PGT-A on TE biopsies (TE Bx/NGS) provides a method of selecting blastocysts with excellent prognosis for establishing clinical pregnancies, minimizing miscarriages and improving live birth rates per ART procedure. However some practitioners distrust the reliability of TE Bx/NGS because mosaicism is seen in normal placentae (derived from the TE) and small numbers of TE cells biopsied may not represent the fetus (derived from the inner cell mass). We examined rebiopsy specimens from the TE and the ICM to determine the reliability of the clinical biopsy to characterize the blastocyst. In particular, we determined concordance between the clinical biopsies and rebiopsy specimens, focusing on 1) chromosomal concordance for disomic results, aneuploid results or mosaic results in the clinical biopsy and 2) clinical concordance (whether biopsy of the ICM was concordant with the initial biopsy's result of "Euploid" vs "NOT euploid") Rebiopsy of blastocysts with known results of clinical PGT-A Results of initial, clinical TE biopsies were obtained from Cooper Genomics. Vitrified blastocysts from patients consenting to research were selected for groups that had no aneuploidy (N = 10), aneuploidy with 1 or 2 aneuploid chromosomes (N = 4) or 1 aneuploid chromosome and 1 mosaic result (N =18). Blastocysts were rewarmed and cells from the TE and ICM were biopsied separately, obtaining as many rebiopsy specimens as possible. Biopsy specimens were subjected to WGA and NGS in our university's core laboratory. NGS results for rebiopsies were compared with results of the clinical biopsy. Rebiopsy chromosomes were considered concordant when the same chromosomal diagnosis was observed Chromosomal concordances were 97.0%, 74.3%, and 13.7% per chromosome, respectively, for disomic (Di), aneuploid (An) and mosaic (Mo) chromosomes in the clinical biopsy. Discordant chromosome results were predominantly mosaic results (2.6%) for Di, mosaic or complementary results (21.6%) for An, or were not seen or non-mosaic aneuploid results (74.5%) for Mo observed for the same chromosome that was seen in the clinical biopsy. These minor discordances can be considered concordant since they mainly confirm the initial results. Counting them as concordant leads to concordances 99.6% for Di, 95.9% for An, and 88.2% for Mo per chromosome. Rebiopsies of inner cell mass were clinically concordant for 100% of the blastocysts (biopsy result of ICM agreed with the clinical result of "euploid" or "not euploid"). Despite small number of biopsied cells (required to avoid damage to the blastocyst) and mosaicism (demonstrated by rebiopsy specimens) the excellent chromosomal concordance for rebiopsy specimens (99.6% and 95.9%) and clinical concordance for ICM biopsies (100%) indicate that TE biopsy/NGS provides excellent accuracy in its assessment of ploidy. Within this non-randomly selected subset of blastocysts, mosaics detected in the clinical biopsy outnumbered mosaics detected only by rebiopsy 2.25:1 (18:8).

Embryonic mosaicism occurs when two or more cell populations with different genotypes are present within the same embryo. New diagnostic techniques for preimplantation genetic screening (PGS), such as next-generation sequencing, have led to increased reporting of mosaicism. The interpretation of mosaicism is complicated because the transfer of some mosaic embryos has resulted in live births. Mosaic embryos may represent a third category between normal (euploidy) and abnormal (aneuploidy). This category of mosaic embryos may be characterized by decreased implantation and pregnancy potential as well as increased risk of genetic abnormalities and adverse pregnancy outcomes. Euploid embryos should be preferentially transferred over mosaic embryos. Genetic counseling is necessary before the transfer of a mosaic embryo is considered. Certain types of mosaic embryos should be preferentially transferred over others. Transfer of embryos with mosaic trisomies 2, 7, 13, 14, 15, 16, 18, and 21 may pose the most risk of having a child affected with a trisomy syndrome; however, the transfer of embryos with mosaic monosomies or other mosaic trisomies are not devoid of risk. Patients must be counseled about the risk of undetected monosomies or trisomies within a biopsy specimen as well as the risk of intrauterine fetal demise or uniparental disomy with the transfer of mosaic embryos. Until more data are available, patients should be encouraged to undergo another cycle to obtain euploid embryos, when possible, rather than transferring a mosaic embryo. (C) 2016 by American Society for Reproductive Medicine.

