Barry Behr, Ph.D., H.C.L.D.

All Publications

• Development and evaluation of a usable blastocyst predictive model using the biomechanical properties of human oocytes. PloS one Meyer, D., Kort, J., Chen, C. H., Zhao, H., Yi, X., Lai, S. Y., Lu, F., Yang, W. J., Hsieh, I. C., Chiang, C. L., Chen, W. M., Huang, J. Y., Camarillo, D., Behr, B. 2024; 19 (5): e0299602

  Abstract

  The purposes of this study were to determine whether biomechanical properties of mature oocytes could predict usable blastocyst formation better than morphological information or maternal factors, and to demonstrate the safety of the aspiration measurement procedure used to determine the biomechanical properties of oocytes.A prospective split cohort study was conducted with patients from two IVF clinics who underwent in vitro fertilization. Each patient's oocytes were randomly divided into a measurement group and a control group. The aspiration depth into a micropipette was measured, and the biomechanical properties were derived. Oocyte fertilization, day 3 morphology, and blastocyst development were observed and compared between measured and unmeasured cohorts. A predictive classifier was trained to predict usable blastocyst formation and compared to the predictions of four experienced embryologists.68 patients and their corresponding 1252 oocytes were included in the study. In the safety analyses, there was no significant difference between the cohorts for fertilization, while the day 3 and 5 embryo development were not negatively affected. Four embryologists predicted usable blastocyst development based on oocyte morphology with an average accuracy of 44% while the predictive classifier achieved an accuracy of 71%. Retaining the variables necessary for normal fertilization, only data from successfully fertilized oocytes were used, resulting in a classifier an accuracy of 81%.To date, there is no standard guideline or technique to aid in the selection of oocytes that have a higher likelihood of developing into usable blastocysts, which are chosen for transfer or vitrification. This study provides a comprehensive workflow of extracting biomechanical properties and building a predictive classifier using these properties to predict mature oocytes' developmental potential. The classifier has greater accuracy in predicting the formation of usable blastocysts than the predictions provided by morphological information or maternal factors. The measurement procedure did not negatively affect embryo culture outcomes. While further analysis is necessary, this study shows the potential of using biomechanical properties of oocytes to predict embryo developmental outcomes.

  View details for DOI 10.1371/journal.pone.0299602

  View details for PubMedID 38696439

  View details for PubMedCentralID PMC11065297

• The first clinical validation of whole-genome screening on standard trophectoderm biopsies of preimplantation embryos. F&S reports Xia, Y., Katz, M., Chandramohan, D., Bechor, E., Podgursky, B., Hoxie, M., Zhang, Q., Chertman, W., Kang, J., Blue, E., Chen, J., Schleede, J., Slotnick, N. R., Du, X., Boostanfar, R., Urcia, E., Behr, B., Cohen, J., Siddiqui, N. 2024; 5 (1): 63-71

  Abstract

  To validate the performance of our laboratory-developed whole-genome screening assay within clinical preimplantation genetic testing environments.Perform a laboratory-developed whole-genome assay on both cell lines and trophectoderm biopsies, subsequently employing the next-generation sequencing procedure to reach a sequencing depth of 30X. Adhere to the Genome Analysis Toolkit best practices for accuracy, sensitivity, specificity, and precision calculations by comparing samples with references. Our assay was then applied to cell lines and biopsies harboring known pathogenic variants, aiming to ascertain these changes solely from the next-generation sequencing data, independent of parental genome information.Clinical laboratory.Coriell cell lines and research embryos with known chromosomal or genetic variants. Research trophectoderm biopsies from a couple that are heterozygous carriers for distinct variants in the same autosomal recessive gene (HOGA1).Not applicable.Accuracy, sensitivity, specificity, and precision were assessed by comparing the samples to their references. For samples with known variants, we calculated our sensitivity to detecting established variants. For the research embryos, noncarrier, carrier, and compound heterozygous states of inherited HOGA1 variants were distinguished independently of parental samples.Amplification of DNA from cell lines and embryos yielded success rates exceeding 99.9% and 98.2%, respectively, although maintaining an accuracy of >99.9% for aneuploidy assessment. The accuracy (99.99%), specificity (99.99%), sensitivity (98.0%), and precision (98.1%) of amplified genome in the bottle (reference NA12878) and embryo biopsies were comparable to results on genomic DNA, including mitochondrial heteroplasmy. Using our assay, we achieved >99.99% sensitivity when examining samples with known chromosomal and genetic variants. This encompassed pathogenic CFTR, BRCA1, and other variants, along with uniparental isodisomies and microdeletions such as DiGeorge syndrome. Our research study identified noncarrier, carrier, and compound heterozygous states within trophectoderm biopsies while simultaneously screening for 1,300 other severe monogenic diseases.To our knowledge, this is the first clinical validation of whole-genome embryo screening. In this study, we demonstrated high accuracy for aneuploidy calls (>99.9%) and genetic variants (99.99%), even in the absence of parental genomes. This assay demonstrates advancements in genomic screening and an extended scope for testing capabilities in the realm of preimplantation genetic testing.

  View details for DOI 10.1016/j.xfre.2024.01.001

  View details for PubMedID 38524212

  View details for PubMedCentralID PMC10958695

• Assessing mail-in cryopreservation for fertility preservation Belarmino, A. R., Morris, J., Kenfield, S. A., Colin, D., Griscom, B., Honig, S., Dupree, J. M., Samplaski, M. K., Halpern, J., Behr, B., Lamb, D., Matthews, W., Smith, J. F. LIPPINCOTT WILLIAMS & WILKINS. 2023

  View details for Web of Science ID 001053772000247

• Previously uncharacterized rectangular bacterial structures in the dolphin mouth. Nature communications Dudek, N. K., Galaz-Montoya, J. G., Shi, H., Mayer, M., Danita, C., Celis, A. I., Viehboeck, T., Wu, G., Behr, B., Bulgheresi, S., Huang, K. C., Chiu, W., Relman, D. A. 2023; 14 (1): 2098

  Abstract

  Much remains to be explored regarding the diversity of uncultured, host-associated microbes. Here, we describe rectangular bacterial structures (RBSs) in the mouths of bottlenose dolphins. DNA staining revealed multiple paired bands within RBSs, suggesting the presence of cells dividing along the longitudinal axis. Cryogenic transmission electron microscopy and tomography showed parallel membrane-bound segments that are likely cells, encapsulated by an S-layer-like periodic surface covering. RBSs displayed unusual pilus-like appendages with bundles of threads splayed at the tips. We present multiple lines of evidence, including genomic DNA sequencing of micromanipulated RBSs, 16S rRNA gene sequencing, and fluorescence in situ hybridization, suggesting that RBSs are bacterial and distinct from the genera Simonsiella and Conchiformibius (family Neisseriaceae), with which they share similar morphology and division patterning. Our findings highlight the diversity of novel microbial forms and lifestyles that await characterization using tools complementary to genomics such as microscopy.

  View details for DOI 10.1038/s41467-023-37638-y

  View details for PubMedID 37055390

• Cost-effectiveness of IVF with PGT-M/A to prevent transmission of spinal muscular atrophy in offspring of carrier couples. Journal of assisted reproduction and genetics Khorshid, A., Boyd, A. L., Behr, B., Zhao, Q., Alvero, R., Bavan, B. 2023

  Abstract

  To evaluate the cost-effectiveness of in-vitro fertilization with preimplantation genetic testing for aneuploidy and monogenic disorders (IVF with PGT-M/A) to prevent transmission of spinal muscular atrophy to offspring of carrier couples.A decision-analytic model was created to compare the cost-effectiveness of IVF with PGT-M/A to unassisted conception with prenatal diagnostic testing and termination (if applicable). IVF with PGT-M/A costs were determined using a separate Markov state-transition model. IVF outcomes data was derived from 76 carriers of monogenic disorders who underwent IVF with PGT-M/A at a single academic REI center. Other probabilities, costs, and utilities were derived from the literature. Costs were modeled from healthcare perspective. Utilities were modeled from the parental perspective as quality-adjusted life-years (QALYs).The incremental cost-effectiveness ratio for IVF with PGT-M/A compared to unassisted conception is $22,050 per quality-adjusted life-year. The average cost of IVF with PGT-M/A is $41,002 (SD: $8,355). At willingness-to-pay thresholds of $50,000 and $100,000, IVF with PGT-M/A is cost-effective 93.3% and 99.5% of the time, respectively.Compared to unassisted conception, IVF with PGT-M/A is cost-effective for preventing the transmission of spinal muscular atrophy to the offspring of carrier couples. These findings support insurance coverage of IVF with PGT-M/A for carriers of spinal muscular atrophy.

  View details for DOI 10.1007/s10815-023-02738-7

  View details for PubMedID 36757555

• The developmental competence of human metaphase I oocytes with delayed maturation in vitro. Fertility and sterility Moon, J. H., Zhao, Q., Zhang, J., Reddy, V., Han, J., Cheng, Y., Zhang, N., Dasig, J., Nel-Themaat, L., Behr, B., Yu, B. 2022

  Abstract

  To evaluate if metaphase I (MI) oocytes completing maturation in vitro to metaphase II ("MI-MII oocytes") have similar developmental competence as the sibling metaphase II (MII) oocytes that reached maturity in vivo.Retrospective cohort study.A total of 1124 intracytoplasmic sperm injection (ICSI) cycles from 800 patients at a single academic center between April 2016 and Dec 2020 with at least one MII oocyte immediately after retrieval and at least one sibling "MI-MII oocyte" that was retrieved as MI and matured to MII in culture before ICSI were included in the study.A total of 7865 MII and 2369 sibling MI-MII retrieved from the same individuals were compared for fertilization and blastocyst formation rates. For patients undergoing single euploid blastocyst transfers (n=406), the clinical pregnancy, spontaneous pregnancy loss rate and live birth rate were compared between the two groups.The fertilization rate was significantly higher in MII oocytes than delayed matured MI-MII oocytes (75.9% vs 56.1%, p<0.001). Similarly, the blastocyst formation rate was higher in embryos derived from MII oocytes as compared to those from MI-MII oocytes (53.8% vs 23.9%; p<0.001). The percentage of euploid embryos derived from MII oocytes was significantly higher than those from MI-MII oocytes (49.2% vs 34.7%; p<0.001). Paired comparison of sibling oocytes within the same cycle showed higher developmental competence of the MII oocytes than MI-MII oocytes. However, the pregnancy, spontaneous pregnancy loss and live birth rate after a single euploid blastocyst transfer showed no statistically significant difference between the two groups (65.7% vs 74.1%: 6.4% vs 5.0%: 61.5% vs 70.0%, respectively, MII vs MI-MII group, p>0.05).Compared to oocytes that matured in vivo and were retrieved as MII, the oocytes that were retrieved as MI and matured to MII in vitro before ICSI showed lower developmental competence, including lower fertilization, blastocyst formation, and euploidy rates. However, euploid blastocysts from either cohort resulted in similar live birth rates, indicating the MI oocytes with delayed maturation can still be useful even though the overall developmental competence was lower than their in vivo matured counterparts.

  View details for DOI 10.1016/j.fertnstert.2022.12.033

  View details for PubMedID 36567036

• Assessment of patients' perceptions towards embryo disposition after donation of embryos to a research biobank. Journal of assisted reproduction and genetics Khorshid, A., Wignarajah, A., Zhang, J., Alvero, R., Lathi, R. B., Behr, B., Murugappan, G. 2022

  Abstract

  PURPOSE: To explore perceptions towards embryo disposition among patients donating excess embryos to a research biobank.METHODS: Cross-sectional study of survey responses collected as part of enrollment in a research biobank. Patients are asked questions regarding the difficulty of their disposition decision, their alternative disposition choice if donation to research was not available, quality of the counseling they received, and if additional counseling throughout their treatment would have been beneficial. Survey responses use 5-point Likert scales, with "1" being lowest/least and "5" being highest/most.RESULTS: A total of 157 men and 163 women enrolled in the biobank. Median scores for difficulty of disposition decision were 3 for females and 2 for males, and for quality of counseling, the median scores were 4 for females and 3 for males. Seventy percent of patients would have chosen to discard their excess embryos had donation to research not been an option. Statistical analyses showed no significant difference in responses based on variations in race, religion, sexual orientation, and infertility diagnoses. Concordance of responses within heterosexual couples was tested and found to be poor to moderate.CONCLUSIONS: Assessing patients' perceptions towards embryo disposition after donation of their excess embryos to a research biobank affords a unique perspective. The difficulty of the disposition decision, the tendency to discard embryos in the absence of a means for donation to research, and the poor agreement between heterosexual partners highlight the importance of donation to research as an accessible disposition option and the need for a personalized approach to counseling and consenting for embryo disposition.

  View details for DOI 10.1007/s10815-022-02659-x

  View details for PubMedID 36401676

• ASSESSMENT OF PATIENTS' ATTITUDES TOWARDS EMBRYO DONATION FOR RESEARCH. Khorshid, A., Wignarajah, A., Alvero, R., Lathi, R. B., Behr, B., Murugappan, G. ELSEVIER SCIENCE INC. 2022: E17-E18

  View details for Web of Science ID 000891942500024

• Implementation of a comprehensive fertility biobanking initiative. F&S science Wignarajah, A., Alvero, R., Lathi, R. B., Aghajanova, L., Eisenberg, M., Winn, V. D., Behr, B., Murugappan, G. 2022; 3 (3): 228-236

  Abstract

  OBJECTIVE: To present the framework of Stanford Fertility and Reproductive Health's comprehensive reproductive biobanking initiatives and the results of the first year of recruitment.DESIGN: Technical description article.SETTING: Academic fertility center.PATIENT(S): Fertility patients >18 years of age.INTERVENTION(S): Enroll the patients interested in research in biobanking protocols.MAIN OUTCOME MEASURE(S): Patient recruitment and sample inventory from September 2020 to September2021.RESULT(S): A total of 253 patients have enrolled in the Stanford Fertility and Reproductive Health biobanking initiatives since September 2020. The current inventory consists of 1,176 samples, including serums, plasmas, buffy coats, endometria, maternal deciduae, miscarriage chorionic villi, and human embryos (zygote, cleavage, and blastocyst stages).CONCLUSION(S): This biobanking initiative addresses a critical, unmet need in reproductive health research to make it possible for patients to donate excess embryos and gametes and preserves, for future research, valuable somatic and reproductive tissues that would otherwise be discarded. We present the framework of this biobanking initiative in order to support future efforts of establishing similar biorepositories.

  View details for DOI 10.1016/j.xfss.2022.01.001

  View details for PubMedID 35977803

• AVERAGE COST OF IN VITRO FERTILIZATION WITH PREIMPLANTATION GENETIC TESTING FOR MONOGENIC DISORDERS AND ANEUPLOIDY PER UNAFFECTED LIVE BIRTH FOR CARRIER COUPLES. Khorshid, A., Boyd, A. H., Behr, B., Zhao, Q., Alvero, R. J., Bavan, B. ELSEVIER SCIENCE INC. 2021: E374-E375

  View details for Web of Science ID 000699951501421

• COST-EFFECTIVENESS OF IN VITRO FERTILIZATION WITH PREIMPLANTATION GENETIC TESTING TO PREVENT TRANSMISSION OF SPINAL MUSCULAR ATROPHY. Khorshid, A., Boyd, A. H., Behr, B., Zhao, Q., Alvero, R. J., Bavan, B. ELSEVIER SCIENCE INC. 2021: E373-E374

  View details for Web of Science ID 000699951501418

• THE COMPETENCY OF SINGLE EUPLOID BLASTOCYSTS FROM IN VITRO-MATURED (IVM) HUMAN OOCYTES WITH PREIMPLANTATION GENETIC TESTING FOR ANEUPLOIDY (PGT-A) AFTER CONTROLLED OVAIAN HYPERSTIMULATION (COH). Moon, J., Zhao, Q., Reddy, V. V., Han, J., Chang, Y., Zhang, N., Dasig, J., Behr, B. R. ELSEVIER SCIENCE INC. 2021: E278-E279

  View details for Web of Science ID 000699951501179

• Fertility technologies and how to optimize laboratory performance to support the shortening of time to birth of a healthy singleton: a Delphi consensus. Journal of assisted reproduction and genetics Coticchio, G., Behr, B., Campbell, A., Meseguer, M., Morbeck, D. E., Pisaturo, V., Plancha, C. E., Sakkas, D., Xu, Y., D'Hooghe, T., Cottell, E., Lundin, K. 2021

  Abstract

  PURPOSE: To explore how the assisted reproductive technology (ART) laboratories can be optimized and standardized to enhance embryo culture and selection, to bridge the gap between standard practice and the new concept of shortening time to healthy singleton birth.METHODS: A Delphi consensus was conducted (January to July 2018) to assess how the ART laboratory could be optimized, in conjunction with existing guidelines, to reduce the time to a healthy singleton birth. Eight experts plus the coordinator discussed and refined statements proposed by the coordinator. The statements were distributed via an online survey to 29 participants (including the eight experts from step 1), who voted on their agreement/disagreement with each statement. Consensus was reached if ≥ 66% of participants agreed/disagreed with a statement. If consensus was not achieved for any statement, that statement was revised and the process repeated until consensus was achieved. Details of statements achieving consensus were communicated to the participants.RESULTS: Consensus was achieved for all 13 statements, which underlined the need for professional guidelines and standardization of lab processes to increase laboratory competency and quality. The most important points identified were the improvement of embryo culture and embryo assessment to shorten time to live birth through the availability of more high-quality embryos, priority selection of the most viable embryos and improved cryosurvival.CONCLUSION: The efficiency of the ART laboratory can be improved through professional guidelines on standardized practices and optimized embryo culture environment, assessment, selection and cryopreservation methodologies, thereby reducing the time to a healthy singleton delivery.

  View details for DOI 10.1007/s10815-021-02077-5

  View details for PubMedID 33599923

• Comparison of aneuploidy rates between conventional invitro fertilization and intracytoplasmic sperm injection in invitro fertilization-intracytoplasmic sperm injection split insemination cycles. F&S reports Deng, J., Kuyoro, O., Zhao, Q., Behr, B., Lathi, R. B. 2020; 1 (3): 277-281

  Abstract

  Objective: To evaluate the influence of insemination methods on outcomes of preimplantation genetic testing for aneuploidy (PGT-A) by assessing PGT-A results in embryos that derived from conventional invitro fertilization (IVF) versus intracytoplasmic sperm injection (ICSI) in sibling oocytes.Design: Retrospective cohort study.Setting: Single academic IVF center.Patients: A total of 118 couples who underwent 131 split insemination cycles from July 2016-July2019.Interventions: In all cycles, sibling oocytes were allocated randomly to conventional IVF or ICSI prior to stripping. Preimplantation genetic testing for aneuploidy was performed via trophectoderm biopsy and next-generation sequencing with 24-chromosome screening.Main Outcome Measures: Rates of euploid, aneuploid, and mosaic embryos per biopsy.Results: A total of 2,129 oocytes were randomized to conventional IVF (n = 1,026) and ICSI (n = 1,103). No difference was observed in the aneuploidy rates (50.3% vs. 45.2%) and percentages of mosaic embryos (1.7% vs. 2.4%) per biopsy between conventional IVF and ICSI sibling oocytes. Percentages of different aneuploidy types and aneuploidies that involved sex chromosome abnormalities (6.2% vs. 7.2%) were similar between the two groups. In the end, the overall chance to have an euploid embryo per allocated oocyte in the two groups was similar (13.2% vs. 15.5%).Conclusions: Blastocysts created with conventional IVF and ICSI using sibling oocytes had similar rates of aneuploidy and mosaicism as examined using 24-chromosome screening. It is unlikely that conventional IVF caused significant contamination during PGT-A. We recommend conventional IVF as the preferred insemination method in PGT-A cycles, and ICSI should be indicated only in cases of male-factor infertility.

  View details for DOI 10.1016/j.xfre.2020.07.006

  View details for PubMedID 34223256

• A preliminary study of sperm identification in microdissection testicular sperm extraction samples with deep convolutional neural networks. Asian journal of andrology Wu, D. J., Badamjav, O., Reddy, V. V., Eisenberg, M., Behr, B. 2020

  Abstract

  Sperm identification and selection is an essential task when processing human testicular samples for in vitro fertilization. Locating and identifying sperm cell(s) in human testicular biopsy samples is labor intensive and time consuming. We developed a new computer-aided sperm analysis (CASA) system, which utilizes deep learning for near human-level performance on testicular sperm extraction (TESE), trained on a custom dataset. The system automates the identification of sperm in testicular biopsy samples. A dataset of 702 de-identified images from testicular biopsy samples of 30 patients was collected. Each image was normalized and passed through glare filters and diffraction correction. The data were split 80%, 10%, and 10% into training, validation, and test sets, respectively. Then, a deep object detection network, composed of a feature extraction network and object detection network, was trained on this dataset. The model was benchmarked against embryologists' performance on the detection task. Our deep learning CASA system achieved a mean average precision (mAP) of 0.741, with an average recall (AR) of 0.376 on our dataset. Our proposed method can work in real time; its speed is effectively limited only by the imaging speed of the microscope. Our results indicate that deep learning-based technologies can improve the efficiency of finding sperm in testicular biopsy samples.

  View details for DOI 10.4103/aja.aja_66_20

  View details for PubMedID 33106465

• Relationship Between Male Age, Semen Parameters and Assisted Reproductive Technology Outcomes. Andrology Kasman, A. M., Li, S., Zhao, Q., Behr, B., Eisenberg, M. L. 2020

  Abstract

  BACKGROUND: Low semen quality often obligates the use of assisted reproductive technology (ART); however, the association between semen quality and ART outcomes is uncertain.OBJECTIVES: To further assess the impact of semen quality on ART outcomes.MATERIALS AND METHODS: A retrospective cohort study was carried out at a single academic reproductive medicine center (January 2012-December 2018). Patients undergoing at least one ART cycle utilizing freshly ejaculated sperm from the male partner were included. We assessed the association between semen quality (as stratified based on WHO 5th edition criteria), paternal age (< or ≥40), and reproductive/perinatal outcomes. To evaluate the differences in ART outcomes by semen parameters and age, generalized estimating equations were applied for rates of fertilization, pregnancy, implantation, miscarriage, live birth, blast formation, gestational age, and normal embryo biopsy.RESULTS: 2063 couples were identified who underwent 4517 ART cycles. Average ages of the male and female partners were 39.8 and 37.7, respectively. Lower pregnancy rates were observed in cycles with lower sperm motility (i.e. <40%; 39.9% vs 44.1%) and total motile count (i.e. <9 million; 38.3% vs 43.5%). When examining only cycles utilizing ICSI, only a lower motility count was associated with a decline in pregnancy rate (39.1% vs 44.9%). No association was identified between semen quality and gestational age or birth weight. Paternal age was not associated with ART outcomes. However, among ART cycles in women <40, aneuploidy rate was higher for older men (p< 0.001). In cycles with women>40, no association between aneuploidy and male age was identified.DISCUSSION: Sperm motility is associated with pregnancy rates, while other semen parameters are not. In cycles in women <40, paternal age is associated with embryo aneuploidy rate.CONCLUSION: Paternal factors are associated with ART outcomes and future studies should explore mechanisms by which semen quality is associated with ART outcomes.

  View details for DOI 10.1111/andr.12908

  View details for PubMedID 32964702

• Preimplantation genetic testing for aneuploidy in poor ovarian responders with four or fewer oocytes retrieved. Journal of assisted reproduction and genetics Deng, J., Hong, H. Y., Zhao, Q., Nadgauda, A., Ashrafian, S., Behr, B., Lathi, R. B. 2020

  Abstract

  PURPOSE: To assess whether preimplantation genetic testing for aneuploidies (PGT-A) at the blastocyst stage improves clinical outcomes compared with transfer of embryos without PGT-A in poor ovarian response (POR) patients.METHODS: Retrospective cohort study of IVF cycles from 2016 to 2019 at a single academic fertility center. IVF cycles with POR and four or fewer oocytes retrieved were stratified into PGT-A (n=241) and non-PGT (n=112) groups. In PGT-A cycles, trophectoderm biopsy, next-generation sequencing with 24-chromosome screening, and single euploid frozen embryo transfer were performed. In non-PGT cycles, fresh or frozen transfer of untested embryos on day 3 or 5 was performed. Main outcomes included live birth rate and miscarriage rate per retrieval.RESULT(S): Patients who underwent PGT-A cycles were significantly less likely to reach embryo transfer compared with those who underwent non-PGT cycles (13.7% vs 70.6%). The live birth rate per retrieval did not differ between the PGT-A and non-PGT groups (6.6% vs 5.4%). Overall, the miscarriage rate was low. The PGT-A group demonstrated a significantly lower miscarriage rate per retrieval (0.4% vs 3.6%) as well as per pregnancy (5.9% vs 40.0%) compared with the non-PGT group. The number needed to treat to avoid one clinical miscarriage was 31 PGT-A cycles.CONCLUSION(S): PGT-A did not improve live birth rate per retrieval in POR patients with four or fewer oocytes retrieved. PGT-A was associated with a lower miscarriage rate; however, a fairly large number of PGT-A cycles were needed to prevent one miscarriage.

  View details for DOI 10.1007/s10815-020-01765-y

  View details for PubMedID 32285297

• RELATIONSHIP BETWEEN SEMEN PARAMETERS AND ASSISTED REPRODUCTIVE TECHNOLOGY OUTCOMES Kasman, A., Li, S., Behr, B., Eisenberg, M. LIPPINCOTT WILLIAMS & WILKINS. 2020: E538

  View details for Web of Science ID 000527010302406

• DEVELOPMENT POTENTIAL AND ANEUPLOIDY RATES IN ZYGOTES DERIVED FROM MII OOCYTES AT STRIPPING VERSUS MI-MII IN VITRO CONVERTED OOCYTES Bavan, B., Zhao, Q., Behr, B. ELSEVIER SCIENCE INC. 2020: E37–E38

  View details for Web of Science ID 000582706800053

• The impact of culture conditions on blastocyst formation and aneuploidy rates: a comparison between single-step and sequential media in a large academic practice. Journal of assisted reproduction and genetics Deng, J., Zhao, Q., Cinnioglu, C., Kayali, R., Lathi, R. B., Behr, B. 2020

  Abstract

  PURPOSE: To compare a single-step medium with a sequential medium on human blastocyst development rates, aneuploidy rates, and clinical outcomes.METHODS: Retrospective cohort study of IVF cycles that used Sage advantage sequential medium (n = 347) and uninterrupted Sage 1-step medium (n = 519) from July 1, 2016, to December 31, 2017, in an academic fertility center. Main outcome measures are blastocyst formation rates per two-pronuclear (2PN) oocyte and aneuploidy rates per biopsy.RESULTS: Of all IVF cycles, single-step medium yielded higher blastocyst formation rate (51.7% vs 43.4%) but higher aneuploidy rate (54.0% vs 45.8%) compared with sequential medium. When stratified by maternal age, women under age 38 had no difference in blastocyst formation (52.2% vs 50.2%) but a higher aneuploidy rate (44.5% vs 36.4%) resulting in a lower number of euploid blastocysts per cycle (2.6 vs 3.3) when using single-step medium compared to sequential medium. In cycles used single-step medium, patients ≥ age 38 had higher blastocyst rate (48.0% vs 33.6%), but no difference in aneuploidy rate (68.8% vs 66.0%) or number of euploid embryos (0.8 vs 1.1). For patients reaching euploid embryo transfer, there was no difference in clinical pregnancy rates, miscarriage rates, or live birth rates between two culture media systems.CONCLUSIONS: Our study demonstrates an increase in aneuploidy in young women whose embryos were cultured in a single-step medium compared to sequential medium. This study highlights the importance of culture conditions on embryo ploidy and the need to stratify by patient age when examining the impact of culture conditions on overall cycle potential.

