In cytometric analysis, the percentage of CCR5-positive unfixed spermatozoa varied from ~10 to ~60%, and it significantly decreased after 5 h capacitation. The percentage of CCR5-positive spermatozoa was increased to more than 90% following fixation and permeabilization, suggesting the existence of large intracellular pools of the receptor. Immunocytochemistry showed positive staining in the anterior region of the sperm head. In ejaculates from male partner of 102 infertile couples, the CCR5 expression rate significantly correlated with sperm count, total sperm number and forward motility, but not with sperm morphology. In stepwise analysis, only forward motility entered into the model; however, this explained only ~8% of the variability in CCR5 expression. Interquartile analysis showed significant differences between the first and fourth quartiles of CCR5 expression for all semen parameters, except morphology.

The percentage of CCR5-positive spermatozoa may vary under conditions associated with changes in membrane architecture and spermatozoa showed large intracellular pools of CCR5. A lower expression of CCR5 in asthenozoospermia seems to be suggested; however, it would only partially contribute to the inter-individual variability in the CCR5 expression. A genetic basis can be hypothesized to explain the variability.

In order to explore this issue further, a cohort of Day 5 single embryo transfers was analysed prospectively for embryological and clinical outcome. All transferred embryos resulted from 8-cell embryos on Day 3, from which subsequently either one cell (group I, n = 182) or two cells (group II, n = 259) were removed, or on which no invasion by means of embryo biopsy was performed (group III, control group, n = 702).

Blastocyst formation was significantly better in group III compared with group II, and similar to group I. Group I and group II did not differ in Day 3 nor in Day 5 embryo development. The overall live birth rate was significantly higher in group I (37.4%, CI 29.0–47.4%) than in group II (22.4%, CI 17.0–28.9%), and comparable to the reference ICSI population (35.0%, CI 30.8–39.7%).

The clinical outcome of 1-cell biopsy was significantly better than that of 2-cell biopsy, even when adjusted for availability of genetically transferable embryos.

Mouse blastocysts were cultured from zygote stage in vitro with and without recombinant mouse GM-CSF (rmGM-CSF), and in vivo developed blastocysts were flushed from Csf2 null mutant and wild-type mice. The effect of GM-CSF on blastocyst expression of stress response and apoptosis genes was evaluated by microarray, qPCR and immunochemistry.

Microarray analysis of the gene transcription profile showed suppression of stress response and apoptosis gene pathways in blastocysts exposed to rmGM-CSF in vitro. qPCR analysis confirmed that rmGM-CSF inhibited expression of heat shock protein (HSP) and apoptosis pathway genes Cbl, Hspa5, Hsp90aa1, Hsp90ab1 and Gas5 in in vitro blastocysts. Immunocytochemical analysis of HSP 1 (HSPA1A/1B; HSP70), BAX, BCL2 and TRP53 (p53) in in vitro blastocysts showed that HSPA1A/1B and BCL2 proteins were less abundant when embryos were cultured with rmGM-CSF. BAX and TRP53 were unchanged at the protein level, but Bax mRNA expression was reduced after GM-CSF treatment. In in vivo developed blastocysts, Csf2 null mutation caused elevated expression of Hsph1 but not other stress response genes.

We conclude that GM-CSF inhibits the cellular stress response and apoptosis pathways to facilitate embryo growth and survival, and the protective effects of GM-CSF are particularly evident in in vitro culture media, whereas in vivo other cytokines can partly compensate for absence of GM-CSF.

The study population, drawn from the two last cycles of the National Survey of Family Growth, consists of women aged 15–24 (n = 2543 in 1995, n = 2157 in 2002). We examined trends in use of ‘contraceptive services’ and ‘other reproductive health services for preventive care’ and tested for changes in the patterns of use of these services over time. Logistic regression models were used to further clarify the factors associated with the use of the two types of services in 2002.

Results show no difference in the overall use of reproductive health services in the past year but did reveal changes in the type of service sought. Use of services for contraception increased by 10 percentage points (39.3% in 1995 to 49.7% in 2002, P < 0.001), although the use of other services remained stable (53.2% in 1995, 50.2% in 2002, P = 0.14). The patterns of use varied over time, exhibiting growing social disparities. In 2002, the use of contraceptive services depended on women's age, number of partners, personal and mother's level of education, and menstrual problems. The use of other reproductive health services for preventive care varied across women's socio-economic background.

This study demonstrates increasing social differentials in the use of reproductive health services for preventive care among young women in the USA between 1995 and 2002, a finding which calls for careful monitoring in the context of limited resources.

Endometrial biopsies, with immunohistochemical nerve fibre detection using protein gene product 9.5 as marker, taken from 99 consecutive women presenting with pelvic pain and/or infertility undergoing diagnostic laparoscopy by experienced gynaecologic laparoscopists, were compared with surgical diagnosis.

In women with laparoscopic diagnosis of endometriosis (n = 64) the mean nerve fibre density in the functional layer of the endometrial biopsy was 2.7 nerve fibres per mm² (±3.5 SD). Only one woman with endometriosis had no detectable nerve fibres. Six women had endometrial nerve fibres but no active endometriosis seen at laparoscopy. The specificity and sensitivity were 83 and 98%, respectively, positive predictive value was 91% and negative predictive value was 96%. Nerve fibre density did not differ between different menstrual cycle phases. Women with endometriosis and pain symptoms had significantly higher nerve fibre density in comparison with women with infertility but no pain (2.3 and 0.8 nerve fibre per mm², respectively, P = 0.005).

Endometrial biopsy, with detection of nerve fibres, provided a reliability of diagnosis of endometriosis which is close to the accuracy of laparoscopic assessment by experienced gynaecological laparoscopists.

This study was registered with the Australian Clinical Trials Registry (ACTR) 00082242 (registered: 12/12/2007). The study was approved by the Ethics Review Committee (RPAH Zone) of the Sydney South West Area Health Service (Protocol number X05-0345) and The University of Sydney Human Research Ethics Committee (Ref. No. 10761) and all women gave their informed consent for participation.

Secretory phase endometrium samples (n = 40), obtained from women with laparoscopically/histologically confirmed minimal–mild endometriosis (n = 20) and from women with a normal pelvis (n = 20) were selected from the biobank at the Leuven University Fertility Centre. Immunohistochemistry was performed to localize neural markers for sensory C, A, adrenergic and cholinergic nerve fibres in the functional layer of the endometrium. Sections were immunostained with anti-human protein gene product 9.5 (PGP9.5), anti-neurofilament protein, anti-substance P (SP), anti-vasoactive intestinal peptide (VIP), anti-neuropeptide Y and anti-calcitonine gene-related polypeptide. Statistical analysis was done using the Mann–Whitney U-test, receiver operator characteristic analysis, stepwise logistic regression and least-squares support vector machines.

The density of small nerve fibres was ~14 times higher in endometrium from patients with minimal–mild endometriosis (1.96 ± 2.73) when compared with women with a normal pelvis (0.14 ± 0.46, P < 0.0001).

The combined analysis of neural markers PGP9.5, VIP and SP could predict the presence of minimal–mild endometriosis with 95% sensitivity, 100% specificity and 97.5% accuracy. To confirm our findings, prospective studies are required.

This prospective, open-label, non-randomized trial included 82 women with pain symptoms caused by rectovaginal endometriosis. Patients received either a combination of letrozole and norethisterone acetate (group L) or norethisterone acetate alone (group N) for 6 months. Changes in pain symptoms during treatment and in the 12 months of follow-up were evaluated. Side effects of each treatment protocol were recorded.

Intensity of chronic pelvic pain and deep dyspareunia significantly decreased during treatment (P < 0.001 versus baseline by 3 months) in both study groups. At both 3- and 6-month assessment, the intensity of chronic pelvic pain (P < 0.001, P = 0.002, respectively) and deep dyspareunia (P < 0.001, P = 0.005, respectively) was significantly lower in group L than group N. At completion of treatment, 63.4% of women in group N were satisfied with treatment compared with 56.1% in group L (P = 0.49). Pain symptoms recurred after the completion of treatment; at 6-month follow-up no difference was observed in the intensity of pain symptoms between the groups. Adverse effects were more frequent in group L than in group N (P = 0.02).

The combination drug regimen was more effective in reducing pain and deep dyspareunia than norethisterone acetate; however, letrozole caused a higher incidence of adverse effects, cost more and did not improve patients' satisfaction or influence recurrence of pain.

In May 2005, we introduced a ‘post-operative OC recommendation’ for patients treated with laparoscopic excision of endometrioma. That is, at the time of the operation, we provided each patient with information about OC, known and possible benefits and risks and let her decide whether to take OC. A retrospective cohort study included 87 patients who underwent a laparoscopy after May 2005. The endometrioma recurrence rate at 24 months was compared between those who used OC for the entire follow-up period OC (n = 34) and all of the others (n = 53). We also performed logistic regression analysis to identify variables associated with recurrence. A before–after study included another 224 patients who underwent a laparoscopy before May 2005 and compared the recurrence rate before and after introduction of the ‘post-operative OC recommendation’.

The recurrence rate in those who used OC for the entire period was significantly lower than in the ‘others’ group (2.9 versus 35.8%, relative risk 0.082, 95% CI 0.012–0.58, P < 0.001). Post-operative OC was determined as an independent variable associated with lower recurrence (OR 0.054, 95% CI 0.007–0.429, P < 0.001). The overall recurrence rate in patients who underwent laparoscopy after the introduction of the ‘post-operative OC recommendation’ was significantly lower than that in patients who received laparoscopy before the introduction (18.6 versus 33.1%, relative risk 0.56, 95% CI 0.32–0.97, P < 0.05).

Post-operative OC use reduces the risk of ovarian endometrioma recurrence after laparoscopic excision. This information will help in appropriate planning of pre- and post-operative management.

This study linked two data sets: Taiwan's birth certificate registry and its National Health Insurance Research Data set. A total of 5627 mothers with uterine leiomyoma and 28 135 unaffected mothers were included for analysis. After adjusting for mother and infant characteristics and monthly family income, log-binominal regression and multivariate regression analyses were conducted to examine the risks of preterm birth, SGA and lower birthweight among mothers with uterine leiomyoma and unaffected mothers.

Women with uterine leiomyoma had a significantly higher percentage of preterm births (10.98 versus 7.78%, P < 0.001) and SGA infants (19.00 versus 17.28%, P = 0.002) than unaffected mothers. The mean birthweights for mothers with and without uterine leiomyoma were 3083 and 3172 g, respectively (P < 0.001). Log-binominal regression models show that the adjusted risk ratios of preterm births and SGA infants for mothers with uterine leiomyoma were 1.32 (95% CI 1.19–1.46) and 1.16 (95% CI 1.08–1.26), respectively, compared with unaffected mothers. After finally adjusting for gestational age and other covariates, a multivariate regression analysis revealed that women with uterine leiomyoma had, on average, a 14.7 g lower birthweight than unaffected mothers (P = 0.022).

We concluded that after adjusting for potential confounders, women with uterine leiomyoma experience a small yet significant increased risk of preterm and SGA infants. We suggest that clinicians intensively monitor women with uterine leiomyoma during pregnancy.

This cohort study includes all patients undergoing TLH for benign pathologies between January 1993 and June 2007 in Cochin university hospital (Paris). Demographic and surgical data were analysed. A comparison between overweight and obese patients versus non-obese patients and multivariate analyses were performed.

Of 1460 patients undergoing TLH, 101 patients (6.9%) had a BMI of 30 or higher and 338 (23.2%) were overweight. After adjustment with respect to the patients’ characteristics and past history (age, parity, past history of laparotomies, previous Cesarean section, menopausal status), no significant difference was found whether in terms of intra-operative (haemorrhage, transfusion, thrombosis, ureter, bladder or bowel injuries) or post-operative complications (hyperthermia, infections, fistula). Concerning the intra- and post-operative characteristics of these patients, only a significantly longer operating time was noted in the case of obesity (RR = 1.80; CI 95%: 1.16–2.81).

In our experience, provided that the operating technique is meticulous, the intra- and post-operative complications are not increased in the case of obesity, although the operating time is longer.

In this large, double-blind, randomized, non-inferiority trial the ongoing pregnancy rates were assessed after one injection of 150 µg corifollitropin alfa during the first week of stimulation and compared with daily injections of 200 IU rFSH using a standard GnRH antagonist protocol.

The study population comprised 1506 treated patients with mean age of 31.5 years and body weight of 68.6 kg. Ongoing pregnancy rates of 38.9% for the corifollitropin alfa group and 38.1% for rFSH were achieved, with an estimated non-significant difference of 0.9% [95% confidence interval (CI): –3.9; 5.7] in favor of corifollitropin alfa. Stratified analyses of pregnancy rates confirmed robustness of this primary outcome by showing similar results regardless of IVF or ICSI, or number of embryos transferred. A slightly higher follicular response with corifollitropin alfa resulted in a higher number of cumulus–oocyte-complexes compared with rFSH [estimated difference 1.2 (95% CI: 0.5; 1.9)], whereas median duration of stimulation was equal (9 days) and incidence of (moderate/severe) ovarian hyperstimulation syndrome was the same (4.1 and 2.7%, respectively P = 0.15).

Corifollitropin alfa is a novel and effective treatment option for potential normal responder patients undergoing ovarian stimulation with GnRH antagonist co-treatment for IVF resulting in a high ongoing pregnancy rate, equal to that achieved with daily rFSH. The trial was registered under ClinicalTrials.gov identifier NTC00696800.

