Assisted Reproductive Technology Methods Research Articles

Reproductive Outcomes After Assisted Conception

The last three decades have witnessed a dramatic increase in the use of assisted reproductive technology (ART) so that now, in developed countries, 1.7% to 4.0% of all children are born after ART. Although absolute risks appear small, data from prospective and retrospective studies indicate increased risks of adverse maternal and perinatal outcomes after ART as compared with spontaneous conception. Recent studies suggest that underlying maternal factors and subfertility play an important role in some of these outcomes rather than the ART procedure itself. A significant risk of assisted conception is multiple pregnancies, but even singleton pregnancies achieved by ART are at a higher risk of hypertensive disease, diabetes, prematurity, low birth weight, and perinatal mortality even after adjusting for confounders. Couples undergoing ART procedures should be counseled in advance regarding increased risks of pregnancy complications and higher rates of obstetric interventions. Although conflicting data exist, studies of children born from ART suggest increased rates of congenital malformations, imprinting disorders (Beckwith-Wiedemann syndrome and Angelman syndrome), and marginally increased risk of cancer. However, the current evidence is inadequate, and prospective long-term studies are needed to eliminate the effect of confounders and draw definite conclusions about the long-term outcomes after ART. The absolute risk of imprinting disorders remains small, and routine screening is not recommended at present. The long-term outcomes after ART are difficult to evaluate because of the variability in ART methods and data reporting, and there is a need for standardized methodology for follow-up after ART.

Clinical characteristics and assisted reproductive technology outcomes in infertile foreign brides: Comparison with native brides in south Taiwan

ObjectiveThis study was conducted to determine if differences in clinical characteristics, embryonic development, and pregnancy outcomes exist between infertile foreign and native brides undergoing assisted reproductive technology (ART) treatment in a foreign-bride-immigrating country of Taiwan. Material and MethodsA retrospective, case-control design was use to recruit consecutive infertile women seeking ART treatment for study. The treatment protocol consisted of controlled ovarian hyperstimulation with down-regulation and gonadotropin and ART method with conventional in vitro fertilization or intracytoplasmic sperm injection. Among the 1083 ART treatment cycles, 97 (9%) were from foreign brides and 986 (91%) from native brides. Records of those who received transfer of fresh embryos obtained by using husband sperm for fertilization were collected for analysis. A case-control design with adjustment of bride age was used to select the records of 86 infertile foreign and 172 native brides for the study. Infertility cause, bride age, retrieved oocyte number and maturity, fertilization rate, embryo score, and post-embryo transfer results were assessed. ResultsThe foreign bride couples had significantly greater infertility due to tubal (36%) and combination (8.1%) factors than native bride couples (20.3% and 4.1%, respectively). Days of gonadotropin stimulation, number of oocytes retrieved, fertilization rate, embryo score, implantation rate, or clinical pregnancy rate, and live birth rate did not differ between the two groups. ConclusionThe cause of infertile foreign bride couples requiring ART treatment had a higher percentage of tubal and combination factors, but similar ART outcomes.

Delivery after assisted reproduction

The aim of the paper was to describe and compare means of deliveries after assisted conception and after spontaneous conception. The rapid spread of assisted reproductive technology (ART) is part of development of modern society. Study is retrospective, descriptive and analytical. Data were collected out of medical charts of patients of Hospital for gynecology and obstetrics CHC Zvezdara, who have delivered babies in this institution after using methods of ART during the years 2001 till 2011. There were 190 ART patients, age ranging from 21 to 56 years, on average 35.6 (± 4.7) years. Majority (94.6%) of women were primiparous, without previous miscarriages. The most frequently used method of ART is IVF/ET (94.8%, of which ICSI was performed in 6.53%). 42.1% of pregnancies were achieved after second attempt of IVF/ET. Pregnancies delivered vaginally lasted on average 38,7±1,2 week of gestation. Majority of premature infants (overall incidence of preamaturety was 20,2%) were born by urgent cesarean section. There were 5 extremely premature infants (2,17%). There was 17,4% of twin pregnancies, and almost all of them (92,5%) we delivered by cesarean section. Average birth weight was 3160g (± 600), and average body length on birth was 51cm (± 3). The most vital infants were born spontaneously (wirh mean Apgar score in the 5th minute of 9,08). During our first ART pregnancy experiences we performed no vaginal deliveries. Further on, evident decreasing trend of cesarean section made place for vaginal deliveries. This ratio in study population slowely approaches to the general population ratio, and for now it is 40:60 (vaginal:cesarean delivery). Current experiences encourage us to deal with ART pregnanices, as if they were spontaniously achieved, and to deliver them respecting obsteric indications.

