Targeted single molecule sequencing methodology for ovarian hyperstimulation syndrome.

Background: One of the most significant issues surrounding next generation sequencing is the cost and the difficulty assembling short read lengths. Targeted capture enrichment of longer fragments using single molecule sequencing (SMS) is expected to improve both sequence assembly and base-call accuracy but, at present, there are very few examples of successful application of these technologic advances in translational research and clinical testing. We developed a targeted single molecule sequencing (T-SMS) panel for genes implicated in ovarian response to controlled ovarian hyperstimulation (COH) for infertility.

Kinase insert domain receptor/vascular endothelial growth factor receptor 2 (KDR) genetic variation is associated with ovarian hyperstimulation syndrome.

Background: The objective of this investigation was to determine if kinase insert domain/vascular endothelial growth factor receptor 2 (KDR/VEGFR2) genetic variation was associated with the development of ovarian hyperstimulation syndrome (OHSS) in patients undergoing controlled ovarian hyperstimulation (COH).

Methods: This was a case-control study of 174 patients who underwent controlled ovarian stimulation. Patient blood samples were genotyped for single nucleotide polymorphisms (SNPs) spanning the KDR locus.

Association between the luteinizing hormone/chorionic gonadotropin receptor (LHCGR) rs4073366 polymorphism and ovarian hyperstimulation syndrome during controlled ovarian hyperstimulation.

Background: The aim of this study was to determine the relationship between a purported luteinizing hormone/chorionic gonadotropin (LHCGR) high function polymorphism (rs4539842/insLQ) and outcome to controlled ovarian hyperstimulation (COH).

Methods: This was a prospective study of 172 patients undergoing COH at the Fertility and IVF Center at GWU. DNA was isolated from blood samples and a region encompassing the insLQ polymorphism was sequenced.

Multinucleation of a sibling blastomere on day 2 suggests unsuitability for embryo transfer in IVF-preimplantation genetic screening cycles.

Objective: To evaluate the impact of multinucleation of a sibling blastomere of day 2 embryos on the rate of aneuploidy detected by day 3 preimplantation genetic screening (PGS) biopsy and the effect on subsequent implantation and pregnancy rates.

Design: Retrospective cohort study.

Setting: University-based IVF center.

Day 2 embryo transfer (ET) and day 3 ET afford similar reproductive outcomes in the poor responder.

A retrospective review of 237 initial, fresh nondonor IVF cycles in which all embryos generated during the cycle were transferred on either day 2 (n = 109) or day 3 (n = 128) were evaluated with regards to reproductive outcomes. Patients who underwent a day 2 ET had similar conception (18% vs. 16%; relative risk [RR], 1.

Profound teratospermia does not influence sex chromosomal aneuploidy rate in in vitro fertilization-preimplantation genetic diagnosis cycles.

Because spermatocyte meiotic error results in embryonic sex chromosomal aneuploidy, it is speculated that teratospermia correlates with increased embryonic sex chromosomal abnormalities. Our findings contradict this theory, suggesting that morphology does not correlate with sex chromosomal genotype.

A novel surgical treatment for cesarean scar pregnancy: laparoscopically assisted operative hysteroscopy.

Objective: To describe laparoscopically assisted hysteroscopy as a unique surgical intervention for a cesarean section scar ectopic pregnancy.

Design: Case report.

Setting: University hospital.

The use of lead follicle diameter to initiate gonadotropin-releasing hormone antagonist does not affect in vitro fertilization clinical pregnancy, implantation, or live birth rates: a prospective, randomized study.

In this prospective, randomized study, waiting for the lead follicle to reach 14 mm before initiating GnRH antagonist administration effectively prevents an LH surge and ovulation during an IVF cycle. Furthermore, delaying GnRH start until the dominant follicle reaches 14 mm neither impacts the clinical pregnancy, implantation, or live birth rates nor increases the incidence of severe ovarian hyperstimulation syndrome.

Does ethnicity influence in vitro fertilization (IVF) birth outcomes?

Objective: To determine if ethnicity influences IVF birth outcome.

Design: Retrospective cohort study.

Setting: University-based IVF program.

Effects of triploidy after intracytoplasmic sperm injection on in vitro fertilization cycle outcome.

Objective: To evaluate the impact on the rates of clinical pregnancy and live birth of polyploidy after intracytoplasmic sperm injection (ICSI).

Design: Retrospective cohort study.

Setting: University-based IVF center.

The influence of sperm morphology on preimplantation genetic diagnosis cycles outcome.

Objective: To evaluate the impact of abnormal sperm morphology on the rates of aneuploidy, implantation, and clinical pregnancy.

Design: Retrospective cohort study.

Setting: University-based IVF center.

Novel follicular-phase gonadotropin-releasing hormone antagonist stimulation protocol for in vitro fertilization in the poor responder.

Poor responders continue to be vexing in infertility therapy. By using GnRH antagonists before ovarian stimulation, we demonstrate an improvement in oocyte, embryo, and zygote yield in patients with a prior poor response.

Second cycle: to hatch or not to hatch?

The objective of this study was to assess the impact of assisted hatching (AH) on pregnancy rate (PR), clinical pregnancy rate (CPR), and implantation rate (IR) after a single failed, fresh, nondonor IVF cycle. Accordingly, we report that patients with one prior implantation failure benefit from AH with improved PR, CPR, and IR in a subsequent cycle.

