Outcomes and cost analysis of single-embryo transfer versus double-embryo transfer.

BACKGROUND: Studies had compared single-embryo transfer to double-embryo transfer with cleavage stage embryos and found that while single-embryo transfer was less costly, it was also associated with a lower live birth rate than double-embryo transfer. A single blastocyst transfer has been shown to improve the live birth rate per cycle compared to single-embryo transfer at cleavage stage. OBJECTIVES: To compare live birth rates and real costs of elective single-embryo transfer to double-embryo transfer and to determine the incremental cost-effectiveness ratio of these two strategies in an unselected pool of women in a single center. DESIGN: Retrospective study. METHODS: We analyzed data of 4232 women who underwent their first fresh in vitro fertilization/intra-cytoplasmic sperm injection cycles with at least two embryos available for transfer in KK Women's and Children's Hospital from 2010 to 2017. RESULTS: Five hundred and sixty-four women underwent elective single-embryo transfer and 3668 women underwent double-embryo transfer. One hundred and fifty-six women who failed to achieve a live birth in their fresh elective single-embryo transfer cycle underwent a sequential thaw single-embryo transfer cycle. Live birth rate of fresh elective single-embryo transfer was significantly higher at 41.3% than that of double-embryo transfer at 32.6%. Cumulative live birth rate for sequential elective single-embryo transfer (fresh elective single-embryo transfer + thaw single-embryo transfer) was 47.9%. After accounting for variables which may affect live birth rates such as age and stage of embryo transfer, the odds of achieving a live birth from double-embryo transfer was 24% lower than that from sequential single-embryo transfer, although not statistically significant. For every live birth gained from an elective single-embryo transfer compared to double-embryo transfer, cost savings were S$20,172 per woman. If a woman had to have a sequential single-embryo transfer after a failed single-embryo transfer in her fresh cycle, cost savings were reduced to S$1476 per woman. CONCLUSION: Single-embryo transfer is a dominant strategy in an unselected population and adopting it in assisted reproductive treatments (ART) can produce cost savings without compromising on live birth rates.

A Case of Pre-Existing Pregnancy During an In-Vitro Fertilization Cycle.

Serum progesterone levels performed prior to oocyte pick-up is commonly used to guide embryo transfer in in-vitro fertilization (IVF) cycles, as elevated levels can negatively influence pregnancy outcomes. However, levels associated with normal pregnancies should trigger clinicians to consider alternative causes such as a pre-existing pregnancy. We report a case of a 37-year-old patient who underwent controlled ovarian hyperstimulation in a gonadotrophin-releasing hormone antagonist cycle while having an undetected early pregnancy. No oocytes were retrieved at oocyte retrieval despite adequate follicular responses. Her serum progesterone level on the day of her trigger injection was 57.8 nmol/L. She was found to have a pregnancy of unknown location, detected 3 weeks after her oocyte retrieval and was subsequently treated with systemic methotrexate.

Risk score to stratify miscarriage risk levels in preconception women.

Spontaneous miscarriage is one of the most common complications of pregnancy. Even though some risk factors are well documented, there is a paucity of risk scoring tools during preconception. In the S-PRESTO cohort study, Asian women attempting to conceive, aged 18-45 years, were recruited. Multivariable logistic regression model coefficients were used to determine risk estimates for age, ethnicity, history of pregnancy loss, body mass index, smoking status, alcohol intake and dietary supplement intake; from these we derived a risk score ranging from 0 to 17. Miscarriage before 16 weeks of gestation, determined clinically or via ultrasound. Among 465 included women, 59 had miscarriages and 406 had pregnancy ≥ 16 weeks of gestation. Higher rates of miscarriage were observed at higher risk scores (5.3% at score ≤ 3, 17.0% at score 4-6, 40.0% at score 7-8 and 46.2% at score ≥ 9). Women with scores ≤ 3 were defined as low-risk level (< 10% miscarriage); scores 4-6 as intermediate-risk level (10% to < 40% miscarriage); scores ≥ 7 as high-risk level (≥ 40% miscarriage). The risk score yielded an area under the receiver-operating-characteristic curve of 0.74 (95% confidence interval 0.67, 0.81; p < 0.001). This novel scoring tool allows women to self-evaluate their miscarriage risk level, which facilitates lifestyle changes to optimize modifiable risk factors in the preconception period and reduces risk of spontaneous miscarriage.