Placental weight in singleton pregnancies with and without assisted reproductive technology: a population study of 536 567 pregnancies

Pregnancies conceived by assisted reproductive technology (ART) are at increased risk of adverse outcomes. Previous studies have suggested increased placental weight and increased placental weight/birthweight ratio in pregnancies associated with adverse outcomes. We therefore studied the association of ART with placental weight and placental weight/birthweight ratio. We included all singleton births in the Medical Birth Registry of Norway during the period 1999-2008 (n = 536 567, including 8259 after ART). We divided placental weight and placental weight/birthweight ratio into quartiles, and calculated the proportions of ART and spontaneous pregnancies in the lowest and the highest quartile by length of gestation. Thereafter, we estimated crude and adjusted odds ratios (ORs) for being in each quartile of placental weight for ART pregnancies with spontaneous pregnancies as the reference. The analyses were repeated with ART pregnancies subgrouped into IVF or ICSI. Mean placental weight was 678.9 g in pregnancies conceived by ART, and 673.0 g in pregnancies after spontaneous conception. ART pregnancies were overrepresented in the highest quartile of placental weight and underrepresented in the highest quartile of birthweight, independent of length of gestation at delivery. Thus, placental weight/birthweight ratio was higher in ART pregnancies. For ART pregnancies, the OR for being in the highest quartile of placental weight was 1.37 (95% confidence interval 1.30-1.45) after adjustment for length of gestation, offspring birthweight, parity, fetal sex, maternal age, pre-eclampsia and diabetes. There was no difference in placental weight/birthweight ratio between IVF and ICSI pregnancies. We found larger placentas and a higher placental weight/birthweight ratio among pregnancies conceived by ART compared with spontaneous pregnancies, and the difference was independent of length of gestation at delivery and ART method.

Herpes simplex virus infection of human spermatozoa correlates with decreased frequency of blastocyst formation and frequency of embryo implantation during in vitro fertilization

We have conducted a comparative analysis of developing human embryos in the course of in vitro fertilization (IVF) as a method of sterility treatment of two groups of patients: herpes simplex virus (HSV) was detected in the fraction of motile sperm of male partners in group I (n = 28) and no HSV was found in sperm in group II (n = 103). W assessed number of fertilized ova, embryos during cleavage, and blastocysts as well as such parameters as frequency of implantation and frequency of pregnancy in IVF cycles. It was established that the presence of HSV in spermatozoa did not affect the efficiency of fertilization or cleavage of zygotes. At the same time, in cases of virus-infected male gametes, the frequency ofblastocyst formation was two times less (p = 0.015), and the frequency of embryo implantation and pregnancy was, on average, five times lower (p = 0.01 andp = 0.016, respectively). Based on the obtained results, a conclusion was made about the negative influence of HSV-virus in male gametes at the early stages of embryo development and the frequency of achievement of pregnancy in respect to the methods of assisted reproductive technologies (ART).

