Assisted Reproduction Center Research Articles

Spontaneous fetal reduction and early pregnancy complications in multiple pregnancies following in vitro fertilization

ObjectiveTo determine the incidence of spontaneous pregnancy reduction and early pregnancy complications among multiple pregnancies conceived via in vitro fertilization (IVF). MethodsSpontaneous pregnancy reduction was assessed among multiple pregnancies conceived after IVF treatment in a private assisted reproduction center in Lagos, Nigeria, between January 2008, and June 2011. The pregnancies were diagnosed at 6–8weeks of gestation by ultrasound scan, which was repeated at 11–14weeks to determine the incidence of spontaneous reduction. ResultsThe incidence of multiple pregnancies was 35.2%. There were 11 (20.0%) twin, 6 triplet (10.5%), and 2 quintuplet (3.6%) pregnancies. Spontaneous reduction occurred in 9 (47.7%) of these pregnancies, including 15.8% of twins, 21% of triplets, and 10.5% of quintuplets. Spontaneous reduction was significant among triplets and quintuplets (P<0.05). Six women (31.5%) older than 36years had spontaneous reduction (P<0.05). Vaginal bleeding occurred in 3 (15.5%) women, 2 (66.7%) of whom had spontaneous abortion. In all instances of monochorionic placentation, spontaneous reduction resulted in spontaneous abortion. ConclusionThe incidence of spontaneous reduction among multiple pregnancies conceived after IVF was high. Vaginal bleeding was a warning sign of eventual abortion. Early pregnancy complications were fewer among pregnancies with dichorionic and trichorionic placentation.

A reliable procedure for decontamination before thawing of human specimens cryostored in liquid nitrogen: three washes with sterile liquid nitrogen (SLN2)

To report a washing procedure, to be performed as frozen specimens are taken out of cryobanks, to minimize the risk of hypothetical culture contamination during thawing. Basic research. Private assisted reproduction center. Two batches of liquid nitrogen (LN(2)) were experimentally contaminated, one with bacteria (Pseudomonas aeruginosa, Escherichia coli, Stenotrophomonas maltophilia) and the other with fungi (Aspergillus niger). Two hundred thirty-two of the most common human gamete/embryo vitrification carriers (Cryotop, Cryoleaf, Cryopette) were immersed in the contaminated LN(2) (117 in the bacteria and 25 in the fungi-contaminated LN(2)). The carriers were tested microbiologically, one group without washing (control) and the other after three subsequent washings in certified ultraviolet sterile liquid nitrogen (SLN(2)). The carriers were randomly allocated to the "three-wash procedure" (three-wash group, 142 carriers) or "no-wash" (control group, 90 carriers) using a specific software tool. Assessment of microorganism growth. In the no-wash control group, 78.6% of the carriers were contaminated by the bacteria and 100% by the fungi. No carriers were found to be contaminated, either by bacteria or fungi, after the three-wash procedure. The three-wash procedure with SLN(2) produced an efficient decontamination of carriers in extreme experimental conditions. For this reason, this procedure could be routinely performed in IVF laboratories for safe thawing of human specimens that are cryostored in nonhermetical cryocontainers, particularly in the case of open or single-straw closed vitrification systems.

Pinopodes, leukemia inhibitory factor, integrin-β3, and mucin-1 expression in the peri-implantation endometrium of women with unexplained recurrent pregnancy loss

To compare pinopode morphology and coverage, and expression of leukemia inhibitory factor (LIF), integrin-β3, and mucin-1 (MUC1) in the peri-implantation endometrium of women with unexplained recurrent pregnancy loss (RPL) versus normal fertile women. Exploratory clinical study. University-affiliated assisted reproductive center. Women undergoing assisted reproduction at centers of reproductive medicine in two university teaching hospitals. Endometrial biopsy samples were obtained from women with RPL (test group, n = 30) and normal fertile women (control group, n = 26) 7 days after the day of luteinizing hormone peak. Pinopode morphology and coverage were assessed by use of scanning electron microscopy. Expression of LIF, integrin β3, and MUC1 were evaluated by immunohistochemical staining. Pinopode morphology and coverage, and the expression of LIF, integrin-β3, and MUC1 in both groups. Pinopode morphology and coverage, and the expression of LIF and integrin-β3 showed no statistically significant differences between the two groups. The expression of MUC1 was statistically significantly reduced in the glandular epithelial cells and endometrial lumen of the RPL group compared with controls. The morphology and the coverage of pinopodes and expression of LIF and integrin-β3 did not change, but MUC1 expression was statistically significantly reduced in women with RPL compared with normal women.