The Effect of Luteinising Hormone Suppression in In Vitro Fertilisation Antagonist Cycles.

Use of GnRH antagonists in IVF stimulation protocols shortens controlled ovarian hyperstimulation (COH) and reduces the risk of ovarian hyperstimulation syndrome (OHSS). However, profound reduction in LH levels has been associated with use of GnRH antagonists. This study aims to determine if LH suppression during GnRH antagonist cycles results in poorer IVF outcomes. This was a prospective pilot longitudinal study where serum LH levels were measured on day 2/3 of the menstrual cycle before COH, 1/2 days following institution of GnRH antagonist and at the day of ovulation trigger. A threshold of LH <0.5 IU/L was used to define profound LH suppression. Data on IVF outcomes was collected. Logistic regression analysis was used to investigate risk factors associated with LH suppression following GnRH antagonist IVF treatment. Ninety-one eligible women were recruited. Women underwent a standard antagonist cycle with Puregon 200u and Ganirelix. No participant had LH <0.5 IU/L prior to GnRH antagonist treatment, and 27 participants (29.7%) had significant LH suppression at either time point. Predictors of profound LH suppression following GnRH antagonist treatment identified (P < 0.20) were age (OR = 0.80, P = 0.013), no previous ovulation induction (OR = 0.26, P = 0.033) and previous GnRH antagonist IVF cycle (OR = 4.32, P = 0.125). Numbers of oocytes, embryos and ongoing pregnancy rates at 12 weeks gestation in patients with and without LH suppression did not differ significantly. We found associations between clinical characteristics and risk of profound LH suppression in women undergoing GnRH antagonist IVF cycles, but no significant differences in IVF and pregnancy outcomes between women with and without significant LH suppression.

Ovarian biomarkers predict controlled ovarian stimulation for in vitro fertilisation treatment in Singapore.

INTRODUCTION: Ovarian biomarkers have been shown to predict responses to controlled ovarian hyperstimulation (COH) during in vitro fertilisation (IVF) in predominantly Caucasian populations, with limited studies performed in Southeast Asian women in Singapore. METHODS: We evaluated the performance of serum anti-Müllerian hormone (AMH), follicle-stimulating hormone and oestradiol levels, antral follicle count (AFC), body mass index, ovarian volume, and age to establish thresholds for the prediction of poor (< 4 oocytes retrieved) and excessive responses (> 19 oocytes retrieved) in 263 women undergoing COH. Univariate and multivariate logistic regression analysis and receiver operating characteristic curves were used to calculate probabilities for poor and excessive responders to COH. RESULTS: 36 (13.7%) and 50 (19.0%) women had poor and excessive response to COH, respectively. An AMH value of 0.69 ng/mL predicted poor ovarian response with positive likelihood ratio (LR) of 2.94, compared to an AFC of ≤ 5 when the positive LR is 2.36. Conversely, an AMH value of ≥ 3.06 ng/mL predicted excessive ovarian response with positive LR of 2.24, compared to an AFC cut-off of ≥ 12 with positive LR of 1.93. CONCLUSION: AMH levels and AFC are equivalent in the prediction of both poor and excessive ovarian response in women undergoing IVF. Our study highlights the importance of establishing population-specific cut-off biomarker values so that protocols can be tailored to optimise IVF treatment.

Laparoscopic Tubal Re-anastomosis or In Vitro Fertilisation in Previously Ligated Patients: A Comparison of Fertility Outcomes and Survey of Patient Attitudes