  View details for DOI 10.1007/s10815-019-01621-8

  View details for PubMedID 31950455

• HIPK4 is essential for murine spermiogenesis. eLife Crapster, J. A., Rack, P. G., Hellmann, Z. J., Le, A. D., Adams, C. M., Leib, R. D., Elias, J. E., Perrino, J. n., Behr, B. n., Li, Y. n., Lin, J. n., Zeng, H. n., Chen, J. K. 2020; 9

  Abstract

  Mammalian spermiogenesis is a remarkable cellular transformation, during which round spermatids elongate into chromatin-condensed spermatozoa. The signaling pathways that coordinate this process are not well understood, and we demonstrate here that homeodomain-interacting protein kinase 4 (HIPK4) is essential for spermiogenesis and male fertility in mice. HIPK4 is predominantly expressed in round and early elongating spermatids, and Hipk4 knockout males are sterile, exhibiting phenotypes consistent with oligoasthenoteratozoospermia. Hipk4 mutant sperm have reduced oocyte binding and are incompetent for in vitro fertilization, but they can still produce viable offspring via intracytoplasmic sperm injection. Optical and electron microscopy of HIPK4-null male germ cells reveals defects in the filamentous actin (F-actin)-scaffolded acroplaxome during spermatid elongation and abnormal head morphologies in mature spermatozoa. We further observe that HIPK4 overexpression induces branched F-actin structures in cultured fibroblasts and that HIPK4 deficiency alters the subcellular distribution of an F-actin capping protein in the testis, supporting a role for this kinase in cytoskeleton remodeling. Our findings establish HIPK4 as an essential regulator of sperm head shaping and potential target for male contraception.

  View details for DOI 10.7554/eLife.50209

  View details for PubMedID 32163033

• Blastocyst formation rate for Asians versus Caucasians and within body mass index categories. Journal of assisted reproduction and genetics Khunte, R. n., Li, M. n., Behr, B. n., Zhao, Q. n., Baker, V. L. 2020

  Abstract

  There are well-documented racial and ethnic disparities for in vitro fertilization (IVF) outcomes, including disparities in clinical pregnancy and live birth rate. Obesity has also been associated with an increase in the risk of infertility and reduction in the efficacy of fertility treatment. However, there are limited data regarding the potential effect of race and obesity on in vitro embryo development. The purpose of this study was to determine whether blastocyst formation rates vary with race and body mass index (BMI).This retrospective analysis included 1134 fresh autologous cycles (N = 8266 embryos), which took place from January 2013 to December 2016. Women were categorized as Caucasian, Asian (not Indian), and Indian (South Asian) and by BMI categories (normal, overweight, and obese). Regression analyses were performed using race and BMI as the primary predictor variables and blastocyst formation as the outcome.Compared to Caucasian, the adjusted OR for blastocyst development was 0.85 (95% CI 0.72-1.00) for Asian women and 1.15 (95% CI 0.95-1.38) for Indian women. Women who were overweight (aOR 0.93; 95% CI 0.77-1.12) or obese (aOR 0.92; 95% CI 0.74-1.12) had similar odds of blastocyst formation comparing to women with normal BMI. Furthermore, analyses examining combined effects of race and BMI revealed no differences in blastocyst formation among Asian or Indian women with varied BMI categories compared to Caucasian women with normal BMI.Blastocyst formation did not differ based on race or BMI.

  View details for DOI 10.1007/s10815-020-01706-9

  View details for PubMedID 32130613

• CORRELATION BETWEEN BLASTOCYST STAGE OF EXPANSION AND CLINICAL OUTCOME: A RETROSPECTIVE ANALYSIS OF 810 SINGLE EUPLOID BLASTOCYST TRANSFER CYCLES AT A SINGLE IVF CENTER. Reddy, V. V., Zhao, Q., Badamjav, O., Dasig, J., Moon, J., Qin, Y., Masoudi, A., Tuyen, K., Behr, B. R. ELSEVIER SCIENCE INC. 2019: E158–E159

  View details for DOI 10.1016/j.fertnstert.2019.07.531

  View details for Web of Science ID 000487821301152

• PILOT UTILIZATION OF CONVOLUTIONAL NEURAL NETWORKS TO IMPROVE THE EFFICIENCY OF FERTILIZATION CHECKS. Reddy, V., Badamjav, O., Meyer, D., Behr, B. ELSEVIER SCIENCE INC. 2019: E5

  View details for DOI 10.1016/j.fertnstert.2019.02.041

  View details for Web of Science ID 000463487700005

• PRELIMINARY APPLICATION OF CONVOLUTIONAL NEURAL NETWORK TO IMPROVE THE EFFICIENCY OF FINDING RARE SPERMATOZOA IN MICRO-TESE SAMPLES. Badamjav, O., Reddy, V., Meyer, D., Eisenberg, M., Behr, B. ELSEVIER SCIENCE INC. 2019: E28

  View details for DOI 10.1016/j.fertnstert.2019.02.076

  View details for Web of Science ID 000463487700040

• At-home sperm testing for epidemiologic studies: Evaluation of the Trak male fertility testing system in an internet-based preconception cohort. Paediatric and perinatal epidemiology Sommer, G. J., Wang, T. R., Epperson, J. G., Hatch, E. E., Wesselink, A. K., Rothman, K. J., Fredriksen, L. L., Schaff, U. Y., Behr, B. n., Eisenberg, M. L., Wise, L. A. 2019

  Abstract

  Semen quality assessment in population-based epidemiologic studies presents logistical and financial challenges due to reliance on centralised laboratory semen analysis. The Trak Male Fertility Testing System is an FDA-cleared and validated at-home test for sperm concentration and semen volume, with a research use only sperm motility test. Here we evaluate the Trak System's overall utility among men participating in Pregnancy Study Online (PRESTO), a web-based study of North American couples planning pregnancy.US male participants aged ≥21 years with ≤6 months of pregnancy attempt time at study enrolment were invited to participate in the semen testing substudy after completing their baseline questionnaire. Consenting participants received a Trak Engine (battery-powered centrifuge) and two test kits. Participants shared their test results via smartphone images uploaded to online questionnaires. Data were then linked with covariate data from the baseline questionnaire.Of the 688 men invited to participate, 373 (54%) provided consent and 271 (73%) completed at least one semen test result. The distributions of semen volume, sperm concentration, motile sperm concentration, total sperm count, and total motile sperm count were similar to 2010 World Health Organization (WHO) semen parameter data of men in the general population. The overall usability score for the Trak System was 1.4 on a 5-point Likert scale (1 = Very Easy, 5 = Difficult), and 92% of participants believed they performed the test correctly and received an accurate result. Lastly, men with higher motile sperm count were more likely to report feeling "at ease" or "excited" following testing, while men with low motile sperm count were more likely to report feeling "concerned" or "frustrated." Overall, 91% of men reported they would like to test again.The Trak System provides a simple and potentially cost-effective means of measuring important semen parameters and may be useful in population-based epidemiologic fertility studies.

  View details for DOI 10.1111/ppe.12612

  View details for PubMedID 31838751

• Orienting Oocytes using Vibrations for In-Vitro Fertilization Procedures Meyer, D., Colon, M., Alizadeh, H., Su, L., Behr, B., Camarillo, D. B., IEEE, Howard, A., Althoefer, K., Arai, F., Arrichiello, F., Caputo, B., Castellanos, J., Hauser, K., Isler, Kim, J., Liu, H., Oh, P., Santos, Scaramuzza, D., Ude, A., Voyles, R., Yamane, K., Okamura, A. IEEE. 2019: 4837–43

  View details for Web of Science ID 000494942303076

• Euploidy in relation to blastocyst sex and morphology JOURNAL OF ASSISTED REPRODUCTION AND GENETICS Wang, A., Kort, J., Behr, B., Westphal, L. M. 2018; 35 (9): 1565–72

  View details for DOI 10.1007/s10815-018-1262-x

  View details for Web of Science ID 000444733000004

• Euploidy in relation to blastocyst sex and morphology. Journal of assisted reproduction and genetics Wang, A., Kort, J., Behr, B., Westphal, L. M. 2018

  Abstract

  PURPOSE: The objective of our study is to assess the relationship of embryo ploidy status in relation to embryo sex, morphological characteristics, and transfer parameters.METHODS: This is a retrospective cohort study at an academic medical center of patients who underwent in vitro fertilization with preimplantation genetic screening (PGS) from 2010 to 2015. Embryos were screened with 24-chromosome preimplantation genetic screening with day 5/6 trophectoderm biopsy. We investigated embryo euploidy in relation to morphology (expansion, inner cell mass, trophectoderm), embryo sex, biopsy day, and blastocyst cohort size. We used multivariate logistic regression to calculate odds ratios of euploidy in relation to these parameters.RESULTS: A total of 1559 embryos from 316 cycles and 233 patients (mean maternal age=37.8±4.2years) were included in the analysis. Six hundred and twenty-eight blastocysts (40.3%) were found to be euploid. Expansion (p<0.001), inner cell mass (ICM) (p<0.01), and trophectoderm grade (p<0.001) were significantly associated with embryo ploidy in bivariate models controlling for maternal age, while embryo sex, biopsy day, and blastocyst cohort size were not associated with embryo ploidy. In a multivariate model, we found that maternal age (p<0.001), higher grade of expansion (p<0.01), and better quality trophectoderm (p<0.001 for A compared to C grade) remained significantly associated with increased embryo euploidy, but ICM grade was no longer significant. Embryo sex was not associated with ploidy status, though male embryos were found to be associated with higher trophectoderm scores (p<0.02).CONCLUSIONS: This is the largest study to date to investigate PGS-tested embryo sex and ploidy status. While maternal age and some morphological parameters (expansion, trophectoderm grade) are associated with euploidy in our cohort, other parameters such as embryo sex, biopsy day, and cohort size are not. Though embryo sex was not associated with euploidy, male embryos were found to be associated with higher trophectoderm grades. Additional investigation in larger studies is warranted.

  View details for PubMedID 30030712

• A MAGNETIC LEVITATION PLATFORM FOR THE ISOLATION OF MATURE SPERM FROM TESE/TESA SAMPLES Durmus, G., Gupta, R., Badamjav, O., Reddy, V., Eisenberg, M. L., Behr, B., Demirci, U. ELSEVIER SCIENCE INC. 2018: E26–E27

  View details for Web of Science ID 000427891800035

• Guidance and Self-Sorting of Active Swimmers: 3D Periodic Arrays Increase Persistence Length of Human Sperm Selecting for the Fittest. Advanced science (Weinheim, Baden-Wurttemberg, Germany) Chinnasamy, T., Kingsley, J. L., Inci, F., Turek, P. J., Rosen, M. P., Behr, B., Tüzel, E., Demirci, U. 2018; 5 (2): 1700531

  Abstract

  Male infertility is a reproductive disease, and existing clinical solutions for this condition often involve long and cumbersome sperm sorting methods, including preprocessing and centrifugation-based steps. These methods also fall short when sorting for sperm free of reactive oxygen species, DNA damage, and epigenetic aberrations. Although several microfluidic platforms exist, they suffer from structural complexities, i.e., pumps or chemoattractants, setting insurmountable barriers to clinical adoption. Inspired by the natural filter-like capabilities of the female reproductive tract for sperm selection, a model-driven design, featuring pillar arrays that efficiently and noninvasively isolate highly motile and morphologically normal sperm, with lower epigenetic global methylation, from raw semen, is presented. The Simple Periodic ARray for Trapping And isolatioN (SPARTAN) created here modulates the directional persistence of sperm, increasing the spatial separation between progressive and nonprogressive motile sperm populations within an unprecedentedly short 10 min assay time. With over 99% motility of sorted sperm, a 5-fold improvement in morphology, 3-fold increase in nuclear maturity, and 2-4-fold enhancement in DNA integrity, SPARTAN offers to standardize sperm selection while eliminating operator-to-operator variations, centrifugation, and flow. SPARTAN can also be applied in other areas, including conservation ecology, breeding of farm animals, and design of flagellar microrobots for diagnostics.

  View details for DOI 10.1002/advs.201700531

  View details for PubMedID 29610725

  View details for PubMedCentralID PMC5827459

• Racial Variation in Semen Quality at Fertility Evaluation. Urology Khandwala, Y. S., Zhang, C. A., Li, S., Behr, B., Guo, D., Eisenberg, M. L. 2017

  Abstract

  To identify racial differences in semen quality among men living in the same geographic area and seeking fertility evaluation.Men obtaining a semen analysis for infertility evaluation or treatment between 2012 and 2016 at a single center were identified, and demographic data including height, weight, body mass index (BMI), and age were described. Mean semen parameters and the proportions of men with suboptimal parameters based on the World Health Organization's fifth edition criteria were also compared based on race. Multivariable regression analysis was conducted incorporating age, BMI, and year of evaluation. Further subanalyses based on BMI were subsequently performed.White men produced greater volumes of semen on average; however, Asian men had higher sperm concentrations and total sperm count. A lower proportion of Asian men compared to white men had semen quality in the suboptimal range for most semen parameters, whereas a higher proportion of white men were found to have azoospermia. Stratification by BMI groups attenuated the observed differences between whites and Asians, yet Asian male semen quality remained higher.Among men evaluated for infertility at a single center, Asians had lower volume but higher sperm concentrations compared with whites, which was influenced by differences in azoospermia prevalence. Although anthropometric and demographic factors attenuated the differences, even after adjustment, the contrasts remained. Our study suggests that racial differences exist in semen quality at the time of infertility evaluation.

  View details for DOI 10.1016/j.urology.2017.03.064

  View details for PubMedID 28522219

• Maternal Endometrial hsa-miR-30d Is Taken Up by the Human Blastocyst and Induces Transcriptional Modifications Shah, M. S., Vilella, F., Behr, B., Reddy, V., Wang, W., Navarro, R., Jimenez, J., Quake, S., Simon, C. SAGE PUBLICATIONS INC. 2017: 227A

  View details for Web of Science ID 000399043900557

• PRELIMINARY STUDY TO INVESTIGATE TELOMERASE REVERSE TRANSCRIPTASE EXPRESSION AMONG HUMAN CUMULUS CELLS. Kort, J. D., Garbuzov, A., Artandi, S. E., Behr, B. ELSEVIER SCIENCE INC. 2017: E9

  View details for Web of Science ID 000397983000012

• Functional topography of the fully grown human oocyte EUROPEAN JOURNAL OF HISTOCHEMISTRY Monti, M., Calligaro, A., Behr, B., Pera, R. R., Redi, C. A., Wossidlo, M. 2017; 61 (1): 32-35

  Abstract

  In vivo maturation (IVM) of human oocytes is a technique used to increase the number of usable oocytes for in vitro fertilization (IVF) and represents a necessity for women with different ovarian pathologies. During IVM the oocytes progress from the germinal vesicle stage (GV) through the metaphase II and during this journey both nuclear and cytoplasmic rearrangements must be obtained to increase the probability to get viable and healthy zygotes/embryos after IVF. As the successful clinical outcomes of this technique are a reality, we wanted to investigate the causes behind oocytes maturation arrest. For obvious ethical reasons, we were able to analyze only few human immature oocytes discarded and donated to research by transmission electron microscopy showing that, as in the mouse, they have different chromatin and cytoplasmic organizations both essential for further embryo development.

  View details for DOI 10.4081/ejh.2017.2769

  View details for Web of Science ID 000396503200006

  View details for PubMedCentralID PMC5304266

• Guidance and Self-Sorting of Active Swimmers via 3-D Periodic Arrays Advanced Science Chinnasamy, T., Kingsley, J. L., Inci, F., Turek, P. J., Rosen, M. P., Behr, B., Tuzel, E., Demirci, U. 2017: 1700531

  Abstract

  Male infertility is a reproductive disease, and existing clinical solutions for this condition often involve long and cumbersome sperm sorting methods, including preprocessing and centrifugation-based steps. These methods also fall short when sorting for sperm free of reactive oxygen species, DNA damage, and epigenetic aberrations. Although several microfluidic platforms exist, they suffer from structural complexities, i.e., pumps or chemoattractants, setting insurmountable barriers to clinical adoption. Inspired by the natural filter-like capabilities of the female reproductive tract for sperm selection, a model-driven design, featuring pillar arrays that efficiently and noninvasively isolate highly motile and morphologically normal sperm, with lower epigenetic global methylation, from raw semen, is presented. The Simple Periodic ARray for Trapping And isolatioN (SPARTAN) created here modulates the directional persistence of sperm, increasing the spatial separation between progressive and nonprogressive motile sperm populations within an unprecedentedly short 10 min assay time. With over 99% motility of sorted sperm, a 5-fold improvement in morphology, 3-fold increase in nuclear maturity, and 2-4-fold enhancement in DNA integrity, SPARTAN offers to standardize sperm selection while eliminating operator-to-operator variations, centrifugation, and flow. SPARTAN can also be applied in other areas, including conservation ecology, breeding of farm animals, and design of flagellar microrobots for diagnostics.

  View details for DOI 10.1002/advs.201700531

  View details for PubMedCentralID PMC5827459

• Hypertension and Male Fertility. The world journal of men's health Guo, D. n., Li, S. n., Behr, B. n., Eisenberg, M. L. 2017; 35 (2): 59–64

  Abstract

  As the age of paternity rises in the developed world, issues of chronic disease may affect prospective fathers. Given the high prevalence of hypertension, researchers have begun to explore the relationship between hypertensive disease and male fertility. The current literature suggests an association between hypertension and semen quality. The use of various antihypertensive medications has also been linked to impaired semen parameters, making it difficult to discern whether the association exists with hypertension or its treatment. Further investigation is warranted to determine whether the observed associations are causal.

  View details for PubMedID 28868816

  View details for PubMedCentralID PMC5583372

• Biomechanics and developmental potential of oocytes and embryos. Fertility and sterility Kort, J. n., Behr, B. n. 2017; 108 (5): 738–41

  Abstract

  The high incidence of multiple embryo transfers is evidence of the need for better methods of embryo selection. Additionally, methods to determine the reproductive competence of unfertilized oocytes are critically needed to inform the growing population of patients undergoing fertility preservation. The ideal method of oocyte and embryo selection would be noninvasive, inexpensive, and able to be incorporated into embryology workflow with minimal disruption. Methods to assess the biomechanical properties of cells offer many of these traits, and there is a growing body of evidence in multiple cell types demonstrating the biomechanical properties of cells are reflective of a cell's intrinsic health. The associations with these properties are not mere coincidence, as many of the biomechanical properties are critical to cellular function. The biomechanical properties of oocytes and embryos undergo a dynamic, characteristic transformation from oocyte maturation through blastocyst formation, lending itself to biomechanical assessment. Many of the assessments made by embryologists, from ease of microinjection during intracytoplasmic sperm injection to degree of blastocyst expansion, are direct proxies for cellular biomechanics. Newer, objective and quantitative methods of biomechanical assessment are being applied to oocyte and embryo selection, with early use supporting their application in assisted reproduction.

  View details for PubMedID 28987788

• Guidance and Self-Sorting of Active Swimmers: 3D Periodic Arrays Increase Persistence Length of Human Sperm Selecting for the Fittest. Adv Sci. Chinnasamy, T., Kingsley, J., Inci, F., Turek, P., Rosen, M., Behr, B., Tüzel , E., Demirci, U. 2017: 1-13
• Relationship between paternal somatic health and assisted reproductive technology outcomes. Fertility and sterility Eisenberg, M. L., Li, S., Wise, L. A., Lynch, C. D., Nakajima, S., Meyers, S. A., Behr, B., Baker, V. L. 2016; 106 (3): 559-565

  Abstract

  To study the association between paternal medical comorbidities and the outcomes of assisted reproductive technology (ART).Retrospective cohort study.Academic reproductive medicine center.We analyzed fresh ART cycles uszing freshly ejaculated sperm from the male partner of couples undergoing ART cycles from 2004 until 2014. We recorded patient and partner demographic characteristics. The cohort was linked to hospital billing data to obtain information on selected male partners' comorbidities identified using ICD-9-CM codes.None.Fertilization, clinical pregnancy, miscarriage, implantation, and live-birth rates as well as birth weights and gestational ages.In all, we identified 2,690 men who underwent 5,037 fresh ART cycles. Twenty-seven percent of men had at least one medical diagnosis. Men with nervous system diseases had on average lower pregnancy rates (23% vs. 30%) and live-birth rates (15% vs. 23%) than men without nervous system diseases. Lower fertilization rates were also observed among men with respiratory diseases (61% vs. 64%) and musculoskeletal diseases (61% vs. 64%) relative to those without these diseases. In addition, men with diseases of the endocrine system had smaller children (2,970 vs. 3,210 g) than men without such diseases. Finally, men with mental disorders had children born at an earlier gestational age (36.5 vs. 38.0 weeks).The current report identified a possible relationship between a man's health history and IVF outcomes. As these are potentially modifiable factors, further research should determine whether treatment for men's health conditions may improve or impair IVF outcomes.

  View details for DOI 10.1016/j.fertnstert.2016.04.037

  View details for PubMedID 27179785

• Human Embryonic Stem Cell Lines with Lesions in FOXP3 and NF1 PLOS ONE Zhu, H., Behr, B., Reddy, V. V., Hughes, M., Pan, Y., Baker, J. 2016; 11 (3)

  Abstract

  Human embryonic stem cells (hESCs) are derived from the inner cell mass (ICM) of blastocyst staged embryos. Spare blastocyst staged embryos were obtained by in vitro fertilization (IVF) and donated for research purposes. hESCs carrying specific mutations can be used as a powerful cell system in modeling human genetic disorders. We obtained preimplantation genetic diagnosed (PGD) blastocyst staged embryos with genetic mutations that cause human disorders and derived hESCs from these embryos. We applied laser assisted micromanipulation to isolate the inner cell mass from the blastocysts and plated the ICM onto the mouse embryonic fibroblast cells. Two hESC lines with lesions in FOXP3 and NF1 were established. Both lines maintain a typical undifferentiated hESCs phenotype and present a normal karyotype. The two lines express a panel of pluripotency markers and have the potential to differentiate to the three germ layers in vitro and in vivo. The hESC lines with lesions in FOXP3 and NF1 are available for the scientific community and may serve as an important resource for research into these disease states.

  View details for DOI 10.1371/journal.pone.0151836

  View details for Web of Science ID 000372582800106

  View details for PubMedCentralID PMC4798423

• MICROFLUIDIC SPERM SORTING DEVICE FOR SELECTION OF FUNCTIONAL HUMAN SPERM FOR IUI APPLICATION Chinnasamy, T., Behr, B., Demirci, U. ELSEVIER SCIENCE INC. 2016: E17–E18

  View details for DOI 10.1016/j.fertnstert.2015.12.063

  View details for Web of Science ID 000373405200031

• Human oocyte developmental potential is predicted by mechanical properties within hours after fertilization. Nature communications Yanez, L. Z., Han, J., Behr, B. B., Reijo Pera, R. A., Camarillo, D. B. 2016; 7: 10809-?

  Abstract

  The causes of embryonic arrest during pre-implantation development are poorly understood. Attempts to correlate patterns of oocyte gene expression with successful embryo development have been hampered by the lack of reliable and nondestructive predictors of viability at such an early stage. Here we report that zygote viscoelastic properties can predict blastocyst formation in humans and mice within hours after fertilization, with >90% precision, 95% specificity and 75% sensitivity. We demonstrate that there are significant differences between the transcriptomes of viable and non-viable zygotes, especially in expression of genes important for oocyte maturation. In addition, we show that low-quality oocytes may undergo insufficient cortical granule release and zona-hardening, causing altered mechanics after fertilization. Our results suggest that embryo potential is largely determined by the quality and maturation of the oocyte before fertilization, and can be predicted through a minimally invasive mechanical measurement at the zygote stage.

  View details for DOI 10.1038/ncomms10809

  View details for PubMedID 26904963

  View details for PubMedCentralID PMC4770082

• Human Embryonic Stem Cell Lines with Lesions in FOXP3 and NF1. PloS one Zhu, H., Behr, B., Reddy, V. V., Hughes, M., Pan, Y., Baker, J. 2016; 11 (3)

  Abstract

  Human embryonic stem cells (hESCs) are derived from the inner cell mass (ICM) of blastocyst staged embryos. Spare blastocyst staged embryos were obtained by in vitro fertilization (IVF) and donated for research purposes. hESCs carrying specific mutations can be used as a powerful cell system in modeling human genetic disorders. We obtained preimplantation genetic diagnosed (PGD) blastocyst staged embryos with genetic mutations that cause human disorders and derived hESCs from these embryos. We applied laser assisted micromanipulation to isolate the inner cell mass from the blastocysts and plated the ICM onto the mouse embryonic fibroblast cells. Two hESC lines with lesions in FOXP3 and NF1 were established. Both lines maintain a typical undifferentiated hESCs phenotype and present a normal karyotype. The two lines express a panel of pluripotency markers and have the potential to differentiate to the three germ layers in vitro and in vivo. The hESC lines with lesions in FOXP3 and NF1 are available for the scientific community and may serve as an important resource for research into these disease states.