This study evaluated whether or not adopting an eSET strategy instead of a DET strategy lowers the probability of having at least one live-born infant in good prognosis couples. Seven hundred and twenty-six couples were divided into two groups. The retrospective arm of the study was undertaken on the first group of couples (n = 483, DET group) and the prospective arm performed on the second group of couples (n = 243, SET group). In these specific populations, the probability of a woman having at least one live-born infant and the probability that one embryo utilized leads to a child were the main outcome measures.

The probability of a woman having at least one live-born infant was 60.5% in the DET group compared with 60.8% in the SET group. The probability of a live-born child per embryo utilized was not significantly different between the SET and the DET groups, 18.9% and 17.6%, respectively. In addition, the cumulative multiple live birth rate was significantly lower in the SET compared with the DET group.

In this observational study, using appropriate cryopreservation techniques, the chance of delivering a live baby, per utilized embryo, in an elective SET strategy is as good as that for DET.

We report a retrospective study in a university tertiary care center. Multivariate analysis and logistic regression were used to identify predictive factors of pregnancy in a series of 450 OD FTET cycles in 198 infertile women between January 1992 and December 2006.

The mean (±SD) recipient age was 35.7 (±4.5). Impaired ovarian function was the main indication for OD. The mean ± SD (range) number of embryos transferred was 1.65 ± 0.5 (1–3). Overall clinical pregnancy, implantation and delivery rates were 30, 18 and 23%, respectively. After univariate analysis, pregnancy rates were significantly higher in recipients under 35 years, in women with a body mass index (BMI) <30 kg/m², in women with an endometrial thickness of ≥8 mm, in amenorrheic women and in women not receiving pituitary down-regulation before endometrial preparation. Using multivariate analysis, the BMI, endometrial thickness and the use of pituitary down-regulation were independent predictors of pregnancy, regardless of age.

This study supports that endometrial thickness of <8 mm, obesity and the use of GnRH analogue pituitary down-regulation before endometrial priming negatively impact pregnancy rates, independently of the recipient's age.

Effect of MICA on the susceptibility to C. trachomatis infection and its association with tubal pathology were investigated in 214 infertile women recruited during the period from 2004 to 2007. Subjects were tested for C. trachomatis antibodies, and were further divided into two groups: those with (n = 42) and without (n = 59) tubal pathology based on laparoscopy results. The relationship between prevalence of C. trachomatis, tubal pathology and MICA allele polymorphisms was analysed.

Women with tubal infertility more often had antibodies to C. trachomatis [66.7 versus 39.1%; odds ratio (OR): 3.12, 95% CI: 1.68–5.78, P = 0.004] than infertile women without tubal pathology. Moreover, allele 008 had a highly negative correlation with C. trachomatis infection (P_c = 0.0036, OR: 2.14), while other allele polymorphisms showed no significant association with the disease. No statistically significant differences were found in the MICA allele frequencies of C. trachomatis-positive women with or without tubal pathology.

The association of a specific MICA allele with C. trachomatis IgG antibodies among women with infertility suggests that the MICA locus might modify host susceptibility to C. trachomatis infection.

A postal questionnaire survey of a random population-based sample of women aged 31–50 years was performed in the Grampian region of Scotland. Questions addressed the following areas: pregnancy history, length of time taken to become pregnant each time, whether medical advice had been sought and self-reported exposure to factors associated with infertility.

Among 4466 women who responded, 400 (9.0%) [95% CI 8.1, 9.8] had chosen not to have children. Of the remaining 4066 women, 3283 (80.7%) [95% CI 79.5, 82.0] reported no difficulties in having children and the remaining 783 (19.3%) [95% CI 18.1, 20.5] had experienced infertility, defined as having difficulty in becoming pregnant for more than 12 months and/or seeking medical advice. In total 398 (9.8%) [95% CI 8.9, 10.7] women had primary infertility, 285 (7.0%) [95% CI 6.2, 7.8] had secondary infertility, 100 (2.5%) [95% CI 2.0, 2.9] had primary as well as secondary infertility. A total of 342 (68.7%) and 208 (73.0%) women with primary and secondary infertility, respectively, sought medical advice and 202 (59.1%) and 118 (56.7%) women in each group subsequently conceived. History of pelvic surgery, Chlamydial infection, endometriosis, chemotherapy, long-term health problems and obesity were associated with infertility. In comparison with a similar survey of women aged 46–50 from the same geographical area, the prevalence of both primary infertility (>24 months) [70/1081, (6.5%) versus 68/710 (9.6%) P = 0.02] and secondary infertility [29/1081 (2.7%) versus 40/710 (5.6%) P = 0.002] were significantly lower.

Nearly one in five women attempting conception sampled in this study experienced infertility, although over half of them eventually conceived. Fertility problems were associated with endometriosis, Chlamydia trachomatis infection and pelvic surgery, as well as obesity, chemotherapy and some long-term chronic medical conditions. There is no evidence of an increase in the prevalence of infertility in this population over the past 20 years.

A survey was conducted among the centres for reproductive medicine that are allowed to handle oocytes and create embryos (B-centres). Data were collected on the nationality of patients and the type of treatment for which they attended during the period 2000–2007.

Sixteen of 18 centres responded to the questionnaire. The flow of foreign patients has stabilized since 2006 at approximately 2100 patients per year. The majority of foreign nationals seeking treatment in Belgium were French women for sperm donation. The next highest group was patients entering the country to obtain ICSI with ejaculated sperm.

There are clear indications that numerous movements are motivated by the wish to evade legal restrictions in one's home country, either because the technology is prohibited or because the patients have characteristics, which exclude them from treatment in their own countries.

In this prospective assessor-blinded cohort study, we included singletons born following controlled ovarian hyperstimulation in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) (COH-IVF; n = 68) or modified natural cycle-IVF/ICSI (MNC-IVF; n = 57) or naturally conceived singletons of subfertile couples (NC; n = 90). Children were assessed with standardized, age-specific and sensitive neurological assessments (TINE and Hempel assessment) at 4, 10 and 18 months. Neurological examination resulted in a neurological optimality score (NOS), a fluency score and a clinical neurological classification. Fluency of movements is easily affected by neurological dysfunction and is therefore a sensitive measure for minimal changes in neuromotor development.

The NOS and the fluency score were similar in COH-IVF, MNC-IVF and NC children. None of the children showed major neurological dysfunction and rates of minor neurological dysfunction at the three ages were not different between the three conception groups.

We found no effects of ovarian hyperstimulation or the in vitro procedure itself on neurological outcome in children aged 4–18 months. The findings of our study are reassuring, nevertheless it should be kept in mind that subtle neurodevelopmental disorders may emerge when children grow older. Continuation of follow-up in older and larger groups of children is therefore still needed.

We analysed a consecutive cohort of 1391 couples, referred to our secondary care hospital between January 2002 and December 2006. Discontinuation rates were studied at six stages. Stage I: immediately after first visit, Stage II: during diagnostic workup, Stage III: after finishing diagnostic workup but before treatment, Stage IV: during or after non-IVF treatment, Stage V: during IVF, Stage VI: after at least 3 cycles of IVF. Reasons to discontinue and spontaneous pregnancy rates after discontinuation were secondary outcomes.

In our cohort 319 couples dropped out of fertility care, 76.8%, [95% confidence interval (CI): 72.2–81.4] on their own initiative and 23.2% (95% CI: 18.6–27.8) on doctor's advice. Percentage (95% CI) of couples discontinuing per stage were: Stage I 6.0% (3.4–8.6), Stage II 3.4% (1.5–5.5), Stage III 35.7% (30.5–41.0), Stage IV 23.5% (18.9–28.2), Stage V 17.9% (13.7–22.1) and Stage VI 13.5% (9.7–17.2). Main reasons for dropout (%, 95% CI) were ‘emotional distress’ (22.3%, 17.7–26.8), ‘poor prognosis’ (18.8%, 14.5–23.1) and ‘reject treatment’ (17.2%, 13.1–21.4). The spontaneous ongoing pregnancy rate after discontinuation was 10% (6.7–13.3).

About half of the couples stopped before any fertility treatment was started and one-third stopped after at least one IVF cycle. The main reasons for withdrawal were emotional distress and poor prognosis. This insight may help to improve quality of patient care by making care more responsive to the needs and expectations of subfertile couples.

A total of 406 women participated in this study (mean age = 22, SD = 2.9). There were 211 participants in the gain condition and 195 in the loss condition.

An analysis of covariance found a main effect for framing (F(1, 402) = 6.3; P < 0.01) after controlling for existing attitudes towards oocyte donation and pre-message intentions to donate. Specifically, participants in the gain-framed condition were significantly more likely to report higher post-message intentions to donate oocytes than participants in the loss condition. However, the framing effect was only observed with British populations and not with women from South East Asia. Further, structural equation modelling analyses revealed lower levels of ‘perceived behavioural control’ (β = –0.420, P < 0.03) and positive attitudes towards ‘the importance of genetic ties between parent and child’ (β = 0.70, P < 0.001) were direct predictors of post-message intentions in the gain (but not loss) frame condition.

Findings obtained from this study indicate that oocyte donation campaigns should consider using gain-framed messages in recruitment appeals and message frames should be matched to the target populations’ perceived level of behavioural control.

The instrument was an author-constructed questionnaire completed online on the Donor Sibling Registry website. Questions assessed women's accounts of medical complications, contact with the infertility clinic through which they had provided ova, and information exchange or contact with people conceived from their ova.

Responses were received from 49.1% of the 287 donors with valid e-mail addresses. The 155 respondents completed the survey an average of 9.4 years after their first donation. Reported medical complications included ovarian hypersensitivity syndrome (30.3%) and infertility (9.6%). Subsequent to ova donation, 2.6% of women reported that they had been contacted by the IVF clinic for medical updates. On the questionnaire, 34.2% of women reported that medical changes they thought would interest donor children; half said that they had attempted to report these changes to the clinic with variable results. Many, who did not report such information, did not realize they could or should. Donors said that they frequently had not sought information about pregnancy outcomes because of confusion about the definition of ‘anonymity’ or ‘confidentiality’.

US-based IVF clinics need to give clearer guidelines to anonymous oocyte donors about follow-up information exchange. Additional long-term studies are needed to ascertain oocyte donors' risks of infertility or cancer.

Human ESCs were differentiated as embryoid bodies (EBs) in vitro. Gene and protein marker expression profiles of EBs in different periods of culture were analysed by quantitative polymerase chain reaction (Q-PCR) and immunolocalization to monitor germ cell development. Secretion of hormones involved in germ cell maturation was measured, to detect the existence of a germ cell niche within EBs.

Q-PCR revealed gene expression profiles consistent with PGC formation and germ cell development. A small population of post-meiotic spermatid cells were identified using sperm-specific antibodies (Protamine 1 and 1.97). Although gene expression profiles characteristic of oocyte development and follicle-like structures were detected, a committed oocyte with extra-cellular zona pellucida was not recognized with zona pellucida-specific monoclonal antibody.

hESCs can form PGCs and post-meiotic spermatids in vitro, however, there remains doubt about oocyte development. Levels of steroid hormones produced by EBs were significant when compared with known values for a similar quantity of human testis, suggesting that hESC may intrinsically create a favourable hormonal niche for spermatogenesis.

DSCs were isolated from human term decidua. The expression of B7-H1/B7-DC and HLA-DR and their alteration following IFN- and/or TNF- stimulation were assessed. DSCs with or without IFN- pretreatment were co-cultured with allogenic CD4⁺ T cells. The effect of PD-1:B7-H1/B7-DC and T cell receptor (TCR):HLA-DR interactions on T cell cytokine production was evaluated by adding blocking antibodies.

DSCs constitutively expressed B7-H1 and B7-DC, as well as small amounts of HLA-DR. Exogenous IFN- and TNF- up-regulated the B7-H1/-DC expression on DSCs, whereas HLA-DR expression was increased only by IFN-. IFN- pretreatment of DSCs stimulated T cell cytokine production through HLA-DR up-regulation. B7-H1 blockade on DSCs strongly enhanced T cell cytokine production (IFN-, TNF- and IL-2), whereas B7-DC blockade had similar but more modest effects. Blockade of both B7-H1 and B7-DC resulted in additive effects.

Our findings support the categorization of human DSCs as non-professional APCs and suggest that PD-1 ligands on DSCs, together with major histocompatibility complex class II, may play a crucial role in the regulation of decidual CD4⁺ T cell cytokine production. This helps to maintain a balanced cytokine milieu at the feto-maternal interface.

Ectopic ESCs were obtained from ovarian chocolate cysts in patients undergoing laparoscopic surgery (n = 22). Eutopic ESCs were isolated from endometrial tissue of cyclic premenopausal women undergoing hysterectomy for fibroids (n = 22). Purified stromal cells were studied in vitro. The number of surviving cells and activation of caspases were assessed by WST-8 assay and immunoblotting. Expression of inhibitor of apoptosis proteins (IAP) family members: cIAP1 (birc2), cIAP2 (birc3), XIAP (birc4), survivin (birc5) were examined using cDNA array and real-time RT–PCR. Effects of gene silencing by small inhibitor RNAs (siRNA) were examined by WST-8-assay, Annexin-V staining and immunoblotting.

After staurosporine (SS) treatment, 55% of eutopic ESCs survived versus 70% of ectopic ESCs. Procaspase-3 or -7 was more intensely activated by SS treatment in eutopic than in ectopic ESCs (P < 0.01). mRNAs for IAP-family genes, such as cIAP-1, XIAP and survivin, were highly expressed in ectopic ESCs before SS treatment. The fold induction of survivin expression after SS treatment was higher in ectopic than eutopic ESCs (2.8 ± 0.27 versus 0.69 ± 0.07, respectively). Survivin gene silencing in SS-treated ectopic ESCs led to an increase of apoptotic cells (P < 0.05, versus control siRNA).