Expression of the alphav integrin adhesion molecule during development of preimplantation human embryos.

Objective: To investigate the expression of alphav integrin in developing human embryos.

Design: Observational study.

Setting: University medical center.

Is blastocyst transfer useful as an alternative treatment for patients with multiple in vitro fertilization failures?

Objective: To determine whether blastocyst transfer is of benefit to patients with multiple IVF failures.

Design: Retrospective cohort study.

Setting: The George Washington University Medical Center.

Response to clomiphene citrate challenge test in normal women through perimenopause.

Objective: To identify the physiological changes that discriminate subgroups of women along the reproductive continuum.

Methods: This prospective study was carried out at the outpatient facility of George Washington University Medical Center and the outpatient clinic of the National Institutes of Health Clinical Center. Twenty-five female subjects were divided into 4 biologically distinct groups: group 1 (n = 4) regular menstrual cycles and under age 35; group 2 (n = 11) regular cycles and over age 35; group 3(n = 6) irregular cycles and over age 35, and group 4(n = 4) menopausal.

Increasing clinical pregnancy rates after IVF/ET. Can immunosuppression help?

Objective: To evaluate the safety and efficacy of immunosuppression as an adjunct to improving the success of in vitro fertilization/embryo transfer (IVF-ET).

Study Design: A randomized, double-blind, placebo-controlled clinical trial.

Results: Seventy-five patients were randomized to receive either prednisone (39 patients, 51%) or placebo (36 patients, 49%).

Basal and stimulated gonadotropin levels in the perimenopause.

Objective: We investigated whether perimenopausal menstrual cycle irregularity is associated with increased gonadotropin immunoactivity, bioactivity, or the bioactivity/immunoactivity ratio at baseline and after short-term stimulation with gonadotropin-releasing hormone.

Study Design: Subjects consisted of four groups: (1) young regular cycling women (< 35 years old), older women (> 35 years) with (2) regular or (3) irregular menstrual cycles, and (4) postmenopausal women. Gonadotropin-releasing hormone stimulation tests (100 micrograms intravenous gonadotropin-releasing hormone) were performed in the National Institute of Mental Health outpatient clinic during the follicular phase of the menstrual cycle or randomly in postmenopausal women.

Immunocytochemical localization of growth factors and their receptors in human pre-embryos and Fallopian tubes.

We utilized indirect immunocytochemistry to demonstrate the presence of growth factors and their receptors in human pre-embryos and Fallopian tubes. In pre-embryos, only transforming growth factor-alpha (TGF-alpha) and the intracellular domain of epidermal growth factor receptor (EGFR) were found at the 4-cell stage. In 8- to 14-cell pre-embryos, TGF-alpha, the intracellular and extracellular domains of EGFR, and insulin-like growth factor-I and its receptor were found.

Achieving multiple-order embryo transfer identifies women over 40 years of age with improved in vitro fertilization outcome.

Objective: To define factors in patients > or = 40 years that may improve outcome and provide prognosis for success in IVF-ET.

Design: Retrospective.

Setting: University infertility center.

Prognostic value of day 3 estradiol on in vitro fertilization outcome.

Objective: To evaluate the prognostic value of day 3 E2 levels, independent of day 3 FSH levels, on responses to ovulation induction and subsequent pregnancy rates (PRs) in IVF-ET patients.

Design: Prospective, observational.

Setting: University-based tertiary care and private reproductive endocrine-infertility units.

Lineage tracing demonstrates that blastomeres of early cleavage-stage human pre-embryos contribute to both trophectoderm and inner cell mass.

We injected a fluorescent lineage tracer (Texas Red-lysine-dextran) into individual blastomeres of donated human diploid 2- to 8-cell pre-embryos and cultured them to blastocysts. Once pre-embryos reached the expanded blastocyst stage, they were fixed and examined in a scanning confocal microscope to identify the location of fluorescent tracer. In successfully injected pre-embryos that developed to expanded blastocysts, we found that randomly injected blastomeres formed both trophectoderm (TE) and inner cell mass (ICM).

Utility of in vitro fertilization at diagnostic laparoscopy.

Objective: To compare stimulation and outcome variables for IVF in stimulated cycles when ova are retrieved during diagnostic infertility laparoscopy versus transvaginal ultrasound (US) directed retrieval and to investigate the presence of unexpected failed fertilization in the diagnostic laparoscopy group, which allows an opportunity to diagnosis an etiology of infertility based on gamete interaction.

Design: Consecutive patients who needed infertility diagnostic laparoscopy and agreed to combination with IVF were compared with concurrent patients undergoing transvaginal US IVF. Male factor screening parameters (semen analysis, sperm penetrating assay) and resultant fertilization were analyzed for these patients.

Antipituitary antibodies in a postpartum woman with partial pituitary deficiency and a normal pituitary MRI scan.

We highlight the diagnostic consideration of lymphocytic adenohypophysitis (LAH) in the postpartum patient with unexplained partial pituitary insufficiency. Further, we emphasize that in patients with a normal magnetic resonance imaging scan, the measurement of antipituitary antibodies (APA) may provide an alternative means of confirming the diagnosis. Finally, although assays of APA are nonstandardized, intensive study and development of a reliable APA assay could promote our understanding of the autoimmune process, eventually facilitating the diagnosis of LAH and eliminating the need for biopsy.