Birthweight of singletons after assisted reproduction is higher after single- than after double-embryo transfer

BACKGROUND: Single-embryo transfer (SET) has proven efficient in reducing multiple pregnancy rates after assisted reproduction technologies (ART). This study compares outcome of singletons after SET and double-embryo transfer (DET). METHODS: We studied 404 SET and 431 DET patients, who delivered a singleton child of >500 g after fresh embryo transfer in a first, second or third cycle. Preterm birth and low birthweight incidences and gestational age and birthweight were compared between both groups. Adjustments were made for maternal age, parity, cycle rank number, treatment indication, ART method, embryo characteristics and sex of the child. RESULTS: Singletons born after DET have a significantly lower birthweight than that after SET (3204.3 +/-617.5 g versus 3324.6+/-509.7 g , P<0.01). Also preterm birth (<37 weeks) [odds ratio (OR) 1.77, 95% confidence interval (CI) 1.06-2.94] and low birthweight (<2500 g) (OR 3.38, 95% CI 1.86-6.12) are significantly more common in DET singletons. CONCLUSIONS: Singleton birth after SET is advantageous compared with DET. This sheds new light on the reasons why singleton births following ART do worse than spontaneously conceived singletons in IVF programs, where double- or multiple-embryo transfer is standard.     

Encouraging results despite complexity of multidisciplinary care of HIV-infected women using assisted reproduction techniques

BACKGROUND: Assisted reproduction technologies can treat infertility for human immunodeficiency virus (HIV) seropositive women. We assessed the efficacy of these techniques in the results and difficulties encountered while conducting our assisted reproduction programme for 49 couples in which at least the woman had HIV infection that was currently under control. METHODS: Treatments included intrauterine insemination (IUI), IVF and ICSI, with ovarian stimulation. Embryos were transferred on day 3 after oocyte retrieval. An elective single transfer was performed, except for patients aged > or = 40 years. RESULTS: The median age of the women was 36 years. Ten IUI, nine IVF, 53 ICSI and 10 frozen-thawed embryo transfers have been performed. No pregnancy occurred following the IUI trials but for the couples with IVF and ICSI attempts the clinical pregnancy rate per embryo transfer was 23.9%. Eight babies have been born leading to a 22.2% take home baby rate per treated couple. Contamination was not observed in any newborn. CONCLUSIONS: Assisted reproduction technologies and particularly ICSI can provide HIV seropositive women with a safe means of mothering children. Results are encouraging when considering the age of the patients and a preferential single embryo transfer.     

Pregnancy rate fluctuations during routine work in an assisted reproduction technology unit

BACKGROUND: Fluctuations in spontaneous pregnancy rates have been observed in the general population. The purpose of this study was to evaluate whether pregnancy rates fluctuate over time in a homogeneous assisted reproduction treatment unit. METHODS: The study sample consisted of 3522 consecutive assisted reproduction cycles conducted from January 1996 to December 1999. Only fresh cycles in women <41 years old were included. All pertinent clinical data were prospectively collected on a computerized database and analysed at the end of the study. RESULTS: Throughout the 4 years of the study, monthly pregnancy rates fluctuated between 10.5 and 34.1% (mean 21.4%) per cycle, and between 13.6 and 41% (mean 27.26%) per transfer. These fluctuations did not follow any specific seasonal pattern. CONCLUSION: During routine work in an assisted reproduction treatment unit, there are random fluctuations in the pregnancy rate. This factor should be considered in studies performed in a specific time-period.     

Life table (survival) analysis to generate cumulative pregnancy rates in assisted reproduction: an alternative method of calculating the cumulative pregnancy rate in assisted reproduction technology

The current method of calculating cumulative pregnancy rate can lead to an overestimation of treatment efficacy, especially over many cycles of assisted reproduction treatment. The choice of scale of passage of time should be dependent upon the types of treatment to be evaluated. The number of treatment cycles to which patients' effort and commitment is directly related may be appropriate where the chance of pregnancy is expected to be significantly higher than non-treatment for them. Limiting the calculation of cumulative pregnancy rate only to the second or third cycle within 1 or 2 years will ensure that most patients are included in the calculation. More research is needed to assess different methods and develop better variables for assessing the efficacy of infertility treatment that can be informative for patients over the course of their treatment.     

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.     