Correlação entre a idade e a contagem dos folículos antrais em mulheres inférteis

To produce age-related nomograms for ovarian antral follicle count (AFC) in infertile women. It was done a cross-sectional study of patients attended in the center of assisted reproduction Fêmina, from March 2010 to October 2011. The patients were submitted to transvaginal ultrasonography from day 2 to day 4 of their menstrual period. Patients included were between 21 to 45 years old, with regular menses, two healthy ovaries, without any evidence of endocrinopathies and who gave written informed consent. Patients excluded were smokers, with galactosemia or ovarian cysts, with antecedents of liver disease, ovarian surgeries or who were treated with chemotherapy or radiotherapy. In order to check the evolution of the AFC in relation to patient age, we used the 5th, 25th, 50th, 75th and 95th percentiles. Linear regression was carried out using these percentiles, permitting us to determine the effect of age on the CFA. A total of 172 patients with a mean age of 32.7 years were included in the trial. The male and tubal factors were the main causes of infertility, accounting for 65% of cases. The age-related nomogram for the 5th, 25th, 50th, 75th and 95th percentiles of AFC revealed that changes were best fitted by a linear function. The percentiles that showed the highest correlations were 25 (r=-0.9; p<0.001), 50 (r=-0.9; p<0.001) and 75 (r=-0.9; p<0.001). A nomogram was constructed correlating age with the different AFC percentiles in infertile women without endocrinopathies. This showed a linear pattern of decline in AFC with age in all percentiles. These nomograms could provide a reference guide for the clinician. However, future validation, with longitudinal data, still is needed.

Role of Endometrial Thickness on Intracytoplasmic Sperm Injection

Problem statement: Endometrial receptivity is a key factor in successful outcome of in vitro fertilization. The endometrial receptivity is determined clinically by estimation of endometrial thickness and pattern by ultrasonography. This study was performed to evaluate the overall effect of endometrial thickness on pregnancy rate, multiple pregnancy rate and miscarriage rate in infertile women undergoing Intracytoplasmic Sperm Injection (ICSI) cycles at Mustasharak medical center, ABHA, Saudi Arabia. Approach: A prospective observational study at Mustasharak assisted reproduction center, ABHA, Saudi Arabia, was performed for women who underwent ICSI between Jan and Dec 2010. On the day of oocytes retrieval; after a true ultrasound longitudinal view of the uterus had been obtained, the endometrial thickness was measured as the maximum thickness between the highly reflective interfaces of the endometrial-myometrial junction. Results: 409 ICSI cycles were included in the study. The highest pregnancy rate was seen in patients who had an endometrial thickness of more than 12 mm, while those who had an endometrial thickness of less than 8 mm had the worst pregnancy rate. Type B endometrial morphology (trilaminar) had a significant higher pregnancy rate than type A endometrial morphology (non trilaminar). Conclusion: This study demonstrated a clear relationship between endometrial thickness on the day of oocytes collection and pregnancy rates following ICSI.