Risk of congenital heart defects associated with assisted reproductive technologies: a population-based evaluation

To estimate the risk of congenital heart defects (CHD) associated with assisted reproductive technologies (ART). We used data from the Paris Registry of Congenital Malformations on 5493 cases of CHD and 3847 malformed controls for which no associations with ART were reported in the literature. Assisted reproductive technologies included inductors of ovulation only, in vitro fertilization, and intracytoplasmic sperm injection. Exposure to ART was higher for cases than controls (4.7 vs. 3.6%, P= 0.008) and was associated with a 40% increase in the maternal age, socioeconomic factors, and year of birth-adjusted odds of CHD without chromosomal abnormalities [adjusted odds ratio (OR) 1.4, 95% confidence interval (CI) 1.1-1.7]. Assisted reproductive technologies were specifically associated with significant increases in the odds of malformations of the outflow tracts and ventriculoarterial connections (adjusted OR 1.7, 95% CI 1.2-2.4) and of cardiac neural crest defects and double outlet right ventricle (adjusted OR 1.7, 95% CI 1.1-2.7). In general, we found specific associations between methods of ART and subcategories of CHD. Cases with CHD were more likely to have been conceived following ART when compared with malformed controls. This higher risk for CHD varied specifically according to the method of ART and the type of CHD and may be due to ART per se and/or the underlying infertility of couples.

64 Risk of Congenital Heart Defects for Children Conceived Following Assisted Reproductive Technologies. A Population Based Evaluation

Background: Children conceived after ART have higher risks for adverse birth outcomes. Insufficient data exist regarding the specific risk for CHD. Objective: To estimate the risk associated with Assisted Reproductive Technologies (ART) for congenital heart defects (CHD), CHD without chromosomal abnormalities and CHD subgroups defined based on anatomo-embryologic criteria. Methods: We used data from the Paris Registry of Congenital Malformations on 5,599 cases of CHD and 3,985 malformed controls including a heterogeneous group of malformations for which no known associations with ART were reported in literature. ART included inductors of ovulation only, in vitro fertilization and intracytoplasmic sperm injection. Results: Exposure to ART (all methods combined) was higher for cases than controls (4.7% vs. 3.6%, p=0.008) and was associated with a 40% increase in the maternal age, socioeconomic factors and year of birth-adjusted odds of CHD without chromosomal abnormalities (Adjusted-OR 1.4, 95%CI 1.1-1.7). ART was specifically associated with significant increases in the odds of malformations of great vessels and outflow tracts (Adjusted-OR 1.7, 95%CI 1.2-2.4), of anomalies of atria and atrial septal defects (Adjusted-OR 1.8, 95%CI 1.1-2.8) and of conotruncal CHD and double outlet right ventricle (Adjusted-OR 1.7, 95%CI 1.1-2.7). In general, we found specific associations between methods of ART and subgroups of CHD. Conclusion: Risk of CHD was higher in children exposed to ART as compared to malformed controls. This association seems to vary according to the method of ART and type of CHD and may be due to ART per se and/or the underlying infertility of couples.

International Federation of Fertility Societies Surveillance 2010: preface

Surveillance is a triennial worldwide compendium of national rules and regulations for assisted reproductive technology. It was last published in 2007.

The hyaluronan binding assay HBA™: a novel and statistically significant predictor of clinical pregnancy (CP) in in vitro fertilization (IVF) and assisted reproductive technology (ART)