Value of serum CA-125 concentrations as predictors of pregnancy in assisted reproduction cycles

The means by which endometrial receptivity influences conception rates in assisted reproductive technology (ART) is poorly understood. As the glycoprotein CA-125 is a product of human endometrium and is measurable in the peripheral circulation, it was investigated whether it might serve as an indicator of endometrial receptivity and predictor of pregnancy. In this prospective study, serum CA-125 concentrations of 75 ART cycles were measured 1 day before and on the day of human chorionic gonadotrophin (HCG) administration, and on the day of oocyte retrieval. These women did not have endometriosis and were induced by long-protocol gonadotrophin-releasing hormone analogue. Pregnancy was achieved in 35 (46.7%) cycles, but not in 40 cycles (53.3%). Serum CA-125 concentrations 1 day before and on the day of HCG administration and on the day of oocyte retrieval were significantly higher in cycles with pregnancy than in those without pregnancy (P < 0.05). It was noted that CA-125 concentrations on the day of oocyte retrieval were the best predictors of pregnancy, with concentrations >10 IU/ml having an accuracy of 86.6% for pregnancy. In conclusion, in intracytoplasmic sperm injection cycles, women with high serum CA-125 concentrations (>10 IU/ml) on the day of oocyte retrieval had very high pregnancy rates.     

Laboratory safety during assisted reproduction in patients with blood-borne viruses

For couples where one or both partners are infected with human immunodeficiency virus or hepatitis C, the doors to receiving fertility care are opening as a result of better antiviral medication, better long-term prognosis and consequent changes in attitude. In line with this, fertility centres electing to treat couples with blood-borne viral (BBV) infection need to re-examine their policies and procedures to ensure the safety of their staff and both non-infected and infected patients during assisted reproduction treatments. At a time when the European Tissue Directive aims to introduce quality standards for assisted reproduction throughout Europe, we highlight the risks involved when treating patients with known BBV infections and argue that safety cannot be met with any certainty unless samples from such patients are handled within a separate high security laboratory or laboratory area, technically adapted to ensure minimal cross-contamination risk to uninfected gametes and embryos.     

Blastocyst development after assisted reproduction using spermatozoa obtained after testicular stem cell transplantation in mice

BACKGROUND: Since its introduction in 1994, testicular stemcell transplantation (TSCT) has been widely used for research.This technique may also become important for preserving fertilityin pre-pubertal cancer patients. Therefore, it is necessaryto investigate the safety aspects of reproduction using spermatozoaobtained after TSCT. In this study, preimplantation developmentof mouse embryos, using spermatozoa obtained after TSCT, wasexamined. METHODS: TSCT-derived spermatozoa were used for IVFand ICSI. Embryos were cultured for five days until they reachedblastocyst stage and were evaluated by differential staining.RESULTS: IVF revealed significantly lower fertilization anddevelopment rates after TSCT-IVF compared to control-IVF. Blastocystsderived from TSCT-IVF had significantly lower inner cell massnumbers (ICMs) and lower ICM/trophectoderm (TE) ratios comparedto control-IVF blastocysts. No differences in fertilizationand development rates were observed between TSCT-ICSI and control-ICSI,and blastocyst quality in the transplanted group was similarto that of the control blastocysts. CONCLUSION: Our study showedthat after TSCT-IVF, fertilization and preimplantation developmentwere disturbed and blastocysts showed reduced ICM and ICM/TEratio. However, after TSCT-ICSI, both fertilization and preimplantationdevelopment were normal and blastocyst formation was comparableto control-ICSI.     

Attitudes towards human reproductive cloning, assisted reproduction and gene selection: a survey of 4600 British twins

BACKGROUND: Surveys have shown opposition to human reproductive cloning (HRC) in many countries. Views of identical (monozygotic, MZ) twins are of particular interest, as they naturally share 100% of their genes. We investigated attitudes of British twins towards HRC in the context of assisted reproduction technologies (ART) and gene selection. METHODS: About 4651 identical and non-identical (dizygotic, DZ) twins expressed their degree of agreement or disagreement to nine statements relating to ART, gene selection and HRC in a self-completion questionnaire. RESULTS: Most subjects (70% and 78% respectively) did not regard the use of medical technologies to treat infertility as interfering with either nature or God's will, despite believing that infertility is not a disease (54%). Attitudes to gene selection and HRC were context dependent, with more favourable views towards preventing serious diseases than towards enhancing traits. About 44% supported a permanent ban of HRC. MZ twins were significantly more likely to agree that HRC should be allowed for medical purposes, such as saving a sibling's life, than were DZ twins. Increasing religiosity generally correlated with more negative attitudes. CONCLUSIONS: Many attitudes are context dependent. More positive views of MZ twins towards HRC could be linked to their experience with being genetically identical.     