  View details for DOI 10.1371/journal.pone.0151836

  View details for PubMedID 26990425

  View details for PubMedCentralID PMC4798423

• Increased body mass index negatively impacts blastocyst formation rate in normal responders undergoing in vitro fertilization JOURNAL OF ASSISTED REPRODUCTION AND GENETICS Comstock, I. A., Kim, S., Behr, B., Lathi, R. B. 2015; 32 (9): 1299-1304

  Abstract

  The aim of this study is to investigate the effect of female BMI and metabolic dysfunction on blastocyst formation rate.This was a retrospective cohort study that was performed in an academic center for reproductive medicine. Patients who were normal weight, overweight with metabolic dysfunction, or obese who had ≥6 oocytes retrieved in a fresh IVF cycle were included in the study. The blastocyst formation rate was calculated from the number of ≥5 cell embryos on day 3 observed in culture until day 5 or day 6. Only good quality blastocysts were included in the calculation as defined by a morphologic grade of 3BB or better.The blastocyst formation rate was significantly better in the normal-weight controls versus overweight/obese patients (57.2 versus 43.6 %, p < 0.007). There was no difference in blastocyst formation between the patients with a BMI 25-29.9 kg/m(2) with metabolic dysfunction and those with a BMI ≥30 kg/m(2).The maternal metabolic environment has a significant impact on embryo quality as measured by blastocyst formation. A decreased blastocyst formation rate is likely a significant contributor to poorer reproductive outcomes in overweight and obese women with infertility.

  View details for DOI 10.1007/s10815-015-0515-1

  View details for Web of Science ID 000362519600002

  View details for PubMedID 26109331

  View details for PubMedCentralID PMC4595387

• Label-free characterization of vitrification-induced morphology changes in single-cell embryos with full-field optical coherence tomography. Journal of biomedical optics Zarnescu, L., Leung, M. C., Abeyta, M., Sudkamp, H., Baer, T., Behr, B., Ellerbee, A. K. 2015; 20 (9): 96004-?

  View details for DOI 10.1117/1.JBO.20.9.096004

  View details for PubMedID 26334977

• Aneuploidy rates and blastocyst formation after biopsy of morulae and early blastocysts on day 5. Journal of assisted reproduction and genetics Kort, J. D., Lathi, R. B., Brookfield, K., Baker, V. L., Zhao, Q., Behr, B. R. 2015; 32 (6): 925-930

  Abstract

  Studies have demonstrated high implantation rates after trophectoderm biopsy of day 5 expanded blastocysts. However, biopsy of cleavage stage embryos may adversely affect embryo development and implantation. No studies have assessed the utility of day 5 morulae and early blastocyst biopsy. This study sought to better understand these slower embryos' aneuploidy rates and implantation potential.This was a retrospective review of all autologous IVF cycles utilizing PGS at a single academic infertility center.The biopsy of day 5 morulae and early blastocysts provided 22 % additional euploid blastocysts available for fresh day 6 transfer compared to day 5 biopsy of only expanded blastocysts. Aneuploidy did correlate with embryo stage on day 5, even after controlling for maternal age, with 16 % of morulae and 35 % of blastocysts being euploid. The majority (83 %) of euploid morulae progressed to the blastocyst stage by day 6. Experience transferring slower developing embryos is limited, but preliminary pregnancy and implantation rates appear similar to euploid embryos biopsied as expanded blastocysts.The biopsy of all non-arrested embryos on day 5 provides genetic information for all blastocysts on day 6, increasing the pool of euploid blastocysts available for fresh transfer and avoiding the need to cryopreserve developmentally competent embryos without genetic information.

  View details for DOI 10.1007/s10815-015-0475-5

  View details for PubMedID 25921084

  View details for PubMedCentralID PMC4491071

• Relationship between semen production and medical comorbidity FERTILITY AND STERILITY Eisenberg, M. L., Li, S., Behr, B., Pera, R. R., Cullen, M. R. 2015; 103 (1): 66-71

  Abstract

  To study the relationship between semen quality and current health status in a cohort of men evaluated for infertility.Cross-sectional study.Fertility clinic.Nine thousand three hundred eighty-seven men evaluated for infertility between 1994 and 2011.None.Charlson comorbidity index, medical diagnoses by organ system.At the time of evaluation, 9,387 men with a mean age of 38 years had semen data available. Of these men, 44% had at least one medical diagnosis unrelated to infertility. When stratifying the cohort by the Charlson comorbidity index (CCI), differences in all measured semen parameters were identified. Men with a higher CCI had lower semen volume, concentration, motility, total sperm count, and morphology scores. In addition, men with diseases of the endocrine, circulatory, genitourinary, and skin diseases all showed significantly higher rates of semen abnormalities. Upon closer examination of diseases of the circulatory system, men with hypertensive disease, peripheral vascular and cerebrovascular disease, and nonischemic heart disease all displayed higher rates of semen abnormalities.The current report identified a relationship between medical comorbidites and male semen production. Although genetics help guide a man's sperm production, his current condition and health play an important role.

  View details for DOI 10.1016/j.fertnstert.2014.10.017

  View details for Web of Science ID 000346911400015

  View details for PubMedID 25497466

• Comparison of epigenetic mediator expression and function in mouse and human embryonic blastomeres HUMAN MOLECULAR GENETICS Chavez, S. L., McElroy, S. L., Bossert, N. L., De Jonge, C. J., Vera Rodriguez, M., Leong, D. E., Behr, B., Westphal, L. M., Pera, R. A. 2014; 23 (18): 4970-4984

  Abstract

  A map of human embryo development that combines imaging, molecular, genetic and epigenetic data for comparisons to other species and across pathologies would be greatly beneficial for basic science and clinical applications. Here, we compared mRNA and protein expression of key mediators of DNA methylation and histone modifications between mouse and human embryos, embryos from fertile/infertile couples, and following growth factor supplementation. We observed that individual mouse and human embryos are characterized by similarities and distinct differences in DNA methylation and histone modification patterns especially at the single-cell level. In particular, while mouse embryos first exhibited sub-compartmentalization of different histone modifications between blastomeres at the morula stage and cell sub-populations in blastocysts, differential histone modification expression was detected between blastomeres earlier in human embryos at the four- to eight-cell stage. Likewise, differences in epigenetic mediator expression were also observed between embryos from fertile and infertile couples, which were largely equalized in response to growth factor supplementation, suggesting that select growth factors might prevent alterations in epigenetic profiles during prolonged embryo culture. Finally, we determined that reduced expression via morpholino technologies of a single histone-modifying enzyme, Rps6ka4/Msk2, resulted in cleavage-stage arrest as assessed by time-lapse imaging and was associated with aneuploidy generation. Taken together, data document differences in epigenetic patterns between species with implications for fertility and suggest functional roles for individual epigenetic factors during pre-implantation development.

  View details for DOI 10.1093/hmg/ddu212

  View details for PubMedID 24821703

• The role of serum testosterone in early pregnancy outcome: a comparison in women with and without polycystic ovary syndrome. Journal of obstetrics and gynaecology Canada : JOGC = Journal d'obstétrique et gynécologie du Canada : JOGC Lathi, R. B., Dahan, M. H., Reynolds-May, M. F., Milki, A. A., Behr, B., Westphal, L. M. 2014; 36 (9): 811-816

  Abstract

  Hyperandrogenic conditions in women are associated with increased rates of miscarriage. However, the specific role of maternal testosterone in early pregnancy and its association with pregnancy outcome is unknown. The purpose of this study was to compare serum testosterone levels during early pregnancy in women with and without polycystic ovary syndrome (PCOS) who either had successful pregnancies or miscarried.We collected serum samples from women attending a university-based fertility centre at the time of their first positive serum beta human chorionic gonadotropin pregnancy test. The samples were subsequently assayed for total testosterone level. We used logistical regression modelling to control for PCOS diagnosis, BMI, and age.Total testosterone levels were available for 346 pregnancies, including 286 successful pregnancies and 78 first trimester miscarriages. We found no difference in total testosterone levels between women who subsequently had an ongoing pregnancy (mean concentration 3.6 ± 2.6 nmol/L) and women with a miscarriage (mean 3.6 ± 2.4 nmol/L). Using the Rotterdam criteria to identify women with PCOS, we also found no differences in serum testosterone between women who had ongoing pregnancies or miscarriages, either with PCOS (P = 0.176) or without PCOS (P = 0.561).Our findings show that early pregnancy testosterone levels do not predict pregnancy outcome, and they call into question the role of testosterone in causing miscarriage in populations of women with PCOS. Further research is needed to elucidate the normal progression of testosterone levels during pregnancy and to investigate further the relationship between PCOS and miscarriage.

  View details for PubMedID 25222360

• The evaluation of pre and post processing semen analysis parameters at the time of intrauterine insemination in couples diagnosed with male factor infertility and pregnancy rates based on stimulation agent. A retrospective cohort study EUROPEAN JOURNAL OF OBSTETRICS & GYNECOLOGY AND REPRODUCTIVE BIOLOGY Luco, S. M., Agbo, C., Behr, B., Dahan, M. H. 2014; 179: 159-162

  Abstract

  To identify pre or post processing semen analysis parameters that may be predictive of successful pregnancy in couples with male factor infertility undergoing intra uterine insemination (IUI). To evaluate the pregnancy rate based on ovulation inducing agent in couples with male factor infertility per the 2010 world health organization criteria treated with IUI.This retrospective study was performed at Stanford University medical center. All couples with male factor infertility fitting inclusion criteria were included over a 2 year period of time. 147 couples with male factor infertility were included and 356 IUIs were analyzed. All subjects in this study had Kruger strict analysis >4% normal forms. Logistic regression analysis was used to control for confounding effects and multiplicity.The overall pregnancy rate was 5.3%. No parameter in either the pre or post analysis predicted pregnancy. Furthermore, it was found that natural cycle and letrazole treatment had similar pregnancy rates (3% and 3%) p=ns. Similar outcomes were also observed between clomiphene citrate and gonadotropin stimulated cycles (7.5% and 6.0%) p=ns.Total motile sperm count which has been found to be a predictor of pregnancy when evaluated in isolation, may be due to a confounding effect. These low pregnancy rates should be considered when deciding whether to suggest IUI and when selecting a protocol for ovulation induction for couples with male factor infertility.

  View details for DOI 10.1016/j.ejogrb.2014.05.003

  View details for Web of Science ID 000340318200030

  View details for PubMedID 24965998

  View details for PubMedCentralID PMC4144991

• Semen quality, infertility and mortality in the USA HUMAN REPRODUCTION Eisenberg, M. L., Li, S., Behr, B., Cullen, M. R., Galusha, D., Lamb, D. J., Lipshultz, L. I. 2014; 29 (7): 1567-1574

  Abstract

  What is the relationship between semen parameters and mortality in men evaluated for infertility?Among men undergoing an infertility evaluation, those with abnormal semen parameters have a higher risk of death, suggesting a possible common etiology between infertility and mortality.Conflicting data exist that suggest either an inverse relationship or no relationship between semen quality and mortality.A study cohort was identified from two centers, each specializing in infertility care. In California, we identified men with data from 1994 to 2011 in the Stanford Reproductive Endocrinology and Infertility semen database. In Texas, we identified men with data from 1989 to 2009 contained in the andrology database at the Baylor College of Medicine Special Procedures Laboratory who were evaluated for infertility. Mortality was determined by data linkage to the National Death Index or Social Security Death Index. Comorbidity was estimated based on calculation of the Charlson Comorbidity Index or Centers for Medicare & Medicaid Services-Hierarchical Condition Categories Model.In all, 11,935 men were evaluated for infertility from 1989 to 2011. During 92 104 person years of follow-up, 69 of 11,935 men died (0.58%). The mean age at infertility evaluation was 36.6 years with a mean follow-up of 7.7 years.Compared with the general population, men evaluated for infertility had a lower risk of death with 69 deaths observed compared with 176.7 expected (Standardized mortality rate 0.39, 95% CI 0.30-0.49). When stratified by semen parameters, however, men with impaired semen parameters (i.e. male factor infertility) had significantly higher mortality rates compared with men with normal parameters (i.e. no male factor infertility). Low semen volume, sperm concentration, sperm motility, total sperm count and total motile sperm count were all associated with higher risk of death. In contrast, abnormal sperm morphology was not associated with mortality. While adjusting for current health status attenuated the association between semen parameters and mortality, men with two or more abnormal semen parameters still had a 2.3-fold higher risk of death compared with men with normal semen (95% CI 1.12-4.65).Our cohort represents infertile men, which may limit generalizability. As comorbidity relied on administrative data, granular information on each man regarding infertility diagnosis and lifestyle factors was unavailable.Men with impaired semen parameters have an increased mortality rate in the years following an infertility evaluation suggesting semen quality may provide a marker of health.This study is supported in part by P01HD36289 from the Eunice Kennedy Shriver National Institute for Child Health and Human Development, National Institutes of Health (to D.J.L. and L.I.L.). The project was also partially supported by an NIH CTSA award number UL1 RR025744. None of the authors has any conflict of interest to declare.

  View details for DOI 10.1093/humrep/deu106

  View details for Web of Science ID 000338126500027

  View details for PubMedCentralID PMC4059337

• Atypical embryo phenotypes identified by time-lapse microscopy: high prevalence and association with embryo development FERTILITY AND STERILITY Wirka, K. A., Chen, A. A., Conaghan, J., Ivani, K., Gvakharia, M., Behr, B., Suraj, V., Tan, L., Shen, S. 2014; 101 (6): 1637-U495

  Abstract

  To characterize atypical dynamic embryo phenotypes identified by time-lapse microscopy, evaluate their prevalence, and determine their association with embryo development.Retrospective multicenter cohort study.Five IVF clinics in the United States.Sixty-seven women undergoing IVF treatment with 651 embryos.Embryo videos were retrospectively analyzed for atypical phenotypes.Identification of four groups of atypical embryo phenotypes: abnormal syngamy (AS), abnormal first cytokinesis (A1(cyt)), abnormal cleavage (AC), and chaotic cleavage (CC). Prevalence and association with embryo morphology and development potential were evaluated.A high prevalence of atypical phenotypes was observed among embryos: AS 25.1% (163/649), A1(cyt) 31.0% (195/639), AC 18% (115/639) and CC 15% (96/639). A high percentage of embryos with atypical phenotype(s) had good quality on day 3 (overall grade good or fair): AS 78.6% (70/89); A1(cyt) 79.7% (94/119), AC 86.4% (70/81), and CC 35.2% (19/54), but the blastocyst formation rates for these embryos were significantly lower compared with their respective control groups: AS 21.5% vs. 44.9%, A1(cyt) 21.7% vs. 44.6%, AC 11.7% vs. 43.1%, and CC 14.0% vs. 42.3%.Embryos exhibiting atypical phenotypes are highly prevalent in human embryos and show significantly lower developmental potential than control embryos.NCT01369446.

  View details for DOI 10.1016/j.fertnstert.2014.02.050

  View details for Web of Science ID 000337364300033

  View details for PubMedID 24726214

• Testosterone Changes Bladder and Kidney Structure in Juvenile Male Rats JOURNAL OF UROLOGY Shortliffe, L. M., Ye, Y., Behr, B., Wang, B. 2014; 191 (6): 1913-1919

  Abstract

  Testosterone affects male development, maturation and aging but limited data exist on testosterone effects on the juvenile genitourinary system. We hypothesized that testosterone has bladder and kidney developmental effects, and investigated this in juvenile male rats.To examine the testosterone effect 21-day-old prepubertal male Wistar rats were divided into 3 groups of 12 each, including sham orchiectomy as controls, and bilateral orchiectomy with vehicle and bilateral orchiectomy with testosterone. Starting at age 28 days (week 0) testosterone enanthate (5 mg/100 gm) or vehicle was injected weekly. Testosterone was measured at study week 0 before injection, and at weeks 1, 6 and 16. Whole bladders and kidneys were evaluated for androgen receptor, bladder collagen-to-smooth muscle ratio, and renal morphometry and immunohistochemistry.Testosterone was not detectable at week 0 in all groups. It remained undetectable at weeks 1, 6 and 16 in the orchiectomy plus vehicle group. Testosterone levels were physiological in controls and rats with orchiectomy plus testosterone but levels were higher in the latter than in the former group. Rats with orchiectomy plus testosterone had increased bladder-to-body and kidney-to-body weight ratios (p<0.01 and <0.05, respectively), and decreased collagen-to-smooth muscle ratio than the orchiectomy plus vehicle and control groups. Rats with orchiectomy plus testosterone had a lower renal total glomerular count (p<0.01) but increased androgen receptor density.In juvenile male rats testosterone was associated with increased bladder and renal mass, and increased bladder smooth muscle. Testosterone associated kidneys also appeared to have fewer but larger glomeruli. These data support an important role for sex hormones in structural and functional development of the bladder and kidney.

  View details for DOI 10.1016/j.juro.2014.01.012

  View details for Web of Science ID 000336531100104

  View details for PubMedID 24518779

• Morphological Assessment of Embryo Viability SEMINARS IN REPRODUCTIVE MEDICINE Abeyta, M., Behr, B. 2014; 32 (2): 114-126

  Abstract

  Morphological assessment is discussed in the context of significant literature at all stages of in vitro development, beginning with the oocyte and culminating at the blastocyst stage. Current evidence is used to debate the inclusion of commonly observed morphological features in grading schemes. The biological rationale behind observed phenomena such as multinucleation and fragmentation are also explored. Current limitations as well as technological advancements that increase our ability to assess viability are highlighted. Particular attention is paid to the relationship between developmental timing and assessment schemes. Failure to standardize assessment timing and inclusion criteria is glaring weaknesses of the literature that currently make consensus unattainable. Mounting evidence suggests that the future of static assessment is very likely to be influenced by information gathered from preimplantation genetic screening and other invasive techniques as well as from continuous monitoring tools such as time lapse.

  View details for DOI 10.1055/s-0033-1363553

  View details for Web of Science ID 000331288800007

  View details for PubMedID 24515906

• Assessment of imaging parameters correlated with the effects of cryopreservation on embryo development Conference on Optical Methods in Developmental Biology II Zarnescu, L., Abeyta, M., Baer, T. M., Behr, B., Ellerbee, A. K. SPIE-INT SOC OPTICAL ENGINEERING. 2014

  View details for DOI 10.1117/12.2040487

  View details for Web of Science ID 000336049000006

• Embryo culture and selection: morphological criteria. Methods in molecular biology (Clifton, N.J.) Hegde, A., Behr, B. 2014; 1154: 501-532

  Abstract

  In this chapter, we have outlined the various morphological criteria for selection of the best embryo at each important milestone encountered in the progress from the oocyte to the blastocyst. As Gerris et al. stated, a combination of one, two, or even three selection points should lead to a more accurate selection of the best embryo, as no one criterion is better than the other. An embryo that fails to meet the entire set of selection criteria must be avoided as culture cannot correct an impaired embryo.

  View details for DOI 10.1007/978-1-4939-0659-8_23

  View details for PubMedID 24782025

• EED and KDM6B Coordinate the First Mammalian Cell Lineage Commitment To Ensure Embryo Implantation MOLECULAR AND CELLULAR BIOLOGY Saha, B., Home, P., Ray, S., Larson, M., Paul, A., Rajendran, G., Behr, B., Paul, S. 2013; 33 (14): 2691-2705

  Abstract

  The first mammalian cell lineage commitment is the formation of the trophectoderm (TE) and the inner cell mass (ICM) lineages during preimplantation development. Proper development of the TE and ICM lineages is dependent upon establishment of specific transcriptional programs. However, the epigenetic mechanisms that functionally contribute to establish TE- and ICM-specific transcriptional programs are poorly understood. Here, we show that proper development of the TE and ICM lineages is coordinated via combinatorial regulation of embryonic ectoderm development (EED) and lysine-specific demethylase 6B (KDM6B). During blastocyst formation, the relative levels of EED and KDM6B expression determine altered polycomb repressor 2 (PRC2) complex recruitment and incorporation of the repressive histone H3 lysine 27 trimethylation (H3K27Me3) mark at the chromatin domains of TE-specific master regulators CDX2 and GATA3, leading to their activation in the TE lineage and repression in the ICM lineage. Furthermore, ectopic gain of EED along with depletion of KDM6B in preimplantation mouse embryos abrogates CDX2 and GATA3 expression in the nascent TE lineage. The loss of CDX2 and GATA3 in the nascent TE lineage results in improper TE development, leading to failure in embryo implantation to the uterus. Our study delineates a novel epigenetic mechanism that orchestrates proper development of the first mammalian cell lineages.

  View details for DOI 10.1128/MCB.00069-13

  View details for Web of Science ID 000320714400005

  View details for PubMedID 23671187

  View details for PubMedCentralID PMC3700131

• Time-lapse microscopy and image analysis in basic and clinical embryo development research REPRODUCTIVE BIOMEDICINE ONLINE Wong, C., Chen, A. A., Behr, B., Shen, S. 2013; 26 (2): 120-129

  Abstract

  Mammalian preimplantation embryo development is a complex process in which the exact timing and sequence of events are as essential as the accurate execution of the events themselves. Time-lapse microscopy (TLM) is an ideal tool to study this process since the ability to capture images over time provides a combination of morphological, dynamic and quantitative information about developmental events. Here, we systematically review the application of TLM in basic and clinical embryo research. We identified all relevant preimplantation embryo TLM studies published in English up to May 2012 using PubMed and Google Scholar. We then analysed the technical challenges involved in embryo TLM studies and how these challenges may be overcome with technological innovations. Finally, we reviewed the different types of TLM embryo studies, with a special focus on how TLM can benefit clinical assisted reproduction. Although new parameters predictive of embryo development potential may be discovered and used clinically to potentially increase the success rate of IVF, adopting TLM to routine clinical practice will require innovations in both optics and image analysis. Combined with such innovations, TLM may provide embryologists and clinicians with an important tool for making critical decisions in assisted reproduction. In this review, we perform a literature search of all published early embryo development studies that used time-lapse microscopy (TLM). From the literature, we discuss the benefits of TLM over traditional time-point analysis, as well as the technical difficulties and solutions involved in implementing TLM for embryo studies. We further discuss research that has successfully derived non-invasive markers that may increase the success rate of assisted reproductive technologies, primarily IVF. Most notably, we extend our discussion to highlight important considerations for the practical use of TLM in research and clinical settings.

  View details for DOI 10.1016/j.rbmo.2012.11.003

  View details for Web of Science ID 000314664400003

  View details for PubMedID 23273754

• Dynamic blastomere behaviour reflects human embryo ploidy by the four-cell stage NATURE COMMUNICATIONS Chavez, S. L., Loewke, K. E., Han, J., Moussavi, F., Colls, P., Munne, S., Behr, B., Pera, R. A. 2012; 3

  Abstract

  Previous studies have demonstrated that aneuploidy in human embryos is surprisingly frequent with 50-80% of cleavage-stage human embryos carrying an abnormal chromosome number. Here we combine non-invasive time-lapse imaging with karyotypic reconstruction of all blastomeres in four-cell human embryos to address the hypothesis that blastomere behaviour may reflect ploidy during the first two cleavage divisions. We demonstrate that precise cell cycle parameter timing is observed in all euploid embryos to the four-cell stage, whereas only 30% of aneuploid embryos exhibit parameter values within normal timing windows. Further, we observe that the generation of human embryonic aneuploidy is complex with contribution from chromosome-containing fragments/micronuclei that frequently emerge and may persist or become reabsorbed during interphase. These findings suggest that cell cycle and fragmentation parameters of individual blastomeres are diagnostic of ploidy, amenable to automated tracking algorithms, and likely of clinical relevance in reducing transfer of embryos prone to miscarriage.

  View details for DOI 10.1038/ncomms2249

  View details for Web of Science ID 000316356700019

  View details for PubMedID 23212380

  View details for PubMedCentralID PMC3535341

• Promotion of Human Early Embryonic Development and Blastocyst Outgrowth In Vitro Using Autocrine/Paracrine Growth Factors PLOS ONE Kawamura, K., Chen, Y., Shu, Y., Cheng, Y., Qiao, J., Behr, B., Pera, R. A., Hsueh, A. J. 2012; 7 (11)

  Abstract

  Studies using animal models demonstrated the importance of autocrine/paracrine factors secreted by preimplantation embryos and reproductive tracts for embryonic development and implantation. Although in vitro fertilization-embryo transfer (IVF-ET) is an established procedure, there is no evidence that present culture conditions are optimal for human early embryonic development. In this study, key polypeptide ligands known to be important for early embryonic development in animal models were tested for their ability to improve human early embryo development and blastocyst outgrowth in vitro. We confirmed the expression of key ligand/receptor pairs in cleavage embryos derived from discarded human tri-pronuclear zygotes and in human endometrium. Combined treatment with key embryonic growth factors (brain-derived neurotrophic factor, colony-stimulating factor, epidermal growth factor, granulocyte macrophage colony-stimulating factor, insulin-like growth factor-1, glial cell-line derived neurotrophic factor, and artemin) in serum-free media promoted >2.5-fold the development of tri-pronuclear zygotes to blastocysts. For normally fertilized embryos, day 3 surplus embryos cultured individually with the key growth factors showed >3-fold increases in the development of 6-8 cell stage embryos to blastocysts and >7-fold increase in the proportion of high quality blastocysts based on Gardner's criteria. Growth factor treatment also led to a 2-fold promotion of blastocyst outgrowth in vitro when day 7 surplus hatching blastocysts were used. When failed-to-be-fertilized oocytes were used to perform somatic cell nuclear transfer (SCNT) using fibroblasts as donor karyoplasts, inclusion of growth factors increased the progression of reconstructed SCNT embryos to >4-cell stage embryos. Growth factor supplementation of serum-free cultures could promote optimal early embryonic development and implantation in IVF-ET and SCNT procedures. This approach is valuable for infertility treatment and future derivation of patient-specific embryonic stem cells.