We demonstrated that survivin plays a critical role in susceptibility of ESCs to apoptosis. Our results indicate that a survivin inhibitor may be effective as a novel treatment for endometriosis.

Expression levels of HOXA-10 mRNA and protein in endometrium were measured during the mid-secretory phase. This study utilized laser capture microdissection, real-time RT–PCR and immunohistochemistry.

HOXA-10 mRNA and protein expression levels in endometrial stromal cells were significantly lower in infertile patients with different types of endometriosis (deep infiltrating endometriosis, ovarian endometriosis and superficial peritoneal endometriosis), with uterine myoma, and unexplained infertility patients as compared with healthy fertile controls. HOXA-10 mRNA expression levels of microdissected glandular epithelial cells were significantly lower than those of microdissected stromal cells, without significant differences among the different groups. No protein expression was detected in glandular epithelial cells. The percentage of patients with altered protein expression of HOXA-10 in stromal cells were significantly higher in patients with only superficial peritoneal endometriosis (100%, 20/20, P < 0.05) compared with the other infertile groups (deep infiltrating endometriosis: 72.7%, 16/22; ovarian endometriosis: 70.0%, 14/20; uterine myoma: 68.8%, 11/16; unexplained infertility: 55.6%, 5/9).

The present findings suggested that altered expression of HOXA-10 in endometrial stromal cells during the window of implantation may be one of the potential molecular mechanisms of infertility in infertile patients, particularly in patients with only superficial peritoneal endometriosis. One of the underlying causes of infertility in patients with only superficial endometriosis may be altered expression of HOXA-10 in endometrial stromal cells.

Thirteen normal (control), 11 early-treated and 6 late-treated PA adult female monkeys had serum AMH levels measured at random times of the menstrual cycle or anovulatory period. Using some of the same animals, basal serum AMH, gonadotrophins and steroids were also measured in six normal, five early-treated and three late-treated PA female monkeys undergoing FSH therapy for IVF during late-reproductive life (>17 years); serum AMH also was measured on day of HCG administration and at oocyte retrieval.

Serum AMH levels in early-treated PA females declined with age to levels that were significantly lower than those of normal (P ≤ 0.05) and late-treated PA females (P ≤ 0.025) by late-reproductive life. Serum AMH levels positively predicted numbers of total/mature oocytes retrieved, with early-treated PA females having the lowest serum AMH levels, fewest oocytes retrieved and lowest percentage of females with fertilized oocytes that cleaved.

Based on these animals, early PA appears to program an exaggerated decline in ovarian reserve with age, suggesting that epigenetically induced hormonal factors during fetal development may influence the cohort size of ovarian follicles after birth.

A population-based health survey (HUNT 1) was conducted during 1984–1986 in Nord-Trøndelag county, Norway, with follow-up from 1995 to 1997 (HUNT 2). The study included 3887 women, <45 years old in HUNT 2. PA was assessed by baseline questionnaire, and fertility and parity by questionnaire at follow-up. Data focused on overall occurrence of infertility in the population (without biological confirmation).

Increased frequency, duration and intensity of PA were associated with increased subfertility, and frequency of PA was associated with voluntary childlessness (P < 0.01). After adjusting for age, parity, smoking, and marital status, women who were active on most days were 3.2 times more likely to have fertility problems than inactive women. Exercising to exhaustion was associated with 2.3 times the odds of fertility problems versus low intensity. Women with highest intensity of PA at baseline had the lowest frequency of continuing nulliparity and highest frequency of having three or more children during follow-up (P < 0.05). Sensitivity analysis including body mass index as confounder did not alter the results. No associations were found between lower activity levels and fertility or parity.

Increased risk of infertility was only found for the small group of women reporting the highest levels of intensity and frequency of PA. Awareness of the possible risks of infertility should be highlighted among non-athletic women who exercise vigorously.

In this population-based cohort study, we included 1 209 151 singleton pregnancies reported to the Medical Birth Registry of Norway between 1984 and 2006 and compared the risk of breech presentation in 8229 ART pregnancies with that in spontaneously conceived pregnancies. Risk ratios (RR), adjusted for maternal age, parity, gestational length and year of birth, were estimated using binominal regression, and we describe differences and time trends in obstetric management for breech and cephalic presentations after ART compared with management of spontaneously conceived pregnancies.

Breech presentation occurred nearly 50% more often in ART singleton pregnancies than in spontaneously conceived singletons [crude RR: 1.48, 95% confidence interval (CI): 1.34–1.64], but after adjustment for potentially confounding factors, the difference was fully attenuated (RR: 0.97, 95% CI: 0.88–1.07). The most important contributors to the attenuation were parity and length of gestation. In general, Caesarean sections and induced deliveries were more likely in ART pregnancies, but over the study period, the proportion of Caesarean sections in ART pregnancies gradually approached that of spontaneously conceived pregnancies.

Increased risk of breech presentation in pregnancies after ART is mediated by lower parity and shorter gestational length. In general, the obstetric management of women with ART pregnancies is gradually approaching the ordinary surveillance of pregnant women.

The authors used data on 41 268 live born singleton boys of mothers who were enrolled into the Danish National Birth Cohort in 1996–2002. During early childhood, 1598 cases of cryptorchidism were identified and 398 of these were orchiopexy verified. Maternal alcohol consumption including number and timing of binge drinking episodes was assessed in two computer-assisted telephone interviews around gestational weeks 17 and 32. Adjusted hazard ratios (HRs) of cryptorchidism were estimated by Cox regression.

Average weekly alcohol consumption as well as frequency of binge drinking at any time during pregnancy was not associated with risk of cryptorchidism. Binge drinking in gestational weeks 7–15 was associated with a slightly increased risk of cryptorchidism with adjusted HRs between 1.03 and 1.66.

Prenatal exposure to alcohol—measured as average intake as well as frequency and timing of binge drinking—was not associated with cryptorchidism. Our findings, however, do not rule out that binge drinking during the suggested male programming window may increase the risk of cryptorchidism.

From nationwide estimates of numbers of live births conceived by IVF (n = 40 330), we estimated the expected numbers of patients with retinoblastoma conceived by IVF in the period 1995–2007. The observed number of retinoblastoma diagnoses in children conceived by IVF was obtained by questionnaires sent to the parents of children with retinoblastoma diagnosed between 1995 and 2005. For non-responders and patients diagnosed after 2005, information was available through the medical files, in which information on fertility treatment has been routinely recorded since 2000. The relative risk (RR) of retinoblastoma among children conceived by IVF was calculated for the total study period (1995–2007) and for the expanded study period (2002–2007).

Of all eligible patients with retinoblastoma (n = 162) diagnosed in the period 1995–2007, seven were conceived by IVF. In the total study period (1995–2007) the risk was significantly elevated [RR = 2.54, 95% confidence interval (CI) = 1.02–5.23]. In the expanded study period (2002–2007), no significantly elevated risk (RR = 1.29, 95% CI = 0.16–4.66) was found.

We found a significantly increased risk of retinoblastoma in children conceived by IVF in the total study period 1995–2007. However, this increased risk was mostly based on the much stronger risk increase observed previously, for 1995–2002. Caution and awareness on the one hand and avoiding unnecessary worries on the other hand are important at this stage of our knowledge.

Informative markers flanking the DMD, IKBKG and NF2 genes were used to construct non-linked haplotypes. Polar bodies 1 (PB1) and 2 (PB2) were biopsied and analyzed to determine allelic association between the mutation and markers in multiplex PCR reactions.

For each family, the first PGD cycle allowed the establishment of linked haplotypes based on homozygous PB1 and PB2 analysis; however, no embryos were available for transfer. Subsequent cycles, when performed, confirmed this linkage. A mutation-free child was born to the family affected with DMD and an ongoing pregnancy (32 weeks) was achieved with the carrier of the IKBKG deletion.

PB analysis for reverse linkage in real-time coupled with the PGD cycle is a powerful tool for diagnosis and linkage between markers and de novo mutations for maternal autosomal dominant or X-linked disorders. Simultaneous amplification of multiple informative markers in conjunction with the mutation allows the building of familial haplotypes and accurate PGD analysis.

A clinic-based case–control study. Seventy-eight patients with non-familial, non-syndromic POF, and 90 controls were recruited to investigate the CAG repeat numbers in exon 1 of the AR gene by PCR and Gene Scan analysis.

The mean CAG repeat length in exon 1 of the AR gene of women with POF was 23.6 ± 3.8, which was significantly higher than controls (20.08 ± 3.45) (P < 0.001). The biallelic mean CAG repeat ranged from 11 to 32 in the control women, compared to 16 to 30 in the POF patients. The 22 CAG repeat allele followed by the 24 CAG repeat allele was found to be at highest frequency (15.38 and 12.8%) in POF cases, although the 19 CAG repeat allele was observed at highest frequency (12.2%) in controls.

The observation suggests that the CAG repeat length is increased in women with POF as compared with controls, and may be pathogenic for POF, at least in a subset of Indian women.

Seventy-two clinicians, basic scientists, epidemiologists and social scientists gathered together at the WHO headquarters in Geneva, Switzerland in December, 2008. Several months in advance, three working groups were established which were responsible for terminology in three specific areas: clinical conditions and procedures, laboratory procedures and outcome measures. Each group reviewed the existing ICMART glossary, made recommendations for revisions and introduced new terms to be considered for glossary expansion.

A consensus was reached on 87 terms, expanding the original glossary by 34 terms, which included definitions for numerous clinical and laboratory procedures. Special emphasis was placed in describing outcome measures such as cumulative delivery rates and other markers of safety and efficacy in ART.

Standardized terminology should assist in analysis of worldwide trends in MAR interventions and in the comparison of ART outcomes across countries and regions. This glossary will contribute to a more standardized communication among professionals responsible for ART practice, as well as those responsible for national, regional and international registries.

Pregnant Sprague Dawley rats were exposed to letrozole alone (0.04 mg/kg), or in combination with estradiol cyclopentylpropionate (ECP) at 0.5, 1 or 2 µg/rat. A control group receiving only the vehicles was also included. Animals were exposed during gestation Days 6–16 (corresponding approximately to 3–10 weeks of gestation in the human). Developmental end-points, including intrauterine mortality, fetal growth, placental weight and incidence of structural abnormalities, were evaluated near term gestation.

Exposure to letrozole alone was lethal for 41% of conceptuses, and caused minor axial skeletal anomalies in 51% of live fetuses. ECP co-administration effectively prevented letrozole-induced embryolethality, but failed to reduce the incidence of axial skeletal alterations.

The obtained results support the concept that inhibition of estrogen biosynthesis represents a critical determinant of letrozole-induced embryonic mortality. A mechanism other than estrogen deprivation appears to underlie the initiation of skeletal anomalies.

In a prospective population-based cohort study, vaginal and cervical swab samples were obtained from 2130 unselected pregnant women undergoing their first and second trimester ultrasound screening. MMP-8 was determined by immunofluorometric assay. Use of antibiotics, history of vaginal bleeding, and history of sexual intercourse were recorded on both occasions. Vaginal smears were obtained for Gram-staining and leukocyte counting. Cervical length was measured by ultrasonography. The main outcome measures were MMP-8 concentrations in the vagina and cervix.

The median (range) MMP-8 concentrations in vaginal and cervical samples were 107.4 µg/l (undetectable–2406.6 µg/l) and 318.3 µg/l (0.1–2074.6 µg/l), respectively, in the first trimester, and 112.5 µg/l (undetectable–2093.4 µg/l) and 344.8 µg/l (0.4–1783.5 µg/l), respectively, in the second trimester. Multiparity and vaginal leukocytosis were both associated with increased MMP-8 concentrations in vaginal and cervical samples in both trimesters. Bacterial vaginosis (BV) was associated with increased vaginal and cervical MMP-8 in the first trimester, but only with increased vaginal MMP-8 in the second trimester. A history of sexual intercourse (in the previous 48 h) was associated with lower MMP-8 concentrations in cervical samples in both trimesters.

MMP-8 concentrations were lower in vaginal samples than in cervical samples, and no difference was found between the first and second trimesters. Multiparity, BV and an elevated leukocyte count in the vagina were associated with increased MMP-8 concentrations. Sexual intercourse had an opposite effect. The study suggests that MMP-8 is a physiologic constituent in lower genital tract fluids, where it may be involved in host response to inflammatory and infectious processes.

A randomised controlled trial in women with RM treated with G-CSF or placebo was conducted in one private reproductive medicine clinic. Sixty-eight women with unexplained primary RM, all with at least four consecutive miscarriages and negative for all clinical investigations, were selected. Patients were randomized for s.c. treatment with G-CSF (n = 35) (1 µg/kg/day) starting on the sixth day after ovulation, or with placebo (n = 33). Patients were randomized using a computer-generated randomization number sequence. Pregnancy outcome (delivery of a healthy baby without major or minor malformations) was the primary outcome measure.

In the group treated with G-CSF, 29 out of 35 (82.8%) women delivered a healthy baby, whereas in the placebo group, this figure was only 16 out of 33 (48.5%) (P = 0.0061, odds ratio = 5.1; 95% confidence interval 1.5–18.4). Significantly higher β-hCG levels were found in gestation weeks 5–9 in women treated with G-CSF versus placebo (P < 0.001).

Our data show that G-CSF may be effective in the treatment of unexplained RM. However, further studies are needed to confirm the effectiveness of this treatment in women with unexplained RM, refractory to conventional treatment.