Perinatal and neonatal outcomes of 494 babies delivered from 972 vitrified embryo transfers

To evaluate the safety of vitrification of embryos by comparing the perinatal outcome of vitrified day 3 ETs with fresh cleavage-stage ETs. Retrospective analysis. Assisted reproduction center. A total of 806 women with 494 infants delivered were included. Supernumerary embryos and embryos of patients at risk of ovarian hyperstimulation syndrome were vitrified using a Cryotop carrier in an open system and were transferred after warming. Implantation and pregnancy rates, obstetric outcomes, neonatal outcomes, and congenital birth defects. A total of 2,543 vitrified embryos were warmed, and 2,375 survived and were transferred. The implantation, pregnancy, miscarriage, and live birth rates were 26.91%, 47.22%, 6.07%, and 38.58%, respectively. In singletons, the mean birth weight was 3,337.44 g, and the mean Apgar scores were 8.91, 9.85, and 9.89 at 1, 5, and 10 minutes, respectively. In multiple gestations, the mean birth weight was 2,556.45 g, and the mean Apgar scores were 8.90, 9.34, and 9.47 at 1, 5, and 10 minutes, respectively. Only seven congenital malformations were observed among the 494 babies. Compared with fresh cleavage-stage ETs, the mean birth weight in the vitrified group was higher in both singleton and multiple gestations. All the other results were similar between the two groups. Compared with fresh ETs, vitrified day 3 ET shows no significant differences in obstetrical and neonatal outcomes. The study suggests that vitrification with Cryotop is an effective and safe method for the cryopreservation of human cleavage-stage (day 3) embryos.

Serum and Follicular Fluid Ghrelin Levels Negatively Reflect Human Oocyte Quality and In Vitro Embryo Development

Adverse outcomes from assisted reproductive techniques may be related in part to the effects of metabolic dysregulation in women undergoing these procedures on oocyte quality and embryo development. Among the various molecules linking metabolic dysfunction with reproductive outcomes is a peptide called ghrelin. It has been hypothesized that ghrelin affects steroidogenesis and ovarian cellular function and may mediate the relationship between metabolism and oocyte quality. Addition of ghrelin in vitro to mice preimplantation embryos in culture prevents embryonic development. Previous studies have suggested that the relationship of ghrelin with oocyte/embryonic development may be mediated by insulin, and that the negative impact of obesity, diabetes, insulin, and other metabolic dysfunction on antiretroviral therapy outcomes may be associated with intrafollicular and serum ghrelin levels. This prospective cohort study investigated the effect of serum and follicular fluid (FF) levels of ghrelin in women undergoing assisted reproduction on ovarian steroidogenesis, oocyte quality, and embryo development. The participants were 31 normal weight women undergoing in vitro fertilization at a university-based center for assisted reproduction. Patients with polycystic ovary syndrome, hypothalamic amenorrhea, or other endocrinopathies were excluded. Successful cleavage was defined as 6 cells or more by day 3 after fertilization. After an overnight fast, serum and FF were collected from preovulatory follicles of each participant (n = 81). Levels of ghrelin, insulin, and ovarian steroids (estradiol, testosterone, and progesterone) were measured. In vitro fertilization outcomes assessed included oocyte maturity and embryo development. FF ghrelin levels correlated significantly with serum levels and were 13% lower (range: 3%–24%). Serum ghrelin levels did not correlate with the body mass index or exogenous gonadotropins. Levels correlated negatively with the cleavage rate and number of viable cleavage stage embryos. Successfully cleaved embryos and those with viable morphology were 3 to 4 times more likely to come from follicles with lower levels of ghrelin than higher levels; the odds ratios were 3.67 (95% confidence interval [CI], 1.02–13.14) and 3.46 (95% CI, 1.01–11.81), respectively. Levels of FF ghrelin were negatively correlated with FF insulin and progesterone, and positively with estradiol. FF insulin levels were 52% higher (absolute difference,1.88; 95% CI,0.31–3.45] mIU/mL) in follicles with successfully cleaved embryos compared with those that were not. These findings suggest that FF ghrelin is not produced de novo in the ovary, but seems to be a serum transudate. FF ghrelin correlates with local steroidogenesis and expression of FF insulin. The effects of serum and FF ghrelin on oocyte/embryo development may be mediated by insulin.