  View details for DOI 10.1371/journal.pone.0049328

  View details for Web of Science ID 000311234000064

  View details for PubMedID 23152897

  View details for PubMedCentralID PMC3495911

• Outcomes of trophectoderm biopsy on cryopreserved blastocysts: a case series REPRODUCTIVE BIOMEDICINE ONLINE Lathi, R. B., Massie, J. A., Gilani, M., Milki, A. A., Westphal, L. M., Baker, V. L., Behr, B. 2012; 25 (5): 504-507

  Abstract

  Preimplantation genetic diagnosis (PGD) is an increasingly common adjunct to IVF. The information gained from PGD may be used to reduce the incidence of chromosomally abnormal pregnancies and augment the current selection process of embryos. As such, patients may choose to utilize PGD in either fresh or cryopreserved IVF cycles. It is a common practice to cryopreserve excess embryos at the blastocyst stage. In these cases, trophectoderm biopsy is the only technique available for PGD. This articles reports this study centre's experience with trophectoderm biopsies of cryopreserved blastocysts in 12 patients who underwent 13 cycles of PGD. The implantation rate per embryo transferred was 46% and the ongoing pregnancy rate per embryo transfer was 63%. The results from this case series demonstrate that trophectoderm biopsy on cryopreserved blastocysts to perform PGD is logistically feasible. In addition, the rate of implantation and ongoing pregnancy were maintained within a reasonable range to justify the procedure. Preimplantation genetic diagnosis (PGD) is an increasingly common adjunct to IVF and is used to evaluate the genetic makeup of the embryo prior to transfer of the embryo into the uterus. The information gained from PGD may be used to identify single-gene disorders that result in genetic disease, reduce the incidence of chromosomally abnormal pregnancies and/or augment the selection process of embryos to be transferred. In order to perform PGD, a biopsy of the embryo is the performed and cells are removed for testing. PGD may be performed in either fresh or frozen (cryopreserved) IVF cycles. Patients who have cryopreserved embryos remaining in storage from a previous fresh cycle may wish to have these embryos tested with PGD. Many embryos are frozen on day 5 of development, referred to as the blastocyst stage. At this stage of development, embryo biopsy is performed via a technique known as 'trophectoderm biopsy', in which 1-3 of the cells destined to become the placenta are removed from the embryo for chromosomal testing. We report our experience with trophectoderm biopsy of frozen blastocysts in 12 patients who underwent 13 cycles of PGD. The implantation rate per embryo transferred was 46% and the ongoing pregnancy rate per embryo transfer was 63%. The results from this case series demonstrate that trophectoderm biopsy on cryopreserved blastocysts to perform PGD is logistically feasible. In addition, the rate of implantation and ongoing pregnancy were maintained within a reasonable range to justify the procedure.

  View details for DOI 10.1016/j.rbmo.2012.06.021

  View details for Web of Science ID 000310639600010

  View details for PubMedID 22985500

• Genome-wide Single-Cell Analysis of Recombination Activity and De Novo Mutation Rates in Human Sperm CELL Wang, J., Fan, H. C., Behr, B., Quake, S. R. 2012; 150 (2): 402-412

  Abstract

  Meiotic recombination and de novo mutation are the two main contributions toward gamete genome diversity, and many questions remain about how an individual human's genome is edited by these two processes. Here, we describe a high-throughput method for single-cell whole-genome analysis that was used to measure the genomic diversity in one individual's gamete genomes. A microfluidic system was used for highly parallel sample processing and to minimize nonspecific amplification. High-density genotyping results from 91 single cells were used to create a personal recombination map, which was consistent with population-wide data at low resolution but revealed significant differences from pedigree data at higher resolution. We used the data to test for meiotic drive and found evidence for gene conversion. High-throughput sequencing on 31 single cells was used to measure the frequency of large-scale genome instability, and deeper sequencing of eight single cells revealed de novo mutation rates with distinct characteristics.

  View details for DOI 10.1016/j.cell.2012.06.030

  View details for Web of Science ID 000306595700018

  View details for PubMedID 22817899

  View details for PubMedCentralID PMC3525523

• Altered subcellular localization of transcription factor TEAD4 regulates first mammalian cell lineage commitment PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Home, P., Saha, B., Ray, S., Dutta, D., Gunewardena, S., Yoo, B., Pal, A., Vivian, J. L., Larson, M., Petroff, M., Gallagher, P. G., Schulz, V. P., White, K. L., Golos, T. G., Behr, B., Paul, S. 2012; 109 (19): 7362-7367

  Abstract

  In the preimplantation mouse embryo, TEAD4 is critical to establishing the trophectoderm (TE)-specific transcriptional program and segregating TE from the inner cell mass (ICM). However, TEAD4 is expressed in the TE and the ICM. Thus, differential function of TEAD4 rather than expression itself regulates specification of the first two cell lineages. We used ChIP sequencing to define genomewide TEAD4 target genes and asked how transcription of TEAD4 target genes is specifically maintained in the TE. Our analyses revealed an evolutionarily conserved mechanism, in which lack of nuclear localization of TEAD4 impairs the TE-specific transcriptional program in inner blastomeres, thereby allowing their maturation toward the ICM lineage. Restoration of TEAD4 nuclear localization maintains the TE-specific transcriptional program in the inner blastomeres and prevents segregation of the TE and ICM lineages and blastocyst formation. We propose that altered subcellular localization of TEAD4 in blastomeres dictates first mammalian cell fate specification.

  View details for DOI 10.1073/pnas.1201595109

  View details for Web of Science ID 000304090600052

  View details for PubMedID 22529382

  View details for PubMedCentralID PMC3358889

• Day 2 Transfer in Clinical ART SEMINARS IN REPRODUCTIVE MEDICINE Behr, B., McElroy, S. 2012; 30 (3): 222-229

  Abstract

  Over the past 20 years, numerous techniques have enhanced assisted reproductive technology outcomes to help couples have >60,000 infants in the United States in 2008. Several different days for embryo transfers have been studied, but debate for the best timing of embryo transfer is still ongoing. With growing concern about multiple gestations and neonatal outcomes, early cleavage stage embryo transfer with novel embryo selection tools may be attractive to some patients and in vitro fertilization programs. In this review, we summarize clinical and basic studies relating to the timing of embryo transfer and highlight the possibilities of safe embryo transfer by combining advanced embryo screening tools with potentially high efficiency and low adverse effects on clinical outcome.

  View details for DOI 10.1055/s-0032-1311524

  View details for Web of Science ID 000303703400009

  View details for PubMedID 22585633

• Testosterone concentrations in early pregnancy: relation to method of conception in an infertile population REPRODUCTIVE BIOMEDICINE ONLINE Lathi, R. B., Moayeri, S. E., Reddy, C. D., Gebhardt, J., Behr, B., Westphal, L. M. 2012; 24 (3): 360-363

  Abstract

  This prospective cohort study of infertility patients compared testosterone concentrations in early pregnancy in infertility patients who conceived naturally or after treatment. Although all groups demonstrated some increase in pregnancy testosterone from baseline concentrations, subjects who conceived following ovulation induction showed a significantly increased rise in testosterone as compared with controls (P<0.01).

  View details for DOI 10.1016/j.rbmo.2011.11.018

  View details for PubMedID 22285241

• Skeletogenic phenotype of human Marfan embryonic stem cells faithfully phenocopied by patient-specific induced-pluripotent stem cells PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Quarto, N., Leonard, B., Li, S., Marchand, M., Anderson, E., Behr, B., Francke, U., Reijo-Pera, R., Chiao, E., Longaker, M. T. 2012; 109 (1): 215-220

  Abstract

  Marfan syndrome (MFS) is a heritable connective tissue disorder caused by mutations in the gene coding for FIBRILLIN-1 (FBN1), an extracellular matrix protein. MFS is inherited as an autosomal dominant trait and displays major manifestations in the ocular, skeletal, and cardiovascular systems. Here we report molecular and phenotypic profiles of skeletogenesis in tissues differentiated from human embryonic stem cells and induced pluripotent stem cells that carry a heritable mutation in FBN1. We demonstrate that, as a biological consequence of the activation of TGF-β signaling, osteogenic differentiation of embryonic stem cells with a FBN1 mutation is inhibited; osteogenesis is rescued by inhibition of TGF-β signaling. In contrast, chondrogenesis is not perturbated and occurs in a TGF-β cell-autonomous fashion. Importantly, skeletal phenotypes observed in human embryonic stem cells carrying the monogenic FBN1 mutation (MFS cells) are faithfully phenocopied by cells differentiated from induced pluripotent-stem cells derived independently from MFS patient fibroblasts. Results indicate a unique phenotype uncovered by examination of mutant pluripotent stem cells and further demonstrate the faithful alignment of phenotypes in differentiated cells obtained from both human embryonic stem cells and induced pluripotent-stem cells, providing complementary and powerful tools to gain further insights into human molecular pathogenesis, especially of MFS.

  View details for DOI 10.1073/pnas.1113442109

  View details for Web of Science ID 000298876500045

  View details for PubMedID 22178754

  View details for PubMedCentralID PMC3252902

• Media composition: growth factors. Methods in molecular biology (Clifton, N.J.) Hegde, A., Behr, B. 2012; 912: 177-198

  Abstract

  Despite the fact that the fundamental principle underlying the most common method of culture media constitution is that of mimicking the natural environment of the preimplantation embryo, one major difference that remains between current embryo culture media and in vivo conditions is the absence of growth factors in vitro. Numerous growth factors are known to be present in the in vivo environment of human and nonhuman preimplantation embryos, often with peak concentrations corresponding to when fertilization and preimplantation embryo growth would occur. Although these growth factors are found in very small concentrations, they have a profound effect on tissue growth and differentiation through attachment to factor-specific receptors on cell surfaces. Receptors for many different growth factors have also been detected in human preimplantation embryos. Preimplantation embryos themselves express many growth factors. The growth factors and receptors are metabolically costly to produce, and thus their presence in the environment of the preimplantation embryo and in the embryo respectively strongly implies that embryos are designed to encounter and respond to the corresponding factors. Studies of embryo coculture also indirectly suggest that growth factors can improve in vitro development. Several animal and human studies attest to a probable beneficial effect of addition of growth factors to culture media. However, there is still ambiguity regarding the exact role of growth factors in embryonic development, the optimal dose of growth factors to be added to culture media, the combinatorial effect and endocrine of growth factors in embryonic development.

  View details for DOI 10.1007/978-1-61779-971-6_11

  View details for PubMedID 22829375

• National collection of embryo morphology data into Society for Assisted Reproductive Technology Clinic Outcomes Reporting System: associations among day 3 cell number, fragmentation and blastomere asymmetry, and live birth rate FERTILITY AND STERILITY Racowsky, C., Stern, J. E., Gibbons, W. E., Behr, B., Pomeroy, K. O., Biggers, J. D. 2011; 95 (6): 1985-1989

  Abstract

  To evaluate the validity of collecting day 3 embryo morphology variables into the Society for Assisted Reproductive Technology Clinic Outcomes Reporting System (SART CORS).Retrospective.National database-SART CORS.Fresh autologous assisted reproductive technology (ART) cycles from 2006-2007 in which embryos were transferred singly (n=1,020) or in pairs (n=6,508) and embryo morphology was collected.None.Relationship between live birth, maternal age, and morphology of transferred day 3 embryos as defined by cell number, fragmentation, and blastomere symmetry. Logistic multiple regressions and receiver operating characteristic curve analyses were applied to determine specificity and sensitivity for correctly classifying embryos as either failures or successes.Live birth rate was positively associated with increasing cell number up to eight cells (<6 cells: 2.9%; 6 cells: 9.6%; 7 cells: 15.5%; 8 cells: 24.3%; and >8 cells: 16.2%), but was negatively associated with maternal age, increasing fragmentation, and asymmetry scores. An area under the receiver operating curve of 0.753 (95% confidence interval 0.740-0.766) was derived, with a sensitivity of 45.0%, a specificity of 83.2%, and 76.4% of embryos being correctly classified with a cutoff probability of 0.3.This analysis provides support for the validity of collecting morphology fields for day 3 embryos into SART CORS. Standardization of morphology collections will assist in controlling for embryo quality in future database analyses.

  View details for DOI 10.1016/j.fertnstert.2011.02.009

  View details for Web of Science ID 000289620900028

  View details for PubMedID 21411078

• Donation of Embryos for Human Development and Stem Cell Research CELL STEM CELL Kalista, T., Freeman, H. A., Behr, B., Pera, R. R., Scott, C. T. 2011; 8 (4): 360-362

  Abstract

  Using donated human embryos for scientific research raises ethical questions about the donation process. We describe a two-stage consent process designed to help couples make informed decisions about embryo disposition. This consent methodology minimizes conflict of interest, respects patient choice, and provides a much-needed resource to patients and the research community.

  View details for DOI 10.1016/j.stem.2011.02.018

  View details for Web of Science ID 000289707100007

  View details for PubMedID 21474099

• IVF Outcomes: Effects of Blood or Mucus on the Tip of a Soft Embryo Transfer Catheter After Embryo Transfer 59th Annual Meeting of the Pacific-Coast-Reproductive-Society Dasig, J., Zhao, Q., Shu, Y., Reddy, V., Gebhardt, J., Behr, B. ELSEVIER SCIENCE INC. 2011: S26–S26

  View details for Web of Science ID 000287733700060

• Day 2 versus day 3 embryo transfer in poor responders: a prospective randomized trial FERTILITY AND STERILITY Shahine, L. K., Milki, A. A., Westphal, L. M., Baker, V. L., Behr, B., Lathi, R. B. 2011; 95 (1): 330-332

  Abstract

  Day 2 embryo transfer has been suggested as a method to improve pregnancy rates in poor responders compared with day 3 transfer. Our prospective randomized controlled trial does not show a difference in outcomes based on day of embryo transfer.

  View details for DOI 10.1016/j.fertnstert.2010.06.093

  View details for PubMedID 20813357

• Inter-laboratory validation of the measurement of follicle stimulating hormone (FSH) after various lengths of frozen storage REPRODUCTIVE BIOLOGY AND ENDOCRINOLOGY Scriver, J., Baker, V. L., Young, S. L., Behr, B., Pastore, L. M. 2010; 8

  Abstract

  Serum follicle stimulating hormone (FSH) levels are used clinically to evaluate infertility, pituitary and gonadal disorders. With increased frequency of research collaborations across institutions, it is essential that inter-laboratory validation is addressed.An inter-laboratory validation of three commercial FSH immunoassays was performed with human serum samples of varying frozen storage length (2 batches of 15 samples each) at -25 degree C. Percentage differences and Bland-Altman limits of agreement were calculated.The inter- and intra-laboratory consistency of FSH values with the same assay manufacturer was much higher after shorter-term storage (frozen for less than 11 months, mean percentage degradation less than 4%) than after long-term storage (2-3 years, mean percentage degradation = 23%). Comparing assay results from different manufacturers, there was similar overall long term degradation as seen with the same manufacturer (-25%), however the degradation was greater when the original FSH was greater than 20 mIU/mL relative to less than 10 mIU/mL (p < 0.001 trend test).The findings suggest that degradation of serum samples stored between 11 months and 2-3 years at -25 degrees C can lead to unstable FSH measurements. Inter-laboratory variability due to frozen storage time and manufacturer differences in assay results should be accounted for when designing and implementing research or clinical quality control activities involving serum FSH at multiple study sites.

  View details for DOI 10.1186/1477-7827-8-145

  View details for Web of Science ID 000285635900001

  View details for PubMedID 21114859

  View details for PubMedCentralID PMC3009700

• Non-invasive imaging of human embryos before embryonic genome activation predicts development to the blastocyst stage NATURE BIOTECHNOLOGY Wong, C. C., Loewke, K. E., Bossert, N. L., Behr, B., De Jonge, C. J., Baer, T. M., Pera, R. A. 2010; 28 (10): 1115-U199

  Abstract

  We report studies of preimplantation human embryo development that correlate time-lapse image analysis and gene expression profiling. By examining a large set of zygotes from in vitro fertilization (IVF), we find that success in progression to the blastocyst stage can be predicted with >93% sensitivity and specificity by measuring three dynamic, noninvasive imaging parameters by day 2 after fertilization, before embryonic genome activation (EGA). These parameters can be reliably monitored by automated image analysis, confirming that successful development follows a set of carefully orchestrated and predictable events. Moreover, we show that imaging phenotypes reflect molecular programs of the embryo and of individual blastomeres. Single-cell gene expression analysis reveals that blastomeres develop cell autonomously, with some cells advancing to EGA and others arresting. These studies indicate that success and failure in human embryo development is largely determined before EGA. Our methods and algorithms may provide an approach for early diagnosis of embryo potential in assisted reproduction.

  View details for DOI 10.1038/nbt.1686

  View details for Web of Science ID 000282870500034

  View details for PubMedID 20890283

• Standardization of grading embryo morphology FERTILITY AND STERILITY Racowsky, C., Vernon, M., Mayer, J., Ball, G., Behr, B., Pomeroy, K. O., Wininger, D., Gibbons, W., Conaghan, J., Stern, J. E. 2010; 94 (3): 1152–53

  Abstract

  Standardization of morphologic assessment for an embryo grading system was developed and is being implemented by the Society for Assisted Reproductive Technology (SART). A recent European consensus conference of embryologists from Europe and America is working toward adopting an embryo classification system modeled similarly to that of SART that, if adopted, would produce a de facto international standard to aid cross-border collaboration.

  View details for DOI 10.1016/j.fertnstert.2010.05.042

  View details for Web of Science ID 000280407900063

  View details for PubMedID 20580357

• Standardization of grading embryo morphology JOURNAL OF ASSISTED REPRODUCTION AND GENETICS Racowsky, C., Vernon, M., Mayer, J., Ball, G., Behr, B., Pomeroy, K. O., Wininger, D., Gibbons, W., Conaghan, J., Stern, J. E. 2010; 27 (8): 437–39

  Abstract

  Standardization of morphological assessment for embryo grading system was developed and is being implemented by the Society for Assisted Reproductive Technology (SART). A recent European consensus conference of embryologists from Europe and America is working toward adopting an embryo classification system modeled similarly to that of SART which, if adopted, would produce a de facto international standard to aid cross border collaboration.

  View details for DOI 10.1007/s10815-010-9443-2

  View details for Web of Science ID 000282475100002

  View details for PubMedID 20532975

  View details for PubMedCentralID PMC2941588

• Parthenogenic Blastocysts Derived from Cumulus-Free In Vitro Matured Human Oocytes PLOS ONE McElroy, S. L., Byrne, J. A., Chavez, S. L., Behr, B., Hsueh, A. J., Westphal, L. M., Pera, R. A. 2010; 5 (6)

  Abstract

  Approximately 20% of oocytes are classified as immature and discarded following intracytoplasmic sperm injection (ICSI) procedures. These oocytes are obtained from gonadotropin-stimulated patients, and are routinely removed from the cumulus cells which normally would mature the oocytes. Given the ready access to these human oocytes, they represent a potential resource for both clinical and basic science application. However culture conditions for the maturation of cumulus-free oocytes have not been optimized. We aimed to improve maturation conditions for cumulus-free oocytes via culture with ovarian paracrine/autocrine factors identified by single cell analysis.Immature human oocytes were matured in vitro via supplementation with ovarian paracrine/autocrine factors that were selected based on expression of ligands in the cumulus cells and their corresponding receptors in oocytes. Matured oocytes were artificially activated to assess developmental competence. Gene expression profiles of parthenotes were compared to IVF/ICSI embryos at morula and blastocyst stages. Following incubation in medium supplemented with ovarian factors (BDNF, IGF-I, estradiol, GDNF, FGF2 and leptin), a greater percentage of oocytes demonstrated nuclear maturation and subsequently, underwent parthenogenesis relative to control. Similarly, cytoplasmic maturation was also improved as indicated by development to blastocyst stage. Parthenogenic blastocysts exhibited mRNA expression profiles similar to those of blastocysts obtained after IVF/ICSI with the exception for MKLP2 and PEG1.Human cumulus-free oocytes from hormone-stimulated cycles are capable of developing to blastocysts when cultured with ovarian factor supplementation. Our improved IVM culture conditions may be used for obtaining mature oocytes for clinical purposes and/or for derivation of embryonic stem cells following parthenogenesis or nuclear transfer.

  View details for DOI 10.1371/journal.pone.0010979

  View details for PubMedID 20539753

• Cryopreservation of Human Oocytes and Embryos REPRODUCTIVE ENDOCRINOLOGY AND INFERTILITY: INTEGRATING MODERN CLINICAL AND LABORATORY PRACTICE Behr, B., Shu, Y., Carrell, D. T., Peterson, C. M. 2010: 689–701

  View details for DOI 10.1007/978-1-4419-1436-1_46

  View details for Web of Science ID 000276968200046

• Normal pregnancy after tetraploid karyotype on trophectoderm biopsy FERTILITY AND STERILITY Krieg, S. A., Lathi, R. B., Behr, B., Westphal, L. M. 2009; 92 (3)

  Abstract

  To report a case of successful pregnancy after trophectoderm biopsy and fluorescence in situ hybridization (FISH) revealed a tetraploid karyotype.Case report.A university medical center.An infertility patient desiring trophectoderm biopsy on frozen blastocysts to facilitate preimplantation genetic screening.Frozen blastocysts were thawed on the evening before transfer. Trophectoderm biopsy was performed the following morning. FISH results were available the same day, and two embryos with tetraploid results were transferred.Chorionic villus sample (CVS) and newborn exam.Normal diploid CVS result and a healthy male infant.Although multiple cells can be analyzed using trophectoderm biopsy, abnormalities in the trophectoderm may not be present in the inner cell mass.

  View details for DOI 10.1016/j.fertnstert.2009.06.007

  View details for Web of Science ID 000283282700007

  View details for PubMedID 19608167

• The Role of Prolactin- and Endometriosis-Associated Infertility OBSTETRICAL & GYNECOLOGICAL SURVEY Wang, H., Gorpludolo, N., Behr, B. 2009; 64 (8): 542-547

  Abstract

  This review will address the current understanding of the relationship between prolactin (PRL) and endometriosis-associated infertility. Although the exact mechanisms of action of hyperprolactinemia in patients with endometriosis-associated infertility have not been clearly established, this report reviews results from relevant studies in the literature. These include serum PRL levels in endometriosis-associated infertility, PRL receptors in ectopic endometriotic tissues, basal PRL levels after TSH and Danazol (isoxazolic derivative of the synthetic steroid 5alpha-ethinyl-testosterone) therapy, peritoneal fluid and nocturnal serum PRL levels in endometriosis, infertility, and luteal phase PRL concentrations in patients with endometriosis.Obstetricians & Gynecologists, Family Physicians.After completion of this article, the reader should be able to explain the relationship between prolactin- and endometriosis-associated infertility, relate endometriosis with infertility, and summarize two ways in which prolactin and endometriosis may be linked in the pathophysiology of infertility.

  View details for Web of Science ID 000268610800017

  View details for PubMedID 19624865

• Pregnancy after trophectoderm biopsy of frozen-thawed blastocyst FERTILITY AND STERILITY Lathi, R. B., Behr, B. 2009; 91 (5): 1938-1940

  Abstract

  To report a case of a successful pregnancy after trophectoderm biopsy and three-probe fluorescent in situ hybridization of a frozen blastocyst.Techniques and instrumentation.A University Medical Center.Infertility patient desiring trophectoderm biopsy on frozen blastocyst for preimplantation testing, from an IVF cycle at a referring IVF program.Frozen blastocysts were thawed the evening before the planned transfer. Trophectoderm biopsy was performed in the morning. The fluorescent in situ hybridization results were obtained the same day; embryo transfer was performed under ultrasound guidance.Serum betahCG and transvaginal ultrasound.Positive betahCG and ongoing pregnancy.Trophectoderm biopsy can be used as a means for testing frozen blastocysts in patients with excess embryos cryopreserved on day 5 or 6 from previously preformed IVF cycles.

  View details for DOI 10.1016/j.fertnstert.2008.02.132

  View details for Web of Science ID 000265969200055

  View details for PubMedID 18371958

• Embryo quality before and after surgical treatment of endometriosis in infertile patients JOURNAL OF ASSISTED REPRODUCTION AND GENETICS Shahine, L. K., Burney, R. O., Behr, B., Milki, A. A., Westphal, L. M., Lathi, R. B. 2009; 26 (2-3): 69-73

  Abstract

  To investigate the hypothesis that surgical treatment of endometriosis in infertile patients may improve pregnancy rates by improving embryo quality.We conducted a retrospective evaluation of 30 infertile patients treated with in vitro fertilization (IVF) before and after surgery for endometriosis. Patients served as their own controls and only cycles with similar stimulation protocols were compared.Using standard visual evaluation, embryo quality on day 3 was similar before and after surgical treatment of endometriosis. Fifty seven percent of patients had stage I-II endometriosis and 43% had stage III-IV disease. No patients had a live birth after the first IVF cycle and 43% of patients had a live birth with the IVF cycle after surgery.Surgical treatment of endometriosis does not alter embryo quality in patients with infertility treated with IVF.

  View details for DOI 10.1007/s10815-008-9287-1

  View details for PubMedID 19214735

• Non-Redundant Prognostic Factors in First-Cycles in In Vitro Fertilization 56th Annual Meeting of the Society-for-Gynecological-Investigation Shahine, L. K., Choi, B., O'Leary, K., Jun, S. H., Westphal, L. M., Behr, B., Wong, W. H., Yao, M. W. SAGE PUBLICATIONS INC. 2009: 279A–279A

  View details for Web of Science ID 000263609801187

• The value of fast blastocoele re-expansion in the selection of a viable thawed blastocyst for transfer 61st Annual Meeting of the American-Society-for-Reproductive-Medicine/51st Annual Meeting of the Canadian-Fertility-and-Andrology-Society Shu, Y., Watt, J., Gebhardt, J., Dasig, J., Appling, J., Behr, B. ELSEVIER SCIENCE INC. 2009: 401–6

  Abstract

  To investigate the role of fast blastocoele re-expansion in the selection of viable thawed blastocysts for transfer.Retrospective study.Academic assisted reproductive program.Transfer cycles were divided into two groups according to the presence or absence of fast re-expanded blastocysts. In group I (124 cycles), all transferred blastocysts had fast re-expanding blastocoele. In group II (113 cycles), no fast re-expanded blastocysts were included in the transfer.Blastocyst survival was defined as >50% of cells remaining intact after thaw and re-expansion after culture in vitro for 2-4 hours before transfer. Blastocysts with >or=50% re-expansion were designated as fast re-expanded blastocysts.Percentage of blastomere loss immediately after thaw, degree of blastocoele re-expansion, and clinical outcomes (pregnancy and implantation rates).The rates of survival and fast blastocoele re-expansion of partially intact blastocysts were significantly reduced as compared with fully intact blastocysts. Significantly higher rates of clinical pregnancy (37.1% vs. 16.8%) and implantation (26.7% vs. 11.3%) were obtained when all transferred blastocysts had fast re-expanding blastocoele as compared with those transfers without fast re-expanded blastocysts included.Our results showed that blastomere loss of thawed blastocyst was associated with a reduced ability to re-expand. As a discriminative morphologic marker of superior embryo viability, a fast re-expanded blastocyst would be given priority for transfer to better utilize the cryopreserved blastocysts.