The study was registered with a ICMJE recognized registry, the Clinical Trial.gov Protocol Registry System, with the number NCT00772122.

Four 4-cell stage embryos were split on Day 2 of preimplantation development and the 16 blastomeres were individually cultured in sequential medium. On Day 3 or 4, the blastomere-derived embryos were plated on inactivated mouse embryonic fibroblasts (MEFs).

Ten out of sixteen blastomere-derived morulae attached to the MEFs, and two produced an outgrowth. They were mechanically passaged onto fresh MEFs as described for blastocyst ICM-derived hESC, and shown to express the typical stemness markers by immunocytochemistry and/or RT–PCR. In vivo pluripotency was confirmed by the presence of all three germ layers in the teratoma obtained after injection in immunodeficient mice. The first hESC line displays a mosaic normal/abnormal 46, XX, dup(7)(q33qter), del(18)(q23qter) karyotype. The second hESC line displays a normal 46, XY karyotype.

We report the successful derivation and characterization of two hESC lines from single blastomeres of four split 4-cell stage human embryos. These two hESC lines were derived from distinct embryos, proving that at least one of the 4-cell stage blastomeres is pluripotent.

In this study, we employed inter-genotype ooplasm transfer followed by ICSI using two mouse strains, C57BL/6 and DBA/2, to explore the influence of foreign ooplasm on paternal pronucleus function. In order to assay for effects on the paternal genome without masking effects of the maternal genome, we examined ooplasm effects in diploid androgenones, which are produced by pronuclear transfer to contain exclusively two paternal sets of chromosomes, in combination with ICSI.

There was no significant effect of intra-strain ooplasm transfer among androgenones made with either C57BL/6 or DBA/2 oocytes. There was a significant negative effect on androgenone blastocyst development with inter-genotype transfer (10% volume) of DBA/2 ooplasm to C57BL/6 oocytes (P < 0.05). The reciprocal inter-genotype ooplasm transfer had no significant effect.

Thus, inter-genotype ooplasm transfer in conjunction with ICSI can alter the function of the paternal genome. However, the effect of foreign ooplasm is restricted to a negative effect, with no evidence of a positive effect. This study provides important new information about the possible consequences of ooplasm donation in human ART.

A systematic search of Medline identified seven studies evaluating post-operative OCP treatment on prevention of endometriosis recurrence.

A reduction in anatomical relapse rate was observed when oral contraceptive therapy was administered for more than 1 year after conservative surgery. Post-operative use of OCP was associated with a reduction in frequency and intensity of dysmenorrhoea recurrence. No association was found between OCP therapy and dyspareunia prevention, although the effect of OCP on chronic pelvic pain was conflicting.

Long-term OCP therapy can be a reliable adjuvant post-operative measure to prevent or reduce frequency/severity of recurrent dysmenorrhoea and anatomical relapse of endometriosis. Since both continuous and cyclic OCP administration regimens seem to have comparable effects, the choice of regimen can be modulated according to patient preferences. The protective effect seems to be related to the duration of treatment.

Pre- and post-treatment endometrium and endometriosis specimens were obtained from 22 women experiencing pain related to endometriosis who were treated with LNG-IUS (n = 11) or GnRHa (n = 11) for 6 months. Changes in the expression of proliferating cell nuclear antigen, Fas, progesterone receptor (PRA) and estrogen receptor (ER-) were analyzed by immunohistochemistry.

The cell proliferation index was significantly reduced in the epithelium and stroma of both the eutopic and the ectopic endometrium after treatment with the LNG-IUS and GnRHa. Only LNG-IUS users showed an increased H-score for Fas in the epithelium of the eutopic and ectopic endometrium (P < 0.05). Expression of ER- and PRA by the glandular epithelium was lower in the eutopic endometrium after both treatments, but this reduction was noted in the ectopic endometrium only after LNG-IUS treatments (P < 0.05). No difference was detected between groups for any of the markers.

LNG-IUS reduced both cell proliferation and the expression of PRA and ER- and increased Fas expression in the eutopic and ectopic endometrium of patients with endometriosis. Some of these actions were not observed with GnRHa.

The uterus and its vasculature were removed from BALB/c mice and transplanted into C57Bl/6 recipient mice at a heterotopic position, with the native uterus left in situ. Both uteri were removed on post-operative day 2 (D2, n = 5), D5 (n = 5) and D10 (n = 6). Immunohistochemistry for neutrophilic granulocytes, macrophages, cytotoxic CD8⁺ T-cells, CD4⁺ T-helper cells and B-cells was performed and cell density was evaluated in both myometrium and endometrium.

Neutrophil density was increased in graft versus native uteri at D5 and D10 in myometrium and D10 in endometrium, and in endometrium was higher in the D5 than D2 graft (all P < 0.05). Infiltration of macrophages occurred from D2 in myometrium and from D5 in endometrium (P < 0.05, graft versus native). Density of CD8⁺ cytotoxic T-cells increased in the graft versus native uteri at D5 in both uterine layers and for the graft versus D2 density (P < 0.01). In contrast, CD4⁺ T-helper cells increased only transiently in graft endometrium at D5 (P < 0.05). Overall CD19⁺ B-cell density was low, with no time-dependent changes in graft myometrium or endometrium.

Acute rejection of an allogeneic uterus transplant in the mouse involves influx of predominately neutrophils, macrophages, CD8⁺ T-cells and CD4⁺ T-cells between D2 and D5 post-operatively.

A prospective observational study was conducted at a university-affiliated private infertility centre. Patients were 745 women, who underwent 1328 IVF/ICSI treatment cycles. Basal FSH, basal LH and combinations of FSH and LH versus treatment data and pregnancy and delivery rates were measured.

Combinations of FSH and LH gave significantly better information than the LH:FSH ratio, or each gonadotrophin alone: highest mean pregnancy rate (39%) was achieved in women with low FSH (<6.7 U/l) and with high LH levels (>4.9 U/l), whereas pregnancy rate was lowest (22%) in women with high FSH and low LH levels. Pregnancy rates were intermediate (27–28%) if FSH and LH were either both low or both high (P for trend = 0.0004). Associations to delivery rates and measures of ovarian response and embryo quality followed the same pattern.

Basal LH modifies and improves the information given by basal FSH alone. Low FSH level combined with high LH probably reflects a well-preserved ovarian reserve and is associated with the highest success rates at IVF/ICSI.

This prospective study included anovulatory women with PCOS undergoing LOD (n = 29) or receiving clomiphene citrate (n = 18). Plasma AMH concentrations were measured before and 1 week after treatment. Further measurements of AMH were made at 3- and 6-month follow-up.

The pretreatment median (range) plasma AMH concentrations were 6.1 (1.0–21.0) and 5.7 (1.3–9.5) ng/ml in women having LOD and clomiphene citrate treatment, respectively. Women who ovulated after LOD (n = 24) had a significantly (P = 0.032) lower pre-operative AMH [5.6 (1.0–21.0) ng/ml] compared with the non-responders [9.0 (6.1–17.1) ng/ml]. Using receiver-operating characteristic curve analysis, AMH was found to be a useful predictor of no ovulation after LOD with area under the curve of 0.804 (P = 0.025). Using a cut-off of 7.7 ng/ml, AMH had a sensitivity of 78% and a specificity of 76% in the prediction of no ovulation after LOD. For all patients (n = 47, clomiphene citrate or LOD), plasma AMH ≥7.7 ng/ml was associated with a reduced chance of ovulation after treatment (P = 0.004). Following LOD, the median AMH concentration significantly (P = 0.003) decreased to 4.7 (0.3–15.1) ng/ml and remained low at 3- and 6-month follow-up.

Pretreatment circulating AMH level seems to be a good predictor of the ovarian response to LOD.

Endometrial samples were obtained from 33 fertile, 26 PCOS, 25 endometriosis and 33 patients diagnosed with unexplained infertility. L-selectin ligands and GlcNAc6ST-2 expression was assessed by immunohistochemistry and immunoblotting.

Immunohistochemical staining of uterine epithelium, from fertile and infertile women, demonstrated differential expression of MECA-79 and HECA-452 epitopes. In fertile women in the secretory phase MECA-79 was more strongly expressed, particularly on the lumen, than in infertile women. HECA-452 staining was significantly stronger in the glands in PCOS and endometriosis patients than in fertile women. GlcNAc6ST-2 expression was reduced in infertile patients, correlating with MECA-79 expression.

This study demonstrated significant differences in expression of L-selectin ligands between fertile and infertile women in natural cycles, and could contribute to patient assessment prior to initiating fertility treatment.

Women were 21–28 years old at tissue cryopreservation. Nine women suffering POF following chemotherapy with or without radiotherapy underwent orthotopic ovarian tissue transplantation. After 12 months of spontaneous cycles without pregnancy, oocyte retrieval was performed in four patients during mildly stimulated or spontaneous cycles. ICSI was performed in all cases, with embryo transfer on day 3. Light and electron microscopy was used to study oocytes and embryos.

Signs of ovarian function restoration (estradiol peak, decreased FSH, follicular development) began 16–26 weeks after reimplantation. Twenty-one oocyte retrieval attempts were made. At least one oocyte was collected in 15 cases, giving an empty follicle rate per retrieval of 29% (6/21). Sixteen oocytes were recovered, of which 6 were abnormal or immature (38%) and 10 (62%) were in metaphase II (MII). Three MII oocytes failed to fertilize, two showed abnormal fertilization and five normal MII oocytes successfully fertilized with subsequent normal embryo development (Grade 2), yielding an embryo transfer rate of 24% per retrieval. No pregnancy occurred.

IVF in women with orthotopically grafted frozen-thawed ovarian tissue involves a higher risk of empty follicles, abnormal or immature oocytes, and low embryo transfer rates.

The follow-up study comprised 233 IVF children aged 8–18 years and 233 spontaneously conceived controls born to subfertile parents. Growth data from birth to 4 years of age, available for 392 children (n = 193 IVF, n = 199 control), were used to study early post-natal growth. Furthermore, early post-natal growth velocity (weight gain) was related to blood pressure and skinfold measurements at follow-up.

We found significantly lower weight, height and BMI standard deviation scores (SDSs) at 3 months, and weight SDS at 6 months of age in IVF children compared with controls. Likewise, IVF children demonstrated a greater gain in weight SDS (P < 0.001), height SDS (P = 0.013) and BMI SDS (P = 0.029) during late infancy (3 months to 1 year) versus controls. Weight gain during early childhood (1–3 years) was related to blood pressure in IVF children (P = 0.014 systolic, 0.04 diastolic) but not in controls. Growth during late infancy was not related to skinfold thickness in IVF children, unlike controls (P = 0.002 peripheral sum, 0.003 total sum). Growth during early childhood was related to skinfold thickness in both IVF and controls (P = 0.005 and 0.01 peripheral sum and P = 0.003 and 0.005 total sum, respectively).

Late infancy growth velocity of IVF children was significantly higher compared with controls. Nevertheless, early childhood growth instead of infancy growth seemed to predict cardiovascular risk factors in IVF children. Further research is needed to confirm these findings and to follow-up growth and development of IVF children into adulthood.

Using aggregated annual in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cycle data in Germany, we evaluated the relationship between changes in the number of cycles in relation to changes in costs faced by consumers following the introduction of a patient co-payment from ‘no fees’ to 1500–2000 by estimating the short-run price-elasticity of demand. The impact of introducing patient co-payments for IVF/ICSI on the likelihood of switching to other low-cost fertility treatments was evaluated using the cross-price elasticity methodology.

The reduction in demand for IVF and ICSI cycles in the year following the introduction of patient co-payments resulted in elasticities of –0.41 and –0.34, respectively. The price-elasticity for the combined reduction of IVF/ICSI in relation to the co-payment was estimated to be –0.36. The cross-price elasticity for clomifene was close to zero (–0.01) suggesting that demand for these interventions are independent of each other and no substitution occurred.

We report price elasticities for IVF and ICSI of –0.41 and –0.34 after introducing a 1500–2000 co-payment. These findings likely represent short-run elasticities that are likely to vary over time as factors that influence the supply and demand for fertility treatments change.

A consecutive cohort of women who had conceived through ART was recruited systematically in early pregnancy. At 3 months post-partum, participants completed postal questionnaires which included a new measure of the degree of difficulty involved in conceiving, the Burden of Infertility and Treatment (BIT) scale.

Of 166 women who participated, 8% had already been admitted to a REPS within 3 months, which is a higher rate compared with other women in the first 12 months (5%). Compared with community samples of new mothers, there was no difference in rate of depression. A higher proportion reported dysregulated infant behaviours (P < 0.0001) and a smaller proportion was breast feeding exclusively (P < 0.0001). Greater difficulty conceiving (higher BIT score) was associated with lower maternal confidence.

Clinical care of the increasing group of women who conceive with ART should include explicit assessment of early post-partum psychological functioning and early intervention if difficulties in managing infant behaviour are reported.

A prospective cohort study of consecutively recruited, new couples in fertility treatment completed questionnaires immediately before initiating treatment (T1) and at 1-year follow-up (T2) (response rates 80%, 88%). The study population included 695 participants (355 women and 340 men) who had no severe depressive symptoms at T1, who had no child together at T1 and whose treatment had failed. We measured severe depressive symptoms with Mental Health Inventory 5 from Short-Form 36, functional aspects of general social relations and of infertility-specific social relations.

