Attitudes of anonymous and identity-release oocyte donors towards future contact with donor offspring.

STUDY QUESTION: What are the attitudes and expectations of past oocyte donors concerning contact with their donor offspring and contact between donor offspring and their own children? SUMMARY ANSWER: The large majority (95%) of open-identity oocyte donors, as well as voluntarily registered donors (registered before the Finnish 2007 ART law), expressed positive or neutral feelings towards contact with their donor offspring and mainly positive expectations towards contact between donor offspring and their own children. WHAT IS KNOWN ALREADY: Although there is a growing support for openness and identity-release programmes in gamete donation, there is not much knowledge on how donors feel about potential contact with their offspring. Most previous studies have investigated donor expectations with a relatively short follow-up time, using small samples or participants in voluntary donor linkage services. STUDY DESIGN, SIZE, DURATION: A retrospective cross-sectional survey of all women who had donated oocytes between 1990 and 2012 at three fertility clinics in Finland was carried out in 2013. A self-administered questionnaire was sent out to a total of 569 former oocyte donors. PARTICIPANTS/MATERIALS, SETTING, METHODS: In total, 428 former oocyte donors answered a questionnaire assessing experiences and attitudes related to donation (response rate 75.2%). In this study, 358 donors who were unknown to the recipient were included. The mean follow-up time after the donation was 11.2 years. Before 2008, donors were non-identifiable but could voluntarily consent to release their identifying information to their donor offspring. After 2008, persons born as a result of gamete donation can, from the age of 18, receive information identifying the donor. Altogether 290 respondents had participated in a donation programme in 1990-2007 (before the Finnish ART-law), and 68 participated after the enactment of the ART-law, enabling us to compare attitudes by type of legislation during donation. MAIN RESULTS AND THE ROLE OF CHANCE: Most voluntarily registered and open-identity donors welcomed or were neutral to potential contact with their donor offspring but were slightly more cautious towards contact between their own children and a donor-conceived child. Open-end comments revealed some ambiguity and uncertainty as to what to expect from such contact and feelings varied from neutral curiosity and interest to desire to meet the donor-conceived child. LIMITATIONS, REASON FOR CAUTION: It is not possible to assess whether the opinions of the study participants is representative of all donors in 1990-2012, as 25% of all contacted former donors did not respond to the survey. WIDER IMPLICATIONS OF THE FINDINGS: This study is one of only a few studies among oocyte donors to evaluate long-term psychosocial consequences of the donation and expectations towards contact with donor offspring, using a large sample. Results from this study show that persisting concerns about adverse outcomes of identity release policies are largely unwarranted, but there is a need to develop counselling practices and material for identity-release donors about how to prepare for and adjust to potential contact with donor offspring. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by grants from the Medical Society Life and Health, and from the Otto A. Malm Foundation. The authors have no competing interests to report. TRIAL REGISTRATION NUMBER: N/A.

Attitudes and disclosure decisions of Finnish parents with children conceived using donor sperm.