Oestradiol valerate pretreatment in GnRH-antagonist cycles: a randomized controlled trial

This randomized controlled trial analyses the ability to control the oocyte retrieval schedule of gonadotrophin-releasing hormone antagonist cycles through the administration of oestradiol valerate during the luteo-follicular transition period prior to the initiation of ovarian stimulation. Eighty-six women undergoing ovarian stimulation for IVF/intracytoplasmic sperm injection were enrolled in the study. The control group (n=42) received a standard ovarian stimulation protocol. In the pretreatment group (n=44), patients were administered oestradiol valerate at a daily dose of 2×2mg from day 25 of the preceding cycle onwards, during 6–10 consecutive days, depending on the day of the week. The primary endpoint was the proportion of patients undergoing oocyte retrieval during a weekend day (i.e. Saturday or Sunday), which was significantly lower in the pretreatment group (1/37, 2.7%) compared with the control group (8/39, 20.5%; P value=0.029). The clinical pregnancy rates per started cycle were similar in the pretreatment group (38.6%) compared with the control group (38.1%). Pretreatment with oestradiol valerate results in a significantly lower proportion of patients undergoing oocyte retrieval during a weekend day and can be a valuable tool for the organization of an assisted reproduction centre.The past decade has seen growing interest in gonadotrophin-releasing hormone (GnRH)-antagonist protocols in an effort to reduce the incidence of potential complications. Nevertheless, GnRH agonists remain the GnRH analogue of choice in many assisted reproduction clinics, since they allow a more flexible and better controlled schedule of oocyte retrievals. The present study prospectively evaluates the efficacy of scheduling assisted reproduction cycles in a gonadotrophin-releasing hormone (GnRH)-antagonist protocol through the flexible pretreatment with oestradiol valerate during the luteo-follicular transition period of the menstrual cycle. Eighty-six women were enrolled in the study. The control group (n=42) received a standard ovarian stimulation protocol, whereas in the pretreatment group (n=44), patients were administered oestradiol valerate (Progynova) at a daily dose of 2×2mg from day 25 of the preceding cycle onwards, during 6–10 consecutive days, depending on the day of the week. After discontinuing the oestradiol valerate pretreatment, ovarian stimulation was initiated using the same protocol as the control group. The primary endpoint was the proportion of patients undergoing oocyte retrieval during weekend days, which was significantly lower in the pretreatment group (1/37, 2.7%) compared with the control group (8/39, 20.5%; P-value=0.029). The clinical pregnancy rates per started cycle were similar in the pretreatment group (38.6%) compared with the control group (38.1%). It can be concluded that pretreatment with oestradiol valerate results in a significant lower proportion of patients undergoing oocyte retrieval during a weekend day and can be a valuable tool for the organization of an assisted reproduction centre.

Hair and urine testing to assess drugs of abuse consumption in couples undergoing assisted reproductive technology (ART)

For the first time in Europe hair and urine testing have been applied to assess drugs of abuse consumption in couples undergoing assisted reproductive technology and the eventual association of toxic habits with other lifestyle, health status and sociodemographic factors was also investigated. Couples attending five assisted reproduction centers in Rome were invited to join the study. When they presented at the Centre for the visit, they were asked to answer a structured questionnaire concerning sociodemographic characteristics and lifestyle habits, and at the same time to provide hair and urine samples. Hair and urine testing for drugs of abuse, urinary profile of principal endogenous steroids involved in fertility process (testosterone, epitestosterone, androsterone, etiocholanolone and dehydroepiandrosterone) and of alcohol and tobacco smoke biomarkers were performed with validated methodologies. Of the 594 enrolled individuals (297 couples), 352 (164 couples and 24 single individuals from the couple) completed the questionnaire and gave both hair and urine samples, apart from 3 bald men, who only gave urine samples. Urine testing showed an overall 4.8% (17 individuals) positivity to drugs of abuse: 4.2% to cannabinoids, 1.4% to cocaine and 0.85% to both drugs. Results of 4cm segment hair samples testing matched those from urine samples. Thus, taking together, results of urine and hair testing confirmed repeated use of cannabis, cocaine and both drugs in 3.7, 0.85 and 0.57% examined individuals, respectively. Drug consumers were in a statistically higher percentage active smokers and alcohol drinkers, less prone to physical activity and with a trend towards higher weight than non consumers. Finally, repeated drug consumption was associated with significant lower concentration of urinary testosterone in males and of urinary dehydroepiandrosterone in females. The findings of the present study confirm the suitability of urine testing to disclose recent drugs of abuse consumption and of hair analysis to verify repeated consumption. Association between different toxic habits and sedentary lifestyle is also substantiated by the obtained results in our cohort of couples attending assisted reproduction centers.