Fifteen per cent of women and 6% of men in unsuccessful treatment reported severe depressive symptoms at T2. Among men, low emotional support, low appreciation and high excessive demands from the partner were significant determinants of the incidence of severe depressive symptoms. Among women and men, low appreciation from the family, many conflicts and high excessive demands from family, friends and neighbours were significant determinants of severe depressive symptoms. The analyses were controlled for gender. Duration of infertility, infertility diagnosis and socio-economic position did not confound the associations.

More women than men report severe depressive symptoms after 1 year of unsuccessful treatment. It is important to be aware of the possible negative impact of relational strain among fertility patients in unsuccessful treatment.

The Symptom Checklist-90-R, a standardized instrument for the measurement of current psychological symptom status, was administered to 120 men upon first presentation to a public health sector infertility clinic (study group) in a tertiary referral centre. The control group comprised 120 men who attended an antenatal clinic with their partner. All men may have previously fathered a child. Raw test scores were converted into standard area T scores and analyzed further.

Participants in the study group differed in their psychological symptom status when compared with controls. Male partners of infertile couples had significantly elevated mean T scores for all nine primary symptom dimensions as well as the three global markers of distress (P < 0.0001 versus control), but these did not exceed the upper range of normal.

When compared with controls, male partners of infertile couples experienced elevated levels of psychological distress, but without, on average, suffering from psychopathology. A comparison with qualitative studies from African countries and with quantitative studies from the Western industrialized world revealed both similarities and differences. Understanding and addressing the male perspective of infertility is an important component of infertility management.

Attachment, appraisal, coping, general well-being, infertility-related stress and relationship satisfaction questionnaires were completed by 98 women and 64 men. Data were analysed by gender comparisons, correlations and path analysis.

Attachment anxiety was associated with well-being in women via appraisal of infertility as a loss and use of self-blame and avoidance (SBA) coping. Attachment anxiety was also linked with infertility-related stress through SBA. In men, attachment anxiety was associated with well-being and infertility-related stress again via SBA coping. Attachment anxiety and avoidance were related to lower relationship satisfaction in women, whereas only the former was important for men.

Attachment patterns link to couples' relationship satisfaction and are associated with adjustment via appraisal and coping. Identification of such patterns may assist in identifying need and tailoring cognitive interventions to individuals. Participants were mainly white and well-educated, and wider generalization cannot automatically be assumed.

We studied ovarian function in women with DM1 (n = 66) and healthy controls (n = 58), all <45 years old. Steroids, gonadotrophins, AMH and inhibin B levels were measured during the follicular phase.

Piece-wise regression analysis demonstrated that AMH levels begin to decrease at 33 years of age in both groups. This age limit was used to compare data in both groups. AMH levels were lower in DM1 women than in controls >33 years (4.1 ± 4.2 versus 9.5 ± 7.9 pmol/l, mean ± SD, P = 0.006). A higher proportion of women with DM1 showed AMH levels in the menopausal range compared with controls (16.7% versus 3.4%, respectively, P = 0.02). For all patients, those with DM1 exhibited lower inhibin B levels than controls (89.3 ± 51.7 versus 113.2 ± 76.0 ng/ml, P < 0.05). FSH and estradiol were similar in both groups. Regression analysis showed an earlier decline in AMH levels in women with DM1 than controls. Even after age adjustment, DM1 was a significant factor for the determination of inhibin B and AMH levels.

Lower AMH levels in women with DM1 during the fourth decade of life suggest the presence of an earlier decline in the ovarian follicle pool in these women. Further studies are needed to evaluate the mechanism of this complication.

Swaledale ewes had their left ovaries removed and either perfused but not cryopreserved (n = 4; control), or perfused and cryopreserved (n = 8; cryopreserved) before autotransplantation sub-cutaneously to the neck by microvascular anastomosis. Right ovaries were removed and fixed as non-grafted controls. Weekly jugular venous blood samples were analysed for plasma FSH, LH, inhibin A and progesterone levels, grafts were scanned transdermally and oestrus was detected. Vascular patency was assessed post-mortem and follicle populations were measured in recovered tissue.

Immediate vascular patency was achieved in all ewes and maintained in 7/8 cryopreserved and 3/4 control grafts. Functional corpora lutea were identified in three ewes (one control; two cryopreserved) 18–25 weeks after grafting. Inhibin A levels indicated resumption of follicular development in four cryopreserved and one control ewes, however, castrate gonadotrophin levels persisted in five cryopreserved and two control ewes. Primordial follicle density was reduced following grafting in both cryopreserved and non-frozen ovaries (P < 0.001).

In conclusion, these results demonstrate successful partial restoration of ovarian function following cryopreservation of the whole ovary and vascular pedicle in a large monovulatory species. The inability to restore full ovarian function was related to loss of primordial follicles rather than vascular patency in both frozen and fresh tissue, suggesting that factors associated with cannulation and perfusion may contribute to this depletion. Further work is therefore needed to elucidate these factors before the procedure could be considered a viable option for fertility preservation.

GIP was purified by liquid chromatograph and identified by mass spectrometry. The interaction of GIP with glycodelin, matrix molecule and spermatozoa were investigated.

Mass spectrometry analysis suggested that GIP contained the N-terminal region of 2-macroglobulin, confirmed by western blot with anti-2-macroglobulin antibody. GIP bound to native but not deglycosylated glycodelin-C in native gel electrophoresis, suggesting that the binding was glycosylation-dependent. GIP did not bind to capacitated and uncapacitated human spermatozoa. The cumulus cells could convert exogenous labeled 2-macroglobulin into GIP in vitro. GIP interacted with hyaluronic acid, a major component of the cumulus matrix. Glycodelin-C bound to hyaluronic acid-coated agarose beads in the presence of GIP. Human spermatozoa acquired the hyaluronic acid–GIP-bound glycodelin-C during incubation in vitro.

The hyaluronic acid–GIP complex formed in the cumulus matrix retains and concentrates glycodelin-C in the cumulus matrix for displacing sperm-bound glycodelin-A and -F and stimulating the zona binding activity of the spermatozoa traversing through the cumulus mass.

CCs from single oocytes of normal responder patients were analysed by DNA microarrays. Concomitantly, estradiol levels on the day of hCG administration, CC morphology, total collected oocyte and metaphase II oocyte number were assessed in relation to LH/hCGR gene expression in CC.

The transcriptome analysis of CC indicated a variable expression of LH/hCGR among the patients and intra-patients. LH/hCGR mRNA expression was negatively correlated with serum estradiol level on the day of hCG administration. Eighty-five genes were significantly modulated between CCs from patients with a high and a low LH/hCGR expression. These genes are involved principally in steroid metabolism and in the ovulation process and include TNFAIP6, a gene expressed during CC–oocyte complex (COC) expansion. There were no significant differences in LH/hCGR gene expression profile between COS protocols.

LH/hCGR is expressed in CC under COS conditions. LH/hCGR expression level is associated with TNFAIP6 gene expression and negatively correlated with serum estradiol level on the day of hCG administration.

The RL95-2 cell line was employed as an in vitro model for receptive endometrium. Cells transfected with gal-3 siRNA or treated with exogenous gal-3 were incubated with or without function-blocking integrinβ1/3 antibody for evaluating synergism of gal-3 and integrins on cell proliferation and adhesion. Proliferation was measured by BrdU incorporation, and adhesion to fibronectin (FN) was determined by an adhesion assay. Integrin expression was analyzed by Flow Cytometry and western blots. Bewo spheroids were co-cultured with the RL95-2 monolayer to mimic the blastocyst–endometrial interaction, and colocalization of gal-3, integrinβ3 and FN at the interface was observed by confocal microscopy.

The knock-down of gal-3 inhibited RL95-2 cell proliferation and adhesion. However, a reduction of proliferation and adhesion was also observed in presence of exogenous gal-3, and this was further reduced by a functional block to integrinβ3. Moreover, gal-3 knock-down significantly increased integrinβ3 expression, however, the colocalization of integrinβ3 and FN was not increased. As expected, the colocalization of integrinβ3 was decreased with the knock-down of gal-3.

This study has provided an in vitro model for the complex interactions between gal-3 and integrinβ3 in the regulation of endometrial cell proliferation and adhesion.

The RNA interference (RNAi) technology was used to specifically knockdown the expression of either AdipoR1 or AdipoR2. Progesterone and estradiol levels in the conditioned media were measured by radioimmunoassay, and determination of cell proliferation by tritiated thymidine incorporation. The levels of adiponectin receptors and proteins involved in the steroidogenesis and in the signalling pathways were examined by western blot.

We generated AdipoR1 (R1) and AdipoR2 (R2) knockdown KGN cell lines. R1 cells were apoptotic and had increased expression levels of cleaved caspase 3 and decreased levels of BAD phosphorylation and PCNA as compared with control or parental KGN cells. R2 cells had similar morphology to control or KGN cells. However, they produced less progesterone and estradiol and expressed lower levels of StAR protein in response to FSH or IGF-1 stimulation compared with control cells. Furthermore, the increase of MAPK ERK1/2 phosphorylation in response to human recombinant adiponectin and FSH was lower in R2 than control cells.

In the human granulosa KGN cell-line, AdipoR1 seems to be involved in the cell survival whereas AdipoR2, through MAPK ERK1/2 activation, may be implicated in the regulation of steroid production.

A group of 207 patients (≤39 years of age), treated by IVF/ICSI, received 200 IU/day rec-FSH from Day 2 of the cycle and daily GnRH antagonist starting on Day 6 of stimulation. The criteria for hCG administration included only the presence of ≥3 follicles of ≥17 mm diameter. One to two embryos were transferred on Day 3 after oocyte retrieval.

The area under the curve (AUC) for E₂ on the day of hCG could not distinguish between pregnant and non-pregnant women (AUC:0.5; 95% confidence interval (CI): 0.42–0.59). No significant difference was observed between the three percentile groups of E₂ values on the day of hCG administration [group 1, lower 25th percentile (<1142 pg/ml); group 2, medium 50th percentile (1142–2446 pg/ml) and group 3, higher 75th percentile (>2446 pg/ml)] for the ongoing pregnancy rates (P = 0.52). On the contrary, the linear regression model showed that higher E₂ values on the day of hCG administration significantly improved the scores of transferred embryos (P = 0.01) as well as the total embryo score (P = 0.02). Yet subgroup analysis only in this high responders group revealed lower E₂ and progesterone levels on the day of hCG in pregnant women compared with the non-pregnant (P = 0.01).

E₂ concentrations on the day of hCG administration in GnRH antagonist cycles are not associated with pregnancy rates. A potential deleterious impact of estradiol on endometrial receptivity is shown for the high responders who have high E₂ levels and improved embryo quality without a concomitant rise in pregnancy rate.

Seventy ICSI patients undergoing controlled ovarian stimulation (COS) in a GnRH-antagonist protocol was randomized into two groups. The control group received a standard treatment with rFSH (Puregon) plus a GnRH-antagonist, daily from Day 6 of stimulation. In the study group, rFSH was discontinued when six follicles ≥12 mm were observed and estradiol levels were >600 ng/l; rFSH was subsequently replaced by low-dose hCG (200 IU/l daily).

Mean values (SD) for dose and duration of rFSH treatment in the control versus low-dose hCG group were 1617 (280) versus 1273 (260) IU rFSH [between-group difference –344, 95% confidence interval (CI) –483 to –205; P < 0.001], and 8.2 (1.6) versus 6.4 (1.3) days (–1.8, –2.6 to –1.1; P < 0.001), respectively. The mean number of metaphase II oocytes of 10.1 versus 8.9 (between-group difference –1.2, 95% CI –3.9 to 1.5) and the ongoing pregnancy rates of 10/35 (29%) versus 13/35 (37%) (between-group difference 8.6%; 95% CI –13.0 to 29.1%; P = 0.45) for control versus hCG, respectively, did not differ.

In this pilot trial, substitution of rFSH by low-dose hCG in the final days of COS leads to a reduction of FSH consumption whereas ICSI outcome, in terms of oocyte yield and ongoing pregnancy rate, remains comparable to the traditional regimen (ClinicalTrials.gov, trial number: NCT00750100).

A total of 232 IVF candidates, 49 of whom had PCOS according to the Rotterdam 2003 consensus criteria, were recruited. AMH levels and ovarian morphology were assessed. The relationships between AMH and insulin resistance and androgenaemia in patients with and without PCOS were studied.

PCOS patients were slightly older than controls (median ages 34 and 30 years, respectively). AMH generally increased with antral follicle count (AFC), insulin, homeostatic model assessment of tissue insulin sensitivity (HOMA-IR), testosterone, free androgen index and luteinising hormone, and decreased with chronological age, homeostatic model assessment of steady state beta cell function (HOMA-B) and serum sex hormone binding globulin (SHBG). For these relationships there were no significant differences in the slopes between PCOS and non-PCOS patients. The ratio of AMH per antral follicle (AMH/AF) was higher in PCOS patients. Both PCOS and non-PCOS groups showed a very similar increase in AMH with increases in AFC, but the PCOS patients had consistently higher AMH across all AFC levels.

These observations indicate that AMH is similarly related to insulin resistance and androgens in women with and without PCOS. This effect appears to be independent of age although an indirect causal effect due to ageing or some other mechanism cannot be ruled out. Excessive granulosa cell activity may be implicated in the abnormal follicular dynamic of the syndrome.

Clinical, metabolic and endocrine parameters were obtained and an oral glucose tolerance test was performed, with calculation of IR indices. The association between thyroid function and IR was evaluated with classification analysis using logistic regression and 10-fold cross-validation to identify a possible TSH threshold for IR. Parameters were then compared between women above and below the TSH threshold using two-sample tests. One-way analyses of covariance were performed to explore whether the impact of TSH on IR is independent of other variables.

