Association between cigarette smoking and ovarian reserve among women seeking fertility care.

INTRODUCTION: This study examined the association of smoking with ovarian reserve in a cross-sectional study of 207 women enrolled in the Louisville Tobacco Smoke Exposure, Genetic Susceptibility, and Infertility (LOUSSI) Study and assessed effect modification by NAT2 acetylator phenotype. METHODS: Information on current smoking status was collected using a structured questionnaire and confirmed by cotinine assay. Serum anti-Müllerian hormone (AMH) levels were used to assess ovarian reserve. Diminished ovarian reserve (DOR) was defined as AMH <1ng/mL. Single nucleotide polymorphisms in the NAT2 gene, which metabolizes toxins found in cigarette smoke, were analyzed to determine NAT2 acetylator status. Linear and logistic regression were used to determine the effects of smoking on ovarian reserve and evaluate effect modification by NAT2. Regression analyses were stratified by polycystic ovary syndrome (PCOS) status and adjusted for age. RESULTS: Current smoking status, either passive or active as measured by urinary cotinine assay, was not significantly associated with DOR. For dose-response assessed using self-report, the odds of DOR increased significantly for every additional cigarette currently smoked (Odds ratio, OR:1.08; 95% confidence interval, 95%CI:1.01-1.15); additionally, every 1 pack-year increase in lifetime exposure was associated with an increased odds of DOR among women without PCOS (OR: 1.08 95%CI: 0.99-1.18). These trends appear to be driven by the heavy or long-term smokers. Effect modification by NAT2 genotype was not established. CONCLUSION: A history of heavy smoking may indicate increased risk of diminished ovarian reserve.

High-fat diet-induced upregulation of exosomal phosphatidylcholine contributes to insulin resistance.

High-fat diet (HFD) decreases insulin sensitivity. How high-fat diet causes insulin resistance is largely unknown. Here, we show that lean mice become insulin resistant after being administered exosomes isolated from the feces of obese mice fed a HFD or from patients with type II diabetes. HFD altered the lipid composition of exosomes from predominantly phosphatidylethanolamine (PE) in exosomes from lean animals (L-Exo) to phosphatidylcholine (PC) in exosomes from obese animals (H-Exo). Mechanistically, we show that intestinal H-Exo is taken up by macrophages and hepatocytes, leading to inhibition of the insulin signaling pathway. Moreover, exosome-derived PC binds to and activates AhR, leading to inhibition of the expression of genes essential for activation of the insulin signaling pathway, including IRS-2, and its downstream genes PI3K and Akt. Together, our results reveal HFD-induced exosomes as potential contributors to the development of insulin resistance. Intestinal exosomes thus have potential as broad therapeutic targets.

Initial hCG Levels in Ectopic Pregnancies After Embryo Transfer: Use of the Bayes Classifier for Risk Assessment.

OBJECTIVE: To determine whether the Bayes classifier can be used to distinguish between an ectopic and intrauterine pregnancy following embryo transfer based on early human chorionic gonadotropin (hCG) levels. STUDY DESIGN: Retrospective chart review of patients undergoing in vitro fertilization and diagnosed with a singleton intrauterine or with an ectopic pregnancy. Blood was drawn for hCG levels between days 12 and 20 after transvaginal oocyte aspiration. Statistical analysis was performed using a mixed effects model and the Bayes classifier. RESULTS: Singleton intrauterine (n=91) and ectopic gestations (n=14) were analyzed. hCG levels increased by 51% daily in both groups, but levels in ectopic pregnancies were only 14% of those from the control group on the same day (p<1×10-15). Using the Bayes classifier, an hCG value <18 IU/L indicated a large probability (>75%) that the pregnancy was ectopic. There was no statistically significant difference in regards to endometrial thickness (p=0.77), fresh or frozen embryo transfer (p=0.53), number of embryos transferred (p=0.13), donor or autologous oocytes (p=0.76), or the day of hCG draw (p=0.13 and 0.43 for first and second measurement). CONCLUSION: The Bayes classifier can be used as a tool to alert the healthcare provider of a possible ectopic gestation.

Ggnbp2 Is Essential for Pregnancy Success via Regulation of Mouse Trophoblast Stem Cell Proliferation and Differentiation.

