão é que os doadores taurus produzem um número maior de embriões que os doadoresindicus.

Técnicas de reproducción asistida en caballos / Assisted reproductive technologies in the horses / Técnicas de reprodução assistida em cavalos

Las técnicas de reproducción asistida son métodos no convencionales por los cuales se obtienen potros deyeguas problema o geriátricas mediante la obtención de oocitos viables. De esta manera, los problemas del úteroo del oviducto que impiden un buen transporte y sobrevivencia de los espermatozoides, adecuada fertilizacióny consecuentemente transporte del embrión, son sobrepasados. Existen primordialmente dos métodos paracolectar oocitos en yeguas – a partir del folículo pre-ovulatorio dominante o de todos los folículos visiblesen los ovarios. Además existen dos métodos para fertilizar los oocitos que han sido obtenidos: in vitro, porinyección intracitoplasmatica de un espermatozoide (ICSI) o in vivo, al transferirlo al oviducto de una yeguareceptora inseminada previamente (Transferencia de oocitos)

Assisted reproductive technology (ART) refers to non-conventional methods for obtaining foals by obtainingisolated oocytes in problematic or geriatric mares. In this way, problems with the mare’s uterus or oviductthat limit proper sperm transport and survival, adequate fertilization, and consequently embryo transportare bypassed. There are two main methods to recover oocytes from a valuable mare – from the dominantpreovulatory follicle, or from all visible follicles on the ovaries. In addition, there are two main methods tofertilize the recovered oocytes: in vitro, by intracytoplasmic sperm injection (ICSI) or in vivo, by transferringit to the oviduct of a previously inseminated recipient mare (Oocyte Transfer)

As técnicas de reprodução assistida são métodos não-convencionais para a obtenção de potros de éguasproblemáticas ou geriátricas, através da retirada de ovócitos viáveis. Assim, os problemas do útero ou do ovídutoque impedem o transporte e a sobrevivência de espermatozoides, a fertilização adequada e, consequentemente,o transporte do embrião são evitados. Existem principalmente dois métodos para coletar ovócitos em éguas– a partir do folículo pré-ovulatório dominante, ou de todos os folículos visíveis nos ovários. Há tambémdois métodos para fertilizar os ovócitos que foram obtidos: in vitro, através de injeção intracitoplasmáticade espermatozoides (ICSI) ou in vivo, transferido-os para o oviduto de uma égua destinatária previamenteinseminada (Transferência de ovócitos).

Influência da interação entre o oócito e as células da granulosa nos resultados dos procedimentos de reprodução assistida / The influence of interaction between oocyte and granulosa cells on the results of procedures in assisted reproduction

A interação oócito-células da granulosa in vivo e sua influência na qualidade oocitária e embrionária tem sido alvo de inúmeros estudos, mas muitas questões ainda necessitam ser esclarecidas. O objetivo deste trabalho foi revisar a importância dessa comunicação, estabelecendo uma relação com a questão da maturação in vitro de oócitos imaturos humanos aplicando esses conhecimentos para definir possíveis marcadores moleculares que poderiam melhorar a seleção de oócitos e, consequentemente, selecionar embriões de boa qualidade para posterior transferência e sucesso de gravidez de pacientes submetidas ao tratamento da infertilidade conjugal. As células da granulosa têm um importante papel na maturação oocitária in vitro e os benefícios da presença dessas células durante essa etapa podem ser atribuídos à formação de um microambiente favorável (bioquímico e metabólico) ao redor do oócito. Foram identificados nesta revisão vários marcadores em potencial nas células do cumulus de oócitos competentes, incluindo vários genes que poderiam ser usados como preditores da competência oocitária, o que pode contribuir para a formulação de critérios mais objetivos e confiáveis para a seleção de oócitos e embriões, e consequente aprimoramento e otimização das técnicas em reprodução humana assistida que são aplicadas nos procedimentos clínicos atuais de fertilização in vitro.

