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Use of a GnRH antagonist in controlled ovarian hyperstimulation for assisted conception in women with polycystic ovary disease: a randomized, prospective, pilot study.

OBJECTIVE: To compare the outcome of using gonadotropin-releasing hormone (GnRH) antagonists versus agonists in women with polycystic ovary disease (PCOD) who underwent controlled ovarian hyperstimulation (COH) for assisted reproductive techniques (ART). STUDY DESIGN: A total of 129 patients with PCOD were randomly allocated to undergo COH with a GnRH antagonist (59 patients) and GnRH agonist (leuprolide acetate) (70 patients) to prevent a premature luteinizing hormone (LH) surge. Assisted fertilization following oocyte retrieval and embryo transfer was performed. RESULTS: None of the cycles were cancelled due to a premature LH surge. There was no significant difference between the antagonist and agonist arms in the number of gonadotropin ampules consumed per cycle. However, in the antagonist arm a shorter duration of ovarian stimulation was recorded as compared to the agonist arm. Although similar numbers of oocytes was retrieved from both groups of patients, the quality of the oocytes, as measured by metaphase 2/total oocyte ratio, was lower in the antagonist arm as compared to the agonist arm. Pregnancy rates were 57.6% and 58.5% in the antagonist and agonist arms, respectively (p > 0.05). Implantation rates were not different (34.0% and 34.6%, respectively). The frequency of ovarian hyperstimulation syndrome also did not differ between the treatment groups (5% and 7.1%, respectively). CONCLUSION: The size of our study, on a specific subgroup of patients, does not allow a reliable conclusion regarding ART outcomefollowing the use of a GnRH antagonist versus agonist. Nevertheless, the protocol with the antagonist gave results that were as good as those of the protocol with the agonist in this PCOD patient population.

Adult↗

American Society for Reproductive Medicine--56th annual meeting.

The scientific program of the first annual meeting of the American Society for Reproductive Medicine (ASRM) of the new millennium was comprehensive, informative and covered most of the current issues in the expanding field of reproductive medicine. The meeting was characterized by many clinical updates, new discoveries in clinical and basic research and significant societal issues in the practice of reproductive medicine. Among the new discoveries, the characterization of two subtypes of progesterone receptors remains an outstanding addition that will impact on the future of breast cancer therapy. Many prospective randomized multicenter clinical trials illustrated the advantages of using gonadotrophin-releasing hormone antagonist instead of the agonist for controlled ovarian hyperstimulation, for patients undergoing assisted reproductive techniques. Antiprogestin medicated intrauterine device and combined hormonal contraceptive patches were introduced to improve the efficacy and reduce side effects of birth control methods. Aromatase inhibitors that have been extensively used for post-menopausal breast cancer therapy were tried successfully for the first time in ovulation induction. Successful egg retrieval from autotransplanted ovarian tissue was reported following successful radiotherapy in a patient with advanced cancer of the cervix. Finally, laparoscopic adnexectomy with complete robotic assistance was reported as an important technical addition in the field of reproductive medicine.

Journal Article↗

Cytotrophoblast stem cell lines derived from human embryonic stem cells and their capacity to mimic invasive implantation events.

BACKGROUND: An effective embryonic-maternal interaction is crucial for successful human pregnancy. Failure of this process is a major cause of infertility and can lead to placental dysfunction resulting in recurrent miscarriage, fetal retardation and pre-eclampsia. Research is severely constrained by ethical and practical considerations; therefore, we aimed to generate cytotrophoblast stem (CTBS) cell lines from human embryonic stem cells (HESCs). METHOD: Beta-HCG was used as a marker of viable trophoblast cells. In defined culture, embryoid bodies were generated from HESCs and selected for trophoblast enrichment by rounds of cellular aggregation and disaggregation. Distinct CTBS cell lines were isolated and characterized. Spheroid cytotrophoblast bodies were generated and their interaction with luteal-phase endometrial stroma was analysed by real-time image analysis. RESULTS: Three CTBS cell lines were derived, which were maintained in the absence of residual HESCs, fibroblast feeder cells or extracellular matrix. CTBS cells displayed typical cytotrophoblast and syncytiotrophoblast characteristics and exhibited further differentiation to invasive endovascular cell phenotype. One cell line was generated with constitutive expression of enhanced green fluorescent protein (eGFP). Spheroid trophoblast bodies mimicked closely the early invasive stages of implantation when incubated with human endometrial stromal preparations in vitro. CONCLUSION: These human CTBS cell lines are a significant new model for investigating human placentation and may have considerable potential in cell therapy applications.