PGD for BRCA testing offers a viable option for those women hoping to decrease disease in and increase survival of their children. We set out to characterize the outcomes of patients undergoing PGD to test for BRCA mutations along with PGS for aneuploidy assessment, and to determine the odds of delivery per retrieval. Retrospective Descriptive Series. All PGD cycles for the BRCA mutation (both BRCA1 and BRCA2) performed at a reference genetics laboratory were queried. Only cycles in which both PGS and PGD were performed were included. All methodologies of PGD and PGS were included in the analysis. Determination of delivery rate was based on an established live birth rate of 65% per euploidic embryo. 48 cycles resulted from the query and 42 cycles were included in the analysis. Twenty nine cycles were PGD for BRCA1 and 13 for BRCA2. The average maternal age was 33.29±4.27 (range 23-40) years with 6.76±4.29 (range 1-17) embryos for biopsy. The overall percentage of affected embryos for BRCA mutations was 52.36%±20.37% of which 40.11% ± 36.51% were found to be aneuploid. No unaffected euploid embryos were available for transfer in 26.19% (n=11) cases. When compared by age, no statistical significance was found in the number of embryos for biopsy, percent affected for BRCA mutation, the number of transferable (unaffected and euploid) embryos available, or chance of live birth following a single embryo transfer per cycle. The rate of aneuploid embryos in BRCA patients appears consistent with previously published rates of age associated aneuploidy(1). BRCA 1 and BRCA2 patients were independently analyzed, showing no statistical difference in any parameters evaluated. Combining PGD for BRCA and PGS for aneuploidy reduces the number of embryos available for transfer but allows for more specific selection of normals. Due to the rate of PGS/PGD normal being 14-29% of embryos, some women may be faced with considering the transfer of an effected male embryo. Patients undergoing embryo selection for BRCA should be well-informed of the odds of finding a PGS/PGD normal embryo, having no embryo to transfer, a male normal PGS carrier and pregnancy.Tabled 1PGD/PGS Results for BRCA Mutations Stratified by Age GroupageBRCA pos (%)Aneuploid Embryos (%)Aneuploid-BRCA pos Embryos (%)Female BRCA pos embryos (%)Male BRCApos (Embryos (%)Embryos for transfer (%)Chance of pregnancy per retrieval + single embryo transfer (%)24-30(n=10)51.51±21.7134.99±20.8455.0±25.355.00±25.3528.7±34.529.08±20.0418.90±13.0331-35(n=21)61.57±22.2938.73±27.8648.2±37.148.17±37.1356.8±35.224.69±22.4616.05±14.60>35 (n=11)54.05±19.5849.0±33.549.0±33.549.03±33.5444.6±33.313.75±13.388.94±8.70p0.4140.0690.8660.8690.1160.2250.225 Open table in a new tab

Patients with primary infertility (PI), secondary infertility (SI), and recurrent pregnancy loss (RPL) often seek in vitro fertilization (IVF) with preimplantation genetic screening (PGS). Next-generation sequencing (NGS) is highly sensitive assay for aneuploidy and mosaicism. The objective of this study is to determine if infertility diagnosis is associated with mosaicism in the first IVF cycle using PGS with NGS. The first IVF cycle of all patients undergoing IVF/PGS with NGS at an academic fertility center from January 2015 to December 2015 was included in the analysis. Pregnancy and infertility history prior to the analyzed cycle was gathered through review of the medical record and categorized into three groups defined based on indication for IVF: PI, as no history of spontaneous conception; SI, having had at least one spontaneous conception; or RPL, two or more consecutive pregnancy losses. Patients with IVF indications other than PI, SI or RPL and those with undocumented pregnancy history were excluded from analysis. Only embryos with true mosaicism were included; all embryos with aneuploidy and mosaicism were categorized aneuploid. Statistical analysis was done with ANOVA and Poisson logistic regression. A total of 316 cycles and 1466 embryos were analyzed. The average age of the population was 38.6 years (± 3.8). The most common cause of infertility was PI, (n=194, 62%), followed by SI (n=74, 23%) and RPL (n=47, 15%), however the no significant difference was found Poisson regression analysis (see Table 1). We also performed an analysis controlling for age, AMH, eggs retrieved and eggs undergoing ICSI which were significant in the univariate analysis. These factors did not alter the significance of association between mosaicism and infertility diagnosis. Post-hoc power to detect a difference between RPL and PI was insufficient at 0.29. No association between cause of infertility and rate of mosaicism was seen in this study. Regardless, mosaicism is more frequently detected with NGS. While some mosaic embryos lead to successful pregnancies and are eligible for transfer in certain circumstances, the mosaicism rate in the patient population as a whole, 18.1%, warrants further investigation. The post-hoc power analysis suggests a larger sample size may be needed to delineate any associations within these diagnoses.Table 1Comparison of IVF/PGS cycle characteristics by cause of infertilityPrimarySecondaryRPLp-value ( Anova*, Poisson regression**)Patients (n)1947447Age (years)38.1 ± 4.040.0 ± 3.339.0 ± 3.6<0.005*AMH (ng/mL)3.84 ± 6.12.03 ± 2.12.06 ± 1.30.02*E2 (pg/mL)53.5 ± 31.464.3 ±50.256.9 ± 41.90.12*FSH (mIU/mL)5.8 ± 2.96.5 ± 2.96.2 ± 2.60.16*# Eggs retrieved14.7 ± 9.112.1 ± 8.013.6 ± 8.2<0.005**# Embryos biopsied4.6 ± 3.64.3 ± 3.74.6 ± 3.70.12**Euploid/Embryo (%)23.5 ± 25.317.9 ± 24.422.5 ± 26.60.45**Mosaic/Embryo (%)17.6 ± 23.113.6 ± 23.520.0 ± 30.40.47**Aneuploid/Embryo (%)58.9 ± 32.568.5 ± 33.857.5 ± 36.90.47**Euploid/Embryo (Adjusted)25.2 ± 11.021.2 ± 9.225.5 ± 10.80.45**Mosaic/Embryo (Adjusted)19.3 ± 5.415.3 ± 4.918.3 ± 4.00.47**Aneuploid/Embryo (Adjusted)55.9 ± 18.563.4 ± 16.655.8 ± 15.70.32** Open table in a new tab