OBJECTIVE: Established andrology assays assist in directing the clinician towards the selection of the optimal ART for each couple facing infertility. The HBA assay assesses the ability of the seminal population to bind to an immobilized substrate of hyaluronan. H-bound (HB) sperm carry reduced levels of chromosomal aberrations and enhanced levels of developmental maturity. This relationship led to the current study examining sperm binding potential, descriptive parameters of the male and female partners and their individual contribution to IVF outcome. The study sought to explore the potential association between the HBA index, morphology (M), total number of motile sperm (MS), ART treatment, age, follicle stimulating hormone (FSH) level, number of dominant follicles (DF) and the likelihood of CP.DESIGN: A retrospective analysis of 253 infertility couples examined each parameter as to its contribution to a positive CP. Data collection followed an IRB-approved protocol. The sample consisted of patients requiring intracytoplasmic sperm injection (ICSI) (n=84), IVF (n=93) and intrauterine insemination (IUI) (n=76).MATERIALS AND METHODS: The HBA score, M, MS, ART treatment, age, FSH and DF were the independent variables (IV) recorded for each patient. The HBA score was calculated as # bound MS/# total MS. With CP as the dependent variable, all IV were entered into a logistic regression model (LRM). Descriptive analyses included ANOVA and Pearson's χ 2 (chi square) using SPSS 15.0.RESULTS: Overall, 11.8% of IUI, 68% of IVF and 72.6% of ICSI patients demonstrated a CP. There was a statistically significant difference in mean HBA scores of IUI and IVF patients (79.87, 78.58, respectively (resp.)) as compared to ICSI patients (62.26, p<0.001). The LRM showed that the HBA score, female age and ART method had a statistically significant role in contributing to CP (p=.003, p=.001, p<.001, resp.). Male age, MS, M and FSH level showed no statistical significance (p>0.05) in contributing to CP.CONCLUSIONS: The use of the HBA index in the evaluation of the infertile couple merits strong consideration in the selection of the most appropriate ART to achieve a CP. The statistically significant value of the HBA index in predicting CP suggests the influence HB-sperm exert on preimplantation embryogenesis. Use of refined criteria for the HBA index may better direct the ART technologies selected for infertile patients thus optimizing CP and improving patient care. OBJECTIVE: Established andrology assays assist in directing the clinician towards the selection of the optimal ART for each couple facing infertility. The HBA assay assesses the ability of the seminal population to bind to an immobilized substrate of hyaluronan. H-bound (HB) sperm carry reduced levels of chromosomal aberrations and enhanced levels of developmental maturity. This relationship led to the current study examining sperm binding potential, descriptive parameters of the male and female partners and their individual contribution to IVF outcome. The study sought to explore the potential association between the HBA index, morphology (M), total number of motile sperm (MS), ART treatment, age, follicle stimulating hormone (FSH) level, number of dominant follicles (DF) and the likelihood of CP. DESIGN: A retrospective analysis of 253 infertility couples examined each parameter as to its contribution to a positive CP. Data collection followed an IRB-approved protocol. The sample consisted of patients requiring intracytoplasmic sperm injection (ICSI) (n=84), IVF (n=93) and intrauterine insemination (IUI) (n=76). MATERIALS AND METHODS: The HBA score, M, MS, ART treatment, age, FSH and DF were the independent variables (IV) recorded for each patient. The HBA score was calculated as # bound MS/# total MS. With CP as the dependent variable, all IV were entered into a logistic regression model (LRM). Descriptive analyses included ANOVA and Pearson's χ 2 (chi square) using SPSS 15.0. RESULTS: Overall, 11.8% of IUI, 68% of IVF and 72.6% of ICSI patients demonstrated a CP. There was a statistically significant difference in mean HBA scores of IUI and IVF patients (79.87, 78.58, respectively (resp.)) as compared to ICSI patients (62.26, p<0.001). The LRM showed that the HBA score, female age and ART method had a statistically significant role in contributing to CP (p=.003, p=.001, p<.001, resp.). Male age, MS, M and FSH level showed no statistical significance (p>0.05) in contributing to CP. CONCLUSIONS: The use of the HBA index in the evaluation of the infertile couple merits strong consideration in the selection of the most appropriate ART to achieve a CP. The statistically significant value of the HBA index in predicting CP suggests the influence HB-sperm exert on preimplantation embryogenesis. Use of refined criteria for the HBA index may better direct the ART technologies selected for infertile patients thus optimizing CP and improving patient care.