Serum and follicular fluid ghrelin levels negatively reflect human oocyte quality and in vitro embryo development

To characterize serum and follicular fluid (FF) ghrelin in women undergoing assisted reproduction and to evaluate ghrelin's impact on ovarian steroidogenesis, oocyte quality, and embryo development. Prospective cohort study. University-based center for assisted reproduction. Normal weight women (n = 31) without endocrinopathies undergoing IVF. Fasting serum and FF from individual preovulatory follicles (n = 81) were collected. Ghrelin, insulin, ovarian steroids, and IVF outcomes including oocyte maturity and embryo development were assessed. The FF ghrelin correlated highly with serum levels and was 13% (3%-24%) lower. Levels were not associated with body mass or exogenous gonadotropins. Serum ghrelin correlated negatively with the cleavage rate and number of viable day 3 embryos. Embryos with successful cleavage and viable morphology came from follicles with lower ghrelin-odds ratios were 3.67 (1.02-13.14) and 3.46 (1.01-11.81), respectively. The FF ghrelin correlated negatively with FF insulin and progesterone, and positively with estradiol. The FF insulin was more than 50% higher (Δ = 1.88 [0.31-3.45] uIU/mL) in follicles with successfully cleaved embryos. The FF ghrelin does not reflect ovarian production and likely represents transudation. Ghrelin in FF correlates with local steroidogenesis and FF insulin expression. Serum and FF ghrelin are reflective of embryo development, possibly through interactions with FF insulin.

First case of transformation for breast fibroadenoma to high-grade malignant cystosarcoma in an in vitro fertilization patient

To evaluate the possible malignant transformation of fibroadenoma of the breast in patients undergoing an IVF cycle. Case report. Assisted reproduction center. A 41-year-old female patient undergoing assisted fertilization treatment. The patient underwent fine needle aspiration biopsy that confirmed fibroadenoma before the IVF attempt. She started a short stimulation protocol with triptorelin and recombinant FSH. After the first unsuccessful IVF attempt, she underwent a second short ovarian stimulation protocol with triptorelin and urinary FSH and she become pregnant. At 17 weeks, due to an increase in volume of the fibroadenoma, an excisional biopsy was performed that showed a malignant phyllode tumor. Then she underwent quadrantectomy. Malignant transformation of breast fibroadenoma. Cytologic examination of the first fine needle aspiration biopsy specimen showed a fibroadenoma of the breast; excisional biopsy showed a high-grade malignant cystosarcoma. Fibroadenoma was transformed into high-grade malignant cystosarcoma after ovarian stimulation in an IVF patient.

Persistent endometrial polyps may affect the pregnancy rate in patients undergoing intrauterine insemination

Abstract Objective To determine whether polypectomy before intrauterine insemination achieved better pregnancy outcomes than no intervention. Study design and setting A prospective comparative study conducted in a private assisted reproduction centre. Subjects and methods A total of 120 women having asymptomatic endometrial polyp (EP) that was diagnosed by 3D ultrasound and color Doppler and undergoing IUI were randomly allocated to one of two pre-treatment groups. Hysteroscopic polypectomy was performed in the study group while in the control group no intervention was done. All patients were scheduled to receive four cycles of IUI in both groups within 12 months duration. The first IUI cycle was planned after three menstrual cycles in both groups. Results No complications after hysteroscopic polypectomy were recorded. A total of 37 pregnancies occurred, 25 cases in the study group which included 57 cases and 12 cases in the control group which included 53 cases. Two cases in the study group and one case in the control group got pregnant during the 3 months waiting period, before performing IUI. Cumulative pregnancy rate in both groups after four IUI cycles was 23 (38.3%) in the study group and 11 (18.3%) in the control group. This difference was statistically significant ( p = 0.015). Conclusion Persistent endometrial polyps are likely to impair reproductive performance in this selected patient group and hysteroscopic polypectomy before IUI can be considered an effective measure especially as it is a safe procedure with a low complication rate.