To assess the reliability of human embryos with previous TE biopsies with an NGS diagnosis of aneuploid-mosaic, aneuploid or euploid by a reference genetics laboratory. Repeat biopsies of these embryos were performed and NGS was repeated in our own laboratory. Prospective translational research study Discarded human embryos diagnosed clinically as aneuploid mosaic (containing both aneuploid and mosaic chromosome complement), aneuploid or euploid by NGS were thawed and re-biopsied to obtain TE and ICM samples. Whole genome amplification using the SurePlex DNA amplification system followed by NGS via the Illumina MiSeq platform was performed. Data was interpreted by BlueFuse Multi software and mosaicism was defined by copy number variation of 30% or more. Statistical analysis was performed using likelihood G test and chi square. 24 embryos from 16 patients were included in the study. 18 embryos had been diagnosed as aneuploid-mosaic, 4 aneuploid and 2 euploid. 96 samples were obtained and NGS was completed in four batches of 24 samples per MiSeq run, composed of 64 TE and 32 ICM biopsies. 24 samples were unable to be interpreted, because of chaotic profiles (16 TE and 8 ICM), thus not included in the analyses. NGS analysis of ICM and TE biopsies showed both were highly concordant with the original diagnosis of aneuploidy (100% and 92.50% p >0.05). ICM and TE both were less concordant with the original diagnosis of mosaicism (31.58% and 17.65% p=0.14). Additional mosaic calls were more frequently found in TE (37.50%) compared to ICM samples (16.67%), p=0.038. The number of cells included in the biopsy did not influence the diagnosis of aneuploidy or mosaicism, however, biopsies with fewer than 5 cells were more likely to result in chaotic profiles (p=0.039). The diagnosis of aneuploidy in a TE biopsy is highly predictive of aneuploidy in both ICM and other samples of the TE. Mosaicism in the TE biopsy is a less reliable predictor of mosaicism in ICM or TE. Mosaicism is more commonly seen in TE than ICM, supporting prior hypotheses that the ICM may be less hospitable to mosaicism than TE. No aneuploid or mosaic embryos resulted in euploid ICMs, eliminating the concern for the the non-transfer of a potentially euploid ICM embryo; thus confirming that a single TE biopsy with an aneuploid result is unlikely to result in a euploid live born.Tabled 1Repeat NGS Results Compared to Initial Clinical TE Biopsy NGS ResultsICMTEpAneuploid Concordant (%)(24/24)100%(37/40) 92.50%>0.05Mosaic concordant (%)(6/19) 31.58%(6/34)17.65%0.136Additional mosaic (%)(4/24)16.67%(18/48)37.50%0.0376Chaotic Profile (%)(8/32)25%(16/64) 25%0.498 Open table in a new tab

Single thawed euploid embryo transfer (STEET) decreases the risk of maternal and neonatal morbidity associated with multiple gestation. [1] If transfer of a euploid embryo fails to progress to a clinical pregnancy, we sought to determine if there are identifiable risk factors for a failed second STEET. Retrospective case-control. All single thawed transfers of embryos designated as euploid by Next Generation Sequencing (NGS) were identified between April 2015 and February 2016 (n=263 transfers, 232 patients). Only patients with a first failed STEET and a subsequent second attempt at elective STEET were included (n=25 patients). Cycle characteristics were compared between those with and without a successful second transfer. Analyses were performed using t-tests and chi square analyses. Clinical pregnancy (fetal heart rate) after NGS diagnosis with STEET is 62.3% in our patient population. After one failed STEET, clinical pregnancy occurred in 56% of patients in the second STEET cycle (p=0.54). Endometrial thickness was significantly increased in patients that achieved clinical pregnancy on the second transfer as compared to those that did not (p=0.01). A lower average age at transfer and higher number of euploid blastocysts remaining was seen in the group achieving pregnancy, but statistical significance was not reached (Table 1).Tabled 1Patients with a second STEET after one failed STEET with diagnosis via NGS from 4/2015 to 2/2016.No Clinical Pregnancy with Second STEET (n=11 patients)Achieved Clinical Pregnancy with Second STEET (n=14 patients)p valueMaternal age at transfer (years)39.2 (+5.1)37.0 (+3.9)0.09Age at vitrification (years)33.1 (+5.0)36.1 (+4.4)0.03Number of days post-retrieval (n)80.7 (+52.8)86.0 (+49.3)0.71Number of euploid blastocysts remaining (n)1.5 (+1.1)2.2 (+1.5)0.07Endometrial echo on cycleday 14 (cm)7.5 (+0.9)8.5 (+1.4)0.01Progesterone level on day of transfer (ng/ml)17.1 (+6.5)18.0 (+5.9)0.87Estradiol level on day of transfer (pg/ml)622.1 (+539.1)492.9 (+573.6)0.43Most common embryo quality (%)3-5Bb (59.1%)3-5 Bb (60.7%)0.91Description of embryo as “fully expanded” on day of transfer (%)31.8%50%0.12 Open table in a new tab Lower pregnancy rates might be expected after an initial failed transfer cycle, as the number of good quality blastocysts available is reduced. However, clinical pregnancy rate with a second STEET following an initial failed cycle was similar to the pregnancy rate seen for all first transfers in our population, providing hope for couples. A thicker endometrial echo and younger maternal age at transfer were associated with increased ability to achieve clinical pregnancy in a second STEET after previous failure, indicating a possible underlying uterine or embryologic factor. Further research is needed to identify other potential contributions to pregnancy failure with STEET in order to increase utilization of single embryo transfer.

To identify if the rate of mosaicism and aneuploidy differ among different IVF centers performing trophectoderm biopsies on blastocysts for Preimplantation Genetic Screening (PGS) with Next Generation Sequencing (NGS). Retrospective analysis. All donor oocyte cycles undergoing PGS with NGS from trophectoderm biopsy specimens, received by a large United States Genetics Laboratory, were queried. Only centers with biopsy specimens from ten or more patients were included in the study. Analysis was limited to cycles from donors between the ages of 18 and 30 years old. Blastocyst biopsy results were classified as being euploid, aneuploid or mosaic. Embryos defined as mosaic showed a 20 percent or greater mosaicism rate within the biopsy specimens. Biopsy results showing mosaicism or mosaicism with a single aneuploidy were classified the same (only 0.97% of all NGS samples run show mosaic with single aneuploidy). Any result with two or more aneuploidies plus mosaicism was classified to be complex abnormal and defined as aneuploid. Statistical analysis was done using ANOVA and linear regression. A total of 268 cycles from 38 different IVF centers resulted from the query. A total of 192 cycles from 9 different IVF centers were included in the analysis. The average age of patients was 25.15± 2.60 years old (range 19-30), with a cumulative total of 1492 blastocysts biopsied. The overall euploid rate was 48.94%±24.61,with an aneuploid rate of 24.65%±20.76 and mosaic rate of 26.41%±25.0. When the 9 centers were compared as a group, there was a significant difference in mosaic rate (p<0.001) and aneuploid rate (p0.018), but not in the euploid rate (p 0.166). Linear regression on all 268 