The interaction of oocyte-granulosa cells in vivo and in vitro and its influence on oocyte and embryo quality has been the subject of numerous studies, but many issues still need to be clarified. The objective of this study was to promote a review about the importance of this communication establishing a connection with the issue of in vitro maturation of immature human oocytes by applying this knowledge to define potential molecular markers that could improve the selection of oocytes and consequently select good quality embryos for later transfer and success of pregnancy in patients undergoing treatment of infertility. The granulosa cells also have an important role in oocyte maturation in vitro and the venefits from the presence of these cells during this process can be atributed to the formation of a favorable micro-environment (biochemical and metabolic) around the oocyte. In this review, we identified several potential markers in the cumulus cells of competent oocytes, including several genes that could be used as predictors of oocyte competence, which contributes for more objective and reliable criteria for the selection of oocytes and embryos, thus improving and optimizing techniques in assisted human reproduction that are applied in current clinical in vitro fertilization.

Aspiração do muco cervical e infusão de jato com meio de cultivo no canal cervical antes da transferência embrionária e resultados de fertilização in vitro / Cervical mucus aspiration and flushing with culture medium cervical canal prior to embryo transfer and in vitro fertilization outcomes

A transferência de embriões é considerada uma etapa fundamental nos ciclos de fertilização in vitro. Os resultados reprodutivos do tratamento são diretamente relacionados à técnica da transferência embrionária. Alguns trabalhos mostraram que o muco cervical poderia afetar os resultados da fertilização in vitro. A aspiração do muco cervical e a limpeza do canal cervical com jato de meio de cultivo foram incorporadas à prática dos serviços de reprodução assistida em todo o mundo. A presente revisão tem como objetivo esclarecer a verdadeira eficácia desses procedimentos no incremento das taxas de gravidez em ciclos de fertilização in vitro.

Embryo transfer is considered an essential stage for in-vitro fertilization cycles. Reproductive outcomes of treatment are directly related to embryo transfer technique. Some papers showed that cervical mucus could affect in-vitro fertilization outcomes. Aspiration of cervical mucus and flushing cervical canal with culture media were incorporated to practice in assisted reproduction technologies centers all over the world. This review aimed to make clear the true efficacy of these procedures with respect to the increase in pregnancy rates of in-vitro fertilization cycles.

Extended culture up to the blastocyst stage: a strategy to avoid multiple pregnancies in assisted reproductive technologies

The aim of this study was to review the experience and outcomes of assisted reproduction cycles with embryos grown up to day 5 of development, comparing different parameters according to the ages of the patients. We retrospectively studied 1,874 assisted reproduction cycles where embryo culture was extended up to the fifth or sixth day of development. All IVF and ICSI cycles were included, comparing, according to patient age, the following rates: blastocyst formation, pregnancy, implantation and abortion. As control, we analyzed cycles with donated oocytes from young donors (OD). The number of embryos reaching the blastocyst stage is similar in all groups of patients. Only the OD group was different in terms of blastocyst formation, pregnancy and implantation rates. Patients over 39 years of age had an abortion rate of 59.1 percent, which is significantly higher than the other groups. Extended embryo culture up to the blastocyst stage can be implemented in programs of assisted reproduction in order to increase the pregnancy rate. The potential of blastocyst implantation is high, allowing us to transfer fewer embryos and reduce the probability of multiple pregnancies.