Cell Differentiation↗

[Short tandem repeat polymerase chain reaction used in prenatal diagnoses of the zygosity and common chromosomal trisomies in multiple pregnancies].

OBJECTIVE: To evaluate the value of short tandem repeat polymerase chain reaction (STR-PCR) in identification of zygosity in multiple pregnancy and detection of the common chromosomal trisomies. METHODS: Amniotic fluid or fetal blood samples were collected from 38 fetuses in 17 multiple pregnant women who had indications for prenatal diagnosis or planed to be performed feticide. Of the 17 cases there were 13 sets of twins (26 fetuses) and 4 sets of triplets (12 fetuses). Parental blood samples were collected. STR-PCR technique was employed to determine the zygosity and detect trisomy 21, trisomy 18, trisomy 13, and trisomy sex chromosomes. Amniotic fluid or blood samples were collected from 18 singleton pregnant fetuses as controls for detecting chromosomal trisomies. RESULTS: All 7 cases with gestation by assisted reproductive techniques were multizygotic. In 7 cases associated with one fetal malformation, only one set of twin was dizygotic. There were no above mentioned trisomies found in the multiple pregnancies while 2 trisomy 21, one trisomy 18 and 13 respectively were detected in the singleton pregnant group. Feticide was performed on 4 multizygotic cases in which 3 were triplets and 1 was twin associated with an abnormal fetus. Following-up showed that there was no any bad effect after procedure. CONCLUSION: STR-PCR can be used in identifying the zygosity correctly and quickly while being used in detecting chromosomal trisomies. It facilitates monitoring multiple pregnancy as well as selecting appropriate methods of feticide.

Adult↗

Management of HIV serodiscordant couples. The clinician point of view.

BACKGROUND: Couples where the male partner is HIV-infected but the female partner is not often express a desire for children. Assisted reproduction techniques (ART) with sperm "wash" can reduce the risk of acquisition of infection for the woman. The information about the characteristics of such couples coming to these clinics for advice and support is scant. OBJECTIVE: To describe the characteristics of a cohort of HIV-discordant couples seeking reproductive assistance in a referral clinic in Barcelona, Spain. METHODS: Demographic, social and HIV related information were collected prospectively according to a standard protocol. Blood tests, a full evaluation for STDs and sperm analysis were performed. RESULTS: Between April 1994 and February 1999, 140 couples contacted the clinic for reproductive advice, 100 couples were fully assessed and were offered reproductive assistance. The vast majority were white, with a history of injecting drug use. Nearly half of the men had been diagnosed during their present relationship. STDs were uncommon and abnormalities among sperm analysis were frequent. CONCLUSION: An increasing number of HIIV infected individuals desire to have children. With the introduction of HAART and the safety of sperm wash in such cases, the need to have children is more frequently expressed. These couples are in general healthy and accept to be included in an ART program.

Adult↗

Neonatal outcome of spontaneous and assisted twin pregnancies.