STUDY QUESTION: What are disclosure intentions and experiences of heterosexual parents with children born after assisted donor insemination (DI) or IVF with donor sperm (dIVF)? SUMMARY ANSWER: Only 16.5% of Finnish DI/dIVF heterosexual parents had told their child of his/her origin; 18% of all children above 3 years of age had received the information. Parents with older children were more unwilling to tell or were more uncertain regarding what to do than parents with younger children. WHAT IS KNOWN ALREADY: In general, 10-35% of parents of DI offspring are willing to inform their child about their conception. Men who need donor sperm to become a father are more secretive than women who need donated oocytes and are less willing to participate in counselling about parenthood. In the past, couples conceiving through gamete donation were adviced to maintain secrecy or sufficient advice on information sharing was not available. Evidence suggests that parental attitudes are moving towards greater openness. In 2007, Finland enacted a law on assisted fertility treatments (1237/2006) stating that gamete donors have to register their identifying information in a registry so that at the age of 18 years, offspring can obtain information about their donor. STUDY DESIGN, SIZE AND DURATION: This retrospective questionnaire study included 139 mothers and 127 fathers with altogether 240 children born after DI or dIVF during 1992-2007. PARTICIPANTS, SETTING AND METHODS: Questionnaires were sent to heterosexual couples who had undergone DI/dIVF treatment at the Väestöliitto Fertility Clinic in Helsinki resulting in live birth (n = 277, 252 mothers and 239 fathers). The parents were asked to report their disclosure intentions towards the child and towards other people about the decision to use donated sperm, their concerns about donor characteristics, their evaluation of the counselling that they received and their views about the current Finnish assisted reproduction law. MAIN RESULTS AND THE ROLE OF CHANCE: The response rate was 55% (139/252) among the mothers and 53% (127/239) among the fathers. Answers provided information on 58% (240/415) of the children born, 91% of whom were at least 3 years old at the time. Of all parents, 16.5% reported that they had already told their child of his/her conception. Of all 240 children, 16.3% had already received information about their conception. The children had been between 3 and 14 years of age (mean 6.8 years) when they were told. Parents of older children were significantly more unwilling to tell their child than parents of younger children (P < 0.005). No difference in disclosure between DI and dIVF emerged. Less than half of the parents (42%) had been satisfied with the psychological support offered to them, with parents of older children having been most dissatisfied. LIMITATIONS, REASONS FOR CAUTION: Although the response rate was relatively high, more than 40% of the parents chose not to participate. As has been shown before, it is likely that those who do not take part are less inclined to disclose and this should be taken into consideration when conclusions are drawn. WIDER IMPLICATIONS OF THE FINDINGS: Our results on disclosure rates are in agreement with previous studies. General attitudes have moved towards greater openness about the use of donated gametes. Furthermore, the availability of psychological counselling before treatment has increased the understanding of the importance of disclosure. People who have become parents after DI or dIVF should also be offered counselling after the child has been born. STUDY FUNDING/COMPETING INTERESTS: This study was supported by grants from the Wilhelm and Else Stockmann Foundation and the Medical Society Life and Health. There are no competing interests to disclose. TRIAL REGISTRATION NUMBER: None.

Does long in vitro culture promote large for gestational age babies?

STUDY QUESTION: Does the length of time during which embryos are cultured in vitro affect the birthweight of the infants? SUMMARY ANSWER: The duration of the embryo culture period is a significant factor in determining the birthweight of the infants. WHAT IS ALREADY KNOWN: IVF children show adverse perinatal outcome when compared with the general population and increased incidence of preterm birth and low birthweight is commonly observed. STUDY DESIGN, SIZE, DURATION: A retrospective cross-sectional cohort study including 1079 infants born after treatment at the Family Federation of Finland Fertility Clinic in Helsinki, between 2000 and 2010. PARTICIPANTS, SETTING, METHODS: All singleton IVF children were included. The gestation- and gender-adjusted birthweights of the babies were analyzed according to mother's age, BMI, and parity, type of treatment (IVF or ICSI), main cause of infertility and embryo culture period. Two outcomes were investigated: the birthweight and the proportion of small and large for gestational age (SGA and LGA) infants. Multiple linear regression analysis was performed to show the significance of each individual factor on determining the birthweight of the babies born. MAIN RESULTS AND THE ROLE OF CHANCE: In the study group as a whole, the distribution of the SGA and LGA babies showed no deviation from the growth charts of the general population. However, when the birthweight of the children was analyzed according to the length of embryo culture from Day 2 to Days 5-6, an increase in the proportion of LGA babies was found (D2 9.4%, D3 11.5%, D5-6 18.8%; D2 n = 871, D3 n = 139, D5-6 n = 69). Multiple linear regression analysis showed that BMI (P < 0.001) and parity (P < 0.001) of the mother, as well as the embryo culture period (P = 0.007) had a significant effect on the birthweight. The value of the adjusted R(2) was 0.437. LIMITATIONS, REASONS FOR CAUTION: Small number of D5-6 infants and a lack of pregnancy-associated factors contributing to birthweight. WIDER IMPLICATIONS OF THE FINDING: This study warrants larger studies to analyze the birthweight of the IVF children, particularly after blastocyst culture. STUDY FUNDING/COMPETING INTEREST: The study was funded by the Family Federation of Finland, Fertility Clinic Helsinki. No competing interests.

