PLCζ disruption with complete fertilization failure in normozoospermia.

PURPOSE: Intracytoplasmic sperm injection (ICSI) is widely used to achieve fertilization in the presence of severe male factor, resulting in high fertilization rates. Nevertheless, 1-3 % of couples experience complete fertilization failure after ICSI. When a male factor is identified, assisted oocyte activation (AOA) can help overcome fertilization failures. The objective of this study is to describe a case of repeated complete fertilization failures after ICSI with donor oocytes, and to investigate the molecular and functional aspects of phospholipase C zeta (PLCζ) protein in the patient semen. METHODS: The patient was a normozoospermic male who had previously fathered, through natural conception, four children by a different partner. Molecular and functional analysis of sperm-specific PLCζ in the patient and control samples by means of gene sequencing, immunocytochemistry, Western blot, mouse oocyte activation test (MOAT), and mouse oocyte calcium analysis (MOCA) were used. RESULTS: PLCζ expression levels and distribution were significantly disrupted, although MOAT and MOCA did not indicate a decrease in activation ability. CONCLUSIONS: Normozoospermic males can have disrupted expression and distribution of PLCζ, and reduced activation ability after ICSI in human oocytes, despite their normal activation potential in functional testing using mouse oocytes. Discrepancy among molecular and functional data might exist, as mutations in the gene sequence may not be the only cause of alteration in PLCζ protein related to activation failures.

Individualized embryo transfer training: timing and performance.

STUDY QUESTION: How long is the individualized training and the stability of competence for the embryo transfer (ET) technique? SUMMARY ANSWER: The embryo transfer technique is easy-to-learn, hardly unlearned, and training should be individualized by monitoring with learning curve-cumulative summation (LC-CUSUM) curves. WHAT IS KNOWN ALREADY: Like many medical procedures, embryo transfer is an operator-dependent technique. Individualized or standardized training of these medical procedures should be monitored to determine when competence is acquired. STUDY DESIGN, SIZE, DURATION: This prospective, monocentric study involving five embryo transfer trainees was carried out between August 2011 and November 2012. PARTICIPANTS/MATERIALS, SETTING, METHODS: The study was carried out in a large private clinic. Five gynaecologist trainees during their first year of assisted reproduction subspecialty performed embryo transfer for patients undergoing either fresh IVF, oocyte donor IVF, or frozen embryo transfer. There were 586 embryo transfers performed in 96 sessions of 3-10 embryo transfers each. An embryo transfer was considered successful if it gave rise to a positive pregnancy test 14 days later. LC-CUSUM and cumulative summation (CUSUM) curves were used to determine when competence was acquired and whether it was maintained over time, respectively. The length of time between two consecutive sessions was assessed for an effect on consolidation of the acquired competence. MAIN RESULTS AND THE ROLE OF CHANCE: We observed that all five trainees became proficient in embryo transfer by procedure 15 (after procedure 15, 9, 7, 13 and 9, respectively). Once competence was achieved, one of the five trainees showed a loss of proficiency. After having acquired competence, the median pregnancy rate per embryo transfer session was significantly lower when the interval between consecutive embryo transfer sessions was ≥10 days compared with <10 days (20.0 versus 46.7%; P = 0.006). LIMITATIONS, REASONS FOR CAUTION: The patient groups included in the study were heterogeneous (IVF, oocyte donor IVF and frozen embryo transfer) and their outcomes are very variable; thus the distribution and proportion of these groups can determine the timing of competence acquisition. Our data show that low numbers of embryo transfer are needed to acquire competence, but since a relative high percentage of embryo transfers in our practice are from oocyte donor IVF, extrapolation of the findings to other clinical context should be done with caution. WIDER IMPLICATIONS OF THE FINDINGS: Personalized embryo transfer training is feasible and useful, allowing clinics, on one hand, to offer a maximum chances of pregnancy with fully trained personnel, and the other hand, to avoid the superfluous and costly overtraining of already proficient trainees. Furthermore, it is advisable to maintain a short interval of time between consecutive embryo transfer sessions after a trainee has acquired competence, to avoid a significant drop in the resulting pregnancy rate. STUDY FUNDING/COMPETING INTEREST(S): This work was supported in part by funding from Fundació Privada EUGIN. There are no conflicts of interest to declare.

Transvaginal versus transabdominal ultrasound guidance for embryo transfer in donor oocyte recipients: a randomized clinical trial.