cycles showed no association between age and rate of euploidy (R=0.023), aneuploidy (R=0.056) or mosaicism (R=0.054); indicating differences in donor ages amongst centers has no affect. There was no significant correlation between the number of blastocysts for biopsy and the percent euploid(R=0.122), aneuploid(R=0.035) or mosaic (R =0.099). The difference in mosaic and aneuploid rateswith NGS amongst donor cycles within different IVF institutions shows that there may be a significant practice or laboratory component affecting outcomes. The lack of association of mosaicism and aneuploidy in this population further indicates environmental factors may play an important role.Tabled 1NGS Results for Donor Cycles aged 18 to 30 years old from IVF Centers with Ten or More CyclesNo. Cycles per Center (a-i)Avg Donor AgeAvg No. Biopsy per PatientNo Amplification (%)aDegraded DNA (%)aEuploid (%)bAneuploid(%)bMosaic (%)ba-1624.00±2.317.06±2.860.88±4.172.65±5.4644.95±28.1132.11±26.9022.94±13.58b-2325.27±2.356.55±3.530.63±1.780.69±4.2652.08±27.7018.06±18.9829.86±26.63c-1222.92±1.447.75±3.144.30±10.762.15±3.7343.68±20.1739.08±19.1817.24±12.20d-4125.50±2.447.30±3.831.03±5.452.40±4.8656.29±25.0726.22±22.8617.48±18.73e-1424.00±2.0410.43±4.110.68±4.451.37±10.6950.35±21.6217.48±10.6932.17±19.79f-1925.42±3.029.21±5.481.14±3.334.00±10.4438.82±20.8827.65±19.8233.53±15.07g-1825.41±3.108.53±4.520.00±0.002.76±11.0447.92±24.6418.06±21.0234.03±21.67h-3626.14±2.588.03±4.740.35±3.331.38±7.8445.49±23.2926.74±17.1327.78±26.31i-1325.31±2.365.31±2.560.00±0.002.90±11.0937.31±22.2014.93±12.1847.76±21.26p0.0050.0550.3510.9870.1690.018<0.001a.Percent expressed as total over total number biopsied b.Percent expressed as total over total embryos with diagnosis given Open table in a new tab

ObjectiveVertebral fracture is the most common clinical manifestation of osteoporosis and is significantly associated with an increased risk of future fractures.1 Bone mineral density has traditionally been the best predictor of fragility fractures, however, lean mass may have a greater contribution to the risk of fracture than previously understood. Dual-energy X-ray absorptiometry (DXA) allows for highly accurate measurements of bone mass as well as both fat and lean body mass. The primary objective of this study is to determine if there is an association between lean body mass and the incidence of vertebral fragility fractures in postmenopausal women. The presence of an association between number of vertebral fractures and T-score, Z-score, body mass index (BMI), muscle mass index (lean mass (kg)/ height (m2)), and fat mass are secondary outcome measures.DesignRetrospective cohort study.Materials and MethodsAll women between the ages of 40 and 100 years, who underwent body composition and bone density testing using DXA scan at the NYU Bone Density and Body Composition Unit from May 2011 to November 2014 were identified. All indications were included. Patients with DXA that did not include a lateral vertebral assessment were excluded. Parametric variables were confirmed by Shapiro-Wilk testing and compared by analysis of variance (ANOVA). Chi-square testing was used for nominal variables.ResultsA total of 358 women met inclusion criteria. The average age was 70.2 years +10.3 (Range 46 to 93 years), average weight was 139.6lbs + 25.9 (Range 90 to 267 lbs) and average body mass index (BMI) was 25.0 + 4.5 kg/m2 (Range 16.7 to 42.3). A total of 124 vertebral fractures were identified in 85 patients (23.7%), with an average of 0.35 (+ 0.7) vertebral fractures per patient. Both lean body mass and Z-score were noted to have an inverse association with number of vertebral fractures (p=0.03 and p=0.02, respectively). Women without vertebral fractures had an average lean mass of 62.4 lbs (+8.5) (average BMI 24.9 +4.5), while women with 3 vertebral fractures had an average lean mass of 59.5 lbs (+8.7) (average BMI 26.0 +4.5). Women with at least one vertebral fracture were more likely to have an average T-score of at least -0.8 (+1.5), but T-score was not found to be significantly associated with number of fractures (p=0.26). Additionally, fat mass (p=0.82), BMI (p=0.19), and muscle mass index (p=0.36) did not prove to be predictive of vertebral fracture in this population.ConclusionsIrrespective of BMI, a lean mass of greater than 62.4lbs was associated with lower incidence of vertebral fracture in our population. These results suggest the importance of assessing lean mass in postmenopausal women, as it is a modifiable risk factor for osteoporotic fractures.Reference:1.Lindsay R, Silverman SL, Cooper C, Hanley DA, Barton I, Broy SB, Licata A, Benhamou L, Geusens P, Flowers K, Stracke H, Seeman E. Risk of new vertebral fracture in the year following a fracture. JAMA 2001;285(3):320. ObjectiveVertebral fracture is the most common clinical manifestation of osteoporosis and is significantly associated with an increased risk of future fractures.1 Bone mineral density has traditionally been the best predictor of fragility fractures, however, lean mass may have a greater contribution to the risk of fracture than previously understood. Dual-energy X-ray absorptiometry (DXA) allows for highly accurate measurements of bone mass as well as both fat and lean body mass. The primary objective of this study is to determine if there is an association between lean body mass and the incidence of vertebral fragility fractures in postmenopausal women. The presence of an association between number of vertebral fractures and T-score, Z-score, body mass index (BMI), muscle mass index (lean mass (kg)/ height (m2)), and fat mass are secondary outcome measures. Vertebral fracture is the most common clinical manifestation of osteoporosis and is significantly associated with an increased risk of future fractures.1 Bone mineral density has traditionally been the best predictor of fragility fractures, however, lean mass may have a greater contribution to the risk of fracture than previously understood. Dual-energy X-ray absorptiometry (DXA) allows for highly accurate measurements of bone mass as well as both fat and lean body mass. The primary objective of this study is to determine if there is an association between lean body mass and the incidence of vertebral fragility fractures in postmenopausal women. The presence of an association between number of vertebral fractures and T-score, Z-score, body mass index (BMI), muscle mass index (lean mass (kg)/ height (m2)), and fat mass are secondary outcome measures. DesignRetrospective cohort study. Retrospective cohort study. Materials and MethodsAll