Ethical and legal aspects of assisted reproduction practice in Asia

This report describes the ethical and legal aspects of assistedreproduction technology (ART) that have been instituted in Asiancountries. The data were collected by a questionnaire circulatedto ART units in Asia. These are Taiwan, Singapore, Korea, Indonesia,Thailand, Japan, Iran, India, Jordan, Malaysia, China, Israel,Hong Kong, Pakistan, Lebanon, Saudi Arabia, and Persian Gulfcountries. According to the survey, there are approximately260 ART centers in Asia (half of which are in Japan). On a globalbasis each ART centre in Asia serves an average population of13 million people. On the other hand, in those Asian countrieswhere the standards of living are relatively high, the availabilityof ART services, including the more sophisticated and costlyART procedures like micromanipulation, is similar to that inthe Western world. In most of the Asian countries practisingART, however, no state registry exists. Taiwan is the only countrythat has specific legislation, and in six other countries somekind of ministerial regulations are practised. We conclude thatART is now practised in 20 countries in Asia. The prevailingrules and cultural heritage in many of these Asian countrieshas a major influence on the implementation of ART in Asia.However, in view of the complicated and sensitive issues involved,and as no supervision on ART clinics exists in most of theseAsian countries, we advocate that some kind of quality controlshould be urgently instituted in all centres practising ART.In this way, it is hoped that the highest standards be attainedfor all parties concerned.     

Genetic risk in micromanipulative assisted reproduction

The empirical data on pregnancy outcome after intracytoplasmicsperm injection (ICSI) are encouraging, but still insufficientto rule out definitely adverse genetic or teratological effects.We analyse potential adverse effects of ICSI by applying generalprinciples of genetic and teratological risk assessment. Therole of gamete micromanipulation as a possible teratogen isdiscussed. We consider whether ICSI patients are at increasedrisk for carrying and propagating genetic lesions of differentkinds. The possible interference of ICSI with genomic imprintingand pre-zygotic sperm selection is reviewed and we discuss thepotentially protective role of DNA repair in oocytes. Consideringthe available empirical data and the conclusions from our theoreticalanalysis it appears that neither lassitude nor over-concernabout adverse effects of ICSI are justified.     

Pentoxifylline: actions and applications in assisted reproduction

Data are presented covering various studies on the use of thephosphodiesterase inhibitor pentoxifylline (PF) in the spermpreparation for procedures in assisted reproduction. Significantimprovements have been shown in the fertilization rate of oocytesalong with a reduced risk of failed fertilization cycles utilizingoligo/asthenozoospermic semen samples. Fertilization is alsoimproved for normozoospermic samples when the acrosome reactionis suboptimal. PF has proven effects on sperm motility, increasingthe proportion of hyperactivated spermatozoa. It can also enhancethe acrosome reaction and this may be the more relevant functionfor clinical prediction. There is a further action as a suppressoror scavenger of reactive oxygen species although higher concentrationsthan that in current clinical use may be required to optimizethis effect. PF should be washed out of the sample used forinsemination to avoid inhibiting the completion of oocyte maturation.     

Babies born after ART treatment cost more than non-ART babies: a cost analysis of inpatient birth-admission costs of singleton and multiple gestation pregnancies

BACKGROUND: Currently, about one-third of infants born after assisted reproductive technology (ART) worldwide are twins or triplets. This study compared the inpatient birth-admission costs of singleton and multiple gestation ART deliveries to non-ART deliveries. METHODS: A cohort of 5005 mothers and 5886 infants conceived following ART treatment were compared to 245 249 mothers and 248 539 infants in the general population. Birth-admission costs were calculated using Australian Refined Diagnosis Related Groups and weighted national average costs (2003-2004 euro). RESULTS: ART infants were 4.4 times more likely to be low birthweight (LBW) compared with non-ART infants, translating into 89% higher birth-admission costs (euro2,832 and euro1,502, respectively). ART singletons were also more likely to be LBW compared with non-ART singletons, translating into 31% higher birth-admission costs (euro1,849 and euro1,415, respectively). After combining infant and maternal admission costs, the average cost of an ART singleton delivery was euro4,818 compared with euro13 890 for ART twins and euro54 294 for ART higher order multiples. Findings were not sensitive to changes in casemix. CONCLUSIONS: The poorer neonatal outcomes of ART singletons compared with non-ART singletons are significant enough to impact healthcare resource consumption. The high costs associated with ART multiple births add to the overwhelming clinical and economic evidence in support of single embryo transfer.     