Elective single-embryo transfer in women aged 40-44 years.

STUDY QUESTION: Is an elective single-embryo transfer (eSET) policy feasible for women aged 40 or older? SUMMARY ANSWER: For older women (aged 40-44 years) with a good prognosis, an eSET policy can be applied with acceptable cumulative clinical pregnancy rates and live birth rates. WHAT IS KNOWN ALREADY: Various studies have shown the effectiveness of eSET in women aged <35 years with high cumulative pregnancy rates and low rates of multiple births. STUDY DESIGN, SIZE, DURATION: This retrospective cohort study included 628 women treated between 2000 and 2009. PARTICIPANTS, SETTING, METHODS: Women aged 40-44 years underwent a fresh cycle of IVF or ICSI treatment with eSET (n = 264) or double-embryo transfer (DET) (n = 364). In the subsequent frozen-thawed embryo transfer cycles, SET/DET was performed in both groups according to the number of embryos available and the opinion of the couple. The study was performed at the Family Federation of Finland Helsinki Fertility Clinic. MAIN RESULTS AND THE ROLE OF CHANCE: In the fresh cycles, the clinical pregnancy rates were 23.5 and 19.5% in the eSET and DET groups, respectively, and live birth rates were 13.6 and 11.0%, respectively. In the fresh cycles with eSET, there were no twin pregnancies, but in the DET group, there were three sets of twins (7.5%). The cumulative clinical pregnancy rates per oocyte retrieval were 37.1 and 24.2% in the eSET and DET groups, respectively (P < 0.001), and the cumulative live birth rates were 22.7 and 13.2%, respectively (P = 0.002). Cumulative twin rates were 6.7% (n = 4) in the eSET group and 8.3% (n = 4) in the DET group (P = 0.726). All of the twin pregnancies in the eSET group resulted from frozen and thawed DET embryo transfer cycles. LIMITATIONS: The characteristics of the two patients groups are not comparable because the suitability of eSET was individually assessed by a clinician based on both clinical prognostic factors and the outcome of IVF or ICSI, i.e. the number and quality of embryos. WIDER IMPLICATIONS OF THE FINDINGS: This study may be generalized to IVF units having experience in eSET and cryopreservation.

Increasing openness in oocyte donation families regarding disclosure over 15 years.

BACKGROUND: Worldwide there is an increasing number of families created by oocyte donation (OD). The aim of this study was to gather information about parents' plans of disclosure to their child and to other people, as well as parents' attitudes and level of satisfaction up to 15 years after their OD treatment. METHODS: A questionnaire with separate material for each partner was sent to all parents (167 mothers, 163 fathers) who had had a child after treatment with donated oocytes at Väestöliitto Fertility Clinics in Helsinki during 1992-2006. These parents had a total of 231 children aged 1-14 years. Parents were asked if they had told or intended to tell their child about his/her origin and how and when they had done so and about the reasons to disclose or not. Other questions were about openness towards other people, concerns about donor characteristics, counselling and feelings towards the child. RESULTS: Of the mothers, 61.1%, and of the fathers, 60.0%, had told or intended to tell the child of his/her conception. Of children over 3 years of age, 26% had already been informed. There was a statistically significant difference between parental telling in different age groups of children (P = 0.011, χ(2)). In the youngest age group (1-3 years), 83.3% of parents were inclined to disclosure compared with 44.4% in the oldest age group (13-14 years). A high proportion of mothers (86.7%) and fathers (71.0%) had told other people about the nature of their child's conception. The majority of parents did not have much concern about the characteristics of the donor. A higher proportion of the mothers (24%) compared with fathers (11%) thought that the psychological support had been insufficient. They thought that discussions with health professionals should be arranged routinely after delivery or when it was time to inform the child. CONCLUSIONS: Parents with young OD children are clearly more inclined to disclosure compared with parents with older children.

