Jared C. Robins

Induction of Vascular Endothelial Growth Factor by IGF-I in Osteoblast-Like Cells Is Mediated by the PI3K Signaling Pathway through the Hypoxia-Inducible Factor-2α

IGF-I is known to stimulate the expression of oxygen- and nutrient-sensitive genes in several cell types. In this study we investigated the signaling pathways and transcriptional mechanisms that mediate IGF-I induction of vascular endothelial growth factor (VEGF) expression in human osteoblast-like cells. IGF-I (50 ng/ml) induced a rapid increase (3-fold) in VEGF mRNA in osteoblasts that was accompanied by an increase in the level of hypoxia-inducible factor-2α (HIF-2α) protein without changes in HIF-2α mRNA expression. These effects were mimicked by chemical inhibition of proteosomal degradation of HIF-2α. Transcriptional activation of a proximal VEGF promoter-luciferase construct was greatly enhanced by cotransfection with an HIF-2α, but not an HIF-1α, construct. IGF-I acutely stimulated Akt phosphorylation, which was abolished by pretreatment of cells with the PI3K inhibitor LY294002. Pretreatment of the cells with LY294002 also greatly attenuated IGF-I induction of HIF-2α and blunted IGF-I-induced VEG...

Embryo morphology score on day 3 is predictive of implantation and live birth rates

Purpose: To determine if embryo cleavage state or morphology on day 3 correlates with implantation or live birth rates. A retrospective cohort study of all fresh embryo transfers over 2 years. Methods: Patients were grouped by the average cleavage state and morphology. Cleavage state groups were: <6, ≥6, <8, and ≥8 cells. Morphology groups by average grade were: group 1 (best) = 1, group 2 = >1≤2, group 3 = >2≤3, and group 4 (worst) = >3≤4. Results: The overall implantation rate for 158 cycles was 28.1% with a live birth rate of 37.3%. Morphologic state was highly predictive of both implantation rate and live birth rate. Implantation rates by group were 54.8% (group 1), 30.4% (group 2), 23.8% (group 3), and 11.1% (group 4). Likewise, live birth rates among groups were 61.5, 39, 20, and 21%, respectively. Cleavage state was not predictive of outcome. Conclusions: Embryo grade is highly predictive of implantation and live birth rate and can be used to determine the number of embryos to transfer. Cleavage state is not predictive of outcome.

Management of the first in vitro fertilization cycle for unexplained infertility: a cost-effectiveness analysis of split in vitro fertilization-intracytoplasmic sperm injection

OBJECTIVE To determine the cost-effectiveness of split IVF-intracytoplasmic sperm injection (ICSI) for the treatment of couples with unexplained infertility. DESIGN Adaptive decision model. SETTING Academic infertility clinic. PATIENT(S) A total of 154 couples undergoing a split IVF-ICSI cycle and a computer-simulated cohort of women <35 years old with unexplained infertility undergoing IVF. INTERVENTION(S) Modeling insemination method in the first IVF cycle as all IVF, split IVF-ICSI, or all ICSI, and adapting treatment based on fertilization outcomes. MAIN OUTCOME MEASURE(S) Live birth rate, incremental cost-effectiveness ratio (ICER). RESULT(S) In a single cycle, all IVF is preferred as the ICER of split IVF-ICSI or all ICSI (

Use of in-cycle antimüllerian hormone levels to predict cycle outcome

58,766) does not justify the increased live birth rate (3%). If two cycles are needed, split IVF/ICSI is preferred as the increased cumulative live birth rate (3.3%) is gained at an ICER of

Multidrug-resistant transport activity protects oocytes from chemotherapeutic agents and changes during oocyte maturation

29,666. CONCLUSION(S) In a single cycle, all IVF was preferred as the increased live birth rate with split IVF-ICSI and all ICSI was not justified by the increased cost per live birth. If two IVF cycles are needed, however, split IVF/ICSI becomes the preferred approach, as a result of the higher cumulative live birth rate compared with all IVF and the lesser cost per live birth compared with all ICSI.

Bioengineering Anembryonic Human Trophoblast Vesicles

OBJECTIVE The goal of this work is to expand the usefulness of antimüllerian hormone (AMH) in predicting in vitro fertilization cycle outcome by demonstrating that AMH concentration obtained in an ongoing treatment cycle predicts both oocyte number and pregnancy. STUDY DESIGN Serum samples were obtained from 190 in vitro fertilization patients at onset of follicle-stimulating hormone stimulation. These were analyzed retrospectively during a single cycle in which clinicians were blinded to the results. Our major outcome measures were the number of oocytes obtained and ongoing pregnancy. RESULTS Patients with an initial AMH concentration of >3 ng/mL were found to produce a mean of 19.8 oocytes and had an ongoing pregnancy rate of 60.3%. In contrast, those with AMH values of ≤1 ng/mL yielded a mean of 6.2 oocytes and had an ongoing pregnancy rate of 23.4% (P < .0001 for both). CONCLUSION Greater AMH serum concentration strongly predicts an increased number of oocytes and ongoing pregnancy (P ≤ .0001).