Extended embryo culture in human assisted reproduction treatments

In order to evaluate the niche of extended embryo culture in an IVF programme, retrospective analysis of non-selected IVF patients, who underwent ovarian stimulation from April 1998 to June 1999 in a single private practice assisted reproductive technology centre, was performed. Embryos were cultured for 48 h in S1/G1.2 medium followed by 48 to 72 h of culture in S2/G2.2 to day 5 or day 6. Only fertilized oocytes exhibiting two pronuclei from donor and non-donor IVF and intracytoplasmic sperm injection (ICSI) cases were examined to determine the relationship between embryo cell number on day 3 and subsequent rate of blastocyst formation. Results indicated that a proportional relationship existed between the number of blastomeres present in day 3 embryos and the rate of blastocyst formation. Fifty-four per cent of embryos that had six cells on day 3 formed blastocysts, while 76% of those embryos with eight cells formed blastocysts. Blastocyst development did not increase further when embryos had more than eight cells on day 3, indicating that embryos with greater cell numbers on day 3 are not always predictive of a greater likelihood of blastocyst formation. Fertilized oocytes exhibiting two pronuclei from donors produced significantly more blastocysts (67%) than those from IVF patients (52%; P < 0.01), and had a significantly higher implantation rate (54%) compared with IVF patients (30%; P < 0.01). Furthermore, blastocyst cryopreservation resulted in significantly higher implantation rates than cryopreserved cleavage stage embryos (P < 0.001).     

GnRH antagonist in assisted reproduction: a Cochrane review

BACKGROUND: In the present systematic review, we wished to compare the efficacy of gonadotrophin-releasing hormone (GnRH) antagonist and GnRH agonist administration for controlled ovarian hyperstimulation in assisted conception. METHODS: Five randomized controlled trials fulfilled the inclusion criteria. In four studies, the multiple low-dose (0.25 mg) antagonist regimen was applied and, in one study, the single high-dose (3 mg) antagonist regimen was investigated. In all trials, reference treatment included a long protocol of GnRH agonist (buserelin, leuprorelin or triptorelin) starting in the mid-luteal phase of the preceding cycle. RESULTS: In comparison with the long protocol of GnRH agonist, the overall odds ratio for the prevention of premature LH surges was 1.76 [95% confidence interval (CI) 0.75-4.16], which was not statistically significant. There were significantly fewer clinical pregnancies in those treated with GnRH antagonists (OR 0.79; 95% CI 0.63-0.99). There was no statistically significant reduction in incidence of severe ovarian hyperstimulation syndrome between the two regimens (relative risk 0.51; OR 0.79; 95% CI 0.22-1.18). CONCLUSIONS: We concluded that the fixed GnRH antagonist protocol is a short and simple protocol with good clinical outcome, but the lower pregnancy rate compared with the GnRH agonist long protocol and the non-significant difference between both protocols regarding prevention of premature LH surge and prevention of severe ovarian hyperstimulation syndrome necessitates counselling subfertile couples before recommending change from GnRH agonist to antagonist. The clinical outcome may be further improved by developing more flexible antagonist regimens, taking into account individual patient characteristics.     

Cystic fibrosis in infertility: screening before assisted reproduction: opinion

Cystic fibrosis (CF) is the most common autosomal recessive disease in Caucasians. In 97-98% of men with CF, bilateral congenital absence of the vas deferens (CBAVD) blocks the transport of spermatozoa resulting in azoospermia. Abnormalities in sperm parameters have also been identified in males with CF. To date, over 800 disease-causing mutations of the CF transmembrane conductance regulator (CFTR) gene have been identified (also called ABCC7). Current legislation suggests that prior to intracytoplasmic sperm injection (ICSI) treatment, men with CBAVD or unexplained oligozoospermia should be considered for screening. If the male is negative with routine screening then the female partner is not screened. This is fundamentally wrong because if the female is screened and is found to be CF positive on routine testing, her partner would then need the fullest possible investigation of the CFTR gene. It is ideal to screen both partners in cases of oligozoospermia. However, if the resources are stretched, then only the female needs to be routinely screened because if she is negative, then the couple's residual risk of having a CF or CBAVD child will be reduced to 1:960. Only when the female is found to be a carrier does the male partner need routine screening followed by full testing for known mutations.     