OBJECTIVE: To compare pregnancy and implantation rates with transvaginal (TV) versus transabdominal (TA) ultrasound-guided embryo transfer (ET). DESIGN: Randomized, clinical trial registered at clinicaltrials.gov (NCT 01137461). SETTING: Private, infertility clinic. PATIENT(S): Three-hundred thirty randomized recipients of donor oocytes. INTERVENTION(S): Embryo transfer using TV (with empty bladder, using the Kitazato ET Long catheter) versus TA ultrasound guidance (with full bladder, using the echogenic Sure View Wallace catheter). MAIN OUTCOME MEASURE(S): Overall pregnancy, clinical pregnancy, implantation, and ongoing pregnancy rates. Duration and difficulty of ET. Patient-reported uterine cramping and discomfort, as evaluated by questionnaire. RESULT(S): No statistically significant differences were observed in clinical pregnancy 50.9% versus 49.4% (95% confidence interval of the difference: -9.2 to +12.2%), implantation 34.5% versus 31.4% (95% CI of the difference: -4 to +10.3%) between the TV and TA ultrasound-guided groups. Transfer difficulty (6% versus 4.2%) and uterine cramping (27.2% versus 18.3%) were not statistically significantly different between treatment groups. Total duration (154±119 versus 85±76 seconds) was statistically significantly higher in the TV ultrasound group. Light to moderate-severe discomfort related to bladder distension was reported by 63% of the patients in the TA ultrasound group. CONCLUSION(S): Transvaginal ultrasound-guided ET yielded similar success rates compared with the TA ultrasound-guided procedure without requiring the assistance of a sonographer. It was associated with increased patient comfort due to the absence of bladder distension.

Triggering with HCG or GnRH agonist in GnRH antagonist treated oocyte donation cycles: a randomised clinical trial.

AIM: To compare donor and recipient outcome after inducing the final oocyte maturation with hCG or GnRH agonist in GnRH-antagonist treated oocyte donation (OD) cycles. METHODS: Two-hundred fifty-seven oocyte donors were enrolled to participate in a clinical trial in a private fertility centre. After stimulation with 225 IU rFSH and Cetrorelix 0.25 mg/day, 212 oocyte donors were randomised with sealed envelopes for triggering with recombinant hCG (Ovitrelle 250 microgr, n = 106) or a GnRH agonist (triptorelin 0.2 mg, n = 106). RESULTS: The number of retrieved COCs (12 +/- 6.3 vs 11.4 +/- 6.4), mature oocytes (8 +/- 4.6 vs 7.5 +/- 4.1), the proportion of mature oocytes (67.2 +/- 20.4% vs 67.1 +/- 20.9%) and fertilisation rates (67.8 +/- 23.5% vs 71.1 +/- 22.1%) were comparable. Clinical, ongoing pregnancy and live birth rates were not statistically different in the corresponding recipient groups. Nine cases of mild and one case of severe OHSS occurred in hCG group, whereas no cases were detected in GnRH agonist group. CONCLUSIONS: The findings of our RCT suggest that donor and recipient outcome are comparable in OD cycles triggered with hCG or a GnRH agonist. Furthermore, the risk of OHSS seems to be reduced considerably, therefore the combination of a GnRH antagonist protocol with GnRH agonist triggering constitutes a safe treatment option for egg-donors.

Endometrial receptivity after oocyte donation in recipients with a history of chemotherapy and/or radiotherapy.

INTRODUCTION: Information is scarce regarding the outcome of oocyte donation (OD) in patients with a history of cancer treatment. Therefore, we conducted a matched controlled analysis on the outcome of OD in these recipients. METHODS: Between January 2000 and November 2005, 33 patients with a history of chemotherapy and/or radiotherapy had an OD cycle. Matching was performed to the chronologically closest patient without a history of cancer therapy by number of days of hormonal stimulation before embryo replacement, number of replaced embryos, day of embryo transfer and origin of sperm. RESULTS: The primary diseases of the patients were Hodgkin's lymphoma (n = 12), non-Hodgkin's lymphoma (n = 3), leukaemia (n = 7), ovarian cancer (n = 6), Ewing's sarcoma (n = 2), breast cancer (n = 1), sympathoblastoma (n = 1) and histiocytosis X (n = 1). Twenty-three patients had undergone chemotherapy and radiotherapy, nine patients chemotherapy only and one radiotherapy only. The mean age of the recipients was 33.1 years [95% confidence interval (CI) 30.9-35.3] and 39.6 (95% CI 37.1-42.1) in the study and control groups, respectively. The average number of received oocytes and transferred embryos, was similar in both groups. Nineteen (57.6%) versus 13 (39.4%) pregnancies resulting in an ongoing pregnancy (i.e. viable at 12 weeks) in 15 (45.4%) versus 9 cycles (27.3%) (NS) were obtained in study and control groups, respectively. Implantation rate in study and control groups was 35.8 versus 17.9%, respectively (P = 0.02). CONCLUSIONS: The results suggest that patients with a history of cancer treatment have a pregnancy rate after OD similar to that in the general population of oocyte recipients.