  View details for DOI 10.1016/j.fertnstert.2007.11.083

  View details for Web of Science ID 000263445300016

  View details for PubMedID 18304536

• Metabolomic assessment of oocyte viability REPRODUCTIVE BIOMEDICINE ONLINE Nagy, Z. P., Jones-Colon, S., Roos, P., Botros, L., Greco, E., Dasig, J., Behr, B. 2009; 18 (2): 219-225

  Abstract

  The aim of the current study was to evaluate whether near-infrared (NIR) spectroscopy-generated metabolomic data obtained from oocyte culture samples would correlate with nuclear maturity status and derived embryo development. A total of 412 oocyte culture samples were collected from 43 patient cycles. Metabolomic profiles of metaphase I and II oocytes were obtained by NIR spectroscopy and were significantly different from each other and from profiles of prophase I (germinal vesicle) oocytes (P +/- 0.001 at the 95% confidence interval). Additionally, NIR spectroscopic analysis of culture medium of oocytes that developed to grade A embryos on day 3 demonstrated significantly higher viability indices (0.62 +/- 0.23) than those that developed to grades C/D (0.42 +/- 0.26; P < 0.006); and on day 5 grade A (0.37 +/- 0.20) was also higher than grades C/D (0.14 +/- 0.21; P < 0.02). Metabolomic profiles of oocytes that resulted in pregnancy had higher viability indices (0.87 +/- 0.27) than those that did not (0.44 +/- 0.17; P < 0.0001). The results of the current study demonstrate that metabolomic profiling from spent culture medium of the oocyte is related to nuclear maturity, is able to predict embryo development at day 3 and day 5 stages, and relates to embryo viability.

  View details for Web of Science ID 000263251700010

  View details for PubMedID 19192342

• Non-invasive assessment of embryo viability by metabolomic profiling of culture media ('metabolomics') 22nd Annual Meeting of the European-Society-of-Human-Reproduction-and-Embryology Nagy, Z. P., Sakkas, D., Behr, B. ELSEVIER SCI LTD. 2008: 502–7

  Abstract

  Increasing the efficiency of the IVF procedure by improving pregnancy/implantation rates and at the same time lowering (or avoiding) the risks of multiple gestations are the primary goals of the current assisted reproductive technology. These aims require a much improved gamete/embryo testing and selection procedure, which, using the current approach of microscopy-based morphology evaluation is unlikely to be achieved. Therefore, alternative or additional, non-invasive techniques have been proposed which may be able to detect alterations of the culture environment surrounding gametes/embryos reflective of the (patho-)physiological processes. One of the most recently applied approaches is to measure metabolomic changes in the culture medium of embryos and oocytes ('exometabolomics'). Initial studies have demonstrated that different types of spectrophotometric tests, including Raman and near-infrared (NIR) techniques, are similarly well capable of detecting specific changes of the 'secretome' (exometabolome). These studies have also demonstrated that metabolomic measurements correlate well with embryo development and morphology assessment. Furthermore, viability index on oocytes/embryos established by metabolomic tests may be a stronger predictor for implantation potential than traditional morphological assessment. Although the results of these initial investigations are promising, further prospective studies are required to define clearly the potential benefits and most relevant applications of this novel non-invasive technology in the field of assisted reproduction.

  View details for Web of Science ID 000259829800007

• Defining Human Embryo Phenotypes by Cohort-Specific Prognostic Factors PLOS ONE Jun, S. H., Choi, B., Shahine, L., Westphal, L. M., Behr, B., Pera, R. A., Wong, W. H., Yao, M. W. 2008; 3 (7)

  Abstract

  Hundreds of thousands of human embryos are cultured yearly at in vitro fertilization (IVF) centers worldwide, yet the vast majority fail to develop in culture or following transfer to the uterus. However, human embryo phenotypes have not been formally defined, and current criteria for embryo transfer largely focus on characteristics of individual embryos. We hypothesized that embryo cohort-specific variables describing sibling embryos as a group may predict developmental competence as measured by IVF cycle outcomes and serve to define human embryo phenotypes.We retrieved data for all 1117 IVF cycles performed in 2005 at Stanford University Medical Center, and further analyzed clinical data from the 665 fresh IVF, non-donor cycles and their associated 4144 embryos. Thirty variables representing patient characteristics, clinical diagnoses, treatment protocol, and embryo parameters were analyzed in an unbiased manner by regression tree models, based on dichotomous pregnancy outcomes defined by positive serum beta-human chorionic gonadotropin (beta-hCG). IVF cycle outcomes were most accurately predicted at approximately 70% by four non-redundant, embryo cohort-specific variables that, remarkably, were more informative than any measures of individual, transferred embryos: Total number of embryos, number of 8-cell embryos, rate (percentage) of cleavage arrest in the cohort and day 3 follicle stimulating hormone (FSH) level. While three of these variables captured the effects of other significant variables, only the rate of cleavage arrest was independent of any known variables.Our findings support defining human embryo phenotypes by non-redundant, prognostic variables that are specific to sibling embryos in a cohort.

  View details for DOI 10.1371/journal.pone.0002562

  View details for PubMedID 18596962

• A comparison of blastocyst formation from one-cell mouse zygotes following an aseptic vitrification system 56th Annual Meeting of the Pacific-Coast-Reproductive-Society Dasig, J., Bertocci, E., Zhao, Q., Shu, Y., Behr, B. ELSEVIER SCIENCE INC. 2008: S7–S7

  View details for Web of Science ID 000255250200009

• Application of custom-made electrofusion pipettes in mouse somatic cell nuclear transfer 56th Annual Meeting of the Pacific-Coast-Reproductive-Society Shu, Y., Rodriguez, R., Kim, S., Behr, B. ELSEVIER SCIENCE INC. 2008: S28–S28

  View details for Web of Science ID 000255250200070

• Derivation of human embryonic stem cells in standard and chemically defined conditions STEM CELL CULTURE Chiao, E., Kmet, M., Behr, B., Baker, J. 2008; 86: 1-?

  View details for DOI 10.1016/S0091-679X(08)00001-0

  View details for Web of Science ID 000256528200001

  View details for PubMedID 18442641

• Normal pregnancy resulting from a non-pronuclear oocyte at the time of examination for fertilization CLINICAL AND EXPERIMENTAL OBSTETRICS & GYNECOLOGY Burney, R. O., Gebhardt, J., Shu, Y., Behr, B., Westphal, L. M. 2008; 35 (3): 170-171

  Abstract

  To report the case of a patient undergoing in vitro fertilization (IVF) in which a non-pronuclear (0PN) oocyte resulted in a normal pregnancy.A 36-year-old woman underwent an IVF-embryo transfer treatment cycle.Four oocytes were retrieved for insemination by IVF. Examination for fertilization revealed two polypronuclearpolygynic and two non-pronuclear oocytes. The non-pronuclear oocytes were observed further for development. One embryo developed from the non-pronuclear cohort and was transferred at the 8-cell stage on day 3. Subsequently, a pregnancy developed, and resulted in the delivery of a healthy term infant.Non-pronuclear oocytes may represent a source of developmentally competent embryos, and further observation of this cohort should be considered, particularly in situations involving a low yield of oocytes at retrieval.

  View details for PubMedID 18754284

• Importance of integrity of blastomere in the re-expansion of cryopreserved blastocysts. 63rd Annual Meeting of the American-Society-for-Reproductive-Medicine Shu, Y., Watt, J., Gebhardt, J., Zhao, Q., Behr, B. ELSEVIER SCIENCE INC. 2007: S314–S314

  View details for Web of Science ID 000249889800899

• Use and outcomes of intracytoplasmic sperm injection for non-male factor infertility 101st Annual Meeting of the American-Urological-Association Kim, H. H., Bundorf, M. K., Behr, B., McCallum, S. W. ELSEVIER SCIENCE INC. 2007: 622–28

  Abstract

  To determine whether intracytoplasmic sperm injection (ICSI) is associated with improved outcomes for non-male factor infertility.We examined the patient characteristics associated with treatment choice-ICSI and conventional in vitro fertilization (IVF)-among patients without a diagnosis of male factor infertility and compared outcomes between the two groups, adjusting for patient characteristics using multivariate regression models.Academic fertility center.We evaluated 696 consecutive assisted reproductive technology (ART) cycles performed for couples with normal semen analysis at the Stanford Reproductive Endocrinology and Infertility Center between 2002 and 2003. We compared patient characteristics, cycle details, and outcomes for ICSI and IVF.Fertilization, pregnancy, and live birth rates.Patient characteristics were similar between the two groups, except the proportion of patients with unexplained infertility (IVF 15.1% vs. ICSI 23.5%), previous fertility (IVF 62.6% vs. ICSI 45.5%), and previous ART cycle (IVF 41.2% vs. ICSI 67.7%). More oocytes were fertilized per cycle for the IVF group (6.6 oocytes versus 5.1 oocytes). Fertilization failure, pregnancy, and live birth rates did not differ between IVF and ICSI. Using logistic regressions, having had previous ART was found to be positively associated with ICSI. Treatment choice of ICSI was not associated with fertilization, pregnancy, or live birth rates.No clear evidence of improved outcomes with ICSI was demonstrated for non-male factor infertility.

  View details for DOI 10.1016/j.fertnstert.2006.12.013

  View details for Web of Science ID 000249751900014

  View details for PubMedID 17445809

• Effects of blastomere loss on developmental competency of frozen-thawed human blastocysts. 63rd Annual Meeting of the American-Society-for-Reproductive-Medicine Shu, Y., Watt, J., Janice, G., Dasig, J., Jensen, J., Behr, B. ELSEVIER SCIENCE INC. 2007: S356–S356

  View details for Web of Science ID 000249889801021

• Unique biomarkers of human oocyte maturation assessed by non-invasive metabolomic profiling. 63rd Annual Meeting of the American-Society-for-Reproductive-Medicine Nagy, Z. P., Behr, B., Roos, P., Dasig, J., Burns, D. ELSEVIER SCIENCE INC. 2007: S4–S4

  View details for Web of Science ID 000249889800011

• Expression of CD9 in frozen-thawed mouse oocytes: preliminary experience FERTILITY AND STERILITY Wen, Y., Quintero, R., Chen, B., Shu, Y., Polan, M. L., Behr, B. 2007; 88 (2): 526-529

  Abstract

  CD9 mRNA and protein expression levels in mouse slow frozen-rapid thawed oocytes were compared with those in fresh oocytes by using comparative quantitative real time reverse transcription-PCR and semiquantitative Western blot, respectively. The expression levels of both CD9 mRNA and protein in the frozen oocytes were significantly lower than those found in the fresh oocytes.

  View details for DOI 10.1016/j.fertnstert.2006.11.130

  View details for Web of Science ID 000248716000044

  View details for PubMedID 17307168

• Identification of unique biomarkers of human oocyte maturation by non-invasive metabolomic profiling 23rd Annual Meeting of the European-Society-of-Human-Reproduction-and-Embryology Behr, B., Nagy, Z. P., Roos, R. P., Dasig, J., Kort, H. I., Burns, D. H. OXFORD UNIV PRESS. 2007: I166–I167

  View details for Web of Science ID 000255322200412

• Risk of monozygotic twinning with blastocyst transfer decreases over time: an 8-year experience FERTILITY AND STERILITY Moayeri, S. E., Behr, B., Lathi, R. B., Westphal, L. M., Milki, A. A. 2007; 87 (5): 1028-1032

  Abstract

  The purpose of our study is to compare the occurrence of monozygotic twinning (MZT) from blastocyst transfer (BT) in our program between an earlier and more recent time period.Retrospective.Academic IVF practice.All pregnancies conceived between March 2002 and December 2005 (N = 932) in our program were compared to pregnancies conceived before March 2002 (N = 554), which were the subject of a previous study.None.The incidence of MZT with day 3 embryo transfer and BT were compared between the study and control groups.During the study period, the rate of MZT was not significantly different for BT at 2.3% (9/385) compared to day 3 embryo transfer at 1.8% (10/547). This rate of 2.3% for BT was significantly lower than the rate of 5.6% (11/197) reported at our institution for BT before March 2002.Our study suggests that the risk of MZT with BT is significantly lower in the more recent time period and is in the range of what is seen with cleavage stage transfer. It is likely that improvements in culture systems as experience is gained with BT played a role.

  View details for DOI 10.1016/j.fertnstert.2006.09.013

  View details for PubMedID 17343858

• Fertilization, embryo development, and clinical outcome of immature oocytes from stimulated intracytoplasmic sperm injection cycles 60th Annual Meeting of the American-Society-for-Reproductive-Medicine Shu, Y., Gebhardt, J., Watt, J., Lyon, J., Dasig, D., Behr, B. ELSEVIER SCIENCE INC. 2007: 1022–27

  Abstract

  To evaluate the fertilization and developmental potential of immature oocytes obtained from controlled ovarian hyperstimulated cycles of patients undergoing intracytoplasmic sperm injection (ICSI).Retrospective study.Academic assisted reproductive technology program.Two hundred patients with at least one mature oocyte and one immature oocyte (study 1), and 44 patients with no mature oocytes (study 2) at time of oocyte denudation.Oocyte denudation was performed immediately after retrieval. Oocytes were cultured in vitro for 4-6 hours before ICSI and then categorized into four groups: group I, metaphase II (MII) oocytes at denudation; group II, in vitro matured MII oocytes; group III, metaphase I (MI) oocytes that did not progress to MII; and group 4, germinal-vesicle (GV) oocytes that converted to MI.Fertilization and embryo development were compared among groups in study 1. Pregnancy and implantation rates were evaluated in study 2.Although the fertilization rate in group III was significantly lower than in groups I and II, no significant difference was found between groups I and II. Day 3 embryos in group I had the highest mean number of blastomeres, proportions of good embryos, and blastocyst formation rate when compared with groups II and III. Two clinical pregnancies were achieved from 26 transfer cycles in study 2, resulting in pregnancy and implantation rates of 7.7% and 4% per transfer cycle, respectively.Although our results show that immature oocytes from stimulated cycles can be normally fertilized and used to increase the number of embryos available for transfer, the increase in number of embryos derived from immature oocytes cannot be efficiently translated into pregnancies and live births. The clinical significance of using immature oocytes in stimulated cycles needs further investigation.

  View details for DOI 10.1016/j.fertnstert.2006.08.110

  View details for Web of Science ID 000246583600005

  View details for PubMedID 17261289

• Effect of reduced oxygen concentrations on the outcome of in vitro fertilization FERTILITY AND STERILITY Kea, B., Gebhardt, J., Watt, J., Westphal, L. M., Lathi, R. B., Milki, A. A., Behr, B. 2007; 87 (1): 213-216

  Abstract

  We compared the effects of two standard oxygen concentrations, physiological (5% O(2), 5% CO(2), and 90% N(2)) and atmospheric (5% CO(2) with the balance as air), on fertilization, embryo development, and pregnancy rate in 106 patients undergoing IVF, excluding donor oocyte cycles and preimplantation genetic diagnosis cycles. The differences in oxygen concentration did not significantly affect fertilization rate, blastocyst formation, or pregnancy rate, but there was a significant difference in mean embryo score between physiological and atmospheric groups on day 3.

  View details for DOI 10.1016/j.fertnstert.2006.05.066

  View details for PubMedID 17081523

• Successful pregnancies after transplantation of frozen-thawed mouse ovaries into chimeric mice that received lethal-dose radiation FERTILITY AND STERILITY Migishima, F., Suzuki-Migishima, R., Quintero, R. B., Yokoyama, M., Behr, B. R. 2006; 86: 1080-1087

  Abstract

  To study whether fecundity was recovered in mice into which umbilical cord blood cells (UCBCs) were transfused after lethal-dose radiation, followed by transplantation of frozen-thawed ovaries.Prospective basic research study.Academic research laboratory.Female C57BL/6 mice as recipients of UCBCs and ovaries, male B6C3F1 mice for mating, and green fluorescent protein (GFP)-transgenic mice: 18.5-day-old fetuses (-/+) for UCBCs and adult GFP mice (+/+) for ovarian tissues.The UCBCs were transfused into each irradiated mouse, with GFP+ ovaries transplanted 4 weeks later. The chimeric mice were mated 3 weeks after ovarian transplantation and were examined 14 to 16 weeks after the transfusion of UCBCs.Percentage of chimerism, number of GFP+ pups.The percentage of chimerism in these mice tends to increase with the radiation dose. The recovery of fecundity was observed in the chimeric mice that were transplanted with fresh and previously vitrified ovaries after exposure to radiation.Even when the exposure dose of radiation administered as pretreatment is lethal, the fecundity of recipients can be maintained if their ovaries are cryopreserved before they are exposed to radiation.

  View details for DOI 10.1016/j.fertnstert.2006.03.023

  View details for Web of Science ID 000241289300007

  View details for PubMedID 16978625

• Preliminary experience on the CD9 expression on frozen-thawed mouse oocytes. 62nd Annual Meeting of the American-Society-for-Reproductive-Medicine (ASRM) Behr, B., Wen, Y., Quintero, R., Chen, B., Polan, M. L. ELSEVIER SCIENCE INC. 2006: S196–S196

  View details for Web of Science ID 000241038500470

• Non-invasive metabolomic profiling of human embryo culture media correlates with pregnancy outcome. Initial results of the metabolomics study group for ART. 62nd Annual Meeting of the American-Society-for-Reproductive-Medicine (ASRM) Seli, E., Sakkas, D., Behr, B., Nagy, P., Kwok, J. S., Burns, D. H. ELSEVIER SCIENCE INC. 2006: S115–S115

  View details for Web of Science ID 000241038500269

• Trophectoderm: A possible indicator of blastocyst survival and re-expansion after cryopreservation. 62nd Annual Meeting of the American-Society-for-Reproductive-Medicine (ASRM) Shu, Y., Gebhardt, J., Watt, J., Lyon, J., Jensen, J., Behr, B. ELSEVIER SCIENCE INC. 2006: S146–S147

  View details for Web of Science ID 000241038500345

• A novel microfluidic device for male subfertility screening JOURNAL OF UROLOGY McCormack, M. C., McCallum, S., Behr, B. 2006; 175 (6): 2223-2227

  Abstract

  To our knowledge no simple, disposable and accurate test of semen quality currently exists. A novel technique to assess the motile sperm concentration of the human ejaculate has been designed and its results are presented.In a micromachined device fluorescently labeled motile sperm traverse a hydrostatic microfluid line to a target detection cuvette. A microfluorometer assesses the fluorescence signal generated by sperm accumulating there throughout a 50-minute study period. A total of 21 semen specimens from men presenting to our university based reproductive endocrinology and infertility center were tested a total of 67 times. Semen parameters determined by computer assisted semen analysis were compared to the signal reported by the microfluidic device.The fluorescence signal increased throughout the data collection period for all samples. Pearson r values relating the device signal to total and progressive motile concentration were 0.79 and 0.80, respectively (each p <0.001). A signal threshold based on the aggregate data were established, correlating with the WHO standard of the normal total motile sperm concentration. As a screening test, the device was 94% sensitive and 97% specific for identifying samples with less than the WHO standard for the total motile concentration, and 96% sensitive and 90% specific when considering the progressive motile concentration.A novel microfluidic device is presented that enables accurate assessment of the motile sperm concentration in human ejaculate compared to computer assisted semen analysis. Its size and design demonstrate the feasibility of applying laboratory on chip technology to male infertility screening.

  View details for DOI 10.1016/S0022-5347(06)00276-X

  View details for Web of Science ID 000237585100067

  View details for PubMedID 16697844

• Regulation of cyclooxygenase activity in cultured endometrial stromal cells by granulocyte-macrophage colony-stimulating factor FERTILITY AND STERILITY Wang, H. B., Wen, Y., Polan, M. L., Boostanfar, R., Feinman, M., Behr, B. 2006; 85: 1118-1124

  Abstract

  To assess the ability of granulocyte-macrophage colony-stimulating factor (GM-CSF) to regulate cyclooxygenase (COX) enzyme activity and prostaglandins (PGs) synthesis, specifically PGE2 production in stromal cells, neither of which have been addressed in the literature.Prospective study.Department of obstetrics and gynecology at a university hospital.Human luteal phase endometrium was obtained from surgical specimens (n = 6) for clinical indications.Confluent stromal cells were stimulated with GM-CSF.Expression of COX mRNA, COX enzyme activity, and PGE2 level in cultured stromal cells.Confluent stromal cell cultures treated with P and E2 for 9 days were stimulated with GM-CSF. After treatment of 12 hours, low-dose GM-CSF (0.001-0.1 ng/mL) increased COX-2 mRNA levels in stromal cell, whereas high dose GM-CSF (1-100 ng/mL) decreased COX-1 and COX-2 mRNA levels. After treatment of 48 hours, low concentrations of GM-CSF (0.001-0.1 ng/mL) increased total COX and COX-2 enzyme activity, whereas high concentrations of GM-CSF (1-100 ng/mL) inhibited COX and COX-2 activity; The PGE2 levels decreased by 31% to 393.3 pg/mL (P < .05) with concentrations of GM-CSF increasing from 1 ng/mL to 100 ng/mL.There appeared to be a biphasic pattern of COX-2 enzyme response to GM-CSF with low concentrations increasing activity and high concentrations inhibiting activity. It is possible that GM-CSF may provide critical regulation of PG production in the preimplantation period.

  View details for DOI 10.1016/j.fertnstert.2005.09.040

  View details for Web of Science ID 000236902300007

  View details for PubMedID 16616083

• Exogenous granulocyte-macrophage colony-stimulating factor promotes follicular development in the newborn rat in vivo HUMAN REPRODUCTION Wang, H. B., Wen, Y., Polan, M. L., Boostanfar, R., Feinman, M., Behr, B. 2005; 20 (10): 2749-2756

  Abstract

  Expression and selective cellular localization of granulocyte-macrophage colony-stimulating factor (GM-CSF) and its receptor in ovarian tissue imply an autocrine/paracrine role in ovarian function. Evidence indicating a functional role for GM-CSF in ovarian follicular cell function has been provided by studies with GM-CSF knockout (GM-/-) mice, which suggest that GM-CSF influences events associated with murine follicular maturation.Immature female rats were treated with GM-CSF, FSH or saline for 5 or 10 days. Ovaries were collected for histologic examination and immunostaining determination of CYP17, a theca cell marker. In addition, ovarian section slides were evaluated by immunofluorescence for CD45, an ovarian leukocyte marker. To investigate the possible mechanism of GM-CSF action on follicular development, theca-interstitial cells (T-I) were separated and cultured. Cells were treated with increasing concentrations of GM-CSF, then evaluated for CYP17 mRNA and protein expression assays.After 10 days of treatment with GM-CSF, the number of small preantral and large preantral follicles was significantly increased compared with the control group (P < 0.05). Similarly, treatment with FSH increased the number of small preantral and large preantral follicles (P < 0.05). CD45 expression measured by immunofluorescence was not different in the three groups, indicating that the distribution of leukocytes was unchanged. In addition, CYP17 was increased in the T-I cells both in vivo and in vitro after GM-CSF treatment.The present results suggest that GM-CSF may play a significant role in follicular development.

  View details for DOI 10.1093/humrep/dei123

  View details for Web of Science ID 000232427600013

  View details for PubMedID 15958400

• Effects of repetitive vitrification on the survival of mouse oocytes. 61st Annual Meeting of the American-Society-for-Reproductive-Medicine/51st Annual Meeting of the Canadian-Fertility-and-Andrology-Society Migishima, F., Shu, Y., Chen, B., Zhao, Y., Polan, M. L., Behr, B. ELSEVIER SCIENCE INC. 2005: S186–S186

  View details for Web of Science ID 000232114600461

• IVF results in de novo DNA methylation and histone methylation at an Igf2-H19 imprinting epigenetic switch MOLECULAR HUMAN REPRODUCTION Li, T., Vu, T. H., Ulaner, G. A., Littman, E., Ling, J. Q., Chen, H. L., Hu, J. F., Behr, B., Giudice, L., Hoffman, A. R. 2005; 11 (9): 631-640

  Abstract

  Recent studies suggest that IVF and assisted reproduction technologies (ART) may result in abnormal genomic imprinting, leading to an increased frequency of Angelman syndrome (AS) and Beckwith-Weidemann syndrome (BWS) in IVF children. To learn how ART might alter the epigenome, we examined morulas and blastocysts derived from C57BL/6J X M. spretus F1 mice conceived in vivo and in vitro and determined the allelic expression of four imprinted genes: Igf2, H19, Cdkn1c and Slc221L. IVF-derived mouse embryos that were cultured in human tubal fluid (HTF) (Quinn's advantage) media displayed a high frequency of aberrant H19 imprinting, whereas in vivo and IVF embryos showed normal maternal expression of Cdkn1c and normal biallelic expression of Igf2 and Slc221L. Embryonic stem (ES) cells derived from IVF blastocysts also showed abnormal Igf2/H19 imprinting. Allele-specific bisulphite PCR reveals abnormal DNA methylation at a CCCTC-binding factor (CTCF) site in the imprinting control region (ICR), as the normally unmethylated maternal allele acquired a paternal methylation pattern. Chromatin immunoprecipitation (ChIP) assays indicate an increase of lysine 4 methylation (dimethyl Lys4-H3) on the paternal chromatin and a gain in lysine 9 methylation (trimethyl Lys9-H3) on the maternal chromatin at the same CTCF-binding site. Our results indicate that de novo DNA methylation on the maternal allele and allele-specific acquisition of histone methylation lead to aberrant Igf2/H19 imprinting in IVF-derived ES cells. We suggest that ART, which includes IVF and various culture media, might cause imprinting errors that involve both aberrant DNA methylation and histone methylation at an epigenetic switch of the Igf2-H19 gene region.