Embryo transfer in angus and brahman recipient cows: effect of two methods of estrus synchronization on induced estrus and pregnancy

Over a 3-year period, 88 Angus and 87 Brahman cows were used as recipients to determine the effects of breedtype and method of estrous synchronization on induced estrus and subsequent pregnancy following embryo transfer. Estrus was synchronized in recipients using either prostaglandin-F2α (PGF; Lutalyse) or progestogen (PRO; Syncro-Mate-B). Recipients were treated (im) with PGF on day -11 (25 mg), 0 (12.5 mg), and 1 (12.5 mg). Recipients were treated with PRO on day -9 (6 mg norgestomet ear implant and 2 mL [im] of 3 mg of norgestomet and 5 mg of estradiol valerate) and the ear implant was removed on day 0. Embryos were randomly transferred to recipients synchronous +24 to -24 h with a 7-d embryo. Both synchronization methods (PGF vs. PRO) were similarly effective in inducing estrus (89.8 vs. 86.7 percent). Percentage of induced estrus was similar between Brahman (91.0 percent) and Angus (85.6 percent). Recipients treated with PGF had a longer (P = 0.001) interval to estrus than PRO treated recipients (77.4 vs. 60.1 h). Estrus response in Angus was earlier (P = 0.043) than in Brahman (65.4 vs. 72.2 h). Neither breed (P = 0.97; Angus 49 percent 35/72 and Brahman 54 percent 38/70) nor estrus synchronization treatment (P = 0.23; PRO 49 percent 35/72 and PGF 54 percent 38/70) affected pregnancy rate. Pregnancy rates in recipients closely synchronized (0 h) was 60.0 percent, within +12 and -12 h was 56.0 percent, and within +24 and -24 h was 51.5 percent. Four-year-old recipients had lower pregnancy rates (34.8 percent) than did 5-year-old (60.9 percent) or greater than 6-year-old (70.0 percent) recipients. These results indicate that PGF, when administered as a split-second dose, is as effective in synchronizing estrus in Angus and Brahman as PRO. Asynchrony of embryo age with recipient stage of cycle up to +24 or -24 h did not significantly affect pregnancy rates.

Durante tres años, vacas receptoras Angus (88) y Brahman (87) se utilizaron para determinar los efectos de raza y método de sincronización sobre las características del celo inducido y la preñez subsecuente a la transferencia embrionaria. El celo fue sincronizado en las receptoras utilizando PGF2α (Lutalyse®) o progesterona (PRO; Syncro-Mate-B). Un grupo de receptoras fue inyectado (im) con PGF los días 11 (25 mg), 0 (12,5 mg), y 1 (12,5 mg). Otro grupo fue tratado con PRO el dia -9 (implante auricular con 6 mg norgestomet e inyectadas (im) con 2 mL con 3 mg de norgestomet y 5 mg de valerato de estradiol), el implante fue removido el día 0. Los embriones fueron transferidos al azar a las receptoras en sincronía +24 a -24 h con embriones de 7-d de edad. Ambos métodos de sincronización (PGF vs. PRO) fueron similares en la inducción del celo (89,8 vs. 86,7 por ciento). El porcentaje de celos inducidos fue similar en Brahman (91,0 por ciento) y Angus (85,6 por ciento). Las receptoras tratadas con PGF presentaron un intervalo al celo más largo (P = 0,001) que aquellas tratadas con PRO (77,4 vs. 60,1 h). El intervalo al estro post-tratamiento fue más corto en la vacas Angus expresaron celo más temprano (P = 0,043) que las Brahman (65,4 vs. 72,2 h). La tasa de preñez no fue afectada por la raza (Angus 49 por ciento 35/72 y Brahman 54 por ciento 38/70; P = 0,97) ni por el tratamiento de sincronización (PRO 49 por ciento 35/72 y PGF 54 por ciento 38/70; P = 0,23). Las tasas de preñez en receptoras de sincronización cercana (0 h) fue del 60,0 por ciento, entre +12 y -12 h fue del 56,0 por ciento, y entre +24 y -24 h fue del 51,5 por ciento. Las receptoras de cuatro años de edad presentaron tasas de preñez menores (34,8 por ciento) que las de cinco (60,9 por ciento) o las de 6 o más años de edad (70,0 por ciento). Los resultados indican que cuando la segunda aplicación de PGF se administra dividida en dos dosis, es tan efectiva para sincronizar...