The Ggnbp2 null mutant embryos died in utero between Embryonic Days 13.5 to 15.5 with dysmorphic placentae, characterized by excessive nonvascular cell nests consisting of proliferative trophoblastic tissue and abundant trophoblast stem cells (TSCs) in the labyrinth. Lethality of Ggnbp2 null embryos was caused by insufficient placental perfusion as a result of remarkable decreases in both fetal and maternal blood vessels in the labyrinth. These defects were accompanied by a significant elevation of c-Met expression and phosphorylation and its downstream effector Stat3 activation. Knockdown of Ggnbp2 in wild-type TSCs in vitro provoked the proliferation but delayed the differentiation with an upregulation of c-Met expression and an enhanced phosphorylation of c-Met and Stat3. In contrast, overexpression of Ggnbp2 in wild-type TSCs exhibited completely opposite effects compared to knockdown TSCs. These results suggest that loss of GGNBP2 in the placenta aberrantly overactivates c-Met-Stat3 signaling, alters TSC proliferation and differentiation, and ultimately compromises the structure of placental vascular labyrinth. Our studies for the first time demonstrate that GGNBP2 is an essential factor for pregnancy success acting through the maintenance of a balance of TSC proliferation and differentiation during placental development.

Live Birth from Electively Cryopreserved Oocytes: A Feasible Option for Couples Opposed to Embryo Cryopreservation A Case Report.

BACKGROUND: This case report describes a relatively novel indication for oocyte cryopreservation. CASE: A couple undergoing infertility treatment at our institution was opposed to embryo cryopreservation for religious reasons. After multiple unsuccessful infer- tility treatment cycles in- cluding ovulation induction combined with' artificial insemination as well as cycles of therapy with in vitro maturation, we were able to offer them fertilization of a limited number of oocytes followed by oocyte cryopreservation. Since our initial fresh embryo transfer was unsuccessful, the thawing of a limited number of these oocytes prevented a second oocyte retrieval. The couple had 3 oocytes thawed and fertilized and had a successful term birth. CONCLUSION: Elective oocyte cryopreservation is a feasible option for successful pregnancy in patients opposed to embryo cryopreservation.

Adipose-Derived Stromal Vascular Fraction Cell Effects on a Rodent Model of Thin Endometrium.

Endometrial dysfunction affects approximately 1% of infertile women, and there is currently no standard therapy for improving fertility treatment outcomes in these patients. In our study, we utilized a rodent model of thin endometrium to test whether intrauterine application of adipose-derived stromal vascular fraction cells (SVF) could improve morphological and physiological markers of endometrial receptivity. Using anhydrous ethanol, endometrial area and gland density were significantly reduced in our model of thin endometrium. Application of SVF was associated with a 29% reduction in endometrial vascular endothelial growth factor (VEGF) expression and significant increases in uterine artery systolic/diastolic velocity ratios and resistance index values, suggesting reduced diastolic microvascular tone. However, no significant improvements in endometrial area or gland density were observed following SVF treatment. 3D confocal imaging demonstrated poor engraftment of SVF cells into recipient tissue, which likely contributed to the negative results of this study. We suspect modified treatment protocols utilizing adjuvant estrogen and/or tail vein cell delivery may improve SVF retention and therapeutic response in subsequent studies. SVF is an easily-obtainable cell product with regenerative capability that may have a future role in the treatment of infertile women with endometrial dysfunction.

Live birth following in vitro maturation of oocytes retrieved from extracorporeal ovarian tissue aspiration and embryo cryopreservation for 5 years.

OBJECTIVE: To report a live birth after in vitro maturation (IVM) of oocytes retrieved from extracorporeal ovarian tissue aspiration in the setting of fertility preservation. DESIGN: Observational study. SETTING: Academic center. PATIENT(S): A 23-year-old woman. INTERVENTION(S): IVM from extracorporeal ovarian tissue aspiration. MAIN OUTCOME MEASURE(S): Live birth after IVM. RESULT(S): A 23-year-old woman conceived with embryos derived from extracorporeal oocyte aspiration followed by IVM, embryo freezing, and frozen embryo transfer. CONCLUSION(S): A healthy live birth from extracorporeal aspiration of immature oocytes, IVM, and a frozen embryo transfer after 5 years was documented. Consideration of this technique should be made as a primary or adjunct intervention in the setting of fertility preservation.

