Phoebe H. Leonard

Fertility drug use and the risk of ovarian tumors in infertile women: a case-control study

OBJECTIVE To assess the influence of infertility and fertility drugs on risk of ovarian tumors. DESIGN Case-control study (Mayo Clinic Ovarian Cancer Study). SETTING Ongoing academic study of ovarian cancer. PATIENT(S) A total of 1,900 women (1,028 with ovarian tumors and 872 controls, frequency matched on age and region of residence) who had provided complete information in a self-report questionnaire about history of infertility and fertility drug use. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Effect of infertility history, use of fertility drugs and oral contraception, and gravidity on the risk of ovarian tumor development, after controlling for potential confounders. RESULT(S) Among women who had a history of infertility, use of fertility drugs was reported by 44 (24%) of 182 controls and 38 (17%) of 226 cases. Infertile women who used fertility drugs were not at increased risk of developing ovarian tumors compared with infertile women who did not use fertility drugs; the adjusted odds ratio was 0.64 (95% CI, 0.37, 1.11). The findings were similar when stratified by gravidity and when analyzed separately for borderline versus invasive tumors. CONCLUSION(S) We found no statistically significant association between fertility drug use and risk of ovarian tumors. Further larger, prospective studies are needed to confirm this observation.

Variability in protein quality used for embryo culture: embryotoxicity of the stabilizer octanoic acid.

OBJECTIVE To screen human serum albumin (HSA) preparations for toxicity and investigate causes of variation. DESIGN Experimental laboratory study. SETTING University-based laboratory. ANIMAL(S) FVB and CF1 mice crossed to create embryos used in experiments. INTERVENTION(S) Mouse embryo assay performed with 5% or 15% HSA (100 mg/mL albumin) from three samples from three separate manufacturers (A, B, C). MAIN OUTCOME MEASURE(S) Blastocyst rates calculated at 96 hours of culture (experiments repeated in triplicate). RESULT(S) The HSA preparations were desalted to remove stabilizers added during HSA processing, then mass spectrometry was used to determine the relative variation in stabilizer concentrations; the effect of the stabilizer octanoic acid on embryo development was tested. At 5% HSA, all samples had blastocyst rates ≥ 70%; at 15% HSA, the blastocyst rates for samples B and C were <50%. Desalting did not affect sample B but did improve the blastocyst rates of sample C. Mass spectrometry revealed high levels of octanoic acid in sample C compared with sample A. The addition of octanoic acid to sample A produced toxicity similar to sample C. CONCLUSION(S) The stabilizer octanoic acid varies by lot and inhibits embryo development. Because octanoic acid is known to cause disruptions in mitochondrial bioenergetics, reduce intracellular pH, and induce oxidative damage in peripheral tissues, its use in embryo culture should be monitored and limited.

Functional impact of Aurora A-mediated phosphorylation of HP1γ at serine 83 during cell cycle progression.

BackgroundPrevious elegant studies performed in the fission yeast Schizosaccharomyces pombe have identified a requirement for heterochromatin protein 1 (HP1) for spindle pole formation and appropriate cell division. In mammalian cells, HP1γ has been implicated in both somatic and germ cell proliferation. High levels of HP1γ protein associate with enhanced cell proliferation and oncogenesis, while its genetic inactivation results in meiotic and mitotic failure. However, the regulation of HP1γ by kinases, critical for supporting mitotic progression, remains to be fully characterized.ResultsWe report for the first time that during mitotic cell division, HP1γ colocalizes and is phosphorylated at serine 83 (Ser83) in G2/M phase by Aurora A. Since Aurora A regulates both cell proliferation and mitotic aberrations, we evaluated the role of HP1γ in the regulation of these phenomena using siRNA-mediated knockdown, as well as phosphomimetic and nonphosphorylatable site-directed mutants. We found that genetic downregulation of HP1γ, which decreases the levels of phosphorylation of HP1γ at Ser83 (P-Ser83-HP1γ), results in mitotic aberrations that can be rescued by reintroducing wild type HP1γ, but not the nonphosphorylatable S83A-HP1γ mutant. In addition, proliferation assays showed that the phosphomimetic S83D-HP1γ increases 5-ethynyl-2´-deoxyuridine (EdU) incorporation, whereas the nonphosphorylatable S83A-HP1γ mutant abrogates this effect. Genome-wide expression profiling revealed that the effects of these mutants on mitotic functions are congruently reflected in G2/M gene expression networks in a manner that mimics the on and off states for P-Ser83-HP1γ.ConclusionsThis is the first description of a mitotic Aurora A-HP1γ pathway, whose integrity is necessary for the execution of proper somatic cell division, providing insight into specific types of posttranslational modifications that associate to distinct functional outcomes of this important chromatin protein.