Cumulative clinical pregnancy rates privileging elective single embryo transfer: a mathematical model from the quebec (Canada) publicly-funded in vitro fertilisation (IVF) program

To estimate the cumulative pregnancy rates in women commencing in vitro fertilisation (IVF) treatment in the new publicly-funded IVF program in Quebec, Canada. Simulation model based on a prospective cohort. Data from fresh IVF cycles performed between August 2010 and February 2011 were submitted to the Canadian ART Register (Quebec) (CARTRQ) by the four assisted reproduction centers in Quebec. Estimation of the cumulative pregnancy rates per cycle started were calculated based on the clinical pregnancy rates of frozen embryo transfer (FET) seen in our clinic during the same time period. From 2403 started IVF cycles, 2247 (93.5%) underwent oocyte retrieval. Embryo transfer was performed in 1,942 (81%) cases, from which 943 (49%) were eSET. The clinical pregnancy rate (CPR) per oocyte retrieval was 26.3%. The multiple pregnancy rate (MPR) was 5.2%; all of which were twins. Frozen embryos were available in 45% of the cases, with significant differences by groups of age (< 35 years: 63.82%, 35-39 years: 41.80%, >40 years: 21.49%; P<0.001). The average number of frozen embryos was 3.8, similarly distributed by groups of age (P=1.0). In our simulation model, cumulative clinical pregnancy rates per oocyte retrieval increased by each cycle of frozen embryo transfer (FET #1: 38%, FET #2: 49.7%, FET #3: 56.9%, FET #4: 64.0%). However it must be noted, only women aged < 35 will have enough frozen embryos to undergo three or more FET. Prior to publicly-funded IVF, the CPR in Quebec was 42.9% with a 27.2% MPR and only 1.6% eSET rate. With the introduction of publicly-funded IVF the eSET rate increased to nearly 50% but a drop in CPR was seen (26.3%) However considering the available frozen embryos the cumulative CPR per oocyte retrieval could be 64%. Differences in the proportion of available frozen embryos according to maternal age are determining factors that influence success.

Working to eliminate multiple pregnancies: a success story in Québec

In August 2010, the provincial government of Québec, Canada introduced funding of assisted reproduction treatment through the provincial health programme. Alongside this benefit, legislation was introduced to control assisted reproduction treatment activities in the province, including restrictions on the number of embryos that could be transferred in any one cycle. The aim of the programme was to transfer a single embryo in every cycle; multiple embryos could be transferred under suboptimal conditions but required physician justification. In the first 3 months of this programme, 1353 cycles of IVF were performed in five Québec assisted reproduction centres, with an overall clinical pregnancy rate of 32% per embryo transfer and 50% of transfers used elective single-embryo transfer (eSET). The multiple-pregnancy rate was only 3.7% per clinical pregnancy. In 2009, prior to the introduction of the programme, eSET was used in only 1.6% of embryo transfers, resulting in a multiple-pregnancy rate of 25.6%. These data demonstrate that providing provincially funded assisted reproduction treatment created an environment in which the aggressive use of eSET was not only possible, but also rapidly implemented. The result was a dramatic drop in multiple-pregnancy rates, approaching those for natural pregnancies. In August 2010, the provincial government of Québec, Canada introduced state-covered assisted reproduction through the provincial health programme. Alongside this, legislation was introduced to control assisted reproduction activities in the province, including the number of embryos that could be transferred in any one cycle with the aim of reducing multiple pregnancies, which are the major negative side effect of assisted reproduction treatment. Multiple embryos could be transferred under suboptimal conditions but required physician justification. In the first 3 months of this programme, 1353 cycles of IVF were performed in the five Québec-based assisted reproduction centres, resulting in an overall clinical pregnancy rate of 32% per transfer and with 50% of transfers of a single embryo. The multiple-pregnancy rate was only 3.7%. Prior to the introduction of the programme, the elective transfer of only one embryo (elective single-embryo transfer or eSET) was used in only 1.6% of cycles in Québec, resulting in a multiple-pregnancy rate of 25.6%. These data demonstrate that having state-sponsored assisted reproduction treatment created an environment in which the aggressive use of eSET was possible, implemented and resulted in a significant drop in multiple-pregnancy rates approaching natural levels of multiple pregnancies.