Laparoscopy vs. laparotomy for embryo transfer to produce transgenic goats (Capra hircus)

This study was performed to produce transgenic Korean native goat (Capra hircus) by laparoscopic embryo transfer (ET) to overcome the limitations of ET performed by laparotomy. Transgenic embryos were produced by DNA pronuclear microinjection of in vivo zygotes. The recipient goats were synchronized for estrus by using an introvaginal progesterone devices as a controlled internal drugreleasing insert (CIDR) for 13 days and injection of 400 IU PMSG 48 h before removal of the insert. Embryos were transferred on day 3 and 4 after removal of the insert. Recipient goats were deprived of feed for 48 h, then suspended in a laparotomy cradle at an angle of 45degrees. After obtaining a sufficient pneumoperitoneum, the laparoscope and forceps were inserted abdominally through 5 mm trocar sleeves. Examination of the ovaries and uterus was performed and then 213 embryos were transferred into the oviducts via the infundibula of 76 recipient goats. To compare pregnancy rates, ET was also performed by laparotomy in 82 recipient goats. The pregnancies in the recipient goats were diagnosed by ultrasound on day 30 after embryo transfer. The pregnancy rate with laparoscopic ET was significantly higher than with ET performed by laparotomy (46.1% vs. 28.6%, p < 0.05). In addition, the pregnancy rates were compared between ovulated and non-ovulated ovaries of the recipient goats in the laparoscopic ET group. No significant difference was observed between the pregnancy rates of ovulated and non-ovulated ovaries (41.3% vs. 33.3%, p < 0.05) suggesting that ET may also be possible in non-ovulated recipients through artificial rupture of Graafian follicles. These results suggest that laparoscopic ET is a highly efficient method for the transfer of goat embryos.

Relationship between pronuclear scoring and embryo quality and implantation potential in IVF-ET / 华中科技大学学报（医学）（英德文版）

To assess the relationship between pronuclear scoring and day-3 embryo quality and pregnancy outcome and to determine the clinical value of pronuclear stage scoring system in human in vitro fertilization-embryo transfer (IVF-ET) program, a pronuclear scoring system was used to score zygotes 16-20 h after insemination during conventional IVF or intracytoplasmic sperm injection (ICSI). The embryos were classified into groups Z1, Z2, Z3 and Z4. Comparisons were made of the rates of arrested embryos and excellent embryos on day 3. Comparisons of pregnancy outcome were made only in those patients in whom cohorts of similarly Z-scored embryos were transferred. The results showed that there were less arrested embryos and more excellent embryos on day 3 in groups Z1 and Z2 than those in group Z3 and Z4. More embryos arrested and less excellent embryos developed in group Z4 than group Z3. The clinical pregnancy rates resulting from the transfer of single pronuclear score homologous embryo types were similar among groups Z1, Z2 and Z3. Implantation rates of group Z1 were higher (P<0.05) than that of group Z3. These findings suggests that pronuclear scoring can predict developmental ability on day 3 and implantation potential. A evaluation that combines the Z-score and day 3 embryo morphology is useful in the determination of the most viable embryos and the number of embryos for transfer.

Efeitos da exposição pré-concepcional de curta duração ao material particulado ambiental sobre o mecanismo reprodutivo feminino / Effects of short-term preconceptional exposure to ambient particulate matter on female reproductive function