Perinatal outcome of children born after frozen and fresh embryo transfer: the Finnish cohort study 1995-2006.

BACKGROUND: The number of children born after frozen embryo transfer (FET) is steadily rising. However, studies on obstetric and perinatal outcomes are limited. Our primary aim was to compare the perinatal health of children born after FET and fresh embryo transfer, and to use data from children born after spontaneous conception as a reference. METHODS: In a register-based cohort study we evaluated the obstetric and perinatal outcomes of children born after FET (n = 2293), fresh embryo transfer (n = 4151) and those born after spontaneous pregnancy (reference group; n = 31 946). Data were collected from the registers of two infertility outpatient clinics, two university hospitals and the Finnish Medical Birth Register (1995-2006). RESULTS: After adjusting for confounding factors the FET group showed decreased risks of preterm birth [adjusted odd ratio (AOR) 0.83, 95% confidence interval (CI) 0.71-0.97], low birthweight (AOR 0.74; 0.62-0.88) and being small for gestational age (AOR 0.63; 0.49-0.83) compared with the fresh embryo transfer group. Mean birthweight was 134 g higher in the FET singletons versus the fresh embryo transfer singletons (P< 0.0001). When FET singletons were compared with the reference group, increased risks of preterm birth (AOR 1.45; 1.25-1.68) and low birthweight (AOR 1.22; 1.03-1.45) and a decreased risk of being small for gestational age (AOR 0.71; 0.54-0.92) were found. No excess of perinatal and infant mortality occurred between the groups. CONCLUSIONS: Embryo freezing does not adversely affect perinatal outcome in terms of prematurity, low birthweight and being small for gestational age versus the fresh embryo transfer and the outcome is similar or even better, particularly regarding fetal growth. Our study, which is one of the largest on FET pregnancies, provides further evidence on the safety of FET.

Cell death and its suppression in human ovarian tissue culture.

In women with premature ovarian failure, fertility may be preserved by ovarian tissue culture in vitro. However, techniques for tissue culture and follicle maturation have remained suboptimal. Our aim was to characterize ovarian tissue degeneration in cultures and to establish a model for cell death research in cultured ovarian tissue. Precise knowledge on the process resulting in cell death in cultured ovarian tissue will ultimately facilitate work aimed at improving long-term culture conditions. Ovarian tissue apoptosis was studied in a serum-free culture model in which nuclear DNA fragmentation was shown to occur within 24 h of the start of the culture. Activation of caspase-3 was detected in some stromal cells and a few oocytes. Since not all of the tissue exhibited signs of apoptosis and since DNA fragmentation increased over time, the tissue probably gradually dies by apoptosis. The antioxidant N-acetyl-L-cysteine (NAC; 25, 50 and 100 mmol/l) was found to inhibit this apoptosis. Thus, apoptosis appears to play a critical role in the degeneration of human ovarian cortical tissue cultures, and this cell death can be suppressed by NAC. The present tissue culture model can be used for identifying components capable of inhibiting cell death in vitro.

The predictive value of pronuclear morphology of zygotes in the assessment of human embryo quality.

BACKGROUND: Recent studies have shown that zygote morphology could be used for the assessment of human embryo quality. Pronuclear (PN) morphology is based on certain distinct features seen in zygotes 16-18 h after fertilization. In the present study PN stage morphology was assessed and combined with a single embryo transfer in order to investigate whether currently used zygote classifications are able to predict embryo quality and implantation rates. METHODS AND RESULTS: Zygotes were analysed according to two different classification systems. In the first, a total of 764 zygotes was analysed according to the degree of polarization of nucleolar precursor bodies (NPB). Zygotes with unpolarized PN (i.e. scattered localization of NPB) showed significantly slower (P < 0.005) cleavage rates (38.9%) than zygotes having at least one pronucleus polarized (57.3% and 54%). However, there was no difference in the pregnancy rate in 105 single embryo transfers between the groups. The appearance of a cytoplasmic halo was related to embryo morphology. Embryos derived from halo-positive zygotes had significantly better (P < 0.05) morphology (60.9%) compared to halo-negative derived embryos (52.2%), but in terms of pregnancy rates no difference was found. A total of 1520 zygotes was analysed according to a second classification system, which was based on the number and distribution of NPB. In the comparative analysis, none of the six different classes produced superior quality embryos or higher pregnancy rates in 144 single embryo transfers. CONCLUSIONS: Our results indicate that there are no significant differences in embryo quality or implantation/pregnancy rates between proposed zygote classes.