Sperm DNA decondensation assay and selection of assisted reproductive technology method

Previously, sperm DNA integrity testing was reported to predict failure of sperm with assisted reproductive technology (ART) regardless of insemination method. Recent publications indicate that the sperm DNA integrity test, SCSA/SDFA, can identify reduced chances for success in some ART methods but not for others. The sperm DNA decondensation (SDD) test, a sperm function test, also previously has been shown to predict failure in all types of subsequent ART attempts. However, recent unpublished data suggest that ICSI can overcome sperm DNA function abnormalities identified by the SDD test. The goal of this retrospective review was to determine if SDD had predictive capability in current IUI and IVF insemination protocols. Retrospective chart review. Within a single fertility practice, all males electing to proceed with SDD testing were identified. ART success, defined as presence of fetal heart beat at 8–10 weeks gestation, was determined for first IUI or IVF attempt after SDD testing. An abnormal SDD score was defined as less than 80% of the control and normal was defined as greater than or equal to 80% of the control. There were no exclusions for female factor or WHO male factor etiology. SDD scores and first IUI or IVF attempt outcome data were available for 58 males. Forty-three males had normal SDD scores and had a 22% success rate with IUI (N = 23) and 35% with IVF (N = 20) attempts. Fifteen males had abnormal SDD scores with a 0% success rate with IUI (N = 6) and IVF (N = 9) attempts. The difference between the 28% success rate of SDD normal patients (N = 43, IUI/IVF combined) and 0% with SDD abnormal patients (N = 15, IUI/IVF combined) was statistically significant using Fischer's exact test (P=0.0325) (Odds Ratio >5). An additional 18 patients underwent IVF with ICSI where 81% (N = 16) of males with normal SDD scores had success and 100% (n = 2) with an abnormal SDD had success, although there was not sufficient statistical power to conclude that the groups were different. There was no significant difference for the average age and FSH levels of female partners between SDD normal and SDD abnormal groups. Overall clinical pregnancy rates for this fertility practice at the time of this study were: IUI, 15%; IVF and ICSI, 53%. Based upon preliminary retrospective data, the SDD may be able to identify those male patients with reduced chances of success with IUI and IVF who may benefit from earlier consideration of ICSI. Further definitive studies are needed.

Establishment of criteria for elective single embryo transfer at day 2 or day 3 by analyzing cases with successful implantation of all embryos transferred

Elective transfer of two good-quality embryos has been used to avoid triplet or high-order multiple pregnancies. However, the rate of twin pregnancies has remained fairly unchanged. In the present study, criteria for elective single embryo transfer (eSET) at day 2 or day 3 were established by analyzing cases with successful implantation of all embryos transferred. A total of 685 fresh or frozen-thawed embryo transfers following in vitro fertilization/intracytoplasmic sperm injection between April 2002 and March 2006 were performed. Only embryo transfers at day 2 or day 3, but not at blastocyst stage, were included. Successful implantation of all embryos transferred was obtained in 17 pregnancy cycles. Thirty-one gestational sacs with fetal heartbeats were obtained by a total of 31 embryo transfers in 17 infertile women. The average age was 32.6 years (23-38), and 14 (82.3%) of the 17 women were <36 years old. Fifteen (88.2%) of the 17 pregnancies were established at the first attempt of assisted reproductive technology (ART). Of the 17 women, eight (47.1%) women were multigravida and four (23.5%) women were multipara. The indications for ART or insemination methods did not seem to be related to the pregnancy results. Twenty-nine (93.5%) of 31 embryos implanted were considered good-quality embryos. Of the 17 fresh embryos transferred at day 2, 15 were at the 4-cell stage and two were at the 5-cell stage. Of the 11 fresh embryos transferred at day 3, one was at the 6-cell stage, two were at the 7-cell stage and eight were at the 8-cell stage. The criteria for eSET at day 2 or day 3 were established as follows: <36 years of age, a first treatment cycle and more than two good-quality embryos developed at least to the 4-cell stage at day 2, or 6-cell stage at day 3. Additionally, the past history of pregnancy or delivery should be considered, as patients positive for such history might have better implantation ability. eSET can be highly recommended to avoid twin pregnancies in subjects with the established criteria.