Um Projeto Temático de Pesquisa foi desenvolvido no Laboratório de Poluição Ambiental do Departamento de Patologia da Faculdade de Medicina da Universidade de São Paulo com o objetivo de avaliar os efeitos da exposição aguda/crônica ao ar ambiente de um grande centro urbano sobre a saúde. Dentro deste projeto, uma linha de pesquisa foi dedicada ao estudo dos efeitos dessa exposição sobre a saúde reprodutiva feminina. Evidências de estudos epidemiológicos e experimentais implicam os fatores ambientais na infertilidade humana e resultado obstétrico adverso. Contudo, poucos estudos foram conduzidos até o presente para avaliar um possível efeito da exposição à poluição ambiental particulada sobre a saúde reprodutiva feminina. Portanto, o objetivo dos projetos da minha linha de pesquisa é fornecer dados que possam demonstrar os possíveis efeitos da exposição pré-concepcional de curta duração às partículas de exaustão do diesel (PED) e à poluição ambiental particulada sobre a função ovariana, o desenvolvimento embrionário inicial e resultado gestacional utilizando um modelo experimental e um epidemiológico. O objetivo do primeiro projeto desta tese foi avaliar os efeitos de dois meios de cultura comerciais no desenvolvimento de oócitos de camundongo fertilizados in vitro até o estágio de blastocisto. Zigotos obtidos de fêmeas de camundongo de 8 semanas de idade submetidas à indução da ovulação foram cultivados in vitro até o estágio de blastocisto em meio simples otimizado enriquecido com potássio (KSOM) ou meio G1/G2. A porcentagem de zigotos que se desenvolveu até o estágio de blastocisto 96 e 120 horas após a inseminação e que sofreu eclosão parcial ou completa no quinto dia de cultivo foi significativamente maior no grupo KSOM. O número médio de células da massa celular interna (MCI) foi 11,7 ± 4,0 e 9,2 ± 5,2 para os zigotos cultivados nos grupos KSOM e G1/G2, respectivamente, mostrando um número significativamente maior de células MCI em blastocistos...

A thematic research project to evaluate the health effects of acute/chronic exposure to ambient air in a large urban center was developed at the Air Pollution Laboratory in the Department of Pathology at the University of São Paulo School of Medicine. Within this project a specific research line was committed to the study of the effects of this exposure on female reproductive health. Evidence from epidemiological and experimental studies implied environmental factors as possible contributors to human infertility and poor obstetric outcome. However, very few studies evaluating a possible effect of exposure to particulate air pollution on female reproductive health have been conducted so far. Thus, the aim of the projects in my research line was to provide data that could show the possible effects of short-term preconceptional exposure to diesel exhaust particles and particulate air pollution on ovarian function, early embryo development and pregnancy outcome using experimental and epidemiological models. The objective of the first project was to examine the effects of two commercial media on the development of mouse ova fertilized in vitro to the blastocyst stage. One-cell embryos obtained from eight-week old superovulated mice were cultured in vitro up to the blastocyst stage in potassium-enriched simplex optimized medium (KSOM) or G1/G2 media. The percentage of zygotes that developed to the blastocyst stage 96 and 120 hours after insemination and that partially or completely hatched by day five of culture was significantly higher in the KSOM group. The mean number of inner cell mass (ICM) cells was 11.7 ± 4.0 and 9.2 ± 5.2 for zygotes cultured in KSOM and G1/G2 groups respectively, revealing a significantly higher cell number in the ICM of blastocysts derived from culture in KSOM medium. I concluded that commercially available KSOM medium is superior to sequential G1/G2 media for culturing one-cell embryos up to the blastocyst stage in...

Associação entre sorologia para Neospora caninum e taxa de prenhez em vacas receptoras de embriões / Association between seropositivity for Neospora caninum and pregnancy rate in bovine receipts submitted to embryo transfer technology

The association between seropositivity for Neospora caninum and pregnancy rate in cows belonging to a surrogate herd submitted to embryo transfer technology was determined. The serological status was evaluated in 275 heifers, aging from 14 to 20-month-old. For N. caninum serology analysis of a monoclonal competitive ELISA test Kit was used, and 81 animals (29.5 percent) showed seropositive. Thus, two groups were randomly formed selecting 33 seropositive heifers and other 33 seronegative animals out of the remaining 194 animals. Seronegative animals were followed up by serological analysis until the end of the trial in order to identify persistently infected individuals. The pregnancy rate was 72.7 percent in the group of N. caninum-positive sera, and, 81.8 percent in the seronegative group.No significant difference was observed between groups according to Chi-square test. No association between N. caninum seropositivity and pregnancy rates in surrogate heifers was found

