Supported by: Burroughs Wellcome Fund Preterm Birth Initiative. P-511 Wednesday, October 22, 2014. DHEA SUPPLEMENTATION RESULTS IN ...
blastocysts which will implant and progress to delivery from those destined to fail. DESIGN: Prospective blinded observational. MATERIALS AND METHODS: Patients with normal ovarian reserve were recruited for participation. Time-lapse imaging utilizing the EevaÒ System (Auxogyn, Menlo Park, CA) was recorded on all embryos through the blastocyst stage. Each blastocyst underwent trophectoderm biopsy for comprehensive chromosomal screening. Euploid blastocysts were transferred and outcome data collected. In the case of two embryo transfers where only one progressed in development, DNA fingerprinting was used to correctly establish the identity of the competent embryo and link this data with time-lapse imaging. RESULTS: 100 transferred embryos were included in the analysis, 52 embryos implanted and progressed through development while 48 did not. The time-lapse imaging cleavage parameters were compared with implantation outcome and no significant difference was noted in any of the following parameters: time from ICSI to start of 1st cytokinesis (p¼0.64) , syngamy (p¼0.78), duration of 1st cytokinesis (p¼0.82), duration of 2cell stage (p¼0.19), duration of 3-cell stage (p¼0.82), duration of 4-cell stage (p¼0.32), time from start of 1st cytokinesis to start of cavitation (p¼0.65). CONCLUSION: Cleavage stage parameters generated from time-lapse imaging do not distinguish outcomes between euploid blastocysts. These parameters do not provide a selection advantage to assist the embryologist in choosing which euploid blastocyst is more likely to implant or deliver. Further investigation is needed to determine if there are additional morphokinetic parameters evaluable through the blastocyst stage that could meaningfully improve embryo selection. At the current time, there are no adjuncts, including time-lapse imaging, which improve selection beyond the combination of traditional morphologic selection at the blastocyst stage with CCS. Supported by: Auxogyn; Equipment.
P-507 Wednesday, October 22, 2014 EMBRYOLOGIST INTERPRETATION OF TIME-LAPSE IMAGING PARAMETERS AT THE BLASTOCYST STAGE DO NOT ALTER SELECTION AMONG TRANSFERRED EUPLOID BLASTOCYSTS. K. H. Hong,a M. D. Werner,a J. M. Franasiak,a E. J. Forman,a,b A. Prodoehl,b K. Upham,b K. Scott,c R. T. Scott, Jr.,a,b a RWJ, Rutgers, Basking Ridge, NJ; bRMA NJ, Basking Ridge, NJ; cRMACT, Norwalk, CT. OBJECTIVE: Time-lapse imaging software has been successfully utilized at the cleavage stage to predict which embryos will blastulate. However, the literature does not contain an analysis of time-lapse imaging parameters during extended culture to the blastocyst stage and clinical outcomes. The goal of the present analysis was to determine if the interpretation of time-lapse images by experienced embryologists offered a selection advantage between high quality euploid blastocysts. DESIGN: Prospective blinded observational. MATERIALS AND METHODS: Patients undergoing IVF < age 42, with normal ovarian reserve, were recruited for participation. Time-lapse imaging with the EevaÒ System (Auxogyn, Menlo Park, CA) was recorded on all embryos through the morning of day 5. All embryos underwent trophectoderm biopsy for comprehensive chromosome screening. Euploid blastocysts were transferred and implantation and pregnancy data was collected. DNA fingerprinting was used to establish the identity of the embryo that implanted and this was linked to the time-lapse imaging. Experienced embryologists, blinded to the results of implantation and traditional grading, viewed all time lapse videos and recorded the time from ICSI to all of the following parameters: compaction, appearance of the blastocoelic cavity, the first expansion, the appearance of the trophectoderm monolayer, and the first post expansion constriction. No automated software was used for this analysis. The average scores amongst embryologists were recorded and T-tests were used for comparison between groups. RESULTS: 5 embryologists individually viewed 100 time lapse videos and recorded parameters as outlined above. Of the 100 embryos included in this analysis, 52% implanted and continued past 9 weeks of gestation and 48% failed to sustain development. No significant differences were noted between competent and failed embryos for any of the following criteria: compaction (p¼0.80), appearance of blastocoelic cavity (p¼0.95), first sign of expansion (p¼0.94), first appearance of trophectoderm monolayer (p¼0.82), and first post expansion constriction (p¼0.35).