INTRODUCTION: We aim to compare live birth rates, cost analysis and a survey of patient attitudes between laparosopic tubal re-anastomosis and in vitro fertilisation (IVF). MATERIALS AND METHODS: A retrospective study was done in a single reproductive medicine and IVF unit in Singapore from January 2011 to December 2016. Previously ligated patients underwent either laparoscopic tubal re-anastomosis or IVF. The primary outcome was first live birth after treatment. Interval to first pregnancy, miscarriage and ectopic pregnancies were also reported. Patients attending the subfertility clinic completed a questionnaire on IVF and tubal re-anastomosis on preferred choice of treatment, before and after reading an information sheet. RESULTS: Twelve patients underwent tubal re-anastomosis while 31 patients underwent IVF treatment. Pregnancy (75.0% vs 35.5%) and live birth (58.3% vs 25.8%) were significantly higher in the tubal surgery group (P <0.05%) after transferring all available embryos in one stimulated IVF cycle. Cost per live birth was lower in the tubal surgery group (SGD27,109 vs SGD52,438). One hundred patients participated in the survey. A majority of patients preferred tubal surgery to IVF (68.2% vs 31.8%) before given information on the procedures, but indicated a preference for IVF (54.6%) to surgery (45.4%) after receiving information on the procedures. CONCLUSIONS: For women less than 40 years of age, desiring fertility after tubal ligation, laparoscopic tubal re-anastomosis offers better live birth rates and cost-effectiveness. Patients in Singapore are equivocal as to their preference after education regarding the choices. Thus, laparoscopic tubal re-anastomosis remains a viable alternative to IVF treatment.

Do men with normal testosterone-oestradiol ratios benefit from letrozole for the treatment of male infertility?

RESEARCH QUESTION: Previous studies of aromatase inhibitors on male infertility have focused on men with low testosterone-oestradiol ratio of less than 10. Can aromatase inhibitors improve spermatogenesis in men with idiopathic male infertility with normal testosterone-oestradiol ratio? DESIGN: Prospective study of men with idiopathic severe oligozoospermia (sperm concentration <5 million/ml) carried out between February 2015 and March 2017. The objective was to assess if semen-analysis parameters improved after treatment with letrozole. Secondary objectives were to monitor the safety of letrozole in men, and to measure the alterations in serum FSH, LH, oestradiol and testosterone levels. RESULTS: Fifteen men with normal testosterone-oestradiol ratio (>10) were treated with letrozole 2.5 mg daily for 4 months. This produced a 5.5-fold increase in sperm concentration (P = 0.0068). All men had increased total serum testosterone and suppressed oestradiol levels after treatment, thus raising the overall testosterone-oestradiol ratio (P < 0.0001). Adverse effects from letrozole were relatively minor and included loss of libido (54%), headaches (25%), fatigue (21%), weakness (13%), loss of hair (8%) and dry mouth (8%). CONCLUSIONS: Letrozole improves sperm concentration and increases testosterone-oestradiol ratio for men with oligozoospermia who have normal testosterone-oestradiol ratio; its role in the treatment of male infertility may be extended to this group of patients. In addition, it is a relatively well-tolerated drug with no serious adverse effects.

Age-related nomograms for antral follicle count and anti-Mullerian hormone for subfertile Chinese women in Singapore.

BACKGROUND: Antral follicle count (AFC) and anti-Mullerian hormone (AMH) are known as the most reliable markers of a woman's ovarian reserve and are related to age. There is currently no specific local age-related centile charts for AFC and AMH. Therefore, we aim to examine the relationship between AFC and AMH with age and construct age-related nomograms among a subfertile Asian population. METHODS: This is a study involving Chinese women who had their AFC and AMH measured as part of their subfertility screening from December 2010 until November 2014 in KK Women's and Children's Hospital, Singapore. Ordinary least squares regression analysis was used to estimate the relationship of AFC and AMH with age, while age-related AFC and AMH nomograms for the 3rd, 10th, 25th, 50th, 75th, 90th and 97th percentiles were produced using the lambda-mu-sigma method. RESULTS: A total of 1,009 women, aged 26 to 44 year-old, were included. On average, the AFC and AMH decreased respectively by 0.79 follicle (95% confidence interval -0.93, -0.64) and 0.38 ng/mL (95% confidence interval -0.43, -0.32) per year of age. The age-related nomograms of AFC showed an approximately linear pattern, inversely correlated with age, regardless of the percentile. For AMH, the pattern is linear for the 75th percentile and below but shows a slightly accelerating decline for the 90th and 97th percentile. Overall, there were large inter-individual variations in AFC and AMH up to about 40 year-old. CONCLUSION: The declines of AFC and AMH over age are mostly linear among subfertile Chinese women in Singapore. The age-related AFC and AMH nomograms could be used as a reference chart by fertility practitioners. However, future validation with longitudinal data is required.