A TSH cut-off value around 2 mIU/l had the best sensitivity and specificity for identifying women with IR. Women with TSH ≥ 2 mIU/l were younger, had a higher body mass index (BMI) and were more insulin-resistant compared with women with TSH < 2 mIU/l. This effect of TSH on IR was independent of age and BMI.

In women with PCOS, a significant association between thyroid function, as reflected by TSH ≥ 2 mIU/l, and IR was found and the association appeared to be independent of age and BMI.

In 31 patients (age <39 years) stimulated with gonadotrophins and GnRH antagonists for intrauterine insemination (IUI) an endometrial biopsy was performed on the first day after the end of menstruation and a second biopsy was performed two (Group 0 + 2, n = 10) or four (Group 0 + 4, n = 11) days after the first biopsy, or on the day of hCG administration (Group 0 + hCG, n = 10). Expression of progesterone (PR) and estrogen (ER) receptor was investigated by immunohistochemistry using monoclonal antibodies.

PR and ER levels were significantly increased in the second versus the first biopsy, in all groups analyzed (P = 0.01), in both stromal and glandular cells. Between the three groups compared, a significant increase in PR expression was observed for glandular cells (P = 0.03), with the highest value observed in Group 0 + 4. Moreover, the increase in PR expression in stromal cells differed between groups (P = 0.01), with the highest value observed in the Group 0 + hCG.

In stimulated cycles for IUI, ER expression in both glandular and stromal endometrial cells, after an initial increase, does not appear to change significantly during the follicular phase. On the contrary, during the same period of time, following an initial rise, PR expression in glandular and stromal cells continues to increase.

Data from the US natality files (1981–2006) were analyzed to determine the impact of changes in birth characteristics on male birth proportion. Male birth proportion was calculated as the number of male births divided by the total number. In separate analyses, trends in male birth proportion from 1981 to 2006 were adjusted for plurality (singleton, multiple), gestational age (<28, 28–32, 33–36, ≥37 weeks) and, from 1989, maternal Hispanic ethnicity. Separate analyses were conducted for white and black births. Log binomial regression was performed to estimate crude and adjusted trends with year as independent variable.

Trends in male birth proportion differed significantly according to plurality among white (P < 0.01), but not black births. Adjustment for gestational age tempered the trends among white singletons (P < 0.0001) and multiples (P < 0.05) but had no effect on trends in black male birth proportion. Adjustment for Hispanic ethnicity had no impact on trends in black male birth proportion and any effect on white births was negated by changes in gestational age trends.

Lack of consistent influences on, or patterns of change in, the proportion of male births between different subpopulations of births suggests that a single mechanism is unlikely to explain the oft-referenced decrease in the overall US sex ratio.

In this retrospective comparative cohort study, we analysed incidence of twin pregnancies in unselected patients undergoing ICSI and PGD (group A; 1992 cycles) with blastocyst transfer at Day 5, versus a period-matched control population of unselected patients undergoing ICSI and blastocyst transfer at Day 5 without PGD (group B; 2429 cycles) from January 2001 to December 2006.

Clinical pregnancy per embryo transfer was established in 618/1992 (31.0%) and 947/2429 (39.0%) in group A versus B, respectively (P < 0.01). Overall MZ twin rate was 29/4421 (0.7%) per embryo transfer and 29/1565 (1.9%) per established clinical pregnancy. The incidence of MZ twinning per established clinical pregnancy did not differ between groups (1.5 versus 2.1%, group A and B, respectively). In group A, seven MZ twins were born versus 19 MZ twins in group B. In group B, one MZ twin pregnancy resulted in two stillbirths. In group A, two MZ twins had severe congenital malformations versus none in group B.

The incidence of MZ twinning was not increased in PGD compared with regular ICSI with blastocyst transfer. This information is useful in counselling patients about potential risks of PGD.

Prospective cohort study, Kaplan–Meier analysis was performed to estimate real (observed) and expected (calculated) cumulative delivery rates, and Cox proportional hazard regression analysis was used to assess the effect of age, number of cumulus oocyte complexes collected, fertility status, parity, genetic category and method of pituitary suppression.

Between 1993 and 2005, 2753 unselected consecutive cycles of ICSI and PGD were carried out in 1498 couples. The cumulative observed delivery rate overall per couple with a maximum of six treatment cycles of ICSI and PGD performed was 29%. The expected cumulative delivery rate (max six cycles) overall was 62%. There were no significant differences in cumulative delivery rates between the different genetic categories (i.e. availability of transferable embryos after PGD of 50 or 75%, chromosomal translocations or aneuploidy screening). The cumulative reproductive outcome in this PGD cohort was also not significantly affected by the fertility status of the couple, their parity or the method of pituitary suppression. However, the age of the patient and the number of oocytes contributed significantly to the reproductive results.

This prospective observational study demonstrates that age has a significantly negative effect on outcome of PGD, due to poor reproductive performance of female partners 40 years of age and older. The number of oocytes collected has a significant and independent effect. The other factors studied did not affect the cumulative reproductive outcome in this PGD cohort.

One hundred and seventy-three embryos diagnosed as chromosomally abnormal, 22 with no PGS result and four degenerated embryos originally diagnosed as normal were fixed and reanalysed by fluorescence in situ hybridization.

In total, 199 embryos were fixed, of which 166 were successfully reanalysed. One hundred and sixty embryos were found to be chromosomally abnormal; 48 of the reanalysed embryos with an initial diagnosis (149) had at least one cell with exactly the same chromosomal constitution shown in the first PGS analysis (34.2%). The reanalysis confirmed the initial overall chromosomally abnormal status of the embryo in 95.9% of the cases. Of all chromosomally abnormal embryos, 4.1% were diagnosed as false positive. The risk for false negative rate was at least 4.1%.

PGS seems to be a good method for selecting against chromosomally abnormal embryos but not for determining an embryo's exact chromosomal constitution.

We assessed presumed pathophysiologies in 74 consecutive POA patients, diagnosed based on elevated age-specific baseline follicle stimulating hormone and/or abnormally low anti-Müllerian hormone levels (<1.5 ng/ml). A genetic etiology was presumed with ≥34 triple CGG expansions on the FMR1 gene. An autoimmune etiology was assumed with at least one abnormality in a laboratory panel, involving antinuclear, antiphospholipid and thyroid antibodies, total immunoglobulin levels and anti-ovarian as well as anti-adrenal autoantibodies. A combined etiology was presumed with both autoimmune and genetic etiologies, and a patient was considered idiopathic when no abnormalities were found.

Twelve of 74 (16.2%) women demonstrated a genetic, 28 (37.8%) an autoimmune, 9 (12.2%) combined and 25 (33.8%) idiopathic etiologies.

Presumed underlying etiologies with POA follow a similar distribution pattern as reported for POF. POA and POF may, therefore, represent a continuum in phenotypical expression of different etiologies of premature ovarian senescence. Like POF, POA should be considered reason to investigate underlying etiologies.

Women pregnant between 16 and 32 weeks gestation participating in a retrospective time to pregnancy (TTP) study were each requested to recruit their eligible (on the basis of age, place of his birth and of his mother’s birth) male partner to complete additional questionnaires, have a physical examination and provide blood and two semen samples.

From 2061 women who completed the TTP questionnaire (response rate, 98%) there were 928 eligible male partners of whom 225 (24%) were responders. There were significant socio-demographic and self-reported exposure differences between responders and non-responders in particular, female professional occupation, knowledge of the fertile phase, pelvic inflammatory disease, non-smoker at time of conception and wine consumption per week were more frequent in the responders. There was no evidence of a bias for the subfertile being more likely to volunteer for the study. Mean TTP for planned pregnancies for responders and non-responders were 3.3 and 3.8 cycles (P = 0.319), respectively, and the cycle specific pregnancy rates were not significantly different after covariate adjustment by Cox regression.

The present study confirms that participation rates are low in studies of semen quality. Although the expected higher participation of subfertile couples was not confirmed, there remains considerable potential for bias and other problems that could invalidate this type of study.

In Experiment 1, DGC and Zeta methods were carried out on 60 unprocessed semen samples. The samples were then assessed by chromomycin A3 staining, acridine orange test, terminal deoxynucleotidyl transferase dUTP nick end labeling assay and the sperm chromatin dispersion test for protamine deficiency and DNA fragmentation. In Experiment 2, sibling oocytes from 30 ICSI candidates were divided into two groups; one group was inseminated with sperm processed by DGC and the second with sperm processed by DGC/Zeta. The outcomes of 30 ICSI cycles were compared between the two groups and also with 34 ICSI candidates whose oocytes were inseminated by DGC-processed sperm.

Both procedures were efficient for the recovery of sperm with normal protamine content and low DNA fragmentation. However, the Zeta method yielded a greater number of sperm with less DNA fragmentation. Fertilization and pregnancy rates were improved following the combined DGC/Zeta procedure. Compared with DGC alone, the pregnancy rate appeared improved but this was not statistically significant (P = 0.091).

Combining DGC and Zeta procedures improves the quality of semen samples which may increase fertilization rates and possibly pregnancy rates.

Immunofluorescence studies showed PLC to be localized in the equatorial region of sperm from fertile men, but sperm deficient in oocyte activation exhibited no specific signal in this same region. Immunoblot analysis revealed reduced amounts of PLC in sperm from infertile men, and in some cases, the presence of an abnormally low molecular weight form of PLC. In one non-globozoospermic case, DNA analysis identified a point mutation in the PLC gene that leads to a significant amino acid change in the catalytic region of the protein. Structural modelling suggested that this defect may have important effects upon the structure and function of the PLC protein. cRNA corresponding to mutant PLC failed to induce calcium oscillations when microinjected into mouse oocytes. Injection of infertile human sperm into mouse oocytes failed to activate the oocyte or trigger calcium oscillations. Injection of such infertile sperm followed by two calcium pulses, induced by assisted oocyte activation, activated the oocytes without inducing the typical pattern of calcium oscillations.

Our findings illustrate the importance of PLC during fertilization and suggest that mutant forms of PLC may underlie certain types of human male infertility.

We conducted a cross-sectional study, recruiting 209 young males from three communities in an endemic malaria area where DDT is sprayed annually. Blood plasma p,p'-DDT and its metabolite p,p'-DDE levels were measured and expressed as lipid adjusted p,p'-DDT and p,p'-DDE values. The sperm chromatin structure assay and Aniline Blue test were used to assess sperm DNA/chromatin integrity.

The lipid adjusted p,p'-DDT mean (±SD) and median concentrations were 109.2 (±106.6) and 83.9 µg/g, respectively; and the lipid adjusted p,p'-DDE mean (±SD) and median concentrations were 246.2 (±218.5) and 177.8 µg/g, respectively. The results point to a weak association between DDT/DDE plasma concentration and the incidence of sperm with chromatin defects.

The results suggest that non-occupational environmental DDT exposure may have a negative impact on sperm chromatin integrity in young South African males.

The genotypes of FAS, FASLG and CASP8 were determined in 620 infertile men. Sperm apoptosis rates were measured by terminal deoxynucleotidyl transferase (Tdt)-mediated dUTP nick end labelling (TUNEL) assay, and the semen quality analysis was performed using computer assisted sperm analysis.

We found that polymorphisms of FAS-670A/G (rs1800682: A>G) and CASP8-652 6N ins/del (rs3834129: –/CTTACT) were associated with sperm apoptosis and semen quality. Individuals with FAS-670GG showed low apoptosis rate and decreased sperm concentration, compared with the FAS-670AA genotype. Similarly, in comparison with the CASP8-652 6N ins/ins genotype, the CASP8-652 6N (ins/del+del/del) genotypes also showed significantly decreased sperm apoptosis rate and poor sperm motility. Other polymorphisms investigated did not appear to affect sperm apoptosis and semen quality.

This study showed for the first time that functional variants of FAS and CASP8 might contribute to the dysfunctional apoptotic mechanism in germ cells, which could result in poor semen quality of ejaculated sperm.

This study involved the follow-up by questionnaires and hospital records of patients with an ongoing pregnancy in a prospective, randomized, double-blind, placebo-controlled trial on the effect of low-dose aspirin during IVF. Aspirin treatment was continued throughout the first trimester of pregnancy. The primary end-point of this follow-up study was the incidence of pregnancy complications. The original trial is registered with the Dutch Trial Register and as an International Standard Randomized Clinical Trial, No. ISRNCTM97507474.

There were 54 patients who had ongoing pregnancies in the original trial; 90.7% returned the questionnaire and all Dutch hospital records were retrieved. A significant difference was found in the incidence of hypertensive pregnancy complications: 3.6% in the aspirin group and 26.9% in the placebo group (P < 0.05), resulting in numbers-needed-to-treat (NNT) of 10.3 to prevent hypertensive complications in one pregnancy after IVF treatment.

The incidence of hypertensive complications was significantly lower in the group of women treated with low-dose aspirin throughout IVF treatment and first trimester of pregnancy. These results suggest a potential benefit of low-dose aspirin during IVF and first trimester to prevent hypertensive pregnancy complications. The findings justify further investigation in placebo-controlled randomized trials.

Serum human chorionic gonadotrophin (hCG), progesterone, inhibin A and activin A levels were measured at 0 and 48 h. Differences in the mean levels and the change in levels over 48 h expressed as a ratio (48/0 h) were examined between the three outcome groups—failing PUL, intrauterine pregnancy (IUP) or ectopic pregnancy. Variables were incorporated into logistic regression models to predict the pregnancy outcomes, which were evaluated using receiver operator characteristic curves.