Epigenetics in assisted reproductive technology.

It has been reported that the rates of epigenetic disorders such as Angelman syndrome (AS) and Beckwith-Wiedemann syndrome (BWS) are high in offspring conceived by assisted reproductive technology (ART). Angelman Syndrome is characterized by intellectual disability and BWS is known as large offspring syndrome (LOS). Weight abnormalities have also been reported in cloned animals. Possible factors underlying these findings include inherent gamete characteristics, influence of invitro culture and peculiarity of ART methods. It is important to conclusively determine whether such epigenetic abnormalities are present in children conceived by ART, so as to consider the health of next generations.

Serum PAPP-A Levels at 10–14 Weeks of Gestation Are Altered in Women After Assisted Conception

The most sensitive and specific first-trimester Down syndrome screening test employs a combination of maternal age, nuchal translucency (NT) thickness, fβhCG, and PAPP-A. First trimester NT measurements do not differ significantly between pregnancies resulting from assisted reproductive technology (ART) and those resulting from spontaneous conception, but maternal blood levels of free beta human chorionic gonadotropin (fβhCG) are higher following in vitro fertilization-embryo transfer (IVF-ET), and levels of pregnancy-associated plasma protein-A (PAPP-A) are lower. Whether or not this results in a higher rate of false positive screening test results is controversial. The investigators compared the screen-positive rates of the first trimester Down syndrome screening test in women with singleton ART pregnancies to those of women whose pregnancies resulted from spontaneous conception. Participants included 130 women who were pregnant after IVF-ET (group 1); 54 who conceived after intracytoplasmic sperm injection (ISCI; group 2); and 914 control women who conceived spontaneously. Fetal NT thickness was estimated at 10–14 weeks’ gestation. Women who conceived via ICSI were 2.8 times as more likely to be screen positive than control women. Although ovarian stimulation in group 1 and group 2 women was associated with significantly lower PAPP-A levels than were present in the control group, women who conceived via IVF-ET were not more likely to be screen positive than control women. Levels of PAPP-A declined significantly as the number of retrieved oocytes or transferred embryos increased, but the levels of other markers were not affected. When a single embryo was transferred, or when ART conception occurred without ovarian stimulation, marker levels were similar to those following spontaneous conception. Although ICSI appears to be associated with a higher rate of false positive first trimester screening test results than that in spontaneously conceived pregnancies, women who conceive by other ART methods are not at increased risk.

Assisted reproductive technologies do not alter mutation frequency or spectrum

Assisted reproductive technologies (ARTs) have now contributed to the birth of >3 million babies worldwide, but concerns remain regarding the safety of these methods. We have used a transgenic mouse model to examine the effects of ARTs on the frequency and spectrum of point mutations in midgestation mouse fetuses produced by either natural reproduction or various methods of ART, including preimplantation culture, embryo transfer, in vitro fertilization, intracytoplasmic sperm injection, and round spermatid injection. Our results show that there is no significant difference in the frequency or spectrum of de novo point mutations found in naturally conceived fetuses and fetuses produced by in vitro fertilization, intracytoplasmic sperm injection, or round spermatid injection. These results, based on analyses of a transgenic mouse system, indicate that with respect to maintenance of genetic integrity, ARTs appear to be safe.

Ethics of Artificial Insemination: An Islamic Perspective

DOI: http://dx.doi.org/10.5915/39-1-5264 This paper aims to address ethical issues related to the assisted reproductive technology method of artificial insemination (AI) from the Islamic perspective. Our evaluation suggests that the Islamic ethical viewpoint of AI is rational, comprehensive, and aims at the greatest good of mankind.