Estimating cumulative live-birth rates after IVF treatment with Kaplan-Meier and competing risk methods.

OBJECTIVE(S): To explore the use of competing risk (CR) as compared to the commonly used Kaplan-Meier (KM) methodology in estimating the cumulative live-birth rate (CLBR) after IVF Treatment in a context of high dropout rates and informative censoring. STUDY DESIGN: We compare the KM and CR methodologies for estimating 2-year CLBR in a retrospective cohort of 2779 patients undergoing 5002 embryo transfers over a period of 9 years, from 2000 to 2008, at KKIVF Centre. RESULTS: We observed a total of 1105 LB (39.8%), and a dropout rate of 44.2% (1228 patients). The overall CLBR is lower with CR compared with KM method (39% vs 52%) after up to nine embryo-transfer cycles over a period of two years. The highest CLBR was achieved for ovulation disorders (57% vs 49%, KM vs CR) followed by male factors (54% vs 43%, KM vs CR), with poorer outcomes from patients with decreased ovarian reserve (37% vs 16%, KM vs CR) and endometriosis (36% vs 25%, KM vs CR). As dropouts in our cohort are generally older and more likely to have poorer ovarian reserves, the CR method, which accounted for these dropouts, is likely to give more meaningful estimation of IVF success rates. CONCLUSION(S): The CR method should be considered as a useful alternative in deriving CLBR for IVF treatment where dropout rates are high and when informative censoring is involved.

Single-tube nonaplex microsatellite PCR panel for preimplantation genetic diagnosis of Hb Bart's hydrops fetalis syndrome.

OBJECTIVE: To develop a single-tube multi-marker assay for improved preimplantation genetic diagnosis (PGD) of deletional and/or non-deletional Hb Bart's hydrops fetalis syndrome, providing haplotype confirmation of deletional status, and maximization of linkage informativity. METHODS: We performed in silico mining to identify novel microsatellites within 1 Mb flanking the alpha-globin gene cluster, and optimized a single-tube assay combining detection of α(0) -thalassemia deletions with multi-marker linkage analysis. We performed validation on 100 single cells prior to clinical PGD application. RESULTS: Of 42 markers encompassing the α-globin gene cluster that were identified in silico, 9 were highly polymorphic (0.68 ≤ polymorphism information content ≤ 0.92; 0.66 ≤ Ho ≤ 0.90; 10 ≤ alleles ≤ 35) and optimized to co-amplify directly from a single cell. A validation analysis of 100 single lymphoblasts yielded 100% amplification success for all markers, and individual marker allele drop-out (ADO) rates of 0-5%. Clinical application of the assay in PGD for Hb Bart's (2 cases/cycles) resulted in a twin pregnancy and healthy live birth of two baby girls. CONCLUSIONS: This single-tube nonaplex microsatellite PCR panel can be applied directly to PGD of most deletional Hb Bart's without the need for deletion-specific customization, and to linkage-based PGD of non-deletional Hb Bart's.

GnRH antagonist pre-treatment: one centre's experience for IVF-ICSI cycle scheduling.

Scheduling gonadotrophin-releasing hormone antagonist (GnRH-ant) cycles for IVF intracytoplasmic sperm injection in patients is a challenge because of unpredictable ovum retrieval procedures on weekends, when less manpower is available. Recently, the use of GnRH-ant pre-treatment to delay an IVF and intracytoplasmic sperm injection (ICSI) cycle showed no negative effect on clinical pregnancy rates. An age-matched, case-control study was conducted to evaluate the effectiveness of such pre-treatment for scheduling purposes. Patients (n = 140) undergoing their first ovarian stimulation for IVF-ICSI were included. Patients starting their stimulation on Tuesdays or Wednesdays were most likely to have their ovum retrieval procedure on Saturdays. Seventy patients received a 3-day course of GnRH-ant before starting stimulation, and were compared with 70 age-matched controls not receiving pre-treatment. The main outcomes were the proportion of ovum retrieval procedures occurring on Saturdays, clinical pregnancy rate and live birth rates. A five-fold reduction in the number of ovum retrievals occurred on Saturdays compared with controls (7.1% versus 34.3%; OR 0.15; 95% CI 0.05 to 0.42; P < 0.001), with no significant differences in clinical pregnancy rate (40.9% versus 37.5%) and live birth rate (27.3% versus 31.3%). GnRH-ant pre-treatment is an effective tool for scheduling of GnRH-ant cycles.