FERTILITY & STERILITYÒ
CONCLUSION: Time-lapse imaging for the designated parameters did not distinguish outcomes amongst euploid blastocysts. Supported by: Auxogyn; Equipment. P-508 Wednesday, October 22, 2014 LIVE BIRTH RATE FOLLOWING IVF/ICSI IN PATIENTS WITH SPERM DNA FRAGMENTATION:A SYSTEMATIC REVIEW AND META-ANALYSIS. G. M. AlKusayer,a,b N. Amily,c A. M. Abou-Setta,d M. A. Bedaiwy.a aDivision of Reproductive Endocrinology and Infertility, University of British Columbia, Vancouver, BC, Canada; bDepartment of Clinical Sciences, Faculty of Medicine, Princess Nora Bint Abdulrahman University, Riyadh, Saudi Arabia; cFaculty of Medicine, University of Ottawa, Ottawa, ON, Canada; dGeorge & Fay Yee Centre for Healthcare Innovation, University of Manitoba, Winnipeg, MB, Canada. OBJECTIVE: Our aim was to examine the association between DNA fragmentation index(DFI)levels and clinical outcomes in couples undergoing in vitro fertilization(IVF)and intracytoplasmic sperm injection(ICSI). DESIGN: Systematic review and meta-analysis. MATERIALS AND METHODS: We searched MEDLINE, EMBASE and the Cochrane Library(inception to April 2014)using the following search terms:‘DNA Fragmentation’or‘DNA damage’combined with ‘IVF’,‘ICSI’, ‘pregnancy’,‘pregnancy loss’,‘miscarriage’or‘live birth’. We included studies investigating the association between sperm DNA fragmentation and clinical outcomes in couples undergoing IVForICSI. The primary outcome was live birth rate. Secondary outcomes were clinical pregnancy and miscarriage rates. DFI cut-off values varied in the assays conditions and threshold values. In three studies the threshold value of high DFI was more than30in both Sperm Chromatin Structure Assay and Sperm Chromatin Dispersion. Two studies used Comet assay with a threshold of more than50and56. One study only reported a threshold of more than or equal to35using theTUNELassay. Study selection, data extraction and quality assessment was conducted independently by2reviewers. Meta-analysis was conducted using a random-effects model. RESULTS: Six cohort studies (2460 couples) meeting the inclusion criteria were identified. Across the studies, there were more live births and clinical pregnancies in couples with low DFI index undergoing IVF/ICSI compared to couples with high DFI index (RR 1.55 [1.01 to 2.39] and RR 1.20 [0.86 to 1.68], respectively). Miscarriage rates were also lower in patients with a low DFI (RR 0.80 [0.56 to 1.13].Overall there was evidence of moderate to high heterogeneity in most analyses not explained by subgroup analyses of the method of fertilization (IVFvs.ICSI), timing of transfer (fresh vs. frozen embryo transfer), or methodological concerns (e.g. unit of analysis). These results were limited by different assays thresholds for the DNA fragmentation levels and the methodological quality of the included studies. CONCLUSION: Our systematic review suggests that couples with lowDNAfragmentation index levels are more likely to have better clinical outcomes compared to those with higherDNAfragmentation followingIVF/ ICSI. P-509 Wednesday, October 22, 2014 AUTOSOMAL RECESSIVE DISEASE RISK IN OFFSPRING OF QUALIFIED SPERM BANK DONORS AND CLIENTS: PROOF OF PRINCIPLE FOR A NOVEL ANALYSIS BASED ON VIRTUAL PROGENY. R. M. Lim,a A. J. Silver,a C. Borroto,a B. R. Spurrier,a R. E. Silver,a A. B. Remis,a A. S. Cohn,a A. Morriss,a L. M. Silver.a,b aGenePeeks, Inc., New York, NY; bPrinceton University, Princeton, NJ. OBJECTIVE: With advances in DNA analysis technologies, multi-gene carrier screening has become accessible to patients. Plummeting costs have also opened the door to large population studies, which provide evidence that nearly everyone is a carrier for one or more recessive diseasecausing mutations. This observation is problematic for genetic testing by sperm banks because carrier status - per se - has no utility in assessing the reproductive disease risk posed by individual sperm donors. Here we examine the potential to reduce reproductive disease risk by redirecting genetic analysis to the simulated diploid genomes created by digitally combining DNA sequences from ‘‘clients’’ and ‘‘donors.’’ DESIGN: Proof of principle for use of a novel computation protocol for revealing pre-conception genetic disease risk.
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MATERIALS AND METHODS: We have developed a novel computational system that simulates the genetics of gamete production and zygote formation to create ‘‘virtual progeny’’ from the DNA sequences of any two people. This process is applied to reproductively pair male and female participants in the ‘‘1000 Genomes Project’’ based on publicly available exome sequences. The output of a single pairing is 1,000 unique virtual progeny genomes. Each digital genome is analyzed for DNA sequence combinations that could cause one or more recessive Mendelian diseases based on clinical annotations and the total expected output of functional gene products from the two copies of each gene. The data are integrated to produce a detailed assessment of reproductive disease risk. RESULTS: Genetic screening of simulated genomes from over 50,000 pairings uncovered reproductive risk far beyond that exposed by carrier screening of individual persons. Based on coding regions interrogated in over 400 genes, all ‘‘donors’’ for the purposes of this investigation were considered healthy reproductive matches for multiple ‘‘clients.’’ CONCLUSION: Virtual progeny analysis can provide a comprehensive approach to the genetic screening of sperm donors in two distinct ways: 1) by incorporating client DNA information as a necessary input into risk analysis and 2) by selectively avoiding sperm donors based on the analysis of recessive genetic disease risk in the virtual offspring of qualified sperm bank donors and clients. Supported by: GenePeeks, Inc.