Comparative study of the psychosocial risks associated with families with multiple births resulting from assisted reproductive technology (ART) and without ART

To determine whether the psychosocial risks associated with multiple births are increased as a consequence of the use of assisted reproductive technology (ART) in comparison with those not resulting from ART. Cross-sectional study. Fertility units of a university hospital and a private hospital. Parents of single children and multiples (twins and triplets) between 6 months and 4 years of age (n=636) were divided into two groups: those who conceived through ART (n = 265) and those who conceived through non-ART (n = 371). Administration of six scales, in office or by mail. Measurement scales of material necessities, social stigma, marital satisfaction, perceived stress, depression, quality of life, and their subscales. The analysis of variance, Student's t-test, and χ(2)-test were applied. No effect was observed based on the use of ART in the psychosocial variables studied. Significant differences were obtained between the groups of single (SB) and multiple births (MB) with respect to material necessities, social stigma, marital satisfaction, depression, and quality of life. Having more than one child per birth, whether resulting from the use of ART or not, increases psychosocial risks for the parents. Assisted reproduction centers should include this information with the obstetric and neonatal risks, stressing the advantages of having one child per birth. Psychosocial risks are not increased as a result of having used ART, but the goal of avoiding the repeated use of ART does influence the desire to transfer the maximum number of embryos possible. Psychological counseling should be included before initiating ART, as well as when a multiple pregnancy is confirmed.

Melatonin protects human spermatozoa from apoptosis via melatonin receptor– and extracellular signal–regulated kinase-mediated pathways

To evaluate whether the protective effect of melatonin on H2O2-induced caspase activation and DNA fragmentation depends on the interaction between melatonin and its surface receptors. Laboratory study. Center for assisted human reproduction at a Spanish hospital. Twenty-one healthy donors. Human spermatozoa were treated with increasing concentrations of hydrogen peroxide (H2O2; 1 μM, 10 μM, 100 μM, 1 mM) and preincubated with 1 mM melatonin. Activation of caspase-3 and -9 as well as DNA fragmentation were examined by fluorescence methods. Our findings showed that H2O2 induced a significant increase in caspase-9 and caspase-3, which was dose independent. Conversely, pretreatment with melatonin reduced H2O2-mediated caspase activation in a dose-dependent way. Moreover, the antiapoptotic effects of melatonin in ejaculated human spermatozoa may involve membrane melatonin receptor MT1. In addition, we found that the survival-promoting pathway extracellular signal-regulated kinase (ERK) is likely to have a role in the protective actions of melatonin in ejaculated human spermatozoa. Finally, we confirmed these results further by demonstrating that melatonin prevention of H2O2-induced DNA fragmentation is dependent on both MT1 receptor and ERK signaling. These results indicate that the stimulation with melatonin triggers a set of events culminating in cell death prevention in ejaculated human spermatozoa.