GnRH agonist trigger and ovarian hyperstimulation syndrome: relook at 'freeze-all strategy'.

A case is reported of early onset ovarian hyperstimulation syndrome (OHSS) after gonadotrophin-releasing hormone agonist (GnRHa) trigger for final oocyte maturation in a GnRH antagonist protocol. The use of GnRHa in place of HCG as a trigger for final oocyte maturation in an antagonist IVF cycle has been proposed as a method for preventing OHSS in predicted high-responders. This approach, however, did not prevent the occurrence of OHSS in our case despite a freeze-all strategy. To the best of our knowledge, this is a possible index case of severe OHSS with GnRHa trigger for oocyte maturation without any luteal HCG rescue for a high responder, despite IVF cycle segmentation.

Social oocyte freezing: a survey among Singaporean female medical students.

AIM: Social oocyte freezing has gained increasing interest worldwide. We conducted a cross-sectional survey on 129 female medical students in Singapore to assess their mindset and attitudes toward fertility and social oocyte freezing. METHODS: An anonymous online survey was conducted among female medical students in Singapore. The desired sample size was 100 participants. Their awareness of the existence of social oocyte freezing was first assessed. An information leaflet was provided subsequently, followed by a more detailed questionnaire. The questions focused on their awareness of age-related fertility decline and their intentions for social oocyte freezing if made available. RESULTS: One hundred and twenty-nine female students participated in the electronic survey, of whom 36.4% had heard of social oocyte freezing. Of these, 70% had personally considered taking up this option. However, after reading the information leaflet, only 48.9% would still consider this option. Of the total, 89.9% considered themselves too old for pregnancy after the age of 35 years, 37.2% would delay family planning for their career, 45.7% would consider social oocyte freezing to postpone family planning for their career, 46.5% would consider oocyte freezing if they had no suitable partners yet, 50.4% may consider freezing their eggs after the age of 30 years and 71.3% may be more amenable to oocyte freezing if government subsidy is available. CONCLUSION: We hypothesize that social oocyte freezing may be a viable option for single young women who wish to delay child-bearing for 'reproductive insurance', so long as this is done with appropriate informed consent with non-directive counseling.

Human embryo cryopreservation: one-step slow freezing does it all?

PURPOSE: Previous studies have shown that a modified one-step slow freezing method with higher sucrose concentration (0.2 M) can achieve higher embryo and blastomere survival rates that are comparable to vitrification. However, no study has evaluated the efficacy of a one-step method using commercial slow freezing kit without altering its composition. This retrospective study examines the effects of using 1.5 M PROH with 0.1 M sucrose (F2 medium) alone in a one-step slow freezing method compared to the conventional two-step method. METHODS: Cleavage stage embryos from 526 thaw cycles previously cryopreserved by either the conventional two-step slow freezing method or the modified one-step method were studied. The embryo and blastomere survival rates, cleavage rate, clinical pregnancy rate and live birth rate were compared between the two groups. RESULTS: The results showed that the embryo survival rate was significantly higher in the modified one-step method compared to the conventional two-step method (86.9 % and 83.1 %, respectively; p = 0.04). Total blastomere survival rate was also significantly increased as a result of the modification (81.0 % versus 76.5 %; p < 0.001). However, there was no statistical difference in the cleavage rates, clinical pregnancy rates (CPR/ET) and live birth rates between the two methods. CONCLUSIONS: Slow freezing using the one-step method is superior to the conventional two-step method in terms of embryo and blastomere survival rates without affecting cleavage rate and clinical outcomes. It can be routinely applied to cleavage stage embryo cryopreservation in IVF centres for greater workflow efficiency.