Y-chromosomal microdeletions among infertile Finnish men.

BACKGROUND: Microdeletions in the Y-chromosome are known to cause a significant proportion of azoo- and oligozoospermia in men. The reported frequency of deletions varies greatly between the studies. Probable reasons for this variation are different selection criteria and number of patients included, and possibly also methodological aspects, whereas the contribution of environmental and genetic factors is not known. The aim of this study was to determine the incidence of Y-chromosome microdeletions among infertile Finnish men. METHODS: Two hundred and one men showing azoospermia (n=68) or severe oligozoospermia (n=133) were included. Multiplex polymerase chain reaction method was used to amplify specific sequence tagged sites (STS) along the Y chromosome. RESULTS: Microdeletions were observed in 18 men (9%), of whom four were azoospermic and 14 oligozoospermic. CONCLUSIONS: The incidence of Y-deletions in the study population of infertile Finnish men falls within the range published in other countries.

Luteal phase start of low-dose FSH priming of follicles results in an efficient recovery, maturation and fertilization of immature human oocytes.

In this prospective study we investigated whether the maturation and fertilization of immature oocytes can be improved by administration of recombinant follicle stimulating hormone (rFSH) starting in the late luteal phase in two groups of women: group 1 (n = 6) women with regular menstrual cycles; and group 2 (n = 6) women with irregular cycles and polycystic ovaries (PCO) on ultrasound examination. Low-dose (37.5 IU) rFSH was commenced 11 days after LH surge during a spontaneous menstrual cycle and on the ninth day of progesterone administration in an irregular cycle. Recombinant FSH was continued until the leading follicle was approximately 10 mm in diameter. The oocytes were retrieved after withdrawing rFSH for 2-5 days. In total, 136 oocytes were recovered (group 1, 67 oocytes; group 2, 69 oocytes). Nine of the oocytes from PCO women were atretic at retrieval. Oocytes complete with cumulus cells were cultured for 44 h in complex tissue culture medium supplemented with gonadotrophins and fetal calf serum. After maturation, the cumulus cells were removed and metaphase II oocytes were injected with spermatozoa. Respectively, the oocyte maturation and fertilization rates were 64 and 72% in group 1, and 78 and 57% in group 2 (not significant). After fertilization, the zygotes (group 1, n = 22; group 2, n = 11) and cleavage stage embryos (group 1, n = 9; group 2, n = 15) were frozen in propanediol. All women except one (11/12) had approximately five zygotes or cleaved embryos frozen. The viability of in-vitro matured frozen-thawed embryos was generally poorer than that (81%) seen after conventional intracytoplasmic sperm injection, with 61% survival in group 1 and 23% in group 2. Fifteen embryo transfers resulted in one miscarriage at 6 weeks gestation. The late luteal start of low-dose rFSH yielded a good number of immature oocytes in women with both regular and irregular cycles. Two out of three of these oocytes matured and fertilized. However, cryosurvival of the zygotes and cleaved embryos was unsatisfactory and thus cryopreservation of in-vitro matured embryos may not be an optimal procedure.

Comparison of two recombinant follicle-stimulating hormone preparations in in-vitro fertilization: a randomized clinical study.