women between the ages of 40 and 100 years, who underwent body composition and bone density testing using DXA scan at the NYU Bone Density and Body Composition Unit from May 2011 to November 2014 were identified. All indications were included. Patients with DXA that did not include a lateral vertebral assessment were excluded. Parametric variables were confirmed by Shapiro-Wilk testing and compared by analysis of variance (ANOVA). Chi-square testing was used for nominal variables. All women between the ages of 40 and 100 years, who underwent body composition and bone density testing using DXA scan at the NYU Bone Density and Body Composition Unit from May 2011 to November 2014 were identified. All indications were included. Patients with DXA that did not include a lateral vertebral assessment were excluded. Parametric variables were confirmed by Shapiro-Wilk testing and compared by analysis of variance (ANOVA). Chi-square testing was used for nominal variables. ResultsA total of 358 women met inclusion criteria. The average age was 70.2 years +10.3 (Range 46 to 93 years), average weight was 139.6lbs + 25.9 (Range 90 to 267 lbs) and average body mass index (BMI) was 25.0 + 4.5 kg/m2 (Range 16.7 to 42.3). A total of 124 vertebral fractures were identified in 85 patients (23.7%), with an average of 0.35 (+ 0.7) vertebral fractures per patient. Both lean body mass and Z-score were noted to have an inverse association with number of vertebral fractures (p=0.03 and p=0.02, respectively). Women without vertebral fractures had an average lean mass of 62.4 lbs (+8.5) (average BMI 24.9 +4.5), while women with 3 vertebral fractures had an average lean mass of 59.5 lbs (+8.7) (average BMI 26.0 +4.5). Women with at least one vertebral fracture were more likely to have an average T-score of at least -0.8 (+1.5), but T-score was not found to be significantly associated with number of fractures (p=0.26). Additionally, fat mass (p=0.82), BMI (p=0.19), and muscle mass index (p=0.36) did not prove to be predictive of vertebral fracture in this population. A total of 358 women met inclusion criteria. The average age was 70.2 years +10.3 (Range 46 to 93 years), average weight was 139.6lbs + 25.9 (Range 90 to 267 lbs) and average body mass index (BMI) was 25.0 + 4.5 kg/m2 (Range 16.7 to 42.3). A total of 124 vertebral fractures were identified in 85 patients (23.7%), with an average of 0.35 (+ 0.7) vertebral fractures per patient. Both lean body mass and Z-score were noted to have an inverse association with number of vertebral fractures (p=0.03 and p=0.02, respectively). Women without vertebral fractures had an average lean mass of 62.4 lbs (+8.5) (average BMI 24.9 +4.5), while women with 3 vertebral fractures had an average lean mass of 59.5 lbs (+8.7) (average BMI 26.0 +4.5). Women with at least one vertebral fracture were more likely to have an average T-score of at least -0.8 (+1.5), but T-score was not found to be significantly associated with number of fractures (p=0.26). Additionally, fat mass (p=0.82), BMI (p=0.19), and muscle mass index (p=0.36) did not prove to be predictive of vertebral fracture in this population. ConclusionsIrrespective of BMI, a lean mass of greater than 62.4lbs was associated with lower incidence of vertebral fracture in our population. These results suggest the importance of assessing lean mass in postmenopausal women, as it is a modifiable risk factor for osteoporotic fractures.Reference:1.Lindsay R, Silverman SL, Cooper C, Hanley DA, Barton I, Broy SB, Licata A, Benhamou L, Geusens P, Flowers K, Stracke H, Seeman E. Risk of new vertebral fracture in the year following a fracture. JAMA 2001;285(3):320. Irrespective of BMI, a lean mass of greater than 62.4lbs was associated with lower incidence of vertebral fracture in our population. These results suggest the importance of assessing lean mass in postmenopausal women, as it is a modifiable risk factor for osteoporotic fractures.

The architecture and structure of the meiotic spindle influence embryo development(1) and risk of aneuploidy in women(2). The factors that disrupt the meiotic spindle remain incompletely understood. Dysfunctional mitochondria produce reactive oxygen species (ROS), which may directly perturb spindles(3). ROS also can disrupt spindles by inducing telomere attrition, since telomeres are essential for spindle formation and are especially susceptible to ROS(4). We studied the impact of ROS, produced by uncoupling mitochondria, on the area and retardance (measure of molecular order) of meiotic spindles and on mean telomere length of individual human oocytes. Prospective, randomized, paired research laboratory intervention. 44 germinal vesicle (GV) stage oocytes were accessioned from 16 patients undergoing IVF/ICSI. GVs from each patient were randomly assigned to control or treatment group. Oocytes in the treatment group were cultured in media containing Carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone (FCCP, 750nM) for one hour to induce mitochondrial stress. Control oocytes were cultured in media without FCCP. GV oocytes from both groups were further cultured to permit meiotic maturation, as indicated by extrusion of the first polar body. Spindles were imaged non-invasively with an orientation independent polarized light microscope (Oosight, Hamilton Thorne, MA, USA). Spindle area and mean retardance were measured with software from the Oosight Imaging System. Mean oocyte telomere length was measured by single cell qPCR. Data were analyzed by Chi-square test and independent t test. Maturation rates of oocytes in the control and FCCP groups were 66.67% and 56.52% respectively. 71.4% of MII oocytes in the control group and 61.5% in the FCCP group had a detectable birefrigent spindle. FCCP decreased the area of the spindles compared to controls (19.84±2.11 sq. microns versus 37.77±4.79, P=0.011). Mean spindle retardance in the FCCP group also was significantly lower than that of controls (1.45±0.11 nm versus 2.19±0.16 nm, P=0.003). Telomere length (T/R ratio) did not differ between treatment and control groups (1.11±1.56 versus 1.29±0.19, P>0.05). Telomere length of oocytes with and without birefringence spindle within the treatment group also did not differ significantly (P>0.05). Meiotic maturation itself is relatively resistant to ROS, consistent with prior studies showing limited cell cycle check point control during oogenesis. However, ROS produced by acute mitochondrial stress disrupts spindle retardance and size. Reactive oxygen, at least acutely, does not induce telomere attrition in human oocytes.