OBJECTIVES: Over the last 10 years, diffusion of assisted reproduction techniques (ovarian stimulation, IVF, GIFT) has led to an increased incidence of multiple pregnancies and consequently, of the related obstetric-neonatal problems. In this study, multiple births have been studied, with particular reference to the twin births occurring in the Gemelli hospital, Rome. The hospital is also a reference centre for obstetric pathologies and infertility treatment. In particular, attention has been focused on neonatal outcome, comparing twins born from spontaneous and assisted pregnancies. STUDY DESIGN: 228 neonates from spontaneous twin pregnancies and 32 from assisted twin pregnancies were taken into consideration with regard to: premature birth, low birth-weight, intrauterine growth retardation, weight discordance, Apgar score, major neonatal diseases, and mortality. RESULTS: Results showed a significant higher incidence of prematurity and low birth-weight, as well as a significant lower gestational age, occurring more frequently in twins resulting from assisted pregnancies than in twins from spontaneous pregnancies. Furthermore, the incidence of severe depression at birth and respiratory disease was significantly higher in twins from assisted pregnancies than in those from spontaneous pregnancies, despite similar gestational age and birth-weight.

Apgar Score↗

The effects of cancer and cancer treatments on male reproductive function.

In male cancer patients, surgery, radiotherapy and chemotherapy can be followed by transient or permanent infertility by affecting ejaculatory or erectile function, or by impairing spermatogenesis. Cancer specialists should, therefore, discuss the impact of different therapies on fertility with their patients prior to treatment, and consider fertility-preserving measures before and during treatment, such as nerve-sparing operative techniques, adequate testicular shielding during radiotherapy and the avoidance of unnecessary gonadotoxic chemotherapy. Pretreatment sperm-cell cryopreservation should be offered, even in cases where the individual's risk of post-treatment infertility might seem minimal or if it might require testicular sperm-cell extraction. Samples that are severely oligospermic should also be cryopreserved. Post-treatment ejaculatory or erectile dysfunction can be reversed pharmacologically, but the success rate varies with the extent of neurologic sequelae of the treatments used. At present there is no established method to stimulate post-treatment impaired spermatogenesis, although currently available assisted reproductive techniques overcome some of the existing problems of infertility in cancer survivors, and ongoing research will hopefully increase these possibilities. A multidisciplinary approach that depends on close cooperation between relevant medical specialists is central to achieving such advances.

Combined Modality Therapy↗

Reproductive assistance to HIV-discordant couples--the German approach.

An estimated 35 000 Germans are infected with HIV, and 75% of these are of childbearing age. Couples with one infected partner are faced with the risk of infection of the healthy partner. By using assisted reproduction techniques (ART) and virus-free sperm it is possible to fulfil the desire for children while minimising the risk of acquisition of infection. Therefore since 1991 in Germany ART is offered to couples of HIV-positive males and seronegative females. However, established programmes in different centres were temporarily suspended due to serious doubts about the reliability and safety of the sperm preparation technique (density gradient centrifugation and swim-up). -. Since 1997 processed sperm samples are routinely tested for HIV by highly sensitive detection methods and cryopreserved. Only if the test is negative are the processed spermatozoa used for homologous insemination, in-vitro fertilization or intracytoplasmatic sperm injection. This procedure removes most medical as well as legal considerations. At present 3 reproductive units in Germany offer reproductive assistance to HIV-discordant couples, and a further 9 institutions plan to establish that procedure. From 1991 to 1999, 54 women were treated by ART. 30 pregnancies occurred and 24 children were born. So far no maternal or infant infection was observed after treatment.

Female↗

[The management of azoospermic patient].