  View details for DOI 10.1093/molehr/gah230

  View details for Web of Science ID 000233361600003

  View details for PubMedID 16219628

• Osmotic behavior of human blastocysts as a potential predictor for survival. 61st Annual Meeting of the American-Society-for-Reproductive-Medicine/51st Annual Meeting of the Canadian-Fertility-and-Andrology-Society Shu, Y., Gebhardt, J., Watt, J., Lyon, J., Jensen, J., Behr, B. ELSEVIER SCIENCE INC. 2005: S186–S187

  View details for Web of Science ID 000232114600462

• Effect of GMCSF on development and gene imprinting in preimplantation mouse embryos. 53rd Annual Meeting of the Pacific-Coast-Reproductive-Society Littman, E. D., Ling, J. Q., Ulaner, G. A., Vu, T. H., Dasig, D., Lyon, J., Giudic, L. C., Hoffman, A. R., Behr, B. ELSEVIER SCIENCE INC. 2005: S8–S8

  View details for Web of Science ID 000229048000005

• The use of recombinant human serum albumin (rHSA) for mouse IVF and embryo culture: Effect of diluent and protein stabilizers. 53rd Annual Meeting of the Pacific-Coast-Reproductive-Society QUINN, P., Carter, D. C., Ye, P., Behr, B. ELSEVIER SCIENCE INC. 2005: S24–S24

  View details for Web of Science ID 000229048000051

• Pregnancies and live births after transfer of cryopreserved hatching or hatched blastocysts. 53rd Annual Meeting of the Pacific-Coast-Reproductive-Society Shu, Y., Gebhardt, J., Watt, J., Lyon, J., Jensen, J., Behr, B. ELSEVIER SCIENCE INC. 2005: S20–S20

  View details for Web of Science ID 000229048000039

• Efficacy of recombinant human serum albumin as a protein source for the development of in vitro cultured human embryos: a pilot study. 53rd Annual Meeting of the Pacific-Coast-Reproductive-Society Lyon, J., QUINN, P., Carter, D. C., Ye, P., Dasig, D., Behr, B. ELSEVIER SCIENCE INC. 2005: S25–S25

  View details for Web of Science ID 000229048000054

• Preliminary experience with low concentration of granulocyte-macrophage colony-stimulating factor: A potential regulator in preimplantation mouse embryo development and apoptosis JOURNAL OF ASSISTED REPRODUCTION AND GENETICS Behr, B., Mooney, S., Wen, Y., Pollan, M. L., Wang, H. B. 2005; 22 (1): 25-32

  Abstract

  To investigate the effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the development of preimplantation mouse embryos.Mouse 2-cell embryos were collected and cultured in P-1 medium supplemented with GM-CSF at different concentrations. Using reverse transcription-polymerase chain reaction, expression Bcl-2 and Bax mRNA in blastocyst were evaluated in the GM-CSF group and control group. Apoptosis detection was performed using the in situ apoptosis detection kit in mouse blastocysts. The statistical significance of the data was analyzed using t-test and chi-square test.The development of blastocyst increased to 89% in the addition of GM-CSF (0.125 ng/mL), compared to controlled group (80%). The number of cells staining for apoptosis was lower in GM-CSF group than that in the control group. Bcl-2 expression was found to be upregulated in blastocysts in the GM-CSF supplemented group compared to the control group.These results suggest that GM-CSF might be an important regulator in embryo development.

  View details for DOI 10.1007/s10815-005-0817-9

  View details for PubMedID 15807219

• Preliminary study of the effect of granulocyte macrophage colony stimulating factor (GMCSF) Supplementation in culture media used for in vitro fertilization embryo development and on Igf2 gene imprinting in preimplantation mouse embryos. Fertility and Sterility Littman, E., Ulaner, G., Ling , J., Vu, T., Giudice, L., Hoffman, A., Behr, B. 2005; 83 (5): S8
• Monozygotic twin birth after the transfer of a cleavage stage embryo resulting from a single pronucleated oocyte JOURNAL OF ASSISTED REPRODUCTION AND GENETICS Dasig, D., Lyon, J., Behr, B., Milki, A. A. 2004; 21 (12): 427-429

  Abstract

  To present a case involving the transfer of a single pronucleated oocyte resulting in a monozygotic twin pregnancy.A descriptive case report of a single patient.The patient conceived and was found to have a monochorionic diamnionic pregnancy which resulted in the birth of normal identical twin boys at 32 weeks of gestation.The case report addresses an issue that has not received proper attention in the literature. It illustrates that observing a single PN in an oocyte at fertilization check should not be an absolute deterrent to transferring the resulting embryo even in an older patient with a high FSH level. This report also suggests that single observations, especially at the assessment of fertilization, in the IVF laboratory are limited when evaluating embryo potential and normalcy.

  View details for PubMedID 15704517

• Correlation between normal sperm head morphology (NSHM) and sperm binding potential to the sperm hyaluronan-binding assay (HBA (TM)). 60th Annual Meeting of the American-Society-for-Reproductive-Medicine Worrilow, K. C., Lyon, J., Belcak, N., Peters, A. J., Johnston, J. B., Behr, B. ELSEVIER SCIENCE INC. 2004: S95–S95

  View details for Web of Science ID 000224010800236

• Significance of immature oocytes in intracytoplasmic sperm injection treatment cycles 60th Annual Meeting of the American-Society-for-Reproductive-Medicine Behr, B., Gebhardt, J., Watt, J., Lyon, J., Dasig, D., Shu, Y. ELSEVIER SCIENCE INC. 2004: S293–S294

  View details for Web of Science ID 000224010800773

• Maturation, embryo development and clinical outcomes in ICSI treatment cycles with completely immature oocytes at the removal of cumulus cells. 60th Annual Meeting of the American-Society-for-Reproductive-Medicine Shu, Y., Gebhardt, J., Watt, J., Lyon, J., Dasig, D., Behr, B. ELSEVIER SCIENCE INC. 2004: S331–S331

  View details for Web of Science ID 000224010800876

• Effects of culture conditions on IVF outcome Serono Symposium on Genesis and Fate of the Preimplantation Empryo Behr, B., Wang, H. ELSEVIER SCIENCE BV. 2004: S72–S76

  Abstract

  Although in vitro fertilization (IVF) success rates have improved over the past decade, multiple pregnancies have become a formidable problem. The solution to this problem seems simple by mandating the reduction in numbers of embryos transferred. However, this is typically not accomplished without a compromise in the pregnancy rate. There have been a number of approaches designed to address high order multiple pregnancies from multi factorial analysis of early cleavage stage embryos to the development of extended culture systems, both of which require manipulations in the culture environment. Manipulations in embryo culture environment may not be benign. Several studies have demonstrated that adverse culture conditions have effects on gene expression and imprinting. Studies have also demonstrated that singleton human IVF babies have lower birth weight and higher incidence of congenital anomalies than natural conception babies. All of these factors need to be considered in relation to long term viability of IVF babies and the Barker hypothesis.

  View details for PubMedID 15196720

• Elective single blastocyst transfer FERTILITY AND STERILITY Milki, A. A., Hinckley, M. D., Westphal, L. M., Behr, B. 2004; 81 (6): 1697-1698

  Abstract

  This report describes our initial experience with elective single blastocyst transfer in 19 patients who had a mean age of 36.3 +/- 2.4 years. The ongoing pregnancy rate, 53% after the fresh embryo transfer and 68% when thaw cycles are included, suggests that single blastocyst transfer has a place in this relatively older patient population.

  View details for DOI 10.1016/j.fertnstert.2003.10.050

  View details for Web of Science ID 000222108800041

  View details for PubMedID 15193500

• The effect of a two-hour, room temperature incubation of human spermatozoa in TEST-yolk buffer on the rate of fertilization in vitro JOURNAL OF ASSISTED REPRODUCTION AND GENETICS Jensen, J. R., Walker, J. H., Milki, A. A., Westphal, L., Behr, B. 2004; 21 (5): 169-173

  Abstract

  To reassess the use of TEST-yolk buffer (TYB) in an in vitro fertilization (IVF) program by comparing fertilization rates achieved in a glucose-free cleavage medium by the standard IVF preparation of sperm versus a 2-h, room temperature incubation of sperm in TYB.Oocytes collected for IVF were randomly split into two groups and inseminated with either TYB-treated sperm or IVF-prepared sperm.Stanford Reproductive Endocrinology and Infertility Center.Fifty couples undergoing IVF with at least 10 mature oocytes.Fertilization rates in vitro.Fertilization rates were significantly higher (p = 0.015) with TYB treatment. The average 2PN fertilization rate was 49.6% (188/379) for the IVF group and 57.4% (221/385) in the IVF with TYB group.A 2-h, room temperature incubation of sperm in TYB produces significantly higher 2PN fertilization rates as compared to standard IVF preparation of sperm in a current generation cleavage medium.

  View details for Web of Science ID 000221941300007

  View details for PubMedID 15279324

  View details for PubMedCentralID PMC3455525

• Igf2 gene imprinting in preimplantation mouse embryos 52nd Annual Meeting of the Pacific-Coast-Reproductive-Society Littman, E. D., Ulaner, G. A., Vu, T. H., Otero, J., Dasig, D., Lyon, J., Gebhardt, J., Giudice, L. C., Behr, B., Hoffman, A. R. ELSEVIER SCIENCE INC. 2004: S14–S15

  View details for Web of Science ID 000220821600015

• Visualization of atypical hatching of a human blastocyst in vitro forming two identical embryos FERTILITY AND STERILITY Behr, B., Milki, A. A. 2003; 80 (6): 1502-1503

  View details for DOI 10.1016/j.fertnstert.2003.07.001

  View details for PubMedID 14667890

• Matrix metalloproteinase and tissue inhibitor of matrix metal loproteinase expression in human preimplantation embryos FERTILITY AND STERILITY Wang, H. B., Wen, Y., Mooney, S., Li, H., Behr, B., Polan, M. L. 2003; 80: 736-742

  Abstract

  To examine human embryos at various stages of preimplantation development for simultaneous expression of matrix metalloproteinases (MMPs) and tissue inhibitor of matrix metalloproteinases (TIMPs).mRNAs of specific MMPs and TIMPs were examined in single human embryos, at different stages of preimplantation development, by reverse transcription-polymerase chain reaction (RT-PCR). Single embryo immunohistochemistry was applied to examine the protein expression.University-affiliated IVF-ET program.Couples, attending the university-affiliated IVF-ET program, electing to donate poor prognosis embryos with anomalous morphology.None.The expression of MMP-1, MMP-2, MMP-9, TIMP-1, TIMP-2, and TIMP-3 in preimplantation embryos.The MMP-2 mRNA was expressed consistently during development from one-cell to blastocyst stage. The TIMP-1 and TIMP-3 mRNAs were detected in embryos at all stages; however, in the later preimplantation developmental stages, an increasing proportion of embryos expressing TIMP-1 and TIMP-3 mRNA were noted. The MMP-1, MMP-9, and TIMP-2 mRNAs were detected in only a minority of human embryos studied. Immunohistochemistry showed MMP-1 and TIMP-1 protein expression in preimplantation embryos.The existence of MMP and TIMP mRNA expression in human preimplantation embryos argues for a role for these metalloproteinases and their inhibitors during the process of implantation in humans.

  View details for DOI 10.1016/S0015-0282(03)00782-9

  View details for PubMedID 14505747

• Blastocyst cryopreservation: An update. Behr, B., Gebhardt, J., Milki, A. A. ELSEVIER SCIENCE INC. 2003: S149

  View details for Web of Science ID 000185672400395

• Elective single blastocyst transfer. Milki, A. A., Littman, E. D., Hinckley, M. D., Westphal, L. M., Gebhardt, J., Behr, B. ELSEVIER SCIENCE INC. 2003: S180

  View details for Web of Science ID 000185672400485

• Case report: Visualization of atypical hatching of a human blastocyst in vitro forming a monozygotic twin. Behr, B., Wang, H. B., Milki, A. A. ELSEVIER SCIENCE INC. 2003: S253

  View details for Web of Science ID 000185672400702

• Comparison of the sex ratio with blastocyst transfer and cleavage stage transfer 58th Annual Meeting of the American-Society-for-Reproductive-Medicine Milki, A. A., Jun, S. H., Hinckley, M. D., Westphal, L. W., Giudice, L. C., Behr, B. SPRINGER/PLENUM PUBLISHERS. 2003: 323–26

  Abstract

  To evaluate the sex ratio in births conceived with blastocyst transfer compared to day 3-ET.A retrospective analysis of IVF patients who became pregnant after blastocyst or cleavage stage transfer at Stanford University Hospital and a literature review were performed.In the day 3-ET group, the male-to-female (M/F) ratio was 157/139 (53%/47%) compared to 97/66 (59.5%/40.5%) in the blastocyst group (P = 0.18). Similar trends have been found in individual studies in the literature but reached statistical significance in only one out of six reports reviewed. The combined data from our study and the literature show a male-to-female ratio of 797/594 (57.3%/42.7%) in blastocyst transfer compared to 977/932 (51.2%/48.8%) in day 3-ET (P = 0.001).Although individual studies may lack power to show an altered sex ratio with blastocyst transfer, the combined data presented in this report do suggest that the M/F ratio is higher with blastocyst transfer compared to cleavage stage transfer.

  View details for PubMedID 12948095

• Granulocyte-macrophage colony-stimulating factor enhances human embryo development to the blastocyst stage: a randomized study. 51st Annual Meeting of the Pacific-Coast-Reproductive-Society Shapiro, B. S., Richter, K. S., Daneshmand, S. T., QUINN, P., Behr, B. ELSEVIER SCIENCE INC. 2003: S15–S16

  View details for Web of Science ID 000182128100028

• The effect of a two hour, room temperature incubation of human spermatozoa in test-yolk buffer on the rate of fertilization in vitro. 51st Annual Meeting of the Pacific-Coast-Reproductive-Society Behr, B., Lyon, J., Watt, J., Dasig, D., Gebhardt, J., Jensen, J. ELSEVIER SCIENCE INC. 2003: S10–S10

  View details for Web of Science ID 000182128100012

• Significance of one pronucleus before fertilization FERTILITY AND STERILITY Westphal, L. M., Rosencrantz, M., Behr, B., Milki, A. A. 2003; 79 (4): 1031-1033

  Abstract

  To describe a case of primary infertility associated with oocytes having one pronucleus before fertilization on repeated IVF attempts.Case report.A university-based assisted reproduction unit.A 30-year-old woman with primary infertility and oocytes containing one pronucleus before fertilization.Oocyte donation.Pregnancy.Conceived triplets after transfer of three embryos using donor oocytes.This patient's infertility was likely associated with an oocyte abnormality, as evidenced by the premature formation of one pronucleus before fertilization. In the future, more studies on the appearance of a single pronucleus before fertilization will be needed to determine its overall significance on fertility.

  View details for DOI 10.1016/S0015-0282(02)04852-5

  View details for PubMedID 12749450

• Effect of ICSI on subsequent blastocyst development and pregnancy rates 50th Annual Meeting of the Pacific-Coast-Reproductive-Society Westphal, L. M., Hinckley, M. D., Behr, B., Milki, A. A. SPRINGER/PLENUM PUBLISHERS. 2003: 113–16

  Abstract

  To investigate whether ICSI (intracytoplasmic sperm injection) results in decreased blastocyst formation and pregnancy compared to IVF (in vitro fertilization).We performed a retrospective analysis of blastocyst transfer (BT) offered routinely to patients under age 40 with > or = three 8-cell embryos on day 3 and compared IVF to ICSI cycles. Sequential media were used with P1 until day 3, then Blastocyst Medium until day 5/6.There were 131 IVF and 75 ICSI cycles. There was no difference in age, number of oocytes, zygotes, 8-cell embryos, blastocysts on days 5 and 6, or embryos transferred. Progression to blastocyst was similar (78% for IVF and 73% for ICSI) as was the viable pregnancy rate (51.4% for IVF and 55% for ICSI). No cycles failed to form blastocysts.The progression to blastocyst and the likelihood of conceiving a viable pregnancy were unaltered by ICSI. Thus it seems appropriate for programs to offer BT to patients undergoing ICSI using the same inclusion criteria applied to their IVF patients.

  View details for PubMedID 12735386

• Incidence of monozygotic twinning with blastocyst transfer compared to cleavage-stage transfer FERTILITY AND STERILITY Milki, A. A., Jun, S. H., Hinckley, M. D., Behr, B., Giudice, L. C., Westphal, L. M. 2003; 79 (3): 503-506

  Abstract

  To evaluate the incidence of monozygotic twinning (MZT) in pregnancies conceived after blastocyst transfer compared to cleavage-stage transfer.Retrospective study.University IVF program.All IVF patients with viable pregnancies conceived during a 4-year period.Blastocyst transfer or day 3 ET.Incidence of MZT assessed by transvaginal ultrasound.There were 11 incidences of MZT in 197 viable pregnancies (5.6%) with blastocyst transfer compared to 7 of 357 viable pregnancies (2%) with day 3 ET. In 10 of 18 pregnancies, MZT was observed in the setting of a higher order multiple gestation (6 of 11 for blastocyst transfer and 4 of 7 for day 3 ET). In the day 3 ET group, assisted hatching or intracytoplasmic sperm injection (ICSI) did not increase MZT (4 of 213, 1.9%) compared to cycles without zona breaching (3 of 144, 2.1%). Similarly, in the blastocyst-transfer group, ICSI did not increase the incidence of MZT (4 of 74, 5.5% for ICSI and 7 of 123, 5.7% for non-ICSI IVF).Compared to day 3 ET, blastocyst transfer appears to significantly increase the incidence of gestations with MZT. This information should be taken into account when counseling patients about the pros and cons of extended culture.

  View details for DOI 10.1016/S0015-0282(02)04754-4

  View details for PubMedID 12620430

• Comparison of blastocyst transfer to day 3 transfer with assisted hatching in the older patient 56th Annual Meeting of the American-Society-for-Reproductive-Medicine Milki, A. A., Hinckley, M. D., Behr, B. ELSEVIER SCIENCE INC. 2002: 1244–47

  Abstract

  To compare cycle outcomes in similar populations of women over 40 who underwent blastocyst transfer compared with women who had day 3 embryo transfer with assisted hatching (ET/AH).Retrospective study. STTING: University hospital-based program.Eighty-six IVF cycles in women ages 40 to 43 years who had more than three eight-cell embryos on day 3. On day 3 of embryo culture, patients chose either to undergo blastocyst transfer or day 3 ET/AH.Pregnancy and cryopreservation rates were recorded.In 48 cycles, blastocyst transfer was performed, and in 38 cycles day 3 ET/AH was performed. There was no statistically significant difference between the blastocyst transfer group and the day 3 ET/AH group with respect to age (41.1 +/- 0.9 years vs. 41.6 +/- 0.8 years), percentage of intracytoplasmic sperm injection cycles (29.2% vs. and 27.6%), number of oocytes (14.9 +/- 5.6 vs. 12.8 +/- 4.0), or number of eight-cell embryos (6.1 +/- 2.2 vs. 5.4 +/- 1.5). Significantly fewers embryos were transferred per cycle with blastocyst transfer (2.6 +/- 1.0) compared with day 3 ET/AH (5.9 +/- 2.0). The viable pregnancy rate was similar in the blastocyst transfer group (29.2%) and in the day 3 ET/AH group (26.3%). Embryos for cryopreservation were available in significantly more cycles in the blastocyst transfer group (52.1%) than in the day 3 ET/AH group (21.1%). Cleavage stage arrest occurred only in one cycle.Blastocyst transfer appears to be as effective as day 3 ET/AH in older patients with good embryos. Higher cryopreservation rate in the blastocyst transfer group may represent an advantage over day 3 ET/AH. Older women may also benefit from the information that extended culture provides them regarding their oocyte quality.

  View details for PubMedID 12477519

• Effect of osteopontin on mouse embryo development in vitro. 58th Annual Meeting of the American-Society-for-Reproductive-Medicine Behr, B., Ben Shlomo, I., Dasig, D., Lyon, J., Polan, M. L., Wang, H. ELSEVIER SCIENCE INC. 2002: S40–S41

  View details for Web of Science ID 000178239400105

• Non-invasive embryo viability assessments: The influence of media constituents during the first culture interval. 58th Annual Meeting of the American-Society-for-Reproductive-Medicine Dasig, D., Gebhardt, J., Lyon, J., Milki, A. A., Watt, J., Behr, B. ELSEVIER SCIENCE INC. 2002: S285–S285

  View details for Web of Science ID 000178239400796

• Regulation of cyclooxygenase activity in human endometrial stromal cells by granulocyte-macrophage colony-stimulating factor in vitro. 58th Annual Meeting of the American-Society-for-Reproductive-Medicine Wang, H. B., Wen, Y., Chen, B., Mooney, S., Behr, B., Polan, M. L. ELSEVIER SCIENCE INC. 2002: S220–S221

  View details for Web of Science ID 000178239400610

• Follicle development in grafted mouse ovaries after cryopreservation and subcutaneous transplantation AMERICAN JOURNAL OF OBSTETRICS AND GYNECOLOGY Wang, H. B., Mooney, S., Wen, Y., Behr, B., Polan, M. L. 2002; 187 (2): 370-374

  Abstract

  Our aim was to determine the impact of freezing, thawing, and subcutaneous transplantation on follicular development in grafted mouse ovaries.The mice were divided into 3 groups: control (group 1), frozen-thawed grafting (group 2), and frozen-thawed grafting with human menopausal gonadotropin injection (group 3). After freezing and thawing, the ovaries were transplanted into the subcutaneous tissue. Two weeks after transplantation, grafted ovaries and blood samples were collected.Ovaries from group 3 contained significantly more follicles (246 +/- 43 follicles) than group 2(P <.05). The pattern and intensity of Cx37 immunohistochemical staining was similar in all groups. Follicle-stimulating hormone concentrations were significantly decreased in group 2 after ovarian grafting.In mice, gonadotropin treatment before subcutaneous grafting improved the survival of growing follicles. Subcutaneous ovarian transplantation may restore ovarian function and could obviate many of the problems that are related to ovarian banking for humans.

  View details for DOI 10.1067/mob.2002.123606

  View details for PubMedID 12193927

• Phospholipase A(2) and cyclooxygenase gene expression in human preimplantation embryos JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM Wang, H. B., Wen, Y., Mooney, S., Behr, B., Polan, M. L. 2002; 87 (6): 2629-2634

  Abstract

  Phospholipase A(2) (PLA(2)) and cyclooxygenase (COX) are two key enzymes in PG synthesis; the latter has two forms, COX-1 and COX-2. mRNA was extracted from single preimplantation embryos and examined for PLA(2), COX-1, and COX-2 gene expression by RT-PCR to investigate whether PLA(2) and COX genes are expressed in human preimplantation conceptuses from zygote to blastocyst stage and to compare COX-1 and COX-2 gene expression within the same stage of embryonic development. Expression of PLA(2), COX-1, and COX-2 was detected in 48, 37, and 45%, respectively, of total embryos examined. COX-1 was expressed in approximately 66% of early human preimplantation embryos from zygote to two-cell stage, whereas COX-2 was expressed in about 58% of later stage embryos from eight-cell to blastocyst stage (P < 0.05). Furthermore, COX-2 mRNA and protein were localized to trophectoderm in blastocyst stage embryos. In conclusion, PLA(2), COX-1, and COX-2 are expressed during early human embryonic development and may contribute to the production of PGs such as PGE(2) in human embryogenesis. COX-1 and COX-2 are differentially expressed, with COX-2 being primarily expressed by trophectoderm in late-stage human preimplantation embryos, which may promote embryonic differentiation and implantation.

  View details for PubMedID 12050227

• Accuracy of day 3 criteria for selecting the best embryos FERTILITY AND STERILITY Milki, A. A., Hinckley, M. D., Gebhardt, J., Dasig, D., Westphal, L. M., Behr, B. 2002; 77 (6): 1191-1195

  Abstract

  To assess the accuracy of day 3 morphologic criteria in identifying the best embryos.Prospective observational study.University IVF program.One hundred cycles in women desiring blastocyst transfer who had > or =3 eight-cell embryos on day 3.On day 3, the embryologist chose the two embryos that would have been transferred that day. On day 5, embryos were examined to determine the best and second-best blastocysts.Accuracy of day 3 picks as measured in culture on day 5, outcome of nontransferred picks, and cryopreservation rate.All cycles reached the blastocyst stage and 73% had cryopreservation. The mean number of blastocysts was 4.8 (3.2 on day 5 and 1.6 on day 6). Neither pick was chosen in 39% of cycles; one pick was transferred in 38%; and both picks were transferred in 23%. Of 116 nontransferred picks, 51 were frozen and 65 arrested, with both picks arresting in 9 cycles. The single best blastocyst was chosen from the picks in 39% of cycles.Morphologic criteria for cleavage-stage embryo selection may fall short when the transfer is limited to two embryos. Culture to blastocyst is warranted in this population to avoid high-order multiples and still be able to choose the two embryos with the highest implantation potential.

  View details for PubMedID 12057727

• Factors relating to a successful cryopreserved blastocyst transfer program 56th Annual Meeting of the American-Society-for-Reproductive-Medicine Behr, B., Gebhardt, L., Lyon, J., Milki, A. A. ELSEVIER SCIENCE INC. 2002: 697–99

  Abstract

  To review the authors' experience in a successful frozen blastocyst program.Retrospective study.University IVF program.Women of all ages undergoing 64 frozen blastocyst nondonor thaw cycles.Thaw cycles with day 5 or day 6 frozen blastocysts replaced into luteal day 5 endometrium in natural or programmed cycles; cryopreservation of blastocysts by Menezo two-step protocol and Testart slow cool program; thawing by two step thaw protocol.Implantation, clinical pregnancy, and delivery rates.The implantation rate was 16% and was similar with day 5 frozen blastocysts and day 6 frozen blastocysts cycles. The clinical pregnancy rate was 36% and the delivery rate was 27%, with no significant difference between day 5 and 6 blastocysts.Blastocyst cryopreservation is a viable option for patients of all ages and complements fresh blastocyst culture and transfer. The presence of good quality blastocysts for freezing on day 5 and day 6 yields comparable results and is critical for the success of thaw cycles.