Prognostic factors in oocyte donation: an analysis through egg-sharing recipient pairs showing a discordant outcome.

OBJECTIVE: To analyze prognostic factors that are associated with a discordant outcome in egg recipients sharing oocytes from the same donor. DESIGN: Matched case-control single-center study. SETTING: Private infertility clinic. PATIENT(S): Four hundred forty-four recipients (222 pairs) sharing oocytes from the same donor and showing a discordant outcome. INTERVENTION(S): Controlled ovarian hyperstimulation of egg donors, oocyte donation, intracytoplasmic sperm injection, and ET in egg recipients. MAIN OUTCOME MEASURE(S): Recipient age, obstetric (gravidity, parity) and gynecologic variables (previous uterine surgery, uterine fibroids, uterine malformations, endometriosis, history of tubal infertility), previous oocyte donation cycles, duration of E(2) replacement, received cumulus-oocyte complexes, mature (MII) oocytes, fertilized oocytes, transferred embryos, mean embryo score, transfer difficulty, and semen parameters. RESULT(S): No significant differences were found in the above-mentioned prognostic factors between the study and control groups. CONCLUSION(S): Recipient- and cycle-related prognostic factors investigated in our study were not associated with a discordant outcome in recipient pairs sharing oocytes from the same donor. Other possible prognostic factors involving oocyte donor heterogeneity, embryo aneuploidy rates, male factor infertility, and endometrial receptivity should be further investigated.

Influence of high-intensity training and of dietetic and anthropometric factors on menstrual cycle disorders in ballet dancers.

Background. Intensity of exercise and low energy consumption, specific type and amount of training, early age at initiation, previous menstrual dysfunctions, low body mass index (BMI) or percentage body fat, pathological feeding habits and psychological stress have been suggested as potential factors accountable for menstrual irregularities in female athletes.Aim. To evaluate the influence of intensive training and of dietetic and anthropometric factors on menstrual cycles in female ballet dancers.Method. A case-control study, in which a structured interview and physical examination were carried out in two groups of teenagers aged between 12 and 18 years. The study included a total of 115 adolescent girls distributed in two groups: dancers (group B, n = 38) and girls of the same age not engaged in any sports activity (group C, n = 77).Results. Early starting high-intensity training delayed the onset of menarche ( p < 0.001). Dancers had a higher prevalence of oligomenorrhea and amenorrhea than control girls ( p = 0.004). Additionally, the dancers had lower scores in anthropometric variables: breast circumference 80 cm vs. 86.6 cm for controls ( p = 0.0001), low weight in 18% of dancers vs. 2.6% of controls ( p = 0.0001), and low height in 18% of dancers vs. 9% of controls ( p = 0.016). In addition, in dancers, low BMI was observed in 21% compared with 13% of controls ( p = 0.0001). Finally, 32% of the dancers were on a weight-control diet while this percentage decreased to 12% for the girls in control group (odds ratio = 3.49, 95% confidence interval = 1.31-9.25).Conclusions. In ballet dancers, high-intensity training was associated with late onset of menarche, menstrual disorders, lower weight and height development, and abnormal feeding behaviors.

Raloxifene inhibits cholesterol aortic content but not atherosclerotic plaque size in oophorectomised cholesterol-fed rabbits.

