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STUDY QUESTION: Are childhood measures of phenotype associated with peri-conception parental, IVF treatment and/or embryonic characteristics of IVF children? SUMMARY ANSWER: Birthweight, childhood body mass index (BMI) and height of pre-pubertal IVF children were strongly associated with peri-conception factors, including follicular and embryonic characteristics. WHAT IS KNOWN ALREADY: A growing number of studies have identified a range of phenotypic differences between IVF and naturally conceived pre-pubertal children; for example, birthweights are lower following a fresh compared with a thawed embryo transfer. STUDY DESIGN, SIZE, DURATION: This retrospective cohort study included IVF children (n = 96) born at term (>37 weeks) after a singleton pregnancy from the transfer of either fresh or thawed embryos in New Zealand. Between March 2004 and November 2008, these children were subjected to clinical assessment before puberty. PARTICIPANTS/MATERIALS, SETTING, METHODS: Clinical assessment provided anthropometric measures of children aged 3.5-11 years old. Peri-conception factors (n = 36) derived retrospectively from parental, treatment, laboratory and embryonic variables (n = 69) were analysed using multiple stepwise regression with respect to standard deviation scores (SDSs) of the birthweight, mid-parental corrected BMI and height of the IVF children. Data from children conceived from fresh (n = 60) or thawed (n = 36) embryos, that met inclusion criteria and had high-quality data with >90% completeness, were analysed. MAIN RESULTS AND THE ROLE OF CHANCE: Embryo treatment at transfer was identified as a predictor of birthweight with thawed embryos resulting in heavier birthweights than fresh embryos [P = 0.02, 95% confidence interval (CI) fresh minus thawed: -1.047 to -0.006]. Birthweight SDS was positively associated with mid-parental corrected BMI SDS (P = 0.003, slope 0.339 ± 0.100). Four factors were related (P < 0.05) to mid-parental corrected height SDS. In particular, child height was inversely associated with the diameter of lead follicles at oocyte retrieval (P < 0.0001, slope -0.144 ± 0.040) and with the quality score of embryos at transfer (P = 0.0008, slope -0.425 ± 0.157), and directly associated with the number of follicles retrieved (P = 0.05, slope 1.011 ± 0.497). Child height was also positively associated with the transfer of a fresh as opposed to thawed embryo (P < 0.001, 95% CI 0.275-0.750). LIMITATIONS, REASONS FOR CAUTION: More than one embryo was transferred in most cycles so mean development and quality data were used. The large number of variables measured was on a relatively small sample size. Large cohorts from multiple clinics using a variety of treatment protocols and embryology methods are needed to confirm the associations identified and ultimately to test these factors as possible predictors of phenotype. WIDER IMPLICATIONS OF THE FINDINGS: This is the first study to directly associate peri-conception measures of IVF treatment with a pre-pubertal child's phenotype. Demonstration that peri-conception measures relate to a pre-pubertal child's phenotype extends the range of factors that may influence growth and development. These findings, if corroborated by larger studies, would provide invaluable information for practitioners, who may want to consider the impact of ovarian stimulation protocols as well as the quality of the embryo transferred on a child's growth and development, in addition to their impact on pregnancy rate. STUDY FUNDING/COMPETING INTERESTS: This work was supported by grants from the National Research Centre of Growth and Development New Zealand (grant 3682065) and the Australasian Paediatric Endocrine Group (APEG; grant 3621994), as well as a fellowship from Fertility Associates New Zealand awarded to M.P.G. In terms of competing interest, J.C.P is a shareholder of Fertility Associates. M.P.G. currently holds the position of Merck Serono Lecturer in Reproductive Biology. W.S.C. and P.L.H. have also received grants and non-financial support from Novo Nordisk, as well as personal fees from Pfizer that are unrelated to the current study. The other authors have no conflict of interest to declare.

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An anonymous questionnaire was circulated to parents of young children to survey exposure to publicity about and attitudes to sperm donation (n = 192 couples; 50% response rate). Of those surveyed, 55% of men and women had seen or heard of the need for sperm donors; for 26% of couples, one or both partners had considered donation, and among half of those (13% of total) no objections to donating were raised. However, only two men had approached the clinic as donors. Objections to donation centred mainly on discomfort at having children outside their family, worry about future contact with donor insemination children, and worry about incest. Women were three times as likely to raise these objections as men. Of all respondents, 20% thought a donor should be used for only one recipient couple, while 50% suggested three or fewer families. Overall, 41% rated having non-identifying information about the recipient couple(s) as an important aspect of sperm donation, although only 9% of men and 17% of women considered personal contact important. While many people are aware of and receptive to the idea of being donors, very few act. Many would prefer that their spermatozoa were used for only one or a few couples about whom they had some information.