  View details for PubMedID 11937118

• CD146 expression in human preimplantation blastocyst. Wang, H., Wen, Y., Mooney, S., Behr, B., Polan, M. L. ELSEVIER SCIENCE INC. 2001: S203–S203

  View details for Web of Science ID 000170863900565

• Vascular endothelial growth factor (VEGF) mRNA splice variants are differentially expressed in human blastocysts 16th Annual Meeting of the European-Society-of-Human-Reproduction-and-Embryology (ESHRE) Krussell, J. S., Behr, B., Milki, A. A., Hirchenhain, J., Wen, Y., Bielfeld, P., Polan, M. L. OXFORD UNIV PRESS. 2001: 57–63

  Abstract

  The aim of our study was to detect and characterize mRNA expression of VEGF isoforms VEGF(121), VEGF(145), VEGF(165), VEGF(189), and VEGF(206) in human blastocysts. We recently demonstrated VEGF mRNA expression during human preimplantation embryo development, and further information regarding the alternatively spliced mRNAs resulting in freely secreted proteins or proteins bound to cell surface heparan-sulphate proteoglycans is needed to better understand the process of angiogenesis during implantation. Human blastocysts unsuitable for transfer obtained from the IVF programme at Stanford University were examined by reverse transcription/hemi-nested polymerase chain reaction for their expression of VEGF mRNA splice variants. VEGF mRNA was expressed in 17 out of 19 (89%) blastocysts. Of the 17 blastocysts, VEGF(121) mRNA was detected in 88%, VEGF(145) mRNA in 100%, VEGF(165) mRNA in 71%, and VEGF(189) mRNA in 24% of blastocysts. There was co-expression of mRNA for VEGF(121) and VEGF(145) only in 29% blastocysts, of mRNA for VEGF(165) and VEGF(145) only in 12%, and of mRNA for VEGF(121), VEGF(145) and VEGF(165) in 59% blastocysts. VEGF(206) mRNA could not be detected. In conclusion, we demonstrated that blastocysts express the mRNAs encoding for the free VEGF proteins, enabling the implanting embryo to immediately induce angiogenesis at the implantation site.

  View details for Web of Science ID 000166460800009

  View details for PubMedID 11134361

• Expression of vascular endothelial growth factor mRNA in human preimplantation embryos derived from tripronuclear zygotes 15th Annual Conference of the European-Society-of-Human-Reproduction-and-Embryology Krussel, J. S., Behr, B., Hirchenhain, J., Wen, Y., Milki, A. A., Cupisti, S., Bielfeld, P., Polan, M. L. ELSEVIER SCIENCE INC. 2000: 1220–26

  Abstract

  To detect the expression of vascular endothelial growth factor (VEGF) mRNA and/or secretion of VEGF protein by human preimplantation embryos.Human preimplantation embryos not suitable for uterine transfer were examined for beta-actin and VEGF mRNA expression. Culture media from normally fertilized and developing preimplantation embryos were assessed for VEGF protein secretion.Clinics and academic research laboratories at the Departments of Obstetrics and Gynecology at the Stanford University, Palo Alto, California and the Heinrich-Heine-University, Düsseldorf, Germany.Couples undergoing IVF by intracytoplasmic sperm injection for various reasons.Six unfertilized oocytes and 33 pathologically fertilized (tripronucleic, 3PN) preimplantation embryos were examined for VEGF mRNA expression, and 16 embryos were examined for VEGF protein secretion.Embryonic expression of VEGF mRNA and VEGF protein as determined by reverse transcription (RT)/nested polymerase chain reaction (PCR) and ELISA.VEGF mRNA and protein could not be detected in unfertilized oocytes. However, 30/33 preimplantation embryos did express VEGF mRNA (11/12 10-to-16-cell embryos, 3/4 morulae, 11/12 early blastocysts, 5/5 hatched blastocysts). The VEGF protein level was below the sensitivity of the ELISA.Production of VEGF may give the embryo the ability to induce neoangiogenesis at the implantation site, thus creating an environment necessary for its survival.

  View details for Web of Science ID 000165897300027

  View details for PubMedID 11119754

• Blastocyst-ET and monozygotic twinning 15th Annual Conference of the European-Society-of-Human-Reproduction-and-Embryology Behr, B., Fisch, J. D., Racowsky, C., Miller, K., Pool, T. B., Milki, A. A. SPRINGER/PLENUM PUBLISHERS. 2000: 349–51

  Abstract

  To examine the rate of monozygotic twinning associated with blastocyst transfer using commercially available, cell-free culture systems with unmanipulated blastocysts.A retrospective analysis was conducted in multiple private and academic infertility centers throughout the United States, of 199 pregnant patients following in vitro fertilization (IVF) blastocyst embryo transfer (ET). Human embryos obtained through standard IVF stimulation protocols were cultured in commercially available, cell-free media systems and transferred as blastocysts. The main outcome measure was the rate of monozygotic twinning.A total of 199 blastocyst-ET pregnancies were achieved during the study period at the fertility centers examined. Monozygotic twinning was noted in 10/199 (5%) of these pregnancies. All were monochorionic diamnionic.Monozygotic twinning previously has been reported following IVF, especially in relation to assisted hatching. While blastocyst transfer has been available for many years using coculture, there have been no published multicenter reports of monozygotic twinning associated with unmanipulated blastocysts. In a multicenter analysis, a definite increase in monozygotic twinning was seen following blastocyst-ET. We believe this phenomenon is real and that this information should be considered when counseling patients for treatment.

  View details for PubMedID 11042833

• Vascular endothelial growth factor (VEGF) mRNA isoforms are differentially expressed in human blastocysts Krussel, J. S., Behr, B., Milki, A. A., Wen, Y., Hirchenhain, J., Biefeld, P., Polan, M. L. OXFORD UNIV PRESS. 2000: 93–93

  View details for Web of Science ID 000088355100228

• Comparison of blastocyst transfer with day 3 embryo transfer in similar patient populations FERTILITY AND STERILITY Milki, A. A., Hinckley, M. D., Fisch, J. D., Dasig, D., Behr, B. 2000; 73 (1): 126-129

  Abstract

  To compare implantation and pregnancy rates (PRs) achieved with blastocyst transfer (BT) and day 3 ET in similar patient populations.Retrospective analysis.Academic infertility center.One hundred consecutive patients <40 years undergoing IVF, each with more than three eight-cell embryos on day 3.Patients used their own eggs for IVF or IVF and intracytoplasmic sperm injection. Embryos were cultured in P1 medium (Irvine Scientific, Santa Ana, CA) until day 3, when they were either transferred or, in the case of embryos for BT, incubated in Blastocyst Medium (Irvine Scientific), followed by transferring on day 5.Implantation and PRs.There were no statistically significant differences in patient age, FSH level, or number of oocytes or zygotes. The BT group had fewer embryos transferred (mean, 2.4) compared with the day 3-ET group (mean, 4.6). The viable PR (cardiac activity at 6-7 weeks was considered indicative of a viable pregnancy) was higher with BT (68%, 34/50) than with day 3 ET (46%, 23/50). The implantation rate was increased with BT (47%, 56 sacs/120 embryos) compared with day 3 ET (20%, 46 sacs/231 embryos).The BT group in our study had higher implantation and PRs compared with the day 3-ET group. Better embryo selection, improved embryo-uterine synchrony, and decreased cervical mucus on day 5 may have accounted for the enhanced outcome. Our data support the use of BT to limit the number of embryos transferred while improving PRs.

  View details for Web of Science ID 000084537500024

  View details for PubMedID 10632426

• Two-blastocyst transfer has similar pregnancy rates and a decreased multiple gestation rate compared with three-blastocyst transfer FERTILITY AND STERILITY Milki, A. A., Fisch, J. D., Behr, B. 1999; 72 (2): 225-228

  Abstract

  To examine the effect of the number of blastocysts transferred on pregnancy and multiple gestation rates.Retrospective study.Academic infertility center.Patients < 40 years undergoing IVF, with FSH levels of < 15 mIU/mL and more than three eight-cell embryos.Embryos were cultured in P1 until day 3 and then transferred to blastocyst medium. A maximum of three blastocysts were transferred.Pregnancy, multiple gestation, and implantation rates.All 55 patients developed blastocysts and underwent ET. Twenty-four patients had three embryos transferred and 29 patients had two embryos transferred. Two patients had only one embryo each for transfer. There was no difference in the viable pregnancy rate between the two-blastocyst transfer and three-blastocyst transfer groups (62% vs. 58%). In the two-blastocyst transfer group, 39% of pregnancies were multiple gestations (all twin gestations), compared with 79% of pregnancies in the three-blastocyst transfer group (50% twin gestations, 29% triplet gestations). The implantation rate was 47% in both groups.A commercially available, sequential culture system is highly effective for producing viable blastocysts. Two-blastocyst transfer eliminated the risk of triplets while maintaining the same high success rates seen with three-blastocyst ET.

  View details for Web of Science ID 000081761300005

  View details for PubMedID 10438984

• Human preimplantation embryos express vascular endothelial growth factor mRNA Kruessel, J. S., Wen, Y., Behr, B., Milki, A. A., Hirchenhain, J., Cupisti, S., Bielfield, P., Polan, M. L. OXFORD UNIV PRESS. 1999: 39–39

  View details for Web of Science ID 000081828000069

• Sibling embryo blastocyst development correlates with the in vitro fertilization day 3 embryo transfer pregnancy rate in patients under age 40 XVI World Congress on Fertility and Sterility/54th Annual Meeting of the American-Society-for-Reproductive-Medicine Fisch, J. D., Milki, A. A., Behr, B. ELSEVIER SCIENCE INC. 1999: 750–52

  Abstract

  To examine the IVF day 3-ET pregnancy rate in patients under 40 with sibling embryo blastocyst development, compared with similar patients without blastocyst formation.Retrospective analysis.Academic infertility center.One hundred twenty-five IVF day 3-ET patients under 40 with sibling embryos for extended culture.Extended culture of nontransferred sibling embryos for blastocyst development.Pregnancy and multiple gestation rates, number of oocytes, embryos formed, and embryos transferred.Thirty-eight percent of patients became pregnant. Forty-eight percent of patients had sibling embryos develop to blastocyst. The blastocyst group had more oocytes retrieved (17.4+/-6.6 versus 14.4+/-5.6), more embryos formed (11.2+/-4.2 versus 8.8+/-3.2), and a higher clinical pregnancy rate (60% versus 18%) than the group without blastocyst development.Blastocyst transfer has been shown to improve implantation rates and reduce the risk of multiple gestations from assisted reproductive technology. Sibling embryo blastocyst development may reflect superior embryo quality, as manifested by increased IVF-ET pregnancy rates. In addition to predicting pregnancy in the current cycle, sibling embryo blastocyst development may provide information about the potential for fresh blastocyst transfer in subsequent cycles and help to identify patients at risk for multiple gestations.

  View details for Web of Science ID 000079355700028

  View details for PubMedID 10202891

• Preliminary clinical experience with human blastocyst development in vitro without co-culture HUMAN REPRODUCTION Behr, B., Pool, T. B., Milki, A. A., Moore, D., Gebhardt, J., Dasig, D. 1999; 14 (2): 454-457

  Abstract

  This preliminary analysis was designed to quantify blastocyst development of supernumerary embryos without the use of feeder cells, conditioned medium or whole serum. Embryos derived from in-vitro fertilization (IVF) that were not transferred or cryopreserved were included in this study. Ova were harvested for IVF after a standard ovarian stimulation with gonadotrophin-releasing hormone agonist/ human menopausal gonadotrophin (GnRHa/HMG) or follicle-stimulating hormone (FSH). Ova were collected and culture in 150 microliters droplets of P1 medium under mineral oil, in groups at 37 degrees C under 5% CO2, 5% O2, 90% N2 (group A) or under 5% CO2 in air (group B) environment. Embryo transfer was performed 72 h post-harvest. Viable embryos not transferred or cryopreserved were placed in blastocyst medium and cultured for an additional 48 h in 5% CO2 in air. Embryos that exhibited an expanded blastocoelic cavity and well-defined inner cell mass at 120 h were counted. Of 838 supernumerary embryos cultured, 448 (53.5%) reached the expanded blastocyst stage by 120 h of culture. Patients were given the option of cryopreservation at that time. The embryos were cryopreserved using a standard protocol with serial addition of glycerol. Embryos reaching the blastocyst stage after more than 120 h of culture were not included. There was no difference in the proportions of blastocyst development between group A, 217/410 (53.5%) and group B, 231/428 (54%). To date, 16 patients have each had up to three thawed blastocysts transferred, out of whom seven became pregnant. This report demonstrates that a simple system of sequential culture generated acceptable, viable blastocyst development (54%) with supernumerary embryos, without the use of feeder cells, conditioned medium or whole serum. Recognizing the differential metabolic requirements of early and late cleavage stage embryos has enabled the application of a glucose/phosphate-free simple culture medium (P1) for up to 72 h of culture and a complex, glucose-containing medium (blastocyst medium) for subsequent blastocyst development.

  View details for Web of Science ID 000079002900034

  View details for PubMedID 10099993

• Blastocyst culture and transfer Human Reproduction Behr B. 1999
• Blastocyst culture and transfer HUMAN REPRODUCTION Behr, B. 1999; 14 (1): 5-6

  View details for Web of Science ID 000078341300003

  View details for PubMedID 10374084

• Enhancement of motility and acrosome reaction in human spermatozoa: differential activation by type-specific phosphodiesterase inhibitors HUMAN REPRODUCTION Fisch, J. D., Behr, B., Conti, M. 1998; 13 (5): 1248-1254

  Abstract

  Inhibition of sperm phosphodiesterase (PDE) has been shown to increase cAMP concentrations and stimulate motility and the acrosome reaction. While several PDE genes exist in mammals, little is known about the physiological role of PDE forms expressed in human spermatozoa. Using type-selective inhibitors, we identified two of the PDE forms expressed in human spermatozoa and studied their involvement in sperm function. Selective inhibitors of calcium-calmodulin-regulated PDE1 (8-methoxy-isobutyl-methylxanthine) and cAMP-specific PDE4 (RS-25344, Rolipram) were used to study PDE forms in human sperm extracts. 8-MeIBMX and Rolipram/RS-25344 inhibited sperm PDE activity by 35-40 and 25-30% respectively. Subcellular fractionation of the sperm homogenate suggests these pharmacologically distinct forms may be located in separate cellular regions. To evaluate the functional significance of different PDE forms, the effect of type-specific PDE inhibition on sperm motility and the acrosome reaction was examined. PDE4 inhibitors enhanced sperm motility over controls without affecting the acrosome reaction, while PDE1 inhibitors selectively stimulated the acrosome reaction. These data indicate at least two distinct PDE types exist in human spermatozoa. Our findings also support the hypothesis that PDE subtypes affect sperm function by regulating separate pools of cAMP and may ultimately offer novel treatments to infertile couples with abnormal semen parameters.

  View details for Web of Science ID 000074105500034

  View details for PubMedID 9647555

• Single blastomeres within human preimplantation embryos express different amounts of messenger ribonucleic acid for beta-actin and interleukin-1 receptor type I 13th Annual Meeting of the European-Society-of-Human-Reproduction-and-Embryology Krussel, J. S., Huang, H. Y., Simon, C., Behr, B., Pape, A. R., Wen, Y., Bielfeld, P., Polan, M. L. ENDOCRINE SOC. 1998: 953–59

  Abstract

  Gaining knowledge about the physiological timetable of gene expression during preimplantation embryo development is crucial, and a better understanding of cytokine and growth factor expression in early embryonic development could lead to improved in vitro culture conditions and enhance in vitro fertilization implantation rates. Our aim was to detect the patterns and levels of two messenger ribonucleic acids [mRNAs; beta-actin and interleukin-1 receptor type I (IL-1R tI)] in single human blastomeres by RT-nested PCR and to compare possible variations in the gene expression both between different embryos and in multiple blastomeres within the same embryo. Single blastomeres from nine human tripronucleic preimplantation embryos were examined by one round of RT and two rounds of nested competitive PCR. Beta-actin mRNA was detected in each blastomere, and IL-1R tI mRNA was found in 72% of the blastomeres examined. Beta-actin was expressed at a level of 511-12185 molecules of complementary DNA/blastomere, and IL-1R tI was expressed at a level of 2-290 molecules of complementary DNA/blastomere. Our results suggest that the mRNA pattern of an embryo cannot be reliably quantitated from the mRNA pattern of a single blastomere and therefore imply limitations for the use of this method for preimplantation diagnosis.

  View details for Web of Science ID 000072403500044

  View details for PubMedID 9506755

• Single blastomeres obtained from human preimplantation embryos express different amounts of mRNA for various genes: implications for future applications of this method for preimplantation diagnosis Kruessel, J. S., Huang, H. Y., Simon, C., Behr, B., Kloodt, A. R., Wen, Y., Bielfeld, P., Polan, M. L. OXFORD UNIV PRESS. 1997: O218–O218

  View details for Web of Science ID A1997YD97600211

• Quantification of hexokinase mRNA in mouse blastocysts by competitive reverse transcriptase polymerase chain reaction 44th Annual Meeting of the Pacific-Coast-Fertility-Society Johnson, M. D., BATEY, D. W., Behr, B. OXFORD UNIV PRESS. 1997: 359–65

  Abstract

  Hexokinase (HX), the enzyme that catalyses the initial reaction in glycolysis, is an important enzyme in glucose metabolism during human and mouse embryonic development. In our previous investigations of the genetic activities of HX, we observed an increased incidence of HX gene expression in blastocysts in comparison with morulae, and variability in the incidence of HX gene expression between embryos at the same developmental stages. These observations prompted us to quantify HX mRNA in mouse blastocysts to define the biological significance of the variable gene transcription. We modified our qualitative reverse transcription-nested polymerase chain reaction (RT-nPCR) assay for HX mRNA in single or groups of embryos to quantify HX mRNA by competitive RT-nPCR. HX mRNA was quantified in cohorts of mouse blastocysts cultured in glucose/phosphate-containing human tubal fluid (HTF) media. These blastocysts expressed HX in minute amounts, averaging 1.95 x 10(-18) g of mRNA. This is the first attempt at quantification of single gene mRNA in preimplantation embryos. Further investigations using similar techniques will enable comparative analyses between embryos to be performed to determine the correlation between specific levels of HX mRNA transcripts in individual embryos and embryonic viability and competence for further development and implantation.

  View details for Web of Science ID A1997XT38600011

  View details for PubMedID 9237264

• Genetic expression of hexokinase and glucose phosphate isomerase in late-stage mouse preimplantation embryos: Transcription activities in glucose/phosphate-containing HTF and glucose/phosphate-free P1 media 42nd Annual Meeting of the Society-for-Gynecologic-Investigation Johnson, M. D., BATEY, D. W., Barro, J. OXFORD UNIV PRESS. 1997: 351–57

  Abstract

  In mouse and human preimplantation development, pyruvate is consumed preferentially during early embryogenesis; however, during the morula and blastocyst stages, glucose is the preferred energy substrate. Studies have suggested that the glycolytic enzymes, hexokinase and glucose phosphate isomerase, are important enzymes in glucose metabolism during these later stages of human and mouse preimplantation development. In order to investigate the genetic activities of these enzymes in late-stage mouse embryos developing in vitro, we analysed hexokinase and glucose phosphate isomerase transcription activities by qualitative RNA assays using reverse transcriptase-nested polymerase chain reaction amplification of individual mouse morulae and early blastocysts incubated in glucose/phosphate-free preimplantation stage one (P1) medium and glucose/phosphate-containing human tubal fluid (HTF) medium. We observed an increased incidence of hexokinase transcripts in the population of blastocysts compared with morulae, and differences in transcript incidence between early blastocysts developing in HTF medium and in P1 medium. In contrast, glucose phosphate isomerase transcripts were consistantly present in all embryos analysed, and appear to be constitutively expressed during late-stage mouse embryogenesis. The different activity patterns of the two glycolytic genes may reflect different mechanisms of gene regulation or differential transcript stability during the later stages of mouse preimplantation development.

  View details for Web of Science ID A1997XT38600010

  View details for PubMedID 9237263

• Blastocyst culture without co-culture: role of embryo metabolism 10th World Congress of In Vitro Fertilization and Assisted Reproduction Behr, B. MONDUZZI EDITORE. 1997: 145–146

  View details for Web of Science ID A1997BJ47A00026

• FACTORS RELATED TO EARLY-PREGNANCY RECOGNITION IN THE WOMAN JOURNAL OF ASSISTED REPRODUCTION AND GENETICS Behr, B., Stone, B., FOOTE, W. D. 1995; 12 (4): 278-282

  Abstract

  While the precise timing of the maternal recognition of pregnancy is not known, it is known that the prevention of return to ovarian cyclicity relies on a conceptus-derived signal.In an attempt to identify the first luteotropic signals detectable in the maternal circulation, a sensitive Leydig cell luteotropin bioassay was employed, and data were compared for nine clinically pregnant and nine nonpregnant patients in an in vitro fertilization program. Blood samples were drawn every other day for 10 days after embryo transfer (ET).The first detectable rise in bioactive luteotropin levels occurred between 6 and 8 days post ET. Serum E2 levels increased on the same days. Differences in luteotropin levels between pregnant and nonpregnant patients were significant between days 6 and 8 (P < 0.0001) and between days 8 and 10 (P < 0.002).Based on morphological studies reported by others, bioactive luteotropic signals identified in this study were detectable in the maternal circulation at about the time of trophoblast lacunae coalescing with maternal uterine blood vessels.