One hundred and forty-one women were classified as PULs: 67 failing PULs (47.5%), 58 IUPs (41.1%) and 16 ectopic pregnancies (11.4%). Activin A levels were not significantly different between the three outcome groups. Inhibin A levels were significantly lower in failing PULs. The logistic regression model based on serum inhibin levels gave an area under the curve (AUC) of 0.88 for failing PUL, 0.87 for IUP and 0.60 for ectopic pregnancy. The model based on serum activin levels gave an AUC of 0.61 for failing PUL, 0.64 for IUP and 0.51 for ectopic pregnancy, and the model based on serum hCG levels gave an AUC of 0.95 for failing PUL, 0.97 for IUP and 0.67 for ectopic pregnancy.

Serum activin A levels at 0 and 48 h are not helpful in predicting the outcome of PULs. Although serum inhibin A levels may be of use in the prediction of failing PULs and IUPs in the PUL populations, they do not perform as well as serum hCG levels.

We developed a simple and effective freezing/thawing method of single dissociated hESCs and hiPSCs in a feeder-free culture in the presence of Rho-associated kinase (ROCK) inhibitor Y-27632.

Exposure to ROCK inhibitor Y-27632 in freezing solution alone does not significantly enhance the post-thaw survival rate of single dissociated hESCs and hiPSCs. However, when ROCK inhibitor was added to both pre- and post-thaw culture media, there was an enhancement in the survival rate, which further increased when ROCK inhibitor was added to Matrigel as well. Under these treatments, hESCs and hiPSCs retained typical morphology, stable karyotype, expression of pluripotency markers and the potential to differentiate into derivatives of all three germ layers after long-term culture.

This method is an effective cryopreservation procedure for single dissociated hESCs in feeder-free culture, which is also applicable for single dissociated hiPSCs using a ROCK inhibitor. The cloning efficiency of hiPSCs and hESCs improves when ROCK inhibitor is added both in Matrigel and in medium in comparison with conventional addition to medium. Therefore, we believe this method would be useful for current and future applications of the pluripotent stem cells.

hESC were derived and cultured on mouse fibroblasts in KO–SR containing medium (serum free media) and passaged mechanically. Our results based on analysis at mRNA (RT–PCR) and protein (fluorescence-activated cell sorting and immunocytochemistry) level show that CD30 is expressed in undifferentiated hESC, even at very early passages, without any correlation with the presence of chromosomal anomalies. We also show that the expression of CD30 is rapidly lost during early spontaneous differentiation of hESC.

We conclude that CD30 expression in hESC cultures is probably a consequence of culture conditions, and that KO–SR may play a role. In addition, the expression of so-called ‘stemness’ markers does not change in undifferentiated hESC during long-term culture or when cells acquire chromosomal abnormalities.

Chromosomal dynamics during the first mitotic division of mouse embryos were analyzed using a new live-cell imaging technology. After imaging, the embryos' developmental capacities were determined.

When ICSI-generated embryos were monitored for their chromosome integrity, some embryos with apparent normal morphology seen by conventional light microscopy had abnormal chromosome segregation (ACS) at the first mitotic division. Chromosomal fragments were misaligned during the first metaphase and formed micronuclear-like structures at the interphase of the 2-cell stage. Similar ACS was also found in mouse embryos produced by microinjecting round spermatids, with even higher frequency. Giemsa staining and immunostaining revealed that these fragments were derived from double-strand DNA breaks in the paternal genome. About half of the embryos with ACS developed into normal-looking morulae or blastocysts and implanted, but almost all of them aborted spontaneously before embryonic day 7.5.

ACS during first mitosis appears to be a major cause of early pregnancy losses in ICSI-generated mouse embryos. Moreover, this novel imaging technology could be applicable as a method for the assessment of embryo quality.

Pregnancy outcome in women after ovum donation aged 40 and above was extracted. Labor and delivery data as well as antenatal records of women carrying twins were compared with those of singletons, as well as to a control group of all twin pregnancies delivered at Sheba Medical Center during 2007.

One hundred and twenty-five women after ovum donation aged ≥40 were studied. Of those, 42 women carried twin pregnancies and 83 carried singletons. The 42 women carrying twins comprised the study group and were compared with 417 control women with twins. Mean maternal age was 49.2 ± 4.3 years. Hypertensive complications (50%), diabetes in pregnancy (31%) and hospitalization in pregnancy (69%) were all extremely high in the study group. Mean gestational age at delivery was lower for the study group compared with controls (35.2 ± 2.3 versus 35.7 ± 2.6 weeks), with 35.7% of infants in the study group born ≤34 weeks gestation compared with 21.8% of controls, (OR: 1.99, 95% CI: 1.02–3.89). Mean birthweight was also significantly lower for study group infants compared with controls, with 77% of study infants born <2500 g compared with only 60% of controls (OR: 2.22, 95% CI: 1.3–3.77).

Pregnancy in advanced maternal age women after ovum donation carrying twins is associated with significant maternal and fetal complications, with increased risks of prematurity and lower birthweight. Possibly, the aged uterus is less suitable for carrying a multifetal pregnancy than a younger uterus. Therefore, the alternative of transferring a single, good-quality embryo should be the preferred option.

A comprehensive literature search was conducted to identify all the English language published observational and randomized studies evaluating the efficacy of medical treatments on pain associated with rectovaginal endometriosis. A combination of keywords was used to identify relevant citations in PubMed, MEDLINE and EMBASE.

A total of 217 cases of medically treated rectovaginal endometriosis were found; 68 in five observational, non-comparative studies, 59 in one patient preference cohort study, and 90 in a randomized controlled trial. An aromatase inhibitor was used in two of the non-comparative studies, vaginal danazol in one, a GnRH agonist in one, and an intrauterine progestin in one. Two estrogen–progestin combinations used transvaginally or transdermally were evaluated in the patient preference study, whereas an oral progestin and an estrogen–progestin combination were compared in the randomized controlled trial. With the exception of an aromatase inhibitor used alone, the antalgic effect of the considered medical therapies was high for the entire treatment period (from 6 to 12 months), with 60–90% of patients reporting considerable reduction or complete relief from pain symptoms.

Despite problems in interpretation of data, the effect of medical treatment in terms of pain relief in women with rectovaginal endometriosis appear substantial.

FINHYST 2006 is a national prospective hysterectomy study in which all hospitals collaborated from 1 January to 31 December 2006. Questionnaires, completed by gynaecologists, covered their experience, patient characteristics and surgical data.

The 5279 hysterectomies distributed by approaches were 44% VHs, 32% LHs and 24% AHs. Less than 2% were subtotal. The main indications for hysterectomy were myomas (33%), uterine prolapse (28%) and menorrhagia (21%). The main indication for VH was not related to uterine prolapse in 39%. Bilateral salpingo-ooforectomy was performed in 36% of AHs, 32% of LHs and 2% of VHs. Antibiotic prophylaxis was used in 97%, and thrombosis prophylaxis in 65%. Haemorrhage was least and operation time shortest with VH, and hospital stay and sick leave were shortest after LH.

In Finland, less invasive approaches comprise 76% of hysterectomies. This trend has resulted nationally in shortening of hospital stay and of convalescence time.

Multicentre randomized controlled trial (RCT) including 228 ovulatory patients scheduled for COS and IUI. Patients were randomized to ‘individual’ (50–100 IU rFSH/day, n = 113) or ‘standard’ (75 IU rFSH/day, n = 115) dose. ‘Individual’ dose was prescribed according to the nomogram, which was based on patients' body weight and antral follicle count. The primary end-point was the proportion of patients with two to three follicles ≥14 mm (maximum two follicles ≥18 mm) on the day of hCG (leading follicle = 18 mm). Primary analysis was made by intention-to-treat.

In the ‘individual’ group, 79/113 (70%) of the patients developed two to three follicles versus 64/115 (56%) in the ‘standard’ group [absolute difference = 14.3 percentage points; 95% confidence interval (CI) 2–26, P = 0.03; absolute difference = 14.4; 95% CI 2–27, P = 0.02, when adjusting for centre]. Among patients with two to three follicles, the proportion of patients with two follicles was 46/79 (58%) in the ‘individual’ group versus 34/64 (53%) in the ‘standard’ group, P = 0.54. Ongoing pregnancy rate was 23/113 (20%) in the ‘individual’ group and 21/115 (18%) in the ‘standard’ group and the rate of multiple gestations was 1/113 (1%) versus 5/115 (4%), P = 0.21.

This RCT is the first to clinically test a dosage nomogram in ovulatory IUI patients' first rFSH treatment cycle. Dosing according to the nomogram was superior to standard dosing.

Trial registration: ClinicalTrials.gov Identifier NCT00374634.

Human secondary follicles were isolated from ovarian tissues obtained from cancer patients and grown in vitro within a bio-engineered culture system for 30 days.

Human ovarian follicles became steroidogenically active, and developed from the early secondary to antral stage in vitro. The follicles contained healthy, growing oocytes that were connected by transzonal projections between the somatic cells and oocyte.

Our data support the notion that human follicle development can be achieved in vitro in a bio-engineered culture system. More studies are required to investigate whether the fully sized oocytes obtained from in vitro grown follicle are competent to resume meiosis and be fertilized.

Aiming to identify global gene profile in RIF patients, gene-array analyses were performed on endometrial samples collected on day 21 of the cycle from fertile women (n = 12) and from RIF patients (n = 20). Validation of cyclin E2, Slug, dickkopf homolog 1 (DKK1), lymphoid enhancer-binding factor 1 (LEF1) and secreted frizzled-related protein 1 (SFRP1) was carried out by real-time PCR.

Gene-array analysis revealed 313 genes exhibiting modified expression levels in RIF patients. Of these, 288 genes (92%) were down-regulated and only 25 genes (8%) were up-regulated. Classification of the down-regulated genes to biological pathways revealed cell cycle, Wnt signaling and cellular adhesion pathways. Real-time PCR validation of cyclin E2, SFRP1 and LEF1 showed significantly lower expression levels in RIF–IVF patients as compared with fertile women. In addition, two up-regulated genes, Slug and DKK1, were also validated. Interestingly, about 8% of the down-regulated genes were estrogen-dependent. Western blot of estrogen receptor revealed low expression of this protein in the RIF group.

The evaluation of the endometrium of RIF patients by gene array analysis demonstrates that the expression of various genes is altered, including those belonging to the cell cycle, Wnt signaling and cellular adhesion pathways.

Women attending their first ICSI cycle at ANDROS Day Surgery for severe male factor infertility were included in the study. An endometrial biopsy was performed during the WOI, in one of the last two cycles before the ICSI cycle. Forty-seven selected gene profiles were analyzed using Low Density Array technology. Only biopsies from women who subsequently became pregnant were evaluated, to exclude any bias regarding embryo viability and embryo transfer difficulties.

Fifteen patients were included in the analysis as they became pregnant after ICSI procedure. Four of 47 selected genes were excluded from the analysis. Of the 43 genes analyzed, only 6 genes (VEGFA, PLA2G2A, ALPL, LIF, NNMT and STC1) showed a statistically uniform expression among patients who subsequently became pregnant. For all the other genes analyzed there were considerable differences in their expression levels amongst women who subsequently became pregnant.

Our data suggest that very few genes, which change their expression level during the WOI, show a quantitative homogeneous expression in endometrially-receptive patients. In conclusion, in this study only six genes showed a homogeneous expression, and are probably involved in embryo implantation mechanisms.

Twenty-nine human first-trimester ovaries from legal abortions [aged 38–64 days post-conception (p.c.)] were collected. Mothers filled out a questionnaire about their smoking habits and delivered a urine sample for cotinine analysis. The ovarian cell numbers were estimated using stereological methods.

A non-linear correlation between the numbers of oogonia and somatic cells in relation to age of the embryo/fetus was shown in 28 ovaries, including the first estimates performed in ovaries younger than 47 days p.c. Prenatal exposure to smoke showed a significant decrease in the number of somatic cells (P ≤ 0.01). The number of oogonia was not significantly associated with prenatal exposure to maternal smoking (P ≤ 0.09). The ratio between the two cell types decreased considerably from 1:45 to 1:23 from 38 to 46 days p.c. and was not affected by smoking.

Oogonia proliferate and/or invade the developing ovary at a much faster relative rate than somatic cells. In utero exposure to maternal smoking significantly reduces the number of somatic cells from Days 38 to 64 p.c. Since oocytes cannot survive without being enclosed by somatic cells in a follicle, reduction in the somatic cells number may have long-range consequences on the number of oocytes available in adult life and on the future fertility of female offspring exposed to smoking in utero.

We established a conditionally immortalized human foreskin fibroblast line that secreted basic fibroblast growth factor (bFGF). These cells were used as feeder cells for hESC culture and induced pluripotent stem (iPS) cell derivation and expansion. This conditional immortalization was performed using lentiviral vector (LV) mediated transduction of Bmi-1 and human telomerase reverse transcriptase genes and the resulting cell line was further modified by LV-mediated transduction of a secreted form of bFGF gene product. Three different laboratories have tested whether this feeder cell line could support the maintenance of four different hESC lines.

Immortalized fibroblasts secreting stable amounts of bFGF supported the growth of all hESC lines, which remained pluripotent and had a normal karyotype for at least 10 passages. Even at high passage (p56), these modified cells, when used as feeders, could support iPS derivation and propagation. Derived iPS cells expressed pluripotency markers, had hESC morphology and produced tissue components of the three germ layers when differentiated in vitro.

These modified fibroblasts are useful as a genetically-defined feeder cell line for reproducible and cost-effective culture of both hESC and iPS cells.