A randomized comparison of two recombinant human follicle-stimulating hormone (recFSH) preparations (Gonal-F and Puregon) in ovarian stimulation for in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) was carried out at the Infertility Clinic of the Family Federation of Finland. A total of 348 women (aged 22-43 years) suffering from infertility due to miscellaneous causes was recruited. Of these, 344 underwent stimulation using equal starting doses (150 IU/day: Gonal-F n = 164, Puregon n = 158 or 300 IU/day: Gonal-F n = 8, Puregon n = 14) after down-regulation with intranasal buserelin from the mid-luteal phase. Similar clinical pregnancy rates were achieved with both preparations; 33.5% per cycle and 37.4% per embryo transfer (24.5% one-embryo and 75.5% two-embryo transfers, n = 147) with Gonal-F (150 IU/day) and 32.9% per cycle and 36.4% per embryo transfer (30.1% one-embryo and 69.9% two-embryo transfers, n = 145) with Puregon (150 IU/day). The ongoing cumulative pregnancy rates after frozen-thawed embryo transfer were 35.4% with Gonal-F and 37.7% with Puregon. Six cycles were cancelled because of a low response (three in each group). Similar numbers of oocytes were obtained in both groups; 13.0 with 150 IU/day and 6.1 with 300 IU/day Gonal-F, and 12.4 with 150 IU/day and 7.1 with 300 IU/day Puregon. The fertilization and cleavage rates and the incidence of moderate or severe ovarian hyperstimulation syndrome (Gonal-F, 2.0% and Puregon, 0.7%) were also similar. Gonal-F and Puregon were equally and highly effective in stimulation for IVF and ICSI.

Successful pregnancy after in-vitro fertilization and transmyometrial embryo transfer in a patient with congenital atresia of cervix: case report.

A case report of a patient with congenital cervical atresia diagnosed at the age of 24 years is given. The attempts to create a neocervix were unsuccessful. Since no signs of retrograde menstruation or haematometra were observed, in agreement with the patient hysterectomy was not performed. At the age of 32 years, a successful pregnancy was achieved after an in-vitro fertilization and transmyometrial embryo transfer. Due to rapidly progressing pre-eclampsia, an elective Caesarean section was performed at 32 weeks gestation. A 1610 g healthy male infant in breech presentation was born. The post-partum period was uneventful.

Recruitment of follicles by recombinant human follicle-stimulating hormone commencing in the luteal phase of the ovarian cycle.

OBJECTIVE: To investigate, in patients who previously had a suboptimal ovarian stimulation cycle, the benefit of starting ovarian stimulation before the onset of menses. DESIGN: Prospective, randomized, controlled study. SETTING: A tertiary referral center for infertility treatment. PATIENT(S): Forty patients undergoing IVF or GIFT from whom only 3-6 oocytes were retrieved in their last cycle. INTERVENTION(S): Recombinant human FSH was administered before the onset of the menstrual period (experimental group) or in the early follicular phase after the onset of menses (control group). MAIN OUTCOME MEASURE(S): The number of oocytes retrieved. RESULT(S): Patients in the experimental group were ready for oocyte retrieval on menstrual cycle day 11 instead of cycle day 14. The number of oocytes retrieved was not significantly different between the two groups. CONCLUSION(S): Poor responders do not benefit from commencing recombinant human FSH therapy in the luteal phase.

Blastocyst development and birth after in-vitro maturation of human primary oocytes, intracytoplasmic sperm injection and assisted hatching.

Immature oocyte recovery followed by in-vitro oocyte maturation and in-vitro fertilization is a promising new technology for the treatment of human infertility. The technology is attractive to potential oocyte donors and infertile couples because of its reduced treatment intervention. Immature oocytes were recovered by ultrasound-guided transvaginal follicular aspiration. Oocytes were matured in vitro for 36-48 h followed by intracytoplasmic sperm injection (ICSI). Embryos were cultured in vitro for 3 or 5 days before replacement. Assisted hatching was performed on a day 5 blastocyst stage embryo. Embryo and uterine synchrony were potentially enhanced by luteinization of the dominant follicle at the time of immature oocyte recovery. Mature oocyte and embryo production from immature oocyte recovery were similar to the previous IVF results of the patients. A blastocyst stage embryo, produced as a result of in-vitro maturation, ICSI, in-vitro culture and assisted hatching, resulted in the birth of a healthy baby girl at 39 weeks of gestation.