Cellular Models of Trophoblast Differentiation

OBJECTIVE To determine the multidrug-resistant transporter (MDR) activity in oocytes and their potential role in oocyte susceptibility to chemotherapy. DESIGN Experimental laboratory study. SETTING University and academic center for reproductive medicine. SUBJECT(S) Women with eggs retrieved for intracytoplasmic sperm injection cycles and adult female FVBN and B6C3F1 mouse strains. INTERVENTION(S) Inhibition of MDR activity in oocytes. MAIN OUTCOME MEASURE(S) Efflux activity of MDRs with the use of quantitative fluorescent dye efflux, and oocyte cell death when exposed to chemotherapy. RESULT(S) Oocytes effluxed fluorescent reporters, and this activity was significantly reduced in the presence of the MDR inhibitor PSC 833. Geminal vesicle oocytes were more efficient at efflux than metaphase 2 oocytes. Human oocytes exposed to cyclophosphamide and PSC 833 showed cell death with the use of two different viability assays compared with control samples and those exposed to cyclophosphamide alone. Immunoblots detected MDR-1 in all oocytes, with the greatest accumulation in the geminal vesicle stage. CONCLUSION(S) Oocytes have a vast repertoire of active MDRs. The implications of this study are that these protective mechanisms are important during oogenesis and that these activities change with maturation, increasing susceptibility to toxicants. Future directions may exploit the up-regulation of these transporters during gonadotoxic therapy.

Follicle curetting at the time of oocyte retrieval increases the oocyte yield

Introduction: Trophoblast cells in vivo form a 3-dimensional structure that promotes complex cell-to-cell interactions that cannot be studied with traditional monolayer culture. We describe a 3-dimensional trophoblast bioreactor to study cellular interactions. Methods: Nonadhesive agarose hydrogels were cast from molds using computer-assisted prototyping. Trophoblast cells were seeded into the gels for 10 days. Morphology, viability, and vesicle behavior were assessed. Results: Trophoblast cells formed uniform spheroids. Serial sectioning on days 3, 7, and 10 revealed central vacuolization with a consistent outer rim 12.3-μ thick. The vesicle configuration has been confirmed with confocal imaging. Electron Microscopic (EM) imaging revealed its ultrastructure. The vesicles migrate across a fibronectin-coated surface and invaded basement membrane. Conclusions: Trophoblast cells cultured in a novel substrate-free 3-dimensional system form trophoblast vesicles. This new cell culture technique allows us to better study placental cell-to-cell interactions with the potential of forming microtissues.

In vitro maturation

Orchestrated trophoblast differentiation is necessary to establish and maintain a normal pregnancy, however the molecular mechanisms that guide this process remain largely unknown. Although early studies of cytotrophoblast differentiation relied on animal models, more recent trophoblast research has involved in vitro models of human tissue. These in vitro models have utilized cultured trophoblast cell lines, primary cell culture, and villous explant cultures-each with its advantages and disadvantages. Traditionally, attempts to develop in vitro models of human placental differentiation have relied on two-dimensional cell culture. Though monolayer culture methods have been refined over time this technique has several limitations, including the inability to study cell-to-cell interactions. Recently, several studies have employed three-dimensional culture methods to overcome many of the limitations of traditional two-dimensional trophoblast culture. These three-dimensional culture systems have an important role in both the study of cytotrophoblast differentiation and development of new therapeutics targeting placenta associated diseases.

Female Fertility : What Every Urologist Must Understand

PurposeTo determine whether follicle curetting at the time of oocyte retrieval increases oocyte yield.MethodsRetrospective review of all patients who underwent oocyte retrieval from July 1, 2003 to June 30, 2005.Main outcome measureNumber of oocytes retrieved. Secondary outcome measures: retrieval time, number of cryopreserved embryos, pregnancy rates, and incidence of ovarian hyperstimulation syndrome.ResultsThere were no differences in patient demographics, antral follicle count, cycle stimulation characteristics, fertilization rates, embryo quantity or quality, embryo cryopreservation rates, clinical pregnancy rates, live birth rates, or ovarian hyperstimulation syndrome between the groups. Retrievals that utilized curetting took three minutes longer. Follicle curetting significantly increased the number of oocytes retrieved, 13.9 ± 0.6 compared to 11.4 ± 0.6 oocytes without curetting (P = 0.003). The quantity of mature oocytes was also increased with curetting (10.3 ± 0.5 versus 8.4 ± 0.5, P = 0.006).ConclusionsThis study demonstrated that follicle curetting significantly increased oocyte yield. While it did not increase live birth rates, this increase in oocyte yield should lead to increased numbers of embryos for selection at transfer and increased embryos for cryopreservation.