Increased risk of early pregnancy loss by profound suppression of luteinizing hormone during ovarian stimulation in normogonadotrophic women undergoing assisted reproduction

The impact of suppressed concentrations of circulating luteinizing hormone (LH) during ovarian stimulation on the outcome of in-vitro fertilization or intracytoplasmic sperm injection treatment in 200 consecutive, normogonadotrophic women (couples) was analysed retrospectively. A standard stimulation protocol with mid-luteal gonadotrophin-releasing hormone (GnRH) agonist down-regulation and ovarian stimulation with recombinant follicle stimulating hormone (FSH) was used in all cases. Blood was sampled from each woman on stimulation days 1 and 8 for analysis of oestradiol and LH in serum. A threshold value of serum LH of 0.5 IU/l on stimulation day 8 (S8) was chosen to discriminate between women with low or 'normal' LH concentrations. Low concentrations of LH on S8 (<0.5 IU/l) were found in 49% (98/200) of the women. This group of women was comparable with the normal LH group with regard to pre-treatment clinical parameters, and to the parameters characterizing the stimulation protocol with the exception of serum oestradiol concentration, which on S8 was significantly lower than in the normal LH group (P < 0.001). The proportion of positive pregnancy tests was similar in the two groups (30% versus 34% per started cycle), but the final clinical treatment outcome was significantly different, with a five-fold higher risk of early pregnancy loss (45% versus 9%; P < 0.005) in the low LH group and consequently a significantly poorer chance of delivery than in the normal LH group. It is concluded that a substantial proportion of normogonadotrophic women treated with GnRH agonist down-regulation in combination with FSH, devoid of LH activity, experience LH suppression, which compromises the treatment outcome. Whether these women would benefit from supplementation with recombinant LH or human menopausal gonadotrophin during ovarian stimulation, remains to be proven in the future by prospective randomized trials.     

Assisted reproduction techniques for HIV serodiscordant couples: 18 months of experience

BACKGROUND: Assisted reproduction techniques can minimize the risk of infection and treat possible sterility associated with serodiscordant couples. METHODS: We assessed the efficacy of these techniques in 57 couples in which at least one partner had human immunodeficiency virus (HIV-1) infection that was currently under control (47 men and 10 women). The semen of seropositive men was prepared and tested for viruses. Assisted reproduction techniques included intrauterine insemination (IUI), IVF and especially ICSI, with ovarian stimulation that used a long agonist protocol and recombinant FSH. Embryos were transferred on day 3 after oocyte retrieval. RESULTS: For couples with seropositive men, five IUI and 49 IVF or ICSI attempts were perfomed, whilst for seropositive women these numbers were three IUI and 12 IVF or ICSI. No pregnancy occurred following the eight IUI trials. Seroconversion was not observed in any partners of seropositive men. Efficacy of treatment for these couples with ICSI was good, the clinical pregnancy rate per embryo transfer was 48.8%. The results for seropositive women were disappointing, with a clinical pregnancy rate per embryo transfer of 9.1%. Fourteen babies from 47 treated couples have so far been born and no pregnancies from IUI. CONCLUSIONS: Assisted reproduction techniques and particularly ICSI provide HIV-1-seropositive men with a safe and highly effective means of fathering children. These techniques may be less effective for seropositive women.     

Kinetics of SRY gene appearance in maternal serum: detection by real time PCR in early pregnancy after assisted reproductive technique

BACKGROUND: Fetal DNA circulating in maternal serum offers a possibility for non-invasive prenatal diagnosis but its kinetics during very early pregnancy is still unclear. In order to clarify this point, the studies on the kinetics of fetal DNA appearance in maternal serum were conducted on patients undergoing assisted reproduction. METHODS: Using a quantitative real time PCR assay, the presence of SRY gene sequences was evaluated in the serum of patients at the onset of pregnancy. RESULTS: Twenty-seven patients were originally studied but first trimester abortion occurred in five cases. Among the 22 ongoing pregnancies, ten were found to bear at least one male fetus and all sera from these women gave positive results for SRY gene detection. The SRY gene was found to be detectable as soon as day 18 after embryo transfer in one case and it had been found in the other nine patients by day 37. CONCLUSIONS: Fetal DNA is found in maternal serum even before the fetal circulation is established, which is highly suggestive that it is released, at least in part, from the trophoblast. Detection of fetal DNA in maternal serum very early in pregnancy may have clinical implications such as with the management of pregnant women carrying a fetus at risk for congenital adrenal hyperplasia.     

Safety issues in assisted reproduction technology: the children of assisted reproduction confront the responsible conduct of assisted reproductive technologies

Neurological sequelae and multiple birth defects have been observed in children conceived by IVF and ICSI. Multiple pregnancy is the most important risk factor. These health problems challenge the responsible practice of medicine. The core values of medicine and the deontology of the profession have been reviewed to define the responsible conduct of research and clinical practice. Professional associations have proposed guidelines to reduce health problems in assisted reproductive technology. Although these health problems could have been prevented, this response by the medical community is nonetheless an important step towards improving responsible medical practices that have become questionable over the years. Professional associations must find out means, not only to implement their guidelines, but also to prevent the recurrence of such episodes in the history of medicine.