Urinary vascular endothelial growth factor concentrations in women undergoing gonadotrophin treatment.

A recently identified cytokine, vascular endothelial growth factor (VEGF, vascular permeability factor) has been implicated in ovarian hyperstimulation syndrome in women undergoing assisted reproduction. We postulate that circulating and urinary VEGF values increase following gonadotrophin stimulation, in parallel with the increased ovarian vascularity. A VEGF radioimmunoassay was developed using iodinated VEGF as tracer, a goat anti-VEGF serum as antiserum and recombinant human VEGF as standard. The specificity of the assay was confirmed by comparing the reverse phase high-performance liquid chromatography (HPLC) pattern of VEGF immunoactivity in urine and urine spiked with recombinant VEGF. Urine was concentrated 5-fold prior to measurement by the radioimmunoassay. VEGF:creatinine ratios in early morning urine samples were used to monitor daily urinary VEGF concentrations based on its high correlation (r = 0.77, P < 0.001) with VEGF concentrations in 24 h urine collections. No diurnal variation in VEGF:creatinine ratios was detected. VEGF:creatinine ratios were determined daily from nine women undergoing gonadotrophin-releasing hormone (GnRH) agonist/gonadotrophin treatment. In a further 16 women, early morning urine samples were collected in the peri-ovulatory period. A significant increase (P < 0.005, n = 25) was observed in VEGF:creatinine ratios following human chorionic gonadotrophin (HCG) administration. VEGF:creatinine ratios correlated poorly (r < 0.34) with plasma oestradiol, follicle number and size. It is concluded that urinary VEGF/creatinine ratios increase following HCG stimulation.

Ultrasonographic appearance of polycystic ovaries is associated with exaggerated ovarian androgen and oestradiol responses to gonadotrophin-releasing hormone agonist in women undergoing assisted reproduction treatment.

While no single biochemical test is diagnostic of polycystic ovary syndrome (PCOS), most patients show a characteristic ovarian ultrasonographic appearance. It has been proposed that a dysfunction of cytochrome P-450c17 alpha in PCOS leads to an increased 17-hydroxyprogesterone (17-OHP) response to a gonadotrophin-releasing hormone (GnRH) agonist-induced gonadotrophin rise. We postulated that this abnormality of steroid metabolism might influence the ovarian response during assisted reproduction treatment. We investigated 106 patients undergoing a short 'boost' stimulation regimen for assisted reproduction treatment, including in-vitro fertilization and gamete intra-Fallopian transfers. The ovarian ultrasound pattern was correlated with serum testosterone, 17-OHP, androstenedione and oestradiol responses, and with the clinical outcome. Polycystic ovaries, defined ultrasonographically as the presence of > or = 10 follicles between 2 and 10 mm diameter in either ovary, were found in 48% of the whole study population. Dexamethasone was given to suppress adrenal androgen secretion. Functional ovarian hyperandrogenism (FOH) was defined as serum testosterone > 0.5 nmol/l after dexamethasone. There was a significantly (P < 0.001) higher prevalence of FOH in patients with polycystic ovaries (23%) compared with normal ovaries (7%). Patients with polycystic ovaries had approximately double the 17-OHP, androstenedione and oestradiol responses to a GnRH agonist as patients with non-polycystic ovaries. Exaggerated 17-OHP and oestradiol responses to GnRH agonist were found in 89% of patients with clinically diagnosed PCOS. The number of oocytes retrieved was positively correlated (r = 0.51, P < 0.001) with the oestradiol responses in all patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Characterization of the binding defect in insulin-like growth factor binding protein-3 from pregnancy serum.