To assess if SART donor oocyte pregnancy rates are consistent with high percentage of aneuploidy and mosaicism in donor oocyte cycles using Preimplantation Genetic Screening (PGS) with Next Generation Sequencing (NGS). Retrospective study and mathematical analysis. All trophectoderm biopsy specimens from donor oocyte cycles received by a genetics laboratory were queried for NGS results. The rate of euploidy, aneuploidy, and mosaicism was calculated. A separate analysis was done to find the ongoing pregnancy rate (OPR) from patients’ first donor single thawed euploid embryo transfers (STEET) from NGS at a single university fertility center from 2/2015 to 3/2016. Lastly, the 2014 SART National Summary data on non-tested donor embryo transfers and their live birth rates was used to calculate the expected live births from untested donor embryo transfers based on euploidy and mosaicism rates from NGS donor data, applying the the OPR from donor STEET data. The calculated expected live birth rate was then compared to the actual live birth rate of untested donor embryo transfer cycles. 268 donor cycles from 38 IVF centers showed a euploid rate of 48.7%±23.8, aneuploid rate of 22.9%±20.4 and mosaic rate of 28.4%±21.9 (Table 1). Thirty two patients were included in the donor NGS STEET cycle analysis. The average age at transfer was 42.7 ±3.9 years with an average donor age of 26.5 ±2.9 years, for an OPR of 62.2% (n=20/32). The 2014 SART National Summary shows 6070 non-PGS donor egg transfers, averaging 1.7 embryos per transfer, for an estimate of 10319 embryos transferred in total. The reported live birth rate of 53.3% (36.9% singletons, 16.2% twins and 0.2% triplets) gives a cumulative 4242 babies delivered (41.1% per embryo). When the euploid rate of 48.7% is applied, 5025 embryos transferred are estimated to have been euploid. The 62.2% OPR of donor STEETs applied to this gives an expected 3126 singleton deliveries (95% CI 2927-3246). The mosaic rate of 28.4% plus data showing an overall approximate 20%1,2 OPR of mosaic embryos adds 586 (95% CI 421-524) singleton births. The total expected babies born is 3712 (95% CI 3505-3877) with a live birth rate of 35.9%(95% CI 34.0-37.6) from donor egg transfers. High rates of aneuploidy and mosaicism in donor oocyte cycles are consistent with the pregnancy rates of untested donor embryo transfers. The use of PGS with NGS can prevent transfer of aneuploid embryos in donor cycles.Tabled 1Donor Oocyte NGS Results from Reference Genetics LaboratoryTotal cycles268Total Trophectoderm Biopsy Specimens for NGS2062Avg age of Donors (years)25.8±2.3Avg age of Recipient (years)43.1±6.3Avg Number Cycles per Center7.1±10.5Avg Number Blastocysts Biopsied per Patient7.7±4.1Euploid (%)48.7± 23.8Aneuploid(%)22.9±20.4Mosaic (%)28.4±21.9No Amplification (%)2.5±7.3Rate of Degraded DNA (%)0.8±4.1 Open table in a new tab

Reactive oxygen species (ROS) are a major cause of aging in all tissues studied, including reproductive tissues. Recent studies demonstrate extensive DNA damage repair capacity in oocytes (1), and genetic variation in DNA damage repair pathways is associated with reproductive lifespan in women (2). We hypothesized that MII oocytes are more resistant to oxidative stress than other stages of development. Prospective, randomized study of a biologic intervention on mouse oocytes and embryos. 80 MII oocytes and 40 cleavage embryos from B6C3F1 mice (Embryotech Laboratories, Inc, USA) were thawed and exposed to oxidative stress induced by Carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone (FCCP, 750nM), which generates ROS by uncoupling mitochondrial electron transport and disrupting mitochondrial function. Oocytes and embryos were randomized into untreated controls, or 5, 10 or 20 hour exposure to FCCP. DNA damage was determined by immuno-fluorescent staining for γ-H2AX, or by mean telomere length, measured by single-cell qPCR. Data were analyzed by chi-square test and one-way ANOVA. 20 hours of exposure to FCCP is lethal for all embryos. Embryos exposed to FCCP for 5 and 10 hours of FCCP show increased γ-H2AX staining (5 hours- 87.5% vs. 25% positive cells P<0.05; 10 hours- 100% vs 25%, P<0.05). However, these same doses and durations of FCCP do not increase DNA damage in oocytes- there is no significant increase of γ-H2AX positive MII oocytes compared to controls (30% in the control group, 40% in the 5-hour group and 30% in the 10-hour group, P>0.05). Significantly higher numbers of γ-H2AX positive MII oocytes are found only in the 20-hour group (100%, P<0.05), a dose which is uniformly lethal to embryos. As previously demonstrated, 45 minutes of 750nM FCCP treatment shortens telomeres in cleavage stage mouse embryos (3). However, MII oocytes exposed to 750nM FCCP for 5, 10, or 20 hours show no statistically significant telomere shortening compared to controls (P>0.05). MIIoocytes are more resistant to oxidative stress than cleavage embryos.