OBJECTIVES: Since first pregnancy after ICSI was achieved in 1992, the treatment and prognosis of severe male factor, i.e azoospermia, has radically changed. The objective of this article is to review the issue from the urologist-andrologist point of view. METHODS: We perform a short revision of the two types of azoospermia and diagnostic tests, and show the algorithm used in our centre for the management of patients with azoospermia. We review the techniques for spermatozoid recovery and surgical treatment of obstructive azoospermia because the role of the urologist is maximum in these aspects. Genetic anomalies in azoospermic patients is one of the issues analyzed in more detail for being one of the most important and of maximum interest currently. RESULTS: Nearly 1% of pregnancies in developed countries are achieved by assisted reproduction techniques, and genetic anomalies among newborns from ICSI cycles have increased to 1.6%, three times normal population. Genetic anomalies are tenfold in azoospermic patients in comparison to general population. We analyze the importance of these studies in patients with secretory azoospermia. We review the most frequent genetic anomalies associated with azoospermia and diagnostic tests employed. Its importance is based on this diagnosis allowing genetic counselling and pre-implant or prenatal diagnosis with the aim of trying to minimize genetic anomalies and disease transmission to next generations. CONCLUSIONS: The diagnosis and treatment of azoospermia continues to be one of the most thrilling challenges in the field of infertility, with promising research lines such as in vitro spermatogenesis from stem cells and autotransplantation of criopreserved cells in patients undergoing radiotherapy and chemotherapy.

Equipment Design↗

Controlling the high order multiple birth rate: the European perspective.

The dramatic increase in the incidence of high order multiple pregnancies that has occurred since the mid-1970s is due to three main factors: increasing female age at conception; increasing use of ovulation induction agents and the introduction of sophisticated assisted reproduction techniques. High order multiple pregnancies are at significantly higher risk of complications than singleton pregnancies, for the fetuses, babies and the mothers. The aim of all fertility treatments is to achieve a healthy live child for each couple seeking treatment. Treatment of infertility by IVF and related techniques provides an ideal opportunity to limit the number of high order multiple pregnancies by reducing the number of embryos transferred to the woman. The realization that high order multiple pregnancy can and should be avoided has increasingly led, in most European countries, to a restriction of the number of embryos for transfer to two or even one, without a significant decrease in a couples' chance of achieving a pregnancy. The experience of European countries in reducing the number of embryos transferred is reviewed and a recommendation made that a responsible attitude to embryo transfer is adopted by all practitioners of assisted reproduction.

Embryo Transfer↗

[Multiple pregnancy].

The incidence of multiple pregnancies has increased considerably over the past 20 years in the western world. The number of twin pregnancies has increased by approximately 25% and the rate of triplet and higher order multiple pregnancies has more than doubled. This is due to the use of ovarian hyperstimulation and assisted reproduction techniques, and to the increasing maternal age. Multiple pregnancy can be considered as the most important adverse outcome of infertility treatment. The main determinant for outcome and management of multiple pregnancies is the chorionicity. This can be determined by ultrasound until 14 weeks of gestation. Twenty percent of all twin pregnancies are monochorionic, and 15% of these develop severe twin to twin transfusion syndrome, which is best treated by LASER separation of the communicating blood vessels. Obstetric complications associated with multiple gestation also occur more frequently in monochorionic twins. They include increased incidence of preterm labor, intrauterine growth restriction and assisted or surgical delivery. Neonatal problems include low birthweight and increased prevalence of congenital malformations. Overall, there has been a decrease in neonatal mortality of twins and triplets over the past 10 years, which is mainly due to the enormous advances in neonatal intensive care.

Cross-Sectional Studies↗

Vasectomy reversal: new techniques and role in the era of intracytoplasmic sperm injection.

The growing popularity of vasectomy as a primary form of birth control has also created an increasing demand for vasectomy reversal (VR). The advent of IVF with intracytoplasmic sperm injection after sperm retrieval has given couples an alternative to traditional VR. Although VR remains the most cost-effective and natural way to conceive after vasectomy, competing interests and market pressures have encouraged the development of this assisted reproductive technique (ART). The success of VR is dependent on several variables. The length of time since vasectomy and an individual surgeon's experience appear to be the most significant factors and are equally important. The success of this procedure is highly contingent on adherence to strict surgical principles of creating a water-tight, tension-free anastomosis, along with the appropriate decision whether to perform either a vasovasostomy or epididymovasostomy at the time of VR.