Since it is not possible to collect FTs from women carrying a healthy pregnancy, we studied tissue collected at the time of hysterectomy for benign disease. Women were injected with hCG in the days leading up to hysterectomy, and pseudopregnancy confirmed by the presence of high serum progesterone levels and the decidualization of the endometrium. FTs from the different stages of the menstrual cycle (n = 24), tubes bearing an ectopic pregnancy (n = 15) and pseudo-pregnant tubes (n = 6) were collected and examined using immunohistochemistry and quantitative RT–PCR.

MUC1 was present at the apical surface of the tubal epithelial cells throughout the menstrual cycle, but intracellular localization was minimal in the follicular phase, increasing to a maximum in the luteal phase. MUC1, including the glycoform recognized by antibody 214D4, was found at the apical surface of tubal epithelium in both the ectopic and pseudo-pregnant groups and the intracellular expression was much stronger in the pseudo-pregnant group than in the ectopic group. The 214D4 epitope was absent from tubal tissue adjacent to ectopic implants.

The decrease in MUC1 expression and altered glycosylation in tubal epithelium from ectopic pregnancy may reflect an increase in receptivity.

We analyzed sperm with flat and round heads from infertile patients with globozoospermia (n = 1) and teratozoospermia (n = 1), and also from GOPC^–/– mice, an animal model of human globozoospermia. Differential interference contrast image analysis, immunocytochemistry with MN13 antibody, transmission electron microscopy and an oocyte activation assay (assessing pronucleus formation) with ICSI were used.

Flat-headed (control) sperm from both a healthy fertile volunteer man and wild-type mice had MN13 and PAS (PT). Flat-headed sperm (<5% of the population) from GOPC^–/– mice also had both MN13 and PAS (PT), and they showed high oocyte activation ability. In contrast, round-headed sperm from a globozoospermia patient (100%) and GOPC^–/– mice (>95% of the population) had neither MN13, nor PAS (PT), nor oocyte activation ability. Oocyte activation was higher in flat- versus round-headed sperm from GOPC^–/– mice (P < 0.05).

Oocyte activation ability may be related to sperm head flatness and presence of MN13 and PAS (PT) in human and mouse sperm. This information is a first step towards the possibility of selecting good-quality sperm with high oocyte activation ability for ICSI.

Two hundred and thirty-seven women with surgically or histologically diagnosed endometriosis of stages III and IV were recruited for this study, and 164 patients with no evidence of endometriosis diagnosed by laparoscopy or laparotomy served as controls. The C627T polymorphism of the IL-2R β was assessed using the TaqMan allelic discrimination assay. ² analysis was used to examine any differences in genotype distributions and allele frequencies of the IL-2R β C627T polymorphism between the endometriosis cases and the controls.

There was no statistically significant difference in the frequency of the IL-2R β C627T polymorphism between the endometriosis patients and the controls (28.7% C/C, 48.1% C/T and 23.2% T/T versus 29.3, 44.5 and 26.2%, respectively, P = 0.72) or in the T allele frequencies (47.3 versus 48.5%, respectively, P = 0.73). Even when the endometriosis cases were subdivided into stages III and IV, no statistically significant differences in genotype distributions or allele frequencies were observed among the three groups.

Contrary to the recent data reported in a Taiwanese population, our results suggest that the C627T polymorphism of the IL-2R β gene may not be associated with the risk of endometriosis in the Korean population.

Women undergoing laparoscopic excision of ovarian endometrioma (n = 52) or other benign ovarian cysts (n = 52) were studied, plus women with non-ovarian endometriosis (n = 11) and healthy controls (n = 27). Serum was collected from all subjects, and peritoneal and cystic fluid from a subset with endometrioma. Follistatin was measured by enzyme-linked immunosorbent assay. The diagnostic accuracy of follistatin to detect endometrioma was evaluated by receiver operating characteristic (ROC) curve and compared with cancer antigen (CA)-125.

Serum follistatin was increased in women with ovarian endometrioma (2080 ± 94 pg/ml) compared with controls (545 ± 49 pg/ml, P < 0.001), other benign ovarian cysts (795 ± 60 pg/ml, P < 0.001) or non-ovarian endometriosis (1271 ± 115 pg/ml, P < 0.001). Cystic fluid showed a higher concentration of follistatin (9850 ± 4461 pg/ml) than peritoneal fluid (1885 ± 261 pg/ml, P < 0.001) and serum (P < 0.001). Follistatin levels detected 48/52 cases of endometrioma (92% sensitivity) at 1433 pg/ml cut-off, corresponding to 92% specificity. CA-125 detected only 44% of endometriomas with 90% specificity. ROC curve comparison showed follistatin was more accurate than CA-125 to discriminate women with endometrioma either from controls or women with other benign ovarian cysts (P < 0.0001).

Serum follistatin is increased in women with endometriosis and allows clear distinction between endometrioma and other benign ovarian cysts. Follistatin has the sensitivity and specificity to become a useful clinical marker of ovarian endometrioma.

Endometrial biopsies were collected at days 2 (pre-receptive) and 7 (receptive) after the urinary luteal hormone surge in the same menstrual cycle from eight fertile women (corresponding to days 16 and 21 of the menstrual cycle). Proteins were extracted and labeled with CyDye DIGE fluorofores and separated using two-dimensional gel electrophoresis.

Image analysis using the DeCyder^TM software followed by protein identification by matrix-assisted laser desorption/ionization-MS and database searching revealed 32 differentially expressed proteins, although only annexin A2 and stathmin 1 were consistently regulated in the two experiments performed. Validation and localization of annexin A2 and stathmin 1 were performed by western blot and immunohistochemistry. Annexin A2 and stathmin 1 were investigated using an endometrial refractoriness model. The results highlight the key potential of these proteins as possible targets for human endometrial receptivity and interception.

This study shows that the human endometrium has a differential proteomic repertoire during the window of implantation. Consequently, we identified annexin A2 and stathmin 1 as differentially expressed molecules in the receptive endometrium.

Two weeks or 1 year after OVX, rats were injected over 3 days with 125 µg/kg of estradiol benzoate (EB), 7.5 mg/kg of the selective ER agonist propylpyrazole triol (PPT), or 15 mg/kg of the selective ER modulator tamoxifen (TX). Controls were given 0.2 ml oil. The last day of ER analog treatment, half of the rats in each group received 25 mg/kg of progesterone (P). The next day, anterior pituitaries were removed and analyzed for PR-AB mRNA and protein. Gonadotrophin secretion in incubated pituitaries was also measured.

(i) PR mRNA expression was higher in young than in middle-aged OVX rats although PR protein was absent in pituitaries from both groups of OVX rats; (ii) activation of ER reduced gonadotroph hypertrophy and increased PR mRNA and protein expression (EB > PPT > TX) more efficiently in young than in middle-aged rats, (iii) ER agonists elicited GnRH-stimulated LH and FSH secretion in young but only FSH secretion in middle-aged OVX rats, (iv) evaluated by peak LH concentrations, GnRH self-priming was observed in both groups of OVX rats and (v) P down-regulated PR protein expression in young, and to a lesser extent, in middle-aged OVX rats, in close association with PR-dependent GnRH self-priming.

Middle-aged OVX rats exhibited clear-cut LH, but not FSH, secretory defects in pituitary sensitivity to estrogen and P.

We examined the association between maternal education, BMI, smoking and offspring's birthweight among women who delivered at term in the Danish National Birth Cohort (n = 75 085).

Compared with mothers with more than 12 years of education, women with 10–12 years of education had babies that were 12 (4–19) g lighter. Mothers with 9 years of education had babies that were 51 (95% CI; 39–62) g lighter. BMI and smoking affected the association between maternal education and birthweight, albeit in different directions. If all mothers had the BMI of the highest educated mothers, the deficits would be larger: –20 (–22 to –19) and –74 (–80 to –68) g, respectively. If all mothers smoked like the highest educated mothers, mothers with a shorter education would have the heaviest babies: the difference would be 9 (2–16) and 23 (11–36) g, respectively. In combination, smoking and BMI all but explained the educational gradient in birthweight at term.

Maternal smoking and BMI are important intermediates of the educational gradient in birthweight at term. As the prevalence of smoking is dropping and the prevalence of obesity is increasing the educational gradient will likely reverse, but it seems unlikely that this will translate into a health advantage for the children of the least educated mothers.

This study analyzed data from a population-based sample of 443 hairdressers and 508 women in other occupations, who responded to a mailed survey. POF was assessed in all eligible participants by self-report of a doctor's diagnosis.

Among 443 hairdressers and 508 women in other occupations, 14 (3.2%) and 7 (1.4%) developed POF, respectively. A non-significant increase in the risk of POF was observed among hairdressers compared with non-hairdressers (adjusted relative risk (RR) 1.90; 95% confidence interval (CI) 0.76, 4.72). When limited to Caucasian women only (~85% of respondents), the increased risk was statistically significant (RR 3.24; 95% CI 1.06, 9.91). Among Caucasian women of 40–55 years of age, hairdressers were more than five times as likely to report POF compared with non-hairdressers (RR 5.58; 95% CI 1.24, 25.22).

Hairdressers may be at increased risk for POF compared with women employed in other occupations.

From January 2006, all couples who had two or more transferable PGD blastocysts biopsied on Day 3 of culture were offered single-blastocyst transfer (SBT) and cryopreservation of surplus blastocyst(s) using a slow-freezing technique. We compared the outcome of 32 cryo-thawed PGD cycles with that of 191 cryo-thawed conventional in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles performed between January 2006 and July 2008. We also compared the outcome of all fresh PGD cycles performed before and after January 2006.

The cryo-thawed blastocyst survival rate was similar between the PGD and IVF/ICSI groups (87% versus 88%, P = 0.94). There was no significant difference in the implantation and clinical pregnancy rates between the two groups (35% versus 29%, P = 0.45 and 34% versus 36%, P = 0.77, respectively). During the same period, the multiple pregnancy rate in the fresh PGD programme dropped from 36% to 10% (OR = 0.20, 95% CI 0.08–0.48, P < 0.001) with no reduction in pregnancy rates.

The survival and implantation potential of biopsied PGD embryos cryopreserved at the blastocyst stage is comparable to that of non-biopsied IVF/ICSI cryopreserved blastocysts. Elective SBT and cryopreservation of surplus blastocysts for later use should extend to include PGD for inherited genetic disorders.

Triplicate sets of pooled metaphase II oocytes or blastocysts were processed for analysis using the Human Genome Survey Microarrays V2.0 (Applied Biosystems).

Of 154 DNA repair genes investigated, 109 were detected in blastocysts and 107 in oocytes. Among differentially expressed DNA repair genes, 40/55 (73%) had lower expression levels in blastocysts compared with oocytes (P < 0.05, fold change >3).

Despite experimental limitations due to culture or freezing and thawing of samples, large numbers of repair genes were detected indicating that all DNA repair pathways are potentially functional in human oocytes and blastocysts. The higher mRNA level for most repair genes in oocytes compared with blastocysts ensures sufficient availability of template until embryonic genome activation.

This study consisted of 299 women with advanced stage endometriosis and 459 control women without endometriosis in a Korean population. Genotyping results of the Glu298Asp polymorphism of the NOS3 were analyzed between the endometriosis and control group.

The genotypic frequencies were significantly different between women with and without endometriosis. The frequency of the non-GG genotype (GT + TT) was higher in the endometriosis group than in the control group (P = 0.001).

These findings suggest that the T allele, encoding aspartic acid, of the Glu298Asp polymorphism of the NOS3 may be associated with advanced stage endometriosis in the Korean population.

This study comprised 344 North Chinese women with endometriosis and 360 healthy women without endometriosis recruited as control. Genotyping of the VEGF gene polymorphisms at –460C/T, –1154G/A, –2578C/A and +936C/T were performed by PCR and restriction fragment length polymorphism analysis.

No significant difference was found in allele and genotype distributions of the –460C/T, +936C/T polymorphisms between patients and controls. However, the frequencies of –1154G/A, –2578C/A genotype and allele were significantly different between the two groups (all P-value <0.013). The –2578A/A, –1154A/A genotypes were found less frequently in patients with endometriosis compared with controls. The haplotype distributions derived from three polymorphisms (–2578C/A, –1154G/A, –460C/T) differed between the two groups (P = 0.000).

The VEGF–460/–1154/–2578 TGC, CAA, TAA and TAC haplotypes were associated with endometriosis. The –1154A and –2578A alleles may be protective against the development of endometriosis in North Chinese women.

We used a dual approach to review the effect of the gr/gr deletion on semen quality. First, we conducted a systematic review and meta-analysis of previous association studies, to compare the prevalence of gr/gr deletions between azoo-/oligozoospermic men and normozoospermic men. Secondly, we studied a cohort of 1041 male partners of subfertile couples unselected for semen quality. We employed a cross-sectional design by screening all men for the gr/gr deletion and comparing the semen quality of men with and without the gr/gr deletion.

Seven studies were included in the meta-analysis. The gr/gr deletion was significantly more prevalent among azoo-/oligozoospermic men than among normozoospermic men (OR 2.4, 95% CI 1.75–3.30). In our cohort, 25 men carried a gr/gr deletion. Men with this genotype had a lower sperm concentration (median 34 x 10⁶/ml versus 53 x 10⁶/ml, P = 0.017), total sperm count (median 108 x 10⁶ versus 152 x 10⁶, P = 0.006) and total motile sperm count (median 20 x 10⁶ versus 50 x 10⁶, P = 0.010) than men without the gr/gr deletion.

Y chromosome gr/gr deletions significantly reduce sperm counts and are thus associated with low semen quality.