Cyanotic congenital heart disease following fertility treatments in the United States from 2011 to 2014

ObjectiveTo examine the risk for cyanotic congenital heart diseases (CCHDs) among live births in the USA, resulting from various forms of infertility treatments.MethodsThis study is a cross-sectional analysis of live births in the USA from 2011 to 2014. Infertility treatments are categorised into two of the following groups on birth certificates: assisted reproductive technology (ART) fertility treatment (surgical egg removal; eg, in vitro fertilisation and gamete intrafallopian transfer) and non-ART fertility treatment (eg, medical treatment and intrauterine insemination). We compared the risk for CCHD in ART and non-ART fertility treatment groups with those infants whose mothers received no documented fertility treatment and were naturally conceived (NC).ResultsAmong 14 242 267 live births from 2011 to 2014, a total of 101 494 live births were in the ART and 81 242 resulted from non-ART fertility treatments. CCHD prevalence in ART, non-ART and NC groups were 393/100 892 (0.39%), 210/80 884 (0.26%) and 10 749/14 020 749 (0.08%), respectively. As compared with naturally conceiving infants, risk for CCHD was significantly higher among infants born in ART (adjusted relative risk (aRR) 2.4, 95% CI 2.1 to 2.7) and non-ART fertility treatment groups (aRR 1.9, 95% CI 1.6 to 2.2). Absolute risk increase in CCHD due to ART and non-ART treatments were 0.03% and 0.02%, respectively. A similar pattern was observed when the analysis was restricted to twins, newborns with birth weights under 1500 g and gestational age of less than 32 weeks.ConclusionsOur findings suggest an increased risk for CCHD in infants conceived after all types of infertility treatment.

216 IN VITRO CAPACITATION OF FROZEN/THAWED STALLION SPERM

The inability of stallion sperm to capacitate in an in vitro setting has limited many equine assisted reproductive technologies such as gamete intrafallopian transfer (GIFT) and IVF. These experiments were conducted to develop methods for artificially capacitating frozen/thawed stallion sperm, using dilauroylphosphatidylcholine (PC-12), calcium ionophore A23187 (IONO), or methyl-β-cyclodextrins (MBC) so that sperm could be used in equine assisted reproductive technologies. Assisted reproductive technologies need to be established to maximize the number of offspring produced with cryopreserved spermfrom stallions with low cryosurvival rates or that have a very limited number of frozen sperm. Low concentrations of lysophosphatidylcholine (LPC) induce the acrosome reaction in capacitated sperm and, therefore, were used to detect fully capacitated sperm treated with PC-12, IONO, or MBC. Ejaculates from 8 stallions were diluted in a modified Tyrode's medium and centrifuged (1000g for 11 min). The sperm pellets were suspended to 400 million sperm mL–1 in EZ-Freezin LE, packaged into 0.5-mL straws, and frozen in liquid nitrogen vapor. Straws were thawed in a 37�C water bath for 30 s and washed through 35% Percoll (400g for 4.5 min) to remove egg yolk particles and seminal plasma. The sperm pellets were suspended in Medium 199 (0.6% BSA and 5 mm calcium chloride) to 50 million sperm mL–1, and stained with propidium iodide and FITC-peanut agglutinin (PNA) (1 mg mL–1 each; viability and acrosome reaction detection, respectively). Sperm were treated with PC-12 (13, 17, 20, 30, 40 µm), IONO (0.5, 1, 1.5, 2 µm), or MBC (0.4, 0.6, 0.8, 1 µm) and incubated in a 37�C water bath (20, 15, 20 min) to artificially capacitate sperm. Sperm were then challenged with LPC (2 or 5 µg), for 10 additional min, and analyzed by flow cytometry to determine the percentages of acrosome-reacted sperm. Data were analyzed by ANOVA and treatments were separated by Student-Newman-Keul's test. The PC-12 (20, 30, 40 µm), IONO (1.5, 2 µm), and MBC (0.4, 0.6, 0.8, 1 µm) artificially capacitated sperm equally well (59 to 70%, 52 to 59%, and 48 to 55%, respectively) and were higher than control sperm (13%, SEM � 9; P < 0.05). Sperm viability was similar at all IONO or MBC concentrations; however, PC-12-treated sperm (17 to 40 µm) had significantly lower sperm viability when compared with control (24 to 28% and 35%, respectively, SEM � 3; P < 0.05). In conclusion, frozen/thawed stallion sperm can be artificially capacitated by treating with PC-12, IONO, or MBC, and this may lead to practical applications for in vitro equine assisted reproductive technologies.

‘Are You Asking Me If We Had Sex To Conceive?’ To Whom Do Parents Of Twins Disclose Mode Of Conception and What Do They Feel About Being Asked?

There are no data on whether parents of twins will disclose mode of conception to researchers or to their children, who will be informants in adulthood. We sent 1600 questionnaires about this via the Victorian branch of the Australian Multiple Birth Association, to be returned anonymously. Parents were asked how their twins were conceived and whether those who used assisted conception would disclose this to researchers studying assisted conception, twin pregnancy or twin children, or to their children. Comments were invited. Altogether 975 (61%) questionnaires were returned and 389 (40%) indicated use of some form of assisted conception: 75 (19%) ovarian stimulation alone, 165 (42%) In Vitro Fertilisation, 132 (34%) Intracytoplasmic Sperm Injection, and 17 (4%) Gamete Intrafallopian Transfer, with 20 reporting use of donor eggs and thirteen donor sperm. Of those using assisted conception, the proportion reporting that they would not, or may not, tell researchers was 5% for assisted conception studies, 6% for twin pregnancy studies, and 7% for studies of twin children, while 7% reported that they would not, or may not, tell their children. From the comments (from 374/975; 38%) it was clear that questions about mode of conception can be offensive to some parents of twins, unless there is a need to know. Further, the question ‘are your twins natural?’ should be avoided. We believe the question ‘Did you need medical help to conceive your twins’, followed up with specific questions, is more acceptable.