Humans↗

Comparison of fertility between intracytoplasmic sperm injection and in vitro fertilization with a partial zona pellucida incision by using a piezo-micromanipulator in cryopreserved inbred mouse spermatozoa.

Cryopreservation of mouse spermatozoa has been widely applied for maintenance of transgenic and knockout lines. However, the fertility of cryopreserved spermatozoa from some inbred strains such as C57BL/6 and BALB/c is extremely poor. We have recently reported that a partial zona-pellucida incision by piezo-micromanipulator (ZIP) significantly improves the fertilization rate and subsequent embryonic development after in vitro fertilization (IVF) using cryopreserved C57BL/6 transgenic mouse spermatozoa and that inbred C57BL/6 mice could be produced by intracytoplasmic sperm injection (ICSI). These findings prompted us to compare the efficiency of fertilization and subsequent embryonic development between ICSI and IVF with ZIP (ZIP/IVF) using cryopreserved spermatozoa. In conventional IVF, BALB/cA, C57BL/6J, and B6C3F1 cryopreserved spermatozoa fertilized 19%, 0%, and 51% of oocytes, respectively. The fertilization rates of manipulated oocytes by ICSI versus ZIP/IVF using cryopreserved BALB/cA spermatozoa were 52% versus 68%, cryopreserved C57BL/6J spermatozoa were 43% versus 63%, and cryopreserved B6C3F1 spermatozoa were 58% versus 82%, respectively. In these strains, fertilization rates for ZIP/IVF were significantly higher (P < 0.05) than for other techniques. However, embryonic development to term for oocytes fertilized by cryopreserved spermatozoa was not significantly different between ZIP/IVF and ICSI in C57BL/6J and B6C3F1. The overall efficiency of mouse production in ZIP/IVF was higher than for ICSI and conventional IVF in C57BL/6J and B6C3F1. Furthermore, ZIP/IVF required approximately 3.3 times less manipulation time than did ICSI. Our results indicate that ZIP is a useful assisted reproductive technique for IVF of ova by cryopreserved spermatozoa and improves production in some mouse strains.

Animals↗

Social aspects in assisted reproduction.

In-vitro fertilization (IVF) and assisted reproductive techniques have become common practice in many countries today, regulated by established legislation, regulations or by committee-set ethical standards. The rapid evolution and progress of these techniques have revealed certain social issues that have to be addressed. The traditional heterosexual couple, nowadays, is not considered by many as the only 'IVF appropriate patient' since deviations from this pattern (single mother, lesbians) have also gained access to these treatments. Genetic material donation, age limitation, selective embryo reduction, preimplantation genetic diagnosis, surrogacy and cloning are interpreted differently in the various countries, as their definition and application are influenced by social factors, religion and law. Financial and emotional stresses are also often described in infertile couples. Information as deduced from the world literature regarding IVF regulation, as well as about the existing religious, cultural and social behaviours towards these new technologies, is presented in this article in relation to the social aspects of assisted reproduction.

Age Factors↗

Maturation and fertilization of equine oocytes.

Equine oocytes obtained either by transvaginal ultrasound-guided follicular aspiration or from slaughterhouse ovaries can be matured in vitro. This generally requires culture in TCM-199 containing serum and hormones for 30 to 36 hours. With this protocol, approximately 50% to 60% of the oocytes are at metaphase-II at the end of the culture period. At least some of these oocytes appear viable based on production of fertilized eggs either through in vitro fertilization or fertilization in vivo of a recipient mare. The success of producing equine embryos in vitro is still extremely low. More than likely the conditions for in vitro oocyte maturation are not optimized, and the techniques for capacitating equine spermatozoa are not adequate. The stallion sperm would appear more difficult to capacitate and many of the approaches used in other species have not worked in horses. To date, the only fertilization that has occurred with in vitro matured oocytes has been with sperm treatments containing the calcium ionophore A23187. Increased success with in vitro production of equine embryos may be gained through the use of assisted reproductive techniques such as partial zona dissection or intracytoplasmic sperm injection.