ObjectiveTo evaluate if the degree of blastocyst expansion following rewarming of euploid embryos in a frozen embryo transfer cycle affects on pregnancy outcomes.DesignRetrospective Cohort Study.Materials and MethodsPatients undergoing a frozen euploid embryo transfer from June 2013 to October 2014 were included. Preimplantation genetic screening using array comparative genomic hybridization designated embryos to be euploid or aneuploid. All embryos had undergone trophectoderm biopsy followed by vitrification. Following our standard FET protocol, vitrified euploid embryos were rewarmed on the day of transfer. Embryologists documented degree of expansion at time of transfer for all frozen embryo transfers. Patients with all causes of infertility, and both autologous and donor cycles were included. Primary outcome included pregnancy, defined by presence of a gestational sac, and clinical pregnancy, defined by live birth or fetal heart beat through cycle day 63. Statistical analysis was done using relative risk and Chi Square.ResultsTabled 1Pregnancy Outcomes by Embryo Expansion at Time of TransferNTotal Pregnant (presence of gestational sac)PercentTotal Clinical Pregnancy (fetal heart beat of Live Birth)PercentExpanded14310674.129566.43Partially Expanded643453.133250.0Collapsed211164.701047.62Total228151137P value0.00470.0392 Open table in a new tab ConclusionsThere is a higher chance of implantation and ongoing pregnancies that lead to live birth in embryos that are re-expanded at time of transfer following the thaw of a euploid embryo. ObjectiveTo evaluate if the degree of blastocyst expansion following rewarming of euploid embryos in a frozen embryo transfer cycle affects on pregnancy outcomes. To evaluate if the degree of blastocyst expansion following rewarming of euploid embryos in a frozen embryo transfer cycle affects on pregnancy outcomes. DesignRetrospective Cohort Study. Retrospective Cohort Study. Materials and MethodsPatients undergoing a frozen euploid embryo transfer from June 2013 to October 2014 were included. Preimplantation genetic screening using array comparative genomic hybridization designated embryos to be euploid or aneuploid. All embryos had undergone trophectoderm biopsy followed by vitrification. Following our standard FET protocol, vitrified euploid embryos were rewarmed on the day of transfer. Embryologists documented degree of expansion at time of transfer for all frozen embryo transfers. Patients with all causes of infertility, and both autologous and donor cycles were included. Primary outcome included pregnancy, defined by presence of a gestational sac, and clinical pregnancy, defined by live birth or fetal heart beat through cycle day 63. Statistical analysis was done using relative risk and Chi Square. Patients undergoing a frozen euploid embryo transfer from June 2013 to October 2014 were included. Preimplantation genetic screening using array comparative genomic hybridization designated embryos to be euploid or aneuploid. All embryos had undergone trophectoderm biopsy followed by vitrification. Following our standard FET protocol, vitrified euploid embryos were rewarmed on the day of transfer. Embryologists documented degree of expansion at time of transfer for all frozen embryo transfers. Patients with all causes of infertility, and both autologous and donor cycles were included. Primary outcome included pregnancy, defined by presence of a gestational sac, and clinical pregnancy, defined by live birth or fetal heart beat through cycle day 63. Statistical analysis was done using relative risk and Chi Square. ResultsTabled 1Pregnancy Outcomes by Embryo Expansion at Time of TransferNTotal Pregnant (presence of gestational sac)PercentTotal Clinical Pregnancy (fetal heart beat of Live Birth)PercentExpanded14310674.129566.43Partially Expanded643453.133250.0Collapsed211164.701047.62Total228151137P value0.00470.0392 Open table in a new tab ConclusionsThere is a higher chance of implantation and ongoing pregnancies that lead to live birth in embryos that are re-expanded at time of transfer following the thaw of a euploid embryo. There is a higher chance of implantation and ongoing pregnancies that lead to live birth in embryos that are re-expanded at time of transfer following the thaw of a euploid embryo.

ObjectiveTo assess if an increase in paternal age has an affect on pregnancy outcomes of frozen euploid embryo transfer cycles.DesignRetrospective Cohort StudyMaterials and MethodsGlobal retrospective IRB was obtained. Patients undergoing STEET from June 2013 to October 2014 were included. Preimplantation genetic screening (PGS) using array comparative genomic hybridization (aCGH) designated embryos to be euploid or aneuploid. All embryos had undergone trophectoderm biopsy followed by vitrification. Patients with all causes of infertility, all indications for PGS, and both autologous and donor cycles were included. Primary outcome included pregnancy, defined by presence of a gestational sac, and clinical pregnancy defined by live birth or presence of fetal heart rate through cycle day 63. Statistical analysis was done using chi-square and linear regression.ResultsTabled 1Pregnancy Outcomes by Paternal Age Groups.Paternal AgeTotalPregnancyPercentClinical PregnancyPercent<35473165.962961.7035-441459666.218860.69≥45523669.233159.61Total24416366.8014860.66P valuep 0.916p 0.977 Open table in a new tab ConclusionsThere is no association between paternal age and pregnancy outcomes in patients undergoing STEET. This provides no support for the notion that older men influence embryo quality, implantation or early gestational development. ObjectiveTo assess if an increase in paternal age has an affect on pregnancy outcomes of frozen euploid embryo transfer cycles. To assess if an increase in paternal age has an affect on pregnancy outcomes of frozen euploid embryo transfer cycles. DesignRetrospective Cohort Study Retrospective Cohort Study Materials and MethodsGlobal retrospective IRB was obtained. Patients undergoing STEET from June 2013 to October 2014 were included. Preimplantation genetic screening (PGS) using array comparative genomic hybridization (aCGH) designated embryos to be euploid or aneuploid. All embryos had undergone trophectoderm biopsy followed by vitrification. Patients with all causes of infertility, all indications for PGS, and both autologous and donor cycles were included. Primary outcome included pregnancy, defined by presence of a gestational sac, and clinical pregnancy defined by live birth or presence of fetal heart rate through cycle day 63. Statistical analysis was done using chi-square and linear regression. Global retrospective IRB was obtained. Patients undergoing STEET from June 2013 to October 2014 were included. Preimplantation genetic screening (PGS) using array comparative genomic hybridization (aCGH) designated embryos to be euploid or aneuploid. All embryos had undergone trophectoderm biopsy followed by vitrification. Patients with all causes of infertility, all indications for PGS, and both autologous and donor cycles were included. Primary outcome included pregnancy, defined by presence of a gestational sac, and clinical pregnancy defined by live birth or presence of fetal heart rate through cycle day 63. Statistical analysis was done using chi-square and linear regression. ResultsTabled 1Pregnancy Outcomes by Paternal Age Groups.Paternal AgeTotalPregnancyPercentClinical PregnancyPercent<35473165.962961.7035-441459666.218860.69≥45523669.233159.61Total24416366.8014860.66P valuep 0.916p 0.977 Open table in a new tab ConclusionsThere is no association between paternal age and pregnancy outcomes in patients undergoing STEET. This provides no support for the notion that older men influence embryo quality, implantation or early gestational development. There is no association between paternal age and pregnancy outcomes in patients undergoing STEET. This provides no support for the notion that older men influence embryo quality, implantation or early gestational development.