La migración de la burbuja de aire postransferencia embrionaria es un evento al azar / Air bubble migration post embrionary tranferir is a random event

Demostrar que la migración de la burbuja de aire postransferencia embrionaria es un evento al azar que no depende de la posición del paciente. Estudio prospectivo multicéntrico. Se practicó transferencia embrionaria a 69 pacientes bajo guía ultrasonográfica. Se cargó el catéter de transferencia con una o dos burbujas junto con el medio conteniendo los embriones usando catéter de Frydman o catéter de Wallace y luego se les hizo seguimiento ecográfico en posición horizontal, 20-25 minutos más tarde y luego a los 90 minutos postransferencia en posición vertical para verificar movimiento y configuración de las burbujas. Centro Médico Docente La Trinidad, Caracas. Nortwestern University Feimberg School of medicine, Chicago, IL. EE.UU. En 50 (72,46 por ciento) de las pacientes se observó movimiento de las burbujas. En posición de pie se demostró una distribución al azar sin efecto visible de la gravedad. Las burbujas de aire se mueven y dividen frecuentemente posterior a la transferencia con la paciente en posición horizontal y de pie, lo que sugiere actividad contráctil uterina. No fue común observar movimiento de burbujas relacionada con la gravedad, lo que sugiere que el descanso horizontal postransferencia embrionaria puede ser innecesario.

To demonstrate that air bubble migration is a random event after embryo transfer regardless of the position of patient. Multicenter prospective study. Sixty nine embryo transfers were performed under ultrasound guidance. Transfer catheter was loaded with one or two air bubbles and medium containing embryos, either Wallace catheter or Frydman catheter were used, ultrasound tracking of air bubble was performed to verify movement and configuration inmediately, 20-25 minutes in horizontal position and 90 minutes on standing position after embryo transfer. Centro Medico Docente La Trinidad, Caracas. Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA. Movement was observed in 50 (72,46 percent) of the patients. Movement was not observed related to gravity while standing up. Air bubbles move and split frequently after embryo transfer with the patient in the horizontal position and on standing, suggestive of active uterine contractions. Gravity related bubble motion was uncommon, suggesting that horizontal rest post embryo transfer may be unnecessary.

Comparison of the outcome of intracytoplasmic sperm injection using fresh and frozen-thawed epididymal spermatozoa obtained by percutaneous epididymal sperm aspiration.

The study was conducted to compare the outcome of intracytoplasmic sperm injection (ICSI) between fresh and frozen-thawed epididymal spermatozoa retrieved by percutaneous epididymal sperm aspiration (PESA) from obstructive azoospermic men. Overall, 53 PESA procedures were performed in 42 obstructive azoospermic men, followed by ICSI procedures with either fresh (n = 40) or frozen-thawed (n = 13) epididymal spermatozoa. Comparing all ICSI cycles with fresh and frozen-thawed epididymal spermatozoa, the fertilization rates (77.4 vs 86.8%) and the cleavage rate (91.3 vs 95.1%) were not statistically different. A total of 64 embryo transfers were performed: 48 embryo transfers after the use of fresh epididymal spermatozoa and 16 embryo transfers after the use of frozen-thawed spermatozoa. The overall pregnancy rate per embryo transfer was also similar between both groups (40.8 vs 40%). The implantation rate per embryo (18.2 vs 12.7%), clinical pregnancy per embryo transfer (36.7 vs 33.3%) and delivery/ongoing pregnancy rate (36.7 vs 33.3%) were not statistically different. In conclusion, there were no significant differences of the outcome of intracytoplasmic sperm injection using fresh and frozen-thawed epididymal spermatozoa obtained by PESA.