Raloxifene, a selective oestrogen receptor modulator, is effective in the treatment of osteoporosis without stimulating the breast and the endometrium. Although it is associated with a decrease of cardiovascular risk markers the effect of these changes on atherogenesis, is not clear. In this study, we aimed to investigate the effect of raloxifene on aorta atherogenesis. A total of 32 cholesterol-fed New Zealand white rabbits were studied for 4 months. Twenty-four rabbits underwent bilateral ovariectomy; of these eight received raloxifene (group OR), eight received oestradiol valerate (group OE) and eight received placebo after sterilisation (group OP). Finally, another eight were sham-operated (non-ovariectomised) and received placebo with a hypercholesterolaemic diet (group SP). After the diet, total levels of cholesterol increased in group SP from 111.25 +/- 34.8 mg/dl to 1112.25 +/- 364.2, in group OP from 122.62 +/- 27.7 mg/dl to 1367.37 +/- 348.4, in group OE from 65.25 +/- 17.01 to 1710.5 +/- 356.2 and in group OR from 108.88 +/- 15.54 mg/dl to 1407.86 +/- 397.7 (no significant differences). At 4 months, in both treated and untreated rabbits, the cholesterol-rich diet caused atherosclerotic lesions affecting 24.51 +/- 16.1% for group SP, 30.47 +/- 12.2% for group OP, 30.31 +/- 18.07% for group OR and 17.91 +/- 10.19 for group OE (P<0.05) of the aortic surface, respectively. Aortic cholesterol expressed as mg of cholesterol/mg aortic weight was found to decrease in raloxifene-treated rabbits: 3.82 +/- 2.14 mg col/aortic mg versus 8.55 +/- 4.63 (group OP) and 11.97 +/- 11.33 (group SP). P<0.001. Raloxifene reduced aortic cholesterol content but not the atherosclerotic plaque extension in cholesterol-fed ovariectomised rabbits.

Tibolone inhibits aortic atherosclerotic lesionformation in oophorectomized cholesterol-fed rabbits.

BACKGROUND: Tibolone is a synthetic steroid effective for the treatment of climacteric symptoms and osteoporosis. Long term treatment with tibolone is associated with a significant decrease in cholesterol levels due to a parallel decrease in high-density lipoprotein. However, the effect of these changes on atherogenesis is not known. OBJECTIVE: To investigate the effect of tibolone therapy on aorta atherogenesis. MATERIAL AND METHODS: Thirty-two New Zealand white rabbits were fed cholesterol-rich feed and studied for four months. The rabbits underwent laparotomy and were randomly assigned to four groups. Twenty-four rabbits underwent bilateral ovariectomy; of these, eight received tibolone (group T), eight received estradiol valerate (group E), eight received placebo after sterilization (group C), and eight were sham operated (group S). RESULTS: After receiving the cholesterol-rich diet, total levels of cholesterol increased in group C from 3.17+/-0.72 mmol/L to 35.36+/-9.01 mmol/L, in group S from 2.88+/-0.9 mmol/L to 28.76+/-9.442 mmol/L, in group E from 1.69+/-0.44 mmol/L to 1.69+/-0.44 mmol/L and in group T from 2.03+/-0.22 mmol/L to 26.33+/-13.45 mmol/L (no significant differences were observed among the groups at the end of the study). At four months, the cholesterol- rich diet caused atherosclerotic lesions in both treated and untreated rabbits, affecting 30.47+/-12.2%, 24.51+/-16.1%, 17.91+/-10.19% and 10.21+/-6.8% of the aortic surface for groups C, S, E and T, respectively (P<0.01 for treated groups). CONCLUSION: The principal result from this study was that treatment with tibolone in cholesterol-fed ovariectomized rabbits reduces aortic atherosclerotic lesion formation and that this reduction is not related to plasma lipid levels.

Use of hormone replacement therapy among Chilean women: a comparison between socioeconomic levels.

OBJECTIVE: To compare the use of hormone replacement therapy (HRT) among Chilean women according to their socioeconomic level. METHODS: A total of 540 women between 50 and 79 years of age were interviewed in Santiago, Chile. Women were allocated into two groups (H, high; L, low), according to their socioeconomic status. RESULTS: Each group consisted of 270 women. The mean age and percentage of menopausal women were similar in both groups. Of the interviewed women, 47% had taken HRT at some time; marked differences between the two groups were observed (L, 15%; H, 79%; < 0.0001). In group H, the percentage of women who had been advised about HRT was close to 88%, whereas, in group L, the percentage was only 24%. Among the women who were informed about HRT, 83% ( = 253) had used it at some time. The percentage of women who used HRT for >2 years was similar in both groups. The main reason for not taking HRT in group H was fear of adverse effects, whereas the main reason for not taking it in group L was the lack of medical advice. CONCLUSIONS: The percentage of women in the low socioeconomic group who use HRT is low. Medical advice is fundamental to increasing HRT use in this group.