ESHRE Task Force on Ethics and Law 10: Surrogacy

This 10th statement of the Task Force on Ethics and Law considers ethical questions specific to varied surrogacy arrangements. Surrogacy is especially complex as the interests of the intended parents, the surrogate, and the future child may differ. It is concluded that surrogacy is an acceptable method of assisted reproductive technology of the last resort for specific medical indications, for which only reimbursement of reasonable expenses is allowed.

Birth defects in children conceived by ICSI compared with children conceived by other IVF-methods; a meta-analysis

Intracytoplasmic sperm injection (ICSI) is a method of assisted reproductive technology that involves the selection of a single sperm cell and the manual injection of this cell into the egg. The lack of relevant experimental studies, the nature of the technology involving non-natural selection of the fertilizing sperm, and possible damage to the egg have caused concern that ICSI could increase the risk of birth defects. Data from available cohort studies comparing ICSI with standard in vitro fertilization (IVF) should be combined to evaluate the risks involved with ICSI. We reviewed more than 2500 titles and abstracts containing keywords related to ICSI and identified 22 scientific articles with data on birth defects among ICSI-births. A total of four peer-reviewed, non-overlapping prospective cohort studies provided reliable and comparable data on birth defects both for children conceived by ICSI and children conceived by standard IVF. These studies included a total of 5395 children born after ICSI. The pooled estimate of the risk of a major birth defect was a 1.12-fold increase after ICSI when compared with standard IVF (risk ratio = 1.12, 95% confidence interval (CI): 0.97-1.28, P = 0.12). There was no marked heterogeneity of risk ratios between these studies (P = 0.10). We found no significantly increased risks after ICSI for any of the categories cardiovascular defects, musculoskeletal defects, hypospadias, neural tube defects, or oral clefts. Our analysis does not indicate that the ICSI-procedure represents significant additional risks of major birth defects in addition to the risk involved in standard IVF. The data was limited, particularly on risks of specific categories of defects.

Looking Farther Afield

Several recent studies have investigated the potential association between assisted reproductive technology (ART) and risk of childhood cancer (e.g., retinoblastoma) or genetic syndromes at high risk of malignancy [e.g., Beckwith-Wiedemann syndrome (BWS); refs. 1, 2]. Most recently, Halliday et al. (3) conducted a case-control study in Victoria, Australia to measure the association between ART and BWS. Of 1.3 million live births during the period 1983 to 2003, 37 cases of BWS were born (confirmed by clinical geneticists) for an overall population prevalence of ∼1 case per 35,500 births. These cases were each matched (age, maternal age, and parity) to four live-born controls. Record linkage was done with the providers of ART services in Victoria to determine which infants were conceived using ART methods. Of the 37 BWS cases, 4 were conceived by ART compared with 1 of 148 matched controls (odds ratio, 17.8; 95% confidence interval, 1.8-432.9). In the context of all 14,894 live births conceived by ART during this time period, the absolute risk of BWS was ∼1 case per 4,000 births or about nine times higher than in the general population. As the authors note, previous reports of ART-associated BWS consistently show hypomethylation of the maternal KvDMR1/LIT1 locus at 11p15.5 (4, 5). In contrast, this mechanism is only observed in ∼45% of the overall BWS population. It is possible that the process of ART preferentially results in maternal allele demethylation, which would have important implications not only for BWS but also for childhood cancer.Genomic imprinting, or the preferential expression of a gene depending on the parent of origin, results from the differential methylation of CpG islands. The imprint is erased during gametogenesis through demethylation and reset later according to the parent of origin of the transmitted chromosome. Demethylation of maternally transmitted alleles is a mechanism by which some genetic disorders and malignancy arise. Although demethylation has been described in the maternal allele (see above), there has been little evidence that this phenomenon occurs in the paternal allele. Marques et al. (6) examined spermatozoan DNA from 123 individuals (27 normozoospermic and 96 oligozoospermic, including 46 moderately affected and 50 severely affected) undergoing routine analysis for infertility. Methylation profiles were studied in two imprinted genes: MEST (maternally imprinted) and H19 (paternally imprinted). For MEST, maternal imprinting was correctly erased in all 123 samples. In contrast, although the H19 paternal imprint was correctly erased in the normozoospermic samples, 8 (17%) of the moderately affected and 15 (30%) of the severely affected men showed incomplete methylation. The authors conclude that abnormal spermatogenesis is associated with a rise in methylation defects at H19. Because the maternal imprint was correctly erased for MEST, this may indicate that changes in DNA methyltransferase activity are responsible for hypospermatogenesis. Importantly, there is coordinated regulation of expression of IGFII (a maternally imprinted gene) by repressor binding sites on the H19 gene (7). Thus, paternal H19 hypomethylation could lead to the inactivation of both IGFII genes, which could have deleterious consequences during fetal development.