P-510 Wednesday, October 22, 2014 CLOMIPHENE CITRATE (CC) ADMINISTRATION CHANGES CERVICAL MUCUS PERMEABILITY IN PATIENTS UNDERGOING FERTILITY TREATMENTS. V. V. Snegovskikh,a,c K. B. SmithDupont,b M. House,c K. Ribbeck,b K. Pagidas.a,c aOB/Gyn, Women and Infants Hospital, Providence, RI; bBiological Engineering, Massachusetts Institute of Technology, Cambridge, MA; cOB/Gyn, Tufts Medical Center, Boston, MA. OBJECTIVE: The properties of cervical mucus(CM) have been investigated for many years,but limited data is available regarding its changes in patients undergoing different types of fertility treatments.In this study we use microfluidics technology to investigate the permeability properties of CM from ovulation induction (OI) with CC and natural cycle patients. DESIGN: Pilot case control study. MATERIALS AND METHODS: Patients presented to our clinic on the day of ovulation for intrauterine insemination(IUI). After informed consent was obtained CM samples were collected via speculum exam using insulin syringe directly from the cervix.Specimens were snap frozen in liquid nitrogen and stored at -80C until use.They did not contain any blood or tissue.Microfluidic devices were fabricated from polydimethylsiloxane (PDMS) as described previously(1) 2-5mL of whole mucus sample was pumped into the channel.Fluorescently labeled positively (PCP) and negatively-charged peptides(NCP) were used as reporters for permeability.The transport peptides were flowed by gravity orthogonally to the mucus channel and the mucus-water interface.Images of fluorescent peptide transport were acquired every 10 sec for at least 15 min using an inverted epifluorescence microscope (IX-71,Olympus, Central Valley,PA)equipped with a 10x objective, a light-emitting diode excitation source and a cooled camera. RESULTS: NCPs showed a similar transport behavior in OI and natural cycle mucus. In both cases the average peptide concentration as a function of distance along the length of the 300 mm microfluidic channel follows a similar trend as observed with phosphate buffer (H7), indicating that peptides diffuse largely unhindered through both types of CM samples. However, the mucus differed in permeability toward PCPs. In a group of OI patients (N¼14) receiving CC100 mg from days 3 to 7 of the cycle we observed a greater concentration of PCPs within 100mm from the mucus barrier compared to natural cycle ovulatory patients (N¼7). This indicates that CM from OI patients may be more adhesive toward positively charged epitopes. CONCLUSION: CM from women undergoing OI with CC has altered permeability properties compared to CM from natural cycle patients.This difference in mucus permeability could be clinically significant, as it may decrease the number of available sperm at the time of ovulation and prove the need of such procedure as IUI. Supported by: Burroughs Wellcome Fund Preterm Birth Initiative.
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ASRM Abstracts
P-511 Wednesday, October 22, 2014 DHEA SUPPLEMENTATION RESULTS IN SUPRAPHYSIOLOGIC DHEA-S SERUM LEVELS THAT INTERFERE WITH PROGESTERONE (P) IMMUNOASSAYS, RESULTING IN SPURIOUS P ELEVATIONS THAT MAY ALTER CLINICAL MANAGEMENT IN IVF. E. J. Forman,a,b J. M. Franasiak,a,b K. Scott,c E. K. Dubell,a M. Fano,a S. Ng,a R. T. Scott, Jr.,a,b aRMANJ, Basking Ridge, NJ; bRWJ, Rutgers, Basking Ridge, NJ; cRMACT, Norwalk, CT. OBJECTIVE: Despite a lack of prospective evidence of benefit, DHEA is being increasingly used for poor responders in an attempt to improve response to stimulation. The impact of the resulting supraphysiologic DHEA-S serum levels on sex steroid assays has not been evaluated. Since late follicular rises in P have been shown to adversely affect outcomes from fresh embryo transfers, even modest alterations in this assay may impact clinical management. This study seeks to determine the relationship between DHEA supplementation and P measurements to characterize the degree of interference with particular immunoassays. DESIGN: Prospective, endocrine. MATERIALS AND METHODS: DHEA-S control reagants with no P present (Advia Centaur, Siemens) were assayed for DHEA-S and P levels. Serum pools were created from IVF patents who were not on DHEA supplementation. Baseline DHEA-S and P was measured in these pools. Increasing amounts of DHEA-S was then added to the pooled serum and P was measured in quadruplicate on 3 different immunoassay instruments (Advia Centaur, Immulite 2000 – Siemens; eCobas 411 – Roche). The mean values were compared using Pearson’s correlation. RESULTS: When measuring the manufacturer’s DHEA-S controls (Centaur), there was a linear increase in the P detected, ranging from 0 ng/ mL in the blank control (no DHEA-S) to 1.5 ng/mL in the high control (DHEA-S >1500 ug/mL). The mean DHEA-S of the pooled serum was 142 ug/mL. The Centaur and eCobas assays had a linear relationship between DHEA-S added and the measured P value (P