Sperm morphology: classification drift over time and clinical implications

OBJECTIVE To assess sperm morphology with Tygerberg (strict) and World Health Organization (WHO) 3rd criteria for intrauterine inseminations (IUI) between two eras to determine if there was a difference in pregnancy rates. DESIGN Retrospective study. SETTING Academic institution. PATIENT(S) 127 couples with 290 IUI treatments during 1996-97 (era 1) and 133 couples with 317 IUI treatments during 2005-06 (era 2). INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Pregnancy rates per cycle and couple. RESULT(S) Average sperm morphology was higher in era 1 than era 2 for both WHO 3rd (37 ± 13% vs. 23 ± 10%) and strict criteria (8.0 ± 5.0% vs. 4.0 ± 3.0%). Pregnancy rates per cycle were 5.9% versus 19.8% in era 1 and 16.7% versus 19.3% in era 2 for couples with WHO 3rd morphology <30% or ≥30%, respectively. Pregnancy rates per cycle were 2.7% versus 15.0% in era 1 and 13.3% versus 14.7% in era 2 for couples with strict morphology ≤4% or >4%, respectively. CONCLUSION(S) There was a strong relationship between morphology and IUI outcome in era 1 that was not present in era 2. These results suggest that classification drift increased the percentage of men diagnosed with teratozoospermia and resulted in a loss of predictive value.

Culture Systems: Mineral Oil Overlay

Mineral oil overlay microdrop is commonly used during in vitro fertilization (IVF) procedures. Though mineral oil appears homogeneous, it is an undefined product that can vary in quality. Here, we describe the history, chemistry, processing, and optimal use of mineral oil for IVF and embryo culture.

The Aurora A-HP1γ pathway regulates gene expression and mitosis in cells from the sperm lineage

BackgroundHP1γ, a well-known regulator of gene expression, has been recently identified to be a target of Aurora A, a mitotic kinase which is important for both gametogenesis and embryogenesis. The purpose of this study was to define whether the Aurora A-HP1γ pathway supports cell division of gametes and/or early embryos, using western blot, immunofluorescence, immunohistochemistry, electron microscopy, shRNA-based knockdown, site-directed mutagenesis, and Affymetrix-based genome-wide expression profiles.ResultsWe find that the form of HP1γ phosphorylated by Aurora A, P-Ser83 HP1γ, is a passenger protein, which localizes to the spermatozoa centriole and axoneme. In addition, disruption in this pathway causes centrosomal abnormalities and aberrations in cell division. Expression profiling of male germ cell lines demonstrates that HP1γ phosphorylation is critical for the regulation of mitosis-associated gene expression networks. In female gametes, we observe that P-Ser83-HP1γ is not present in meiotic centrosomes of M2 oocytes, but after syngamy, it becomes detectable during cleavage divisions, coinciding with early embryonic genome activation.ConclusionsThese results support the idea that phosphorylation of HP1γ by Aurora A plays a role in the regulation of gene expression and mitotic cell division in cells from the sperm lineage and in early embryos. Combined, this data is relevant to better understanding the function of HP1γ in reproductive biology.

Surgical Management of Polycystic Ovary Syndrome: A Contemporary Viewpoint on Place of Ovarian Surgery in PCOS Management

Polycystic ovary syndrome (PCOS) is the most common cause of ovulatory dysfunction and resultant infertility. Although the mainstay in management is predominantly medical therapy directed at overcoming diverse symptomatology, surgical management of PCOS was one of the earliest therapeutic options available to treat the disease. Soon after their original description of the disorder, Stein and Leventhal serendipitously discovered that ovarian wedge resection resulted in symptomatic improvement and correction of ovulatory defect of PCOS. The initial enthusiasm soon dampened and ovarian wedge resection subsequently fell into disfavor secondary to sequelae of ovarian cortical trauma that included pelvic adhesions and detriment to ovarian reserve. Recent innovations in the surgical approach, however, offer a potential for benefit for patients unresponsive to conventional medical therapy; focal targeted theca cell destruction utilizing laparoscopic ovarian drilling (LOD) or ultrasound-guided ovarian follicle aspiration, can correct the endocrine milieu of PCOS, albeit transiently, while mitigating serious adverse effects that were common with the conventional wedge resection approach. These beneficial endocrine effects usually result in amelioration of ovulatory dysfunction and infertility. Minimally invasive surgical management of PCOS in the appropriately selected patient is a cost-effective therapeutic option that has comparable success rates to ovulation induction in the management of infertility without incurring any additional treatment related risk of multiple pregnancy. Alongside thecal decompression, interventions such as LOD, bariatric surgical procedures are increasingly being utilized to address obesity and accompanying co morbidities that are highly prevalent in populations of women with PCOS. This chapter focuses on ovarian surgical interventions that can be of relevance to the management of PCOS in a subgroup of patients, whereas Chapter 16 addresses the relevance of bariatric surgical interventions in PCOS.