Risks of birth defects and other adverse outcomes associated with assisted reproductive technology.

Approximately 1 million children world-wide have been born through assisted reproductive technology (ART),1 which helps many motivated couples give birth to healthy infants. Children conceived through ART comprise as many as 1% to 2% of total births in some countries.2,3 High rates of multiple births, with attendant complications of prematurity and low birth weight, are well documented. Concerns are now emerging about associated increased risks for congenital anomalies. Use of newer techniques may bring additional hazards, especially those requiring more biologic manipulation than artificial insemination and other, older ART methods. Outcomes data through ART registries permit risk estimates, although some data ascertainment is incomplete, especially in the United States. This report reviews ART outcomes, with focus on the potential risks of birth defects and their possible mechanisms, as well as limitations of current risk assessments. Moreover, it suggests ways to improve ART tracking and outcomes. ART is broadly defined as conception through any procedure or medication assisting in achieving pregnancy, including ovulation induction and intrauterine insemination. The Centers for Disease Control and Prevention (CDC) defines ART more narrowly as “any procedure that entails the handling of both eggs and sperm or of embryos for the purpose of establishing a pregnancy.”4 These procedures include in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). In IVF, fertilization is achieved by mixing eggs and sperm together in the laboratory. In ICSI, fertilization consists of direct microinjection of 1 sperm into 1 egg. In both, fertilized eggs are implanted after several days of ex vivo cell growth. Techniques of surgical embryo transfer include gamete intrafallopian transfer and zygote intrafallopian transfer. Emerging techniques involve additional gamete and embryo manipulation such as in vitro oocyte maturation.1 Complete data for the numbers of ART-related births, specific ART methods used, and birth …

Physical and mental development of children after in vitro fertilization and embryo transfer.

Objective: To evaluate the physical and mental development of children after in vitro fertilization (IVF) and frozen embryo transfer (FET). Methods: Between July 1995 and November 2003, 506 patients delivered 658 babies after IVF and ET treatment at our clinic (intracytoplasmic sperm injection (ICSI), 418; conventional IVF, (C-IVF) 125; FET, 115). A survey of the physical and mental developmental of the children was conducted by mailing questionnaires to parents. Comparisons were made between three treatment procedures, and development of singleton, twin and triplet delivery. Results: The response rate was 73.4% (483/658) for 324 children born after ICSI, 78 born after C-IVF, and 81 born after FET. The height and weight of assisted reproductive technology (ART) children at birth were significantly lower than that of naturally conceived babies (ART children: natural delivery; 46.8cm, 49.0cm and 2524g, 3040g, respectively). However, there was no significant difference between the singletons alone and naturally conceived children irrespective of the ART method. In addition, mental development was the same between singletons and naturally conceived children. The ART group tended to delay body development such as 'holding their head up', 'sitting up', 'crawl' to moving growth in multiple births. Conclusion: The physical and mental development of twins or triplets was significantly more delayed than that of naturally conceived babies, but had improved to a similar extent of the singletons after the age of 6months. (Reprod Med Biol 2004; 3: 63-67).