  View details for Web of Science ID A1995RK91900013

  View details for PubMedID 7580026

• The effect of a two-hour, room temperature incubation of human spermatozoa in Test-Yolk Buffer on the rate of fertilization in vitro. 51st Annual Pacific Coast Reproductive Society Conference Behr, B., Lyon, J., Watt, J., Dasig, D., Gebhardt, J., Jensen, J. 2003
• Non-invasive embryo viability assessments: The influence of media constituents during the first culture interval. 58th Annual American Society for Reproductive Medicine Conference Dasig , D., Gebhardt , J., Lyon, J., Milki , A., Watt, J., Behr, B. 2002
• Does the orientation of the first polar body during ICSI affect fertilization and embryo development rates? 52nd Annual Pacific Coast Reproductive Society Conference Tan , T., Tran, C., Ivankhenko, V., Behr , B. 2004
• Incidence of monozygotic twinning with blastocyst transfer compared to cleavage stage transfer. 58th Annual American Society for Reproductive Medicine Conference Milki, A., Jun, S., Hinckley, M., Behr, B., Giudice, L., Westphal, L. 2002
• Effects of osteopontin on mouse embryo development in vitro. 58th Annual American Society for Reproductive Medicine Conference Behr , B., Shlomo, I., Dasig, D., Lyon, J., Polan, M. 2002
• The gap junction gene Connexin 37 is upregulated by low level GM-CSF in mouse preimplantation embryos. 58th Annual American Society for Reproductive Medicine Conference Behr, B., Dasig, D., Gebhardt , J., Wang, H., Yen, W., Polan, M. 2002
• Granulocyte-macrophage colony-stimulating: a regulator in preimplantation embryo development and apoptosis? (Prize Paper Award) 18th Annual European Society of Human Reproduction & Embryology Conference Wang , H., Dasig, D., Gebhardt, J., Polan , M., Behr, B. 2002
• A comparison of aneuploidy rates between Asian and Caucasian patients. 30th Annual European Society of Human Reproduction and Embryology Meeting Behr, B., Smotrich, D., Gaona, M., Hamic , A., Wang, X., Kort, J. 2014
• Ovarian hyperstimulation syndrome (ohss) does not compromise embryonic development and pregnancy outcome in a subsequent frozen blastocyst transfer cycle. Annual American Association of Bioanalysts Conference Shu, Y., Behr, B. 2009
• Effect of biopsy day on donor oocyte 24 chromosome preimplantation genetic screening (PGS) pregnancy rates with fresh embryo transfers (ET). 68th Annual American Society of Reproductive Medicine Conference Behr, B., Botes, A., Kettler , C., Gaona, M., Wang, S., Smotrich, D. 2012
• Granulocyte-macrophage colony-stimulating factor enhances human embryo development to the blastocyst stage: a randomized study. 51st Annual Pacific Coast Reproductive Society Conference Shapiro, B., Richter, K., Daneshmand , S., Quinn, P., Behr, B. 2003
• Preimplantation genetic diagnosis after density gradient separation of the X-and –Y bearing human spermatozoa. 51st Annual Pacific Coast Reproductive Society Conference Boostanfar, R., Ivakhnenko, V., Potter, D., Feinman, M., Behr, B. 2003
• Empty zona pellucida as a container for preservation of blastomere in preimplantation genetic diagnosis. 51st Annual Pacific Coast Reproductive Society Conference Behr, B., Ivakhnenko, V., Tran , C., Tan, T. 2003
• Treatment with granulocyte-macrophage colony-stimulating factor promotes in vivo follicular development of immature rat. 51st Annual Pacific Coast Reproductive Society Conference Wang, H., Mooney , S., Wen, Y., Klein, C., Polan, M., Behr, B. 2003
• Pilot clinical study to predict IVF outcomes using embryo mechanical parameters. 72nd Annual American Society for Reproductive Medicine Conference Yanez, L., Sedan, O., Baker, V., Behr, B., Camarillo, D. 2016
• Does euploid embryo ranking by trophectoderm cell mitochondrial DNA content correspond with ranking by blastocyst morphology within an individual patient’s cohort of blastocysts? 72nd Annual American Society for Reproductive Medicine Conference Kort, J., Lathi, R., Behr, B. 2016
• Non-contact, label-free 3D imaging of developing embryos to enhance viability prediction. 69th Annual American Society of Reproductive Medicine Conference Zarnescu, L., Sudkamp, H., Behr, B., Baer, T., Ellerbee, A. 2013
• Leveling the playing field – embryo transfers after trophectoderm biopsy have similar outcomes in fresh and frozen cycles. 69th Annual American Society of Reproductive Medicine Conference Gustin, S., Zhao, Q., Behr, B., Lathi, R. 2013
• To Biopsy or Not to Biopsy: Non-Blastocysts on Day 5. 69th Annual American Society of Reproductive Medicine Conference Kort, J., Zhao, Q., Lathi, R., Baker, V., Behr, B. 2013
• Human Embryo Donation for Research: RENEW Biobank Experience. 61st Annual Pacific Coast Reproductive Society Conference Trivedi , D., Cromer, R., Dasig, J., Suarez, M., Behr, B. 2013
• Development and validation of an automated computer vision algorithm for real-time embryo viability prediction. 68th Annual American Society of Reproductive Medicine Conference Loewke , K., Moussavi, F., Maddah, M., Ivani, K., Behr, B., Suraj, V. 2012
• High implantation rates with blastocyst biopsy, array comparative genome hybridization (aCGH) and day-6 (D6) replacement. 68th Annual American Society of Reproductive Medicine Conference Behr, B., Tormasi , S., Anderson, S., Glassner, M., Welch, C., Smotrich, D. 2012
• The rate of euploid embryos in egg donors compared to infertile patients. 68th Annual American Society of Reproductive Medicine Conference Botes, A., Smotrich, D., Sherri, W., Kettler, C., MacAdam, R., Behr, B. 2012
• The interchangeability of HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) and MOPS (3-(N-morpholino)propanesulfonic acid) based solutions for vitrification and subsequent warming. 68th Annual American Society of Reproductive Medicine Conference Abeyta, M., Zhao, Q., Gebhardt , J., Suarez, M., Reddy, V., Behr, B. 2012
• A magnetic levitation platform for the isolation of mature sperm from TESE/TESA samples. 66th Annual Pacific Coast Reproductive Society Conference Durmus, G., Gupta, R., Badamjav, O., Reddy, V., Eisenberg, M., Behr, B., Demirci, U. 2018
• Do embryo time-lapse parameters predict euploid embryo transfer outcomes? 73rd Annual American Society for Reproductive Medicine Conference Drejza, M., Kort, J., Behr, B. 2017
• Oocyte telomerase levels correlate with blastocyst development. 73rd Annual American Society for Reproductive Medicine Conference Kort, J., Garbuzov, A., Arand, A., Behr, B., Artand, S. 2017
• The effect of embryo biopsy on perinatal outcomes: An analysis of SART CORS. 73rd Annual American Society for Reproductive Medicine Conference Kort, J., Behr, B., Wantman, E., Baker, V. 2017
• Preliminary study to investigate telomerase reverse transcriptase expression among human cumulus cells. 66th Annual Pacific Coast Reproductive Society Conference Kort, J., Garbuzov, A., Artandi , S., Behr, B. 2017
• Concurrent genome and transcriptome analysis from a single trophectoderm biopsy. (Prize Paper Award.) 66th Annual Pacific Coast Reproductive Society Conference Chiang, H., Kort, J., Behr, B., Snyder, M. 2017
• Culturing cryopreserved cleavage stage embryos to blastocysts is a cost-effective method of utilizing frozen embryos. 60th Annual Meeting of the American Society for Reproductive Medicine Feinman, M., Boostanfar, R., Le, A., Potter, D., Khoury, C., Behr, B. 2004
• Accuracy of morphologic assessment of blastocysts in selecting genetically normal embryos. 60th Annual American Society for Reproductive Medicine Conference Dasig, D., Behr , B., Lyon, J., Gebhardt , J., Watt, J., Milki, A. 2004
• Significance of immature oocytes in ICSI treatment cycles. 60th Annual American Society for Reproductive Medicine Conference Behr, B., Gebhardt, J., Watt , J., Lyon, J., Dasig, D., Shu, Y. 2004
• Igf2 gene imprinting in preimplantation mouse embryos. (Prize exchange paper) 20th Annual ESHRE/ASRM Conference Littman , E., Ulaner, G., Vu, T., Otero, J., Dasig, D., Lyon, J., Gebhardt, J., Giudice , L., Behr, B., Hoffman, A. 2004
• Advantages of the use of the non-contact laser in the preimplantation genetic diagnosis. 52nd Annual Pacific Coast Reproductive Society Conference Ivankhenko, V., Tran , C., Tan, T., Behr, B. 2004
• Matrix metalloproteinase and tissue inhibitor of matrix metalloproteinase expression in human preimplantation embryos. 59th Annual American Society for Reproductive Medicine Conference Wang , H., Wen, Y., Mooney, S., Li, H., Behr, B., Polan, M. 2003
• Does blastocyst transfer alter the sex ratio? One program’s experience and review of the literature. 58th Annual American Society for Reproductive Medicine Conference Milki , A., Jun, S., Hinckley, M., Westphal, L., Behr, B., Giudice, L. 2002
• Regulation of cyclooxygenase activity in human endometrial stromal cells by granulocyte-macrophage colony-stimulating factor in vitro. 58th Annual American Society for Reproductive Medicine Conference Wang, H., Wen, Y., Chen , B., Mooney, S., Behr, B., Polan, M. 2002
• Behavior of air-dried and rehydrated human spermatozoa injected into cytoplasts. 58th Annual American Society for Reproductive Medicine Conference Ivakhnenko , V., Amet, T., Behr , B. 2002
• Embryo cryopreservation: A two-year experience with three programmable cryopreservation units. 58th Annual American Society for Reproductive Medicine Conference Behr, B., Tan, T., Khoury, C., Anh, L., Ivakhnenko, V., Tran , C. 2002
• Granulocyte-macrophage colony-stimulating: A regulator in preimplantation embryo development and apoptosis? (Prize Exchange Paper) 50th Annual Pacific Coast Reproductive Society Conference Wang, H., Dasig, D., Gebhardt , J., Polan, M., Behr, B. 2002
• ICSI does not compromise blastocyst development. 50th Annual Pacific Coast Reproductive Society Conference Westphal , L., Hinckley, M., Behr, B., Milki, A. 2002
• Culture environment and its effects on ATP levels in preimplantation mouse blastocysts. 57th Annual American Society for Reproductive Medicine Conference Dasig , D., Fisch , J., Behr, B. 2001
• CD146 expression in human preimplantation blastocyst. 57th Annual American Society for Reproductive Medicine Conference Wang, H., Wen, Y., Mooney, S., Behr , B., Polan , M. 2001
• The Use of an Innovative Culture System to Maintain a Variety of Atmospheric Conditions. 57th Annual American Society for Reproductive Medicine Conference Behr, B., Dasig , D., Gebhardt , J. 2001
• Accuracy of day three criteria for selecting the best embryos. 57th Annual American Society for Reproductive Medicine Conference Milki , A., Hinckley, M., Gebhardt, J., Dasig , D., Westphal, L., Behr, B. 2001
• Morphological dynamic of the nucleoli in human pronuclear oocytes. 64th Annual American Society for Reproductive Medicine Conference Ivankhnenko , V., Kolb, B., Nelson, J., Tourgeman, D., Wilcox, J., Behr, B. 2008
• Importance of integrity of blastomere in the re-expansion of cryopreserved blastocysts. 63rd Annual American Society for Reproductive Medicine Conference Shu, Y., Watt , J., Gebhardt , J., Zhao, Q., Behr, B. 2007
• Early pregnancy testosterone levels in miscarriages compared to ongoing pregnancies. 54th Annual Pacific Coast Reproductive Society Conference Lathi, R., Reddy, C., Dahan, M., Milki, A., Gebhardt, J., Behr, B., Westphal, L. 2006
• Extended post-thaw culture is an efficient way to reduce the number of frozen embryos in storage. 54th Annual Pacific Coast Reproductive Society Conference Khoury, C., Frederick, J., Potter, D., Behr, B. 2006
• A short freezing program with propanediol: preclinical testing with mouse embryos. 54th Annual Pacific Coast Reproductive Society Conference Lyon, J., Shu, Y., Behr, B. 2006
• Comparison of transvaginal ultrasound guided and traditional blind embryo transfer on pregnancy rates outcome. 54th Annual Pacific Coast Reproductive Society Conference Behr, B., Boostanfar, R., Wang, H., Feinman, M. 2006
• Microsort and PGD: Separating the boys from the girls. 54th Annual Pacific Coast Reproductive Society Conference Ivankhenko, V., Anh, L., Feinman, M., Behr, B., Potter, D., Boostanfar, R. 2006
• Does number of oocytes matter in PGD cycles? 54th Annual Pacific Coast Reproductive Society Conference Tran, C., Tan, T., Ivakhnenko, V., Behr, B. 2006
• Outcomes after 2 chromosomes probe panel vs. multiple probe panels for PGD with FISH. 54th Annual Pacific Coast Reproductive Society Conference Tran, C., Ivakhnenko, V., Tan, T., Behr, B. 2006
• Preliminary study of the effect of TEST-Yolk buffer on human spermatozoa prior to ICSI. 54th Annual Pacific Coast Reproductive Society Conference Gebhardt , J., Jensen, J., Watt, J., Lyon, J., Shu, Y., Behr, B. 2006
• Effects of oocyte cryopreservation on a specific oolemma bound protein, CD9: a potential functional marker after cryopreservation. 54th Annual Pacific Coast Reproductive Society Conference Wen, Y., Chen, B., Polan, M., Behr, B. 2006
• Pregnancy loss and aneuploidy rate in fresh versus frozen blastocyst transfer. 61st Annual American Society for Reproductive Medicine Conference Moayeri, S., Burney, R., Milki, A., Westphal, L., Behr, B., Lathi, R. 2005
• Effects of GMCSF on development and gene imprinting in preimplantation mouse embryos. 53rd Annual Pacific Coast Reproductive Society Conference Littman, E., Ling, J., Ulaner, G., Vu, T., Dasig, D., Lyon, J., Giudice, L., Hoffman, A., Behr, B. 2005
• Comparison between two different human oocyte cryopreservation protocols on survival, fertilization and embryo development. 53rd Annual Pacific Coast Reproductive Society Conference Behr, B., Le, A., Khoury, C., Boostanfar, B., Feinman, M., Frederick, J. 2005
• Effects of reduced oxygen concentrations on IVF outcomes. (Prize Poster Award) 53rd Annual Pacific Coast Reproductive Society Conference Kea, B., Gebhardt , J., Westphal, L., Lathi, R., Watt , J., Behr, B. 2005
• Pregnancies and live births after transfer of cryopreserved hatching or hatched blastocysts. 53rd Annual Pacific Coast Reproductive Society Conference Shu, Y., Gebhardt , J., Watt, J., Lyon, J., Jensen, J., Behr , B. 2005
• Does euploid embryo ranking by trophectoderm cell mitochondrial DNA content correspond with ranking by blastocyst morphology within an individual patient’s cohort of blastocysts? 72nd Annual American Society for Reproductive Medicine Conference Kort, J., Lathi, R., Behr, B. 2016
• Microfluidic sperm sorting device for selection of functional human sperm for IUI application. 64th Annual Pacific Coast Reproductive Society Conference Chinnasamy, T., Behr, B., Demirci, U. 2016
• The relationship between a man’s somatic health and ART outcomes. 71st Annual American Society for Reproductive Medicine Conference Eisenberg, M., Li, S., Behr, B., Nakajima, S., Baker, V. 2015
• Blastocyst Implantation is Correlated with Outputs from Automated Time-lapse Analysis by the EEVA Test. 71st Annual American Society for Reproductive Medicine Conference Liebermann , J., Bartolucci, A., Troup , S., Wagner Coughlin, C., Yee, B., Behr, B. 2015
• Computer-automated Time-lapse Test Results are Predictive of Pregnancy Following Blastocyst Transfer. 71st Annual American Society for Reproductive Medicine Conference Behr, B., Beltsos, A., Yee, B., Kingsland , C., Benadiva, C., Liebermann, J. 2015
• Non-invasive Technology Combining Time-lapse Imaging and Statistical Modeling: Bringing Automation into the Lab to Improve Blastocyst Selection. 71st Annual American Society for Reproductive Medicine Conference Behr, B., Tan, L., Conaghan , J., Liebermann, J., Bartolucci, A., Chen, A. 2015
• Shared Oocyte Donation Program Produces High Clinical Pregnancy Rate-A Three Year Follow Up Study. 63rd Annual Pacific Coast Reproductive Society Conference Khoury, C., Frederick, J., Coffler, M., Sills, E., Behr, B., Potter, D. 2015
• Is there an Association between Gonadotropin Dosing and Aneuploidy Rates? 70th Annual American Society for Reproductive Medicine Conference Kort, J., Zolton, J., Behr, B., Lathi, R., Baker, V. 2014
• Mechanical Biomarkers of Oocyte Maturation. 70th Annual American Society for Reproductive Medicine Conference Zarnescu, L., Han, J., Behr, B., Reijo Pera, R., Camarillo, D. 2014
• Assessing the Effects of Vitrification on Embryo Viability using Label-Free Imaging. 70th Annual American Society for Reproductive Medicine Conference Zarnescu, L., Abeyta, M., Baer, T., Behr, B., Ellerbee, A. 2014
• Paternal Age and Total Motile Sperm Parameters Are Not Correlated with Embryo Aneuploidy Rates. 62nd Annual Pacific Coast Reproductive Society Conference Kort, J., Zhao, Q., Behr, B. 2014
• Increased Body Mass Index Negatively Impacts Blastocyst Formation Rate in Patients Undergoing In Vitro Fertilization. 62nd Annual Pacific Coast Reproductive Society Conference Comstock, I., Behr, B., Lathi, R. 2014
• Day 5 “Early Blastocysts”: Worthy of a Biopsy in IVF Cycles with PGS? 62nd Annual Pacific Coast Reproductive Society Conference Kort, J., Lathi, R., Baker, V., Zhao, Q., Behr, B. 2014
• Are Anti-Müllerian Hormone Levels Predictive of Aneuploidy Rates? 62nd Annual Pacific Coast Reproductive Society Conference Kort, J., Lathi, R., Zhao, Q., Behr, B., Baker, V. 2014
• Office administration of single does GnRH antagonist: minimizing patient injections utilizing a simple and highly effective IVF protocol. 61st Annual American Society for Reproductive Medicine Conference Boostanfar , R., Feinman, M., Le, A., Wang, H., Behr, B. 2005
• Outcome parameters of day 5 versus day 6 frozen blastocysts. 64th Annual American Society for Reproductive Medicine Conference Zhao, Q., Gebhardt , J., Shu, Y., Watt, J., Dasig, J., Behr, B. 2008
• Non-invasive imaging for the detection of human embryonic aneuploidy at the blastocyst stage. 68th Annual American Society of Reproductive Medicine Conference Friedman , B., Chavez, S., Behr, B., Lathi, R., Baker, V., Reijo Pera, R. 2012
• Cumulative Pregnancy Rates Following Fresh And Frozen-Thawed Embryo Transfer Cycles Using A Recombinant Follicle-Stimulating Hormone (rFSH)/Gonadotropin-Releasing Hormone (GnRH) Antagonist Protocol. Pacific Coast Reproductive Society Annual Meeting 60th Anniversary Celebration Doody, K., Behr, B., Grob, P., IJzerman-Boon, P., Gordon, K. 2012
• Oocyte Defects Leading to Triploid Miscarriages After Donor Oocyte IVF. Pacific Coast Reproductive Society Annual Meeting 60th Anniversary Celebration Marshall, F., Behr, B., Lathi, R. 2012
• Spontaneous Abortion Rates Following Frozen Embryo Transfer: Preliminary Experience Comparing Slow and Vitrified Cryopreservation Methods. Pacific Coast Reproductive Society Annual Meeting 60th Anniversary Celebration Dasig, J., Zhao, J., Reddy, V., Gebhardt , J., Suarez, M., Behr, B. 2012
• Autovaccine Targeting for Endometriosis By Inducing CTL: a Pilot Study. Pacific Coast Reproductive Society Annual Meeting 60th Anniversary Celebration Wang, H., Feng, D., Behr, B. 2012
• Is Oocyte Maturation Rate A Predictor of IVF Outcome? Pacific Coast Reproductive Society Annual Meeting 60th Anniversary Celebration Behr, B., Gebhardt, J., Dasig, J., Reddy, V., Suarez , M., Baker, V. 2012
• Impact of FISH and 24 Chromosome PGS on Donor Oocyte Pregnancy Rate. 67th Annual American Society of Reproductive Medicine Conference Behr, B., Ross, R., Batzofin, D., Wang, S., Smotrich, D. 2011
• Non-Invasive Assessment of Embryo Viability Using Novel Automated Imaging Technology. (First Prize Winner) 67th Annual American Society of Reproductive Medicine Conference Behr, B., Chavez, S., Loewke, K., Reijo Pera, R. 2011
• Cumulative Pregnancy Rates Following Fresh and Frozen-Thawed Embryo Transfer Cycles Using a Recombinant Follicle-Stimulating Hormone (rFSH)/Gonadotropin-Releasing Hormone (GnRH) Antagonist Protocol. 59th Annual Pacific Coast Reproductive Society Conference Doody, K., Behr, B., Grob, P., IJzerman-Boon, P., Gordon, K. 2011
• The effects of mosaicism on all-chromosome PGS/D. 25th Annual European Society of Human Reproduction and Embryology Conference Rabinowitz, M., Johnson, D., Cinnioglu, C., Gemelos, G., Banjevie, M., Alper, M., Ross, R., Smotrich, D., Potter, D., Behr, B. 2009
• Corifollitropin alfa versus recombinant FSH treatment for COS in a GnRH antagonist protocol: equal efficacy in terms of oocyte and embryo quality. 25th Annual European Society of Human Reproduction and Embryology Conference Behr, B., Heijnen, E. 2009
• A statistical method for ranking embryos by likelihood of containing normal cells. 57th Annual Pacific Coast Reproductive Society Conference Johnson, D., Shah, N., Potter, D., Behr, B., Rabinowitz, M. 2009
• Pregnancy after trophectoderm biopsy of frozen-thawed blastocyst. 56th Annual Pacific Coast Reproductive Society Conference Lathi, R., Behr , B. 2008
• Novel technology for simultaneous reliable measurement of multiple alleles and copy number across 24 chromosomes in single human blastomeres. 56th Annual Pacific Coast Reproductive Society Conference Johnson, D., Rabinowitz, M., Cinnioglu , C., Keller, J., Banjevic, M., Singer, J., Baner, J., Sheena, J., Behr, B. 2008
• Efficacy of preimplantation genetic screening on PGS cycles versus cancelled PGS cycles. 57th Annual Pacific Coast Reproductive Society Conference Mekonnen, Z., Lathi, R., Baker, V., Behr, B. 2009
• Microsort® improves per cycle pregnancy rates in patients undergoing in vitro fertilization/pre-implantation genetic diagnosis for gender selection. 64th Annual American Society for Reproductive Medicine Conference Potter, D., Khoury, C., Frederic, J., Boostanfar, R., Tourgeman , D., Behr, B. 2008
• Elective single-embryo transfer in frozen blastocyst transfer cycles. 64th Annual American Society for Reproductive Medicine Conference Shu, Y., Watt, J., Gebhardt, J., Zhao, Q., Milki, A., Behr, B. 2008
• A comparison of starting temperatures for slow freezing cryopreservation of blastocysts. 64th Annual American Society for Reproductive Medicine Conference Gebhardt, J., Zhao, Q., Watt, J., Shu, Y., Dasig, J., Behr, B. 2008
• Reliable concurrent calling of multiple genetic alleles and 24-chromosome ploidy without embryo freezing using parental support technology (PS). 64th Annual American Society for Reproductive Medicine Conference Rabinowitz, M., Johnson, D., Salzman, J., Banjevic, M., Cinnioglu, C., Behr, B. 2008
• Allocation of custom-made electrofusion pipettes in mouse somataic cell nuclear transfer. 56th Annual Pacific Coast Reproductive Society Conference Shu, Y., Rodriguez, R., Kim, S., Behr, B. 2008
• Stability of human zonae pellucidae during extended culture in vitro. 56th Annual Pacific Coast Reproductive Society Conference Ivankhnenko , V., Tan, T., Kolb, B., Tourgeman, D., Wilcox, J., Behr, B. 2008
• Comparison between day 3 transfer and day 5 transfer over a period of 6 years. 56th Annual Pacific Coast Reproductive Society Conference Khoury, C., Behr, B., Potter, D., Frederick , J. 2008
• Comparison of blastocyst transfer with and without preimplantation genetic screening in both IVF patients and oocyte donors. 56th Annual Pacific Coast Reproductive Society Conference Khoury, C., Frederick, J., Potter, D., Behr, B. 2008
• Reduced oocyte damage rates after ICSI with blunted needle. 56th Annual Pacific Coast Reproductive Society Conference Ivakhnenko, V., Tran, C., Kolb, B., Nelson, J., Wilcox, J., Behr, B. 2008
• A comparison of blastocyst formation from one-cell mouse zygotes following an aseptic vitrification system. 56th Annual Pacific Coast Reproductive Society Conference Dasig, J., Bertocci, F., Zhao, Q., Shu, Y., Behr, B. 2008
• Live birth of a normal healthy baby following replacement of refrozen blastocysts: previously thawed, hatched on day three and cultured to the blastocyst stage and then refrozen. 63rd Annual American Society for Reproductive Medicine Conference Khoury, C., Frederick, J., Potter, D., Behr, B. 2007
• Effects of blastomere loss on developmental competency of frozen thawed human blastocysts. 63rd Annual American Society for Reproductive Medicine Conference Shu, Y., Watt, J., Gebhardt, J., Dasig, J., Jensen, J., Behr, B. 2007
• Osmotic behavior of human blastocysts as a potential predictor for survival. 61st Annual American Society for Reproductive Medicine Conference Shu , Y., Gebhardt, J., Watt, J., Lyon, J., Jensen, J., Behr, B. 2005
• Post-thaw embryo compaction is predictive of the success of frozen-thawed transfer. 61st Annual American Society for Reproductive Medicine Conference Wang, H., Le, A., Boostanfar, R., Feinman, M., Behr, B. 2005
• Correlation between normal sperm head morphology (NSHM) and sperm binding potential to the sperm hyaluronan-binding assay (HBA(TM)). 60th Annual American Society for Reproductive Medicine Conference Worrilow, K., Lyon , J., Belcak, N., Peters, A., Johnston, J., Behr, B. 2004
• Freezing at lower starting temperature (-6°C) improves human embryo viability and pregnancy rate. 60th Annual American Society for Reproductive Medicine Conference Tran , C., Le, A., Tan, T., Tran, A., Ivakhnenko , V., Behr, B. 2004
• Single point determination of day 2 embryos yield comparable implantation rate when compared to sequential assessment. 60th Annual American Society for Reproductive Medicine Conference Tan, T., Tran, C., Ivakhnenko, V., Behr, B. 2004
• Maturation, embryo development and clinical outcomes in ICSI treatment cycles with completely immature oocytes at the removal of cumulus cells. 60th Annual American Society for Reproductive Medicine Conference Shu, Y., Gebhardt, J., Watt, J., Lyon, J., Dasig, D., Behr, B. 2004
• Elective single blastocyst transfer: cumulative outcomes from the fresh and subsequent frozen cycles. 60th Annual American Society for Reproductive Medicine Conference Milki, A., Hinckley, H., Lathi, R., Westphal, L., Behr, B. 2004
• Effects of osteopontin on apoptosis and DNA repair during mouse embryo development in vitro. 60th Annual American Society for Reproductive Medicine Conference Lyon, J., Wang, H., Dasig, D., Behr, B. 2004
• Standardization of transvaginal ultrasound guided embryo transfer post fellowship: a case control study. 52nd Annual Pacific Coast Reproductive Society Conference Boostanfar, R., Feinman , M., Gao , C., Le, A., Behr, B. 2004
• Selection of day 2 embryos with mononucleated blastomeres as a simple predictor of implantation potential and pregnancy outcome. 59th Annual American Society for Reproductive Medicine Conference Tan , T., Tran, C., Ivakhnenko, V., Behr, B. 2003
• Case report: visualization of atypical hatching of a human blastocyst in vitro forming a monozygotic twin. 59th Annual American Society for Reproductive Medicine Conference Behr, B., Wang, H., Milki, A. 2003
• Elective single blastocyst transfer. 59th Annual American Society for Reproductive Medicine Conference Milki, A., Littman, E., Hinckley, H., Westphal , L., Gebhardt, J., Behr, B. 2003
• Blastocyst cryopreservation: an update. 59th Annual American Society for Reproductive Medicine Conference Behr, B., Gebhardt, J., Milki, A. 2003
• Optimal day for embryo transfer? Reply of the authors. 59th Annual American Society for Reproductive Medicine Conference Milki , A., Hinckley, M., Behr, B. 2003
• Comparison of laser versus acid Tyrode’s assisted hatching on pregnancy rates in frozen embryo transfers. 51st Annual Pacific Coast Reproductive Society Conference Tran, C., Tan, T., Ivankhenko, V., Behr, B. 2003
• Relationship between cleavage rate and male-to-female sex ratios in preimplantation genetic diagnosis cycles. 61st Annual American Society for Reproductive Medicine Conference Lyon, J., Ivakhnenko, V., Behr, B. 2005
• Preimplantation genetic diagnosis in embryos created from oocyte donation. 61st Annual American Society for Reproductive Medicine Conference Nelson, J., Potter, D., Wilcox, J., Frederick, J., Kolb, B., Behr, B. 2005
• Does the incidence of monozygotic twinning with blastocyst transfer decrease over time? 61st Annual American Society for Reproductive Medicine Conference Moayeri , S., Behr, B., Lathi, R., Westphal, L., Giudice , L., Milki, A. 2005
• Effects of repetitive vitrification on the survival of mouse oocytes. 61st Annual American Society for Reproductive Medicine Conference Migishima, F., Shu, Y., Chen, B., Zhao, Y., Polan, M., Behr, B. 2005
• Regulation of CYP17 expression in theca-interstitail cells by GM-CSF. 53rd Annual Pacific Coast Reproductive Society Conference Wang, H., Le, A., Boostanfar, R., Feinman, M., Behr, B. 2005
• Extended post-thaw culture improves frozen embryo transfer success rates. 21st Annual European Society of Human Reproduction and Embryology Conference Potter, D., Khoury, C., Feinman, M., Frederick, J., Behr, B. 2005
• Preliminary experience comparing human recombinant hyaluronidase to bovine hyaluronidase in mouse in vitro fertilization. 53rd Annual Pacific Coast Reproductive Society Conference Behr , B., Lyon, J., Littman, E., Dasig, D. 2005
• Efficacy of recombinant human serum albumin as a protein source for the development of in vitro culture human embryos: a pilot study. 53rd Annual Pacific Coast Reproductive Society Conference Lyon, J., Quinn, P., Carter, D., Ye, P., Dasig, D., Behr, B. 2005
• The use of recombinant human serum albumin (rHSA) for mouse IVF and embryo culture: effect of diluent and protein stabilizers. 53rd Annual Pacific Coast Reproductive Society Conference Quinn, P., Carter, D., Ye, P., Behr, B. 2005
• Laser zona pellucida removal (LZPR): We now have a choice. 53rd Annual Pacific Coast Reproductive Society Conference Ivakhnenko , V., Tran, C., Tan, T., Behr, B. 2005
• A novel microfluidics device for male infertility screening. 21st Annual European Society of Human Reproduction and Embryology Conference McCormack, M., Aravanis, A., Pyle, J., Behr, B. 2005
• The clinical outcomes of cryopreserved pronuclear embryos from standard IVF and ICSI. 53rd Annual Pacific Coast Reproductive Society Conference Wang, H., Le, A., Boostanfar, R., Feinman, M., Behr, B. 2005
• The effect of cryopreservation on human sperm hyaluronan binding activity. American Urological Association Annual Meeting McCormack, M., Behr, B., McCalllum, S. 2005
• Testosterone Up-regulates androgen receptor and changes bladder mass and collagen to smooth muscle ratio. American Urological Association Annual Meeting Wang, B., Youxin, Y., Behr, B., Shortliffe, L. 2005