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Fifty-three couples embarking on donor insemination (DI) replied to an anonymous questionnaire asking what they wanted to know about the donor should they become pregnant. Fifty-one percent were definitely and 32% probably going to tell a child of its donor origins. Items most frequently listed for themselves and on behalf of a child were--interests/sports (57%), physical attributes (41%), occupation (37%) and family background (26%). Forty-two percent of the women and 28% of the men thought the child should have access to the identity of the donor eventually. Thirty-eight donors were surveyed by a questionnaire; 68% were agreeable to their identity being available to a donor insemination child when the child reached maturity. When the category of identifiable donor was created, 20 of 36 (56%) new donors and donors still donating chose this option.

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Serum IgG and IgA antibodies to Chlamydia trachomatis were measured in 102 in vitro fertilisation (IVF) patients with tubal disease and in 102 infertility patients without tubal disease to assess the impact of chlamydial infection on the demand for IVF treatment in New Zealand. Sixty-five percent of the IVF patients had IgG titres greater than or equal to 1:64, compared to 24% of the controls; 29% of the IVF patients had IgA titres greater than or equal to 1:32 compared to 0% of the controls. The proportion of IVF patients with IgG antibodies increased from 39% for those from socioeconomic group 1 to 79% of those from groups 4 to 6. The difference in prevalence of IgG antibodies between the tubal and nontubal patients suggests that about 40% of the tubal infertility currently being treated by IVF has chlamydia as a causal agent. The presence of IgG or IgA antibodies did not seem to affect the chance of pregnancy in the IVF programme, nor the chance of subsequent miscarriage.

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Reproductive and social histories of the first 100 patients attending the in vitro fertilisation (IVF) programme at National Women's Hospital, Auckland, have been studied. The average age at first treatment was 31.6 (SD 3.9) for women, and 34.2 (4.6) for men. The couples had been married 7.6 (3.3) years and had experienced 6.7 (3.2) years infertility. It was a second marriage for 16. The husbands had on average a higher social classing than the population (class: number [population]--1:16 (7%), 2:18 (14%), 3:42 (28%), 4:19 (29%), 5:4 (14%), 6:1 (8%); but this bias diminished in the next 159 couples. Nine couples withdrew before their quota of cycles, 7 from stress. Tubal disease was the cause of infertility in 93, but in 69 its origin was untraceable. In 22 it could be attributed to pelvic inflammatory disease (eight associated with IUCDs and 2 with sexually transmitted disease) and in two to sterilisation. Although 59 women had a history of having conceived, only 34 were parous, and only 11 had a child of the current union.

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Oxytocin-like immunoreactivity, estradiol, and progesterone were measured in follicular fluid collected during oocyte collection in an in vitro fertilization program in which clomiphene citrate was used to stimulate follicular development. Follicles which yielded morphologically normal embryos after fertilization of the oocyte had oxytocin concentrations ranging from less than 10 to 600 ng/liter. Oxytocin concentrations did not differ between follicles from 12 pregnancy cycles (median, 169; N = 21) and follicles from 12 nonpregnancy cycles (median, 110; N = 18). Oxytocin concentrations were correlated negatively with progesterone concentrations (Spearman's rank correlation coefficient r = -0.50; P = 0.001). In cycles with some follicles having progesterone concentrations less than 10 and some greater than 10 mumol/liter, oxytocin concentrations were higher in the less progestogenic follicles in 15 of 16 cases.

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Prepubertal ewe lambs were treated with empty or filled melatonin implants. The implants were placed s.c. at birth and pituitary responsiveness to various doses of LHRH, LH/FSH pulsatility and prolactin and melatonin secretion were examined at 10, 19, 28, 36 and 45 weeks of age. Control animals (N = 10) showed no consistent alteration in pituitary responsiveness to LHRH during development. Ewes treated with melatonin (N = 10) had puberty onset delayed by 4 weeks (P less than 0.03) but no effect of melatonin on LH or FSH response to LHRH injection was observed at any stage of development. In the control and melatonin-treated ewe lambs the responses to LHRH injection were lower during darkness than during the day at all stages of development. No consistent differences in LH or FSH pulsatility were observed between treatment groups or during development. Prolactin concentrations, however, failed to decrease at the time of puberty (autumn) in the melatonin-treated group. Melatonin-treated ewe lambs maintained normal rhythmic melatonin production which was superimposed on a higher basal concentration and showed the same increase in melatonin output with age as the control ewes. These results indicate that the delayed puberty caused by melatonin implants is not due to decreased pituitary responsiveness to LHRH or to dramatic changes in basal LH or FSH secretion.