Minimising twins in in vitro fertilisation: a modelling study assessing the costs, consequences and cost-utility of elective single versus double embryo transfer over a 20-year time horizon

To assess the cumulative costs and consequences of double embryo transfer (DET) or elective single embryo transfer (eSET) in women commencing in vitro fertilisation (IVF) treatment aged 32, 36 and 39 years. Microsimulation model. Three assisted reproduction centres in Scotland. A total of 6153 women undergoing treatment at one of three Scottish IVF clinics, between January 1997 and June 2007. A microsimulation model, populated using data inputs derived from a large clinical data set and published literature, was developed to compare the costs and consequences of using eSET or DET over multiple treatment cycles. Disability-free live births; twin pregnancy rate; women's quality-adjusted life-years (QALYs); health service costs. Not only did DET produce a higher cumulative live birth rate compared with eSET for women of all three ages, but also a higher twin pregnancy rate. Compared with eSET, DET ranged from costing an additional £ 27,356 per extra live birth in women commencing treatment aged 32 years, to costing £ 15,539 per extra live birth in 39-year-old women. DET cost ∼ £ 28,300 and ∼ £ 20,300 per additional QALY in women commencing treatment aged 32 and 39 years, respectively. Considering the high twin pregnancy rate associated with DET, coupled with uncertainty surrounding QALY gains, eSET is likely to be the preferred option for most women aged ≤ 36 years. The cost-effectiveness of DET improves with age, and may be considered cost-effective in some groups of older women. The decision may best be considered on a case-by-case basis for women aged 37-39 years.

Experimental contamination assessment of a novel closed ultravitrification device

To evaluate the safety of a new ultravitrification closed device. Ultravitrification research. Private assisted reproduction center. Microorganisms (bacteria). A styrofoam container holding 1,000 mL of liquid nitrogen (LN2) was contaminated with Pseudomonas aeruginosa and Escherichia coli. Forty closed devices (Ultravit) and 20 open devices (Cryotop) loading approximately 0.5 μL of antibiotic-free medium were plunged into this contaminated LN2 for 5-10 seconds and then inoculated into selective culture dishes. Colony-forming units were analyzed and counted after an overnight incubation at 37°C. Detection of micro-organisms in different devices after incubation. There was no contamination in any of the closed devices, whereas in 45% of open devices these bacteria were present. With this study we demonstrated, in an experimental model using contaminated LN2, that this new closed device is a safe system that does not allow cell contact with LN2, avoiding cell contamination.

Paternal body mass index is associated with decreased blastocyst development and reduced live birth rates following assisted reproductive technology

To determine the relationship between paternal body mass index (BMI), embryo development and pregnancy, and live birth outcomes after assisted reproductive technology (ART). Retrospective analysis of ART cycles. Major assisted reproduction center. Three hundred five couples undergoing ART in a private fertility clinic. No intervention was undertaken in patients involved in this study. Live birth outcomes and clinical pregnancy rates. No significant relationship between paternal BMI and early embryo development was found. However, increased paternal BMI was associated with decreased blastocyst development, clinical pregnancy rates and live birth outcomes. To our knowledge, this is the first report linking increased paternal BMI and clinical pregnancy and live birth rates after ART treatment. Further work to elucidate the mechanisms involved is required.

Live birth after anonymous donation of twice-cryopreserved embryos that had been stored in liquid nitrogen for a cumulative storage time of approximately 13.5 years

To report a live birth after transfer of anonymously donated, twice-cryopreserved embryos that had been stored in liquid nitrogen for approximately 13.5 years. Case report. A private assisted reproduction center. A 44-year-old recipient of donated cryopreserved embryos. Anonymous donation of cryopreserved blastocysts for procreation. Live birth after thawing and replacement of re-cryopreserved blastocysts. Fourteen pronuclear-stage embryos and four cleavage-stage embryos were cryopreserved during a primary IVF cycle. In two separate cycles, one cycle for the primary patient and a subsequent cycle for the first embryo donor recipient, the 18 embryos were thawed and grown to the blastocyst stage for transfer. Supernumerary blastocysts (n = 5) not replaced at either of these two thaw cycles were re-cryopreserved and subsequently donated to another embryo donor recipient. Five blastocysts survived the thaw and three were transferred, resulting in a live birth. The embryos were cryopreserved for a cumulative storage time of approximately 4,909 days (13.4 years). The longevity (viability) of cryopreserved embryos maintained in liquid nitrogen remains to be determined; cryopreserved embryo donation for procreation should not be overlooked, regardless of the length of time that embryos remain in cryostorage.