Animals↗

A proteomic analysis of mammalian preimplantation embryonic development.

Genetic studies on the mammalian preimplantation embryo are providing a wealth of information regarding gene expression. However, changes in the transcriptome do not always reflect cellular function or the complexity and diversity of the mammalian proteome with post-translational modifications or protein-protein interactions. To elucidate embryonic cellular function, a detailed understanding at the protein level is necessary. The aim of this study was to generate protein profiles of mammalian embryos throughout development, and to investigate the effects of oxygen concentration on the embryonic proteome. A protocol was developed to analyse small groups of embryos (n = 5) by time-of-flight mass spectrometry. F1 mice zygotes were cultured in G1/G2 sequential media with recombinant albumin (2.5 mg/ml) in 6% CO(2) and O(2) concentrations of either 5% or 20%. In vivo-developed embryos were flushed from the reproductive tract (day 4). Protein profiles were generated for all embryonic samples and statistical analysis revealed 32 potential proteins/biomarkers with significant changes (P < 0.05). Embryos generated under 5% O(2) more closely resembled in vivo-developed embryos. Under 20% O(2) conditions, embryos showed down-regulation of 10 proteins/biomarkers (masses between 4 to 20 kDa) (P < 0.05) confirming the pathological effects of oxygen during embryonic development. These data demonstrate for the first time the complexity of the mammalian preimplantation proteome. The unique protein profiles of in vivo-developed embryos and a panel of selected biomarkers represent optimal cellular function, against which comparisons can be made to facilitate improvements in mammalian assisted reproduction techniques procedures.

Animals↗

Value of ultrasound during the menstrual cycle and early pregnancy.

Ultrasound is a recently introduced technique that allows repeated noninvasive imaging of the uterus, ovaries and associated structures. It offers a relatively simple method for the repeated examination of the abdominal organs including growing fetuses, and is increasingly being used to measure blood flow in the identification of disease states. The clinical applications of real time pelvic ultrasonography are principally in the field of gynaecology to assess anatomical normality, and to monitor the progress of natural and stimulated cycles in assisted reproductive techniques. Once infertility treatment has been successful, ultrasound has proved to have a great value, when combined with established endocrinological parameters, to monitor the development of pregnancy, and the normality of fetal growth.

Female↗

The use of intravenous albumin in patients at high risk for severe ovarian hyperstimulation syndrome.

Previous experiences in subjects with other forms of third space fluid accumulation have shown that albumin is efficacious in preventing and correcting haemodynamic instability. Using a similar approach in an effort to increase the serum oncotic pressure and to reverse the leakage of fluids from the intravascular space, high risk subjects for severe ovarian hyperstimulation syndrome (SOHS) were treated with albumin. In a recent large study two high risk factors were identified, i.e. the number of oocytes and levels of serum oestradiol. Thirty-six women undergoing assisted reproductive techniques who presented both these factors, received intravenous albumin at a dose of 5% in Ringers lactate in doses of 500 ml during oocyte retrieval and 500 ml immediately thereafter in the recovery room. Daily measurements of urine output, serum and urine electrolytes, weight, abdominal girth, and haematocrit prior to and after oocyte retrieval revealed normal serum and urine electrolyte levels, and no signs of haemoconcentration. No patient in this study developed SOHS, and of course none had to be hospitalized. Vaginal ultrasound performed in the majority of the subjects revealed < or = 100 ml of peritoneal fluid 48-72 h after oocyte retrieval. The only complication from the use of intravenous albumin was the appearance of a 'flu-like condition' (low grade temperature, nausea and muscle pains) developed by 12 women between days 3 and 5 after oocyte collection. Intravenous albumin had thus prevented the development of severe ovarian hyperstimulation syndrome in an assisted reproduction programme.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