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In preliminary experiments, pH measurements were made with pH paper and with a pH electrode on cervical mucus in situ and after aspiration. Paper and electrode gave comparable results with aspirated mucus. Measurements made on mucus in situ and then after aspiration were concordant only when pH paper, but not the pH electrode, was used to measure pH in situ. All spermatozoa were immotile in the post-coital test when the pH of the mucus was below 6. A pH below 6 was found in 38% (23 out of 61) of post-coital tests in which there were fewer than 1% of motile spermatozoa (19% of all tests). The male partners of women who had a mucus pH greater than 6 but had less than 1% of spermatozoa motile in the post-coital test, had slightly poorer quality semen than the partners of women with mucus pH less than 6, or in which there were motile spermatozoa in the mucus.

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Antisera were raised in rabbits against Pro-Leu-Gly-NH2 (PLG)-bovine thyroglobulin conjugates. Specificity of the antisera towards oxytocin was evaluated by a radioimmunoassay as well as by immunohistochemical procedures on hypothalamic tissue and hormone bound to Sepharose beads. It was concluded that immunization with PLG can raise antisera that are specific to oxytocin.

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The pregnancy rates after artificial insemination by donor semen (AID) have been compared with pregnancy rates in normal fertile women to assess the efficiency of AID. To do this, the curve y = a(1-(1-b)x) was fitted to life-tabled cumulative pregnancy rates. The equation describes a model in which the parameter a is the proportion of women who are potentially fertile under the conditions of treatment, and in which the parameter b is the pregnancy rate per cycle (or fecundity) of these fertile women. For 259 AID patients with no previous pregnancy a was 65% while for 57 AID patients with a previous pregnancy after AID 'a' was 99.9%. The values of b were similar for the two groups of patients, being 20% and 22% respectively. Women without fertility problems who had become pregnant after discontinuing oral contraception provided the reference group. Since only pregnant women were selected, a was 100% by definition. The values of b for the reference group were 22% for 100 primigravid women and 20% for 100 multigravid women. Only 65% of the AID patients were potentially fertile with AID, but those that were fertile became pregnant at the same rate as normal women who discontinued oral contraception.

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Semen quality and karyotype were screened in all men offering to be donors for an artificial insemination (AID) program. The criteria for accepting or rejecting semen have now been set with respect to this sample of the population. There was no evidence of differences between the pregnancy rates of the accepted donors. One of 172 potential donors with a clear medical history had a chromosomal abnormality, 4 had pericentric inversions of chromosome 9, and 14 had other heterochromatic variants. Of the recipients of AID, 5 of 196 women had chromosomal abnormalities, and 12 had heterochromatic variants.

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Three media for human semen were tested--(1) HAMS F10 supplemented with fructose, glycine and glycerol (MHAMS), (2) another chemically defined medium, but specifically designed for human semen (HSPM) and (3) medium containing egg yolk (EYCM). Twenty ejaculates of donor quality were divided into three aliquots, each aliquot was mixed with a different medium, the solutions were vapour frozen, and stored at -196 degrees C. Motility and quality of motility were measured 0.5 h and 3 h after thawing. Average cryosurvivals 0.5 h after thawing for MHAMS, HSPM and EYCM were 44.1 (s.d. = 17.0)%, 56.0 (s.d. = 15.3)% and 66.0 (s.d. = 10.8)%, respectively; the proportions of ejaculates with cryosurvival below 50% were 11/20, 5/20 and 1/20 respectively. The results indicated that the EYCM medium had clear advantages over other cryoprotective solutions for use in a spermatozoa freezing programme.

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The rise in concentrations of FSH and LH in serum seen 24 h after castration was suppressed by the administration of an extract of bull seminal plasma or testosterone propionate at the time of castration. Whereas testosterone propionate preferentially suppressed LH, the seminal plasma extract suppressed FSH and LH equally. Small doses of bull seminal plasma extract and testosterone, that had little effect separately, acted synergistically to supress levels of FSH and LH to those found in intact animals, while combinations of larger doses had little further effect. This selective interaction suggests how inhibin and testosterone might together regulate concentrations of FSH and LH in the blood of the male rat.

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Bull seminal plasma administered to male rats at the time of castration inhibited the rise in the levels of FSH and LH otherwise seen in the serum 24 h later. That the gonadotrophin-inhibiting activity was also present in the seminal plasma from vasectomized bulls suggests that it was not of testicular origin. Although the substance with gonadotrophin-inhibiting activity was a protein, it may be chemically distinct from inhibin.

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Chicken posterior pituitary tissue was partially fractionated by extraction with 0.1 M HCl followed by gel exclusion chromatography. Two fractions, B and C, bound oxytocin. Two components of fraction B and at least 1 component of fraction C had properties characteristic of mammalian neurophysins: they appeared to be synthesized in the hypothalamus, were depleted upon osmotic stress, were rich in residues of cysteine and the 2 components of B bound to [8-lysinel]-vasopressin coupled to Sepharose.

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