To identify if pregnancy outcomes differ in patients that make multiple euploid embryos from a single in vitro fertilization (IVF) cycle in comparison to patients who make only one. Retrospective Cohort Study Global retrospective IRB was obtained. All patients who underwent IVF with preimplantation genetic screening (PGS) with array comparative genomic hybridization (aCGH), with at least one euploid embryo were included. The first single euploid frozen embryo transfer cycle following the IVF cycle was included. All causes of infertility, all indications for PGS, autologous and donor cycles were included. Patients who elected to undergo two or more euploid embryo transfer cycles were excluded. The primary outcome of the study included pregnancy (presence of a gestational sac) and clinical pregnancy (presence of fetal heart beat through cycle day 63 or a live birth). Statistical analysis was done using ANOVA, t-test and a linear regression model. A total of 327 single euploid embryo transfers were included. The average age of patients included in the study was 37.37 ± 4.90. The overall pregnancy rate was 68.50%(224/327), of which 61.47% (201/327) led to ongoing clinical pregnancies or live births. The most common number of euploid embryos per cohort was one (n=108) with the highest number being 20 (n=1). There was no difference in pregnancy rates when embryos from cycles with multiple euploid embryos were compared to cycles with only one euploid embryo (RR 0.9965, CI 0.8872-1.119, p 0.9457). Linear regression showed no difference was found in pregnancy rates when the percentage of euploid embryos per cohort was compared as well (R =0.0905). There was a significant difference in pregnancy rates in embryos of higher-grade inner cell mass, however when controlled for embryo grades no difference was noted in pregnancy rates in lower numbered euploid embryo cohorts.Tabled 1Pregnancy Outcomes by Number of Euploid Embryos per IVF cycle.Number of EuploidTota Patients (n = 327)Average Number of Eggs RetrievedTotal Pregnant (presence of gestational sac)PercentTotal Clinical Pregnancies (Fetal heart beat or live birth)Percent110812.32 +/- 5.757065656028415.60 +/- 7.236274556534215.79 +/- 6.912969256043019.30 +/- 7.632273217051622.56 +/- 9.921063106361322.38 +/- 8.9196975471327.50 +/- 5.096465388 or more2130.0 +/- 10.7816761362p valuep 0.552p 0.552p 0.676 Open table in a new tab The number or percentage of euploid embryos per IVF cycle, is not associated with the pregnancy outcome of STEET.

To assess using time lapse microscopy (TLM) if post warming LZB improves the ability of the blastocyst to completely exit the zona pellucida (hatching). Prospective Cohort study. Three groups were included in the study; 1) non biopsied slow frozen blastocysts, 2) non biopsied vitrified blastocysts and 3) biopsied aneuploid blastocysts, as determined by trophectoderm biopsy (TE) and array comparative genomic hybridization (aCGH) performed prior to vitrification. All blastocysts in group 3 had an existing zona breach prior to cryopreservation. All blastocysts were warmed using standard protocol. Each group was subdivided into a control arm with no additional zona pellucida (ZP) manipulation, and a test arm subjected to LZB following warming. This post warming LZB creates an opening larger (approximately 1/3 of the zona) than that routinely created for TE biopsy. Time lapse imaging was reviewed by a single observer. Primary outcomes were number of successfully hatched blastocysts and timing of hatching events. A total of 144 blastocysts donated for research (IRB #H6902) were included in the study. Survival rate post warming was 96%: An additional 7 embryos were zona free following warming and were excluded from analysis. Hence, 131 blastocysts were observed using TLM. Group 1 consisted of 21 with LZB and 22 controls. Group 2 consisted of 21 with LZB and 24 controls. Group 3 consisted of 22 with LZB and 21 controls. A total of 55 (41.98%) warmed blastocysts completed hatching within 81 hours, while 76 (58%) did not complete hatching despite exhibiting viability. There was no difference in patient ages among the blastocysts subjected to LZB (34.58 ± 4.89) and controls (35.95 ± 4.24) p=0.0895 or within groups. Groups 1& 2 (non biopsied) showed a difference in the LZB groups versus controls in the number of blastocysts that completed hatching. Group 3 (previously biopsied) blastocysts showed no difference and were able to complete hatching, likely due to the preexisting breach in the ZP created for the TE biopsy. (Table 1).Tabled 1Outcomes of Post Warm LZB vs Control Groups.GroupLazer Zona Breaching (LZB)% Completed Hatching (N)P (completed)Hours to Start of HatchingP (start)Hours to Complete HatchingP (end)Group 1+66.7% (21).0000322 ± 90.15438 ± 120.161-4.5% (22)619Group 2+76.2% (21).000000918 ± 50.14026 ± 90.732-4.2% (24)2733Group 3+54.5% (22)0.88216 ± 130.33420 ± 210.603-52.3% (21)11 ± 935 ± 23 Open table in a new tab TLM revealed that non-biopsied embryos have difficulty completing zona hatching, indicating that zona hardening may play a role. In the absence of LZB, implantation may be impeded due to inability to escape from the ZP. In contrast, previously biopsied blastocysts did not show a difference in hatching rates whether subjected to LZB post warming or not.