During pregnancy, insulin-like growth factor (IGF) binding protein-3 (IGFBP-3) undergoes proteolysis, rendering it undetectable by radioligand binding techniques. This study examines the physical and functional defect in pregnancy IGFBP-3. Ternary complex formation has been measured by the binding of the acid-labile subunit of the circulating IGFBP-3 complex, which also requires IGF-I or IGF-II binding. IGF-depleted pregnancy IGFBP-3, prepared by size-exclusion chromatography at low pH, could not form a ternary complex in the presence of [Tyr60]IGF-I or of an IGF-I analogue extensively altered in the A-domain, whereas analogues altered in the C- or D-domains complexed as well as native IGF-I. After purification by immunoaffinity chromatography, non-pregnancy and pregnancy IGFBP-3 formed ternary complexes with IGF-I equally well, although the pregnancy-proteolysed protein appeared degraded to approximately 30 kDa. On analysis by affinity labelling, cross-linked ternary complexes containing non-pregnancy or pregnancy IGFBP-3 were predominantly 135-140 kDa, with an additional complex of 110-115 kDa in the pregnancy preparation. After reverse-phase h.p.l.c., affinity-isolated pregnancy IGFBP-3 was inactive, whereas the protein from non-pregnancy serum retained activity. Thus pregnancy-proteolysed IGFBP-3 is altered in its specificity for IGF analogues, and is more labile than non-pregnancy IGFBP-3, but shows little impairment in normal IGF binding or ternary complex formation.

Growth hormone and insulin-like growth factor regulate insulin-like growth factor-binding protein-1 in Laron type dwarfism, growth hormone deficiency and constitutional short stature.

Insulin-like growth factors (IGFs) mediate the effects of growth hormone (GH), and the insulin-like growth factor-binding proteins (IGFBPs) modulate the actions of IGFs in tissues. We studied the circulating levels of IGFBP-1 in 6 children and 9 adults with Laron type dwarfism (LTD), in 11 children and 21 adults with growth hormone deficiency (GHD), and in 8 children with constitutional short stature. Compared with the situation in healthy children, the basal serum IGFBP-1 concentration was 5.4-fold higher in LTD children, 4.1-fold higher in GHD children, and 3.8-fold higher in children with short stature (p < 0.02 vs controls in all groups). In adult patients with multiple pituitary hormone deficiency (MPHD), the IGFBP-1 concentration was 2-fold elevated, but it was normal in adult LTD patients. Intravenous (N = 10) or subcutaneous (N = 9) administration of IGF-I (75 micrograms.kg-1 and 150 micrograms.kg-1, respectively) in LTD children resulted in a rapid 50-60% fall in serum insulin (p < 0.02), a decline in blood glucose and a concomitant 40-60% rise of IGFBP-1 levels (p < 0.05). Treatment for seven days with IGF-I (150 micrograms.kg-1 x d-1) resulted in a decrease by 34% and 44% of serum IGFBP-1 level in two out of three children with LTD. After prolonged GH therapy, the IGFBP-1 level fell in GHD children by 29% (p < 0.05), in GHD adults by 52% (p < 0.02) and in children with constitutional short stature by 17% (p < 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)

Expression of insulin-like growth factor binding protein-1 mRNA in human fetal kidney.

Expression of insulin-like growth factor binding protein-1 (IGFBP-1) messenger RNA (mRNA) was studied in tissues of human fetuses from 15 to 23 weeks of gestation. Northern blot analysis revealed IGFBP-1 mRNA in the fetal liver and kidney but not in other fetal tissues, including the brain, heart, lung, skeletal muscle and spleen. Studies by in situ hybridization histochemistry showed that, in all fetal kidneys tested, the IGFBP-1 mRNA was localized preferentially to the epithelial cells of the collecting ducts, as well as to the cells of developing glomeruli and in the subcapsular nephrogenic mesenchyme. Less intense labeling for IGFBP-1 mRNA was seen in the connective tissue stroma of the medullary pyramids. A weak signal was detected in the mature glomeruli, and in the cells of the medullary mesenchyme and capsular connective tissue. IGFBP-1 protein was detected by immunoperoxidase staining mostly around small blood vessels but not in the respective endothelium. The protein was also present in many epithelial cells of the collecting ducts and in stromal connective tissue. These results show that the predominant sites of IGFBP-1 transcription in the developing kidney are those with most active differentiation.