Successful increase in uterine volume and subsequent pregnancy in a patient with a history of radiation and chemotherapy.

A 26-year-old woman with a history of cranial radiation and chemotherapy desired pregnancy. Pelvic ultrasound scanning demonstrated a small uterine volume of 7 mL. After 25 weeks of estrogen therapy, her uterine volume increased to 37 mL. The patient had an uncomplicated pregnancy with the use of donor oocytes and delivered a term healthy daughter.

Amenorrhea and "man hands".

A 36-year-old multiparous woman presented with amenorrhea and hyperhidrosis of 6 months' duration. Physical examination revealed large, stubby fingers and toes with soft tissue thickening. Serum growth hormone and insulin-like growth factor-1 (IGF-1) levels were elevated and a pituitary macroadenoma was found on computerized tomography (CT).

Adipose tissue and reproduction in women.

Adipose tissue has been viewed as the primary source of stored energy, but with the discovery of novel adipose tissue gene products, i.e., adipokines, another equally important role has emerged. Adipose tissue is a key endocrine organ involved in multiple processes, including glucose homeostasis, steroid production, immunoregulation, hematopoesis, and reproduction. The distribution of adipose tissue may also have a significant impact on reproductive function.

Age and a single day-14 beta-HCG can predict ongoing pregnancy following IVF.

The current study was undertaken to investigate the use of beta human chorionic gonadotrophin (beta-HCG) concentration and other significant factors to predict the likelihood of an IVF pregnancy progressing to detection of cardiac activity by ultrasound, and to create data tables which can be used for patient counselling. A retrospective data analysis was undertaken of 1374 IVF cycles performed from January 1997 to July 2007, resulting in 662 pregnancies. Maternal age (P = 0.0005), day-14 (P < 0.001) and day-16 (P < 0.001) post-oocyte aspiration beta-HCG concentrations were found to be significant in predicting pregnancy outcome. Multiple logistic regression modelling revealed that the most accurate predictive model used a single day-14 beta-HCG concentration and maternal age. Day-14 and day-16 beta-HCG concentrations were highly correlated, with the addition of a day-16 concentration adding no additional predictive value. Ongoing pregnancy rates were proportional to day-14 beta-HCG concentration and inversely proportional to maternal age. The multiple pregnancy incidence increased proportionally with the initial beta-HCG concentration. Thus, for the counselling of patients following IVF, a single day-14 post-oocyte-aspiration beta-HCG concentration and maternal age are most predictive of the pregnancy continuing to detection of cardiac activity by ultrasound.

Endometriosis markers: immunologic alterations as diagnostic indicators for endometriosis.

Because endometriosis, a chronic disease affecting 7% to 10% of women, is associated with immunologic aberrations, the authors hypothesize that the presence of specific immune alterations may be diagnostic. Autoantibodies were assayed by Western immunoblotting using antigens derived from the plasma membrane, cytosol, and nucleus from endometrial and ovarian cells. Natural killer (NK) activity was defined by levels of signaling protein zeta and induction of interferon (IFN)-gamma following exposure to patients' sera. Patients with endometriosis exhibited autoantibodies reactive with cellular proteins; endometrial membrane proteins exhibited the greatest reactivity, followed by nuclear antigens. In all subcellular fractions, patients with stage 3 endometriosis exhibited significantly more immunoreactivity than did stage 2 patients, which was greater than that observed in stage 1 patients. The stage-associated increased reactivity resulted from both recognition of additional proteins and enhanced reactivity with shared proteins. Patient sera suppressed NK zeta expression, which resulted in suppression of NK IFN induction. Alterations in autoreactivity and NK activity are observed in endometriosis and may be useful as diagnostic markers, even in early stage disease.

Pregnancy-linked suppression of TcR signaling pathways by a circulating factor absent in recurrent spontaneous pregnancy loss (RPL).

Evidence suggests that maternal cell-mediated immunity is suppressed during pregnancy and that failure to suppress immune responses can result in partial or total rejection of the fetus. The molecular events associated with suppression of maternal T-cell activation mediated by circulating pregnancy-associated 14 kDa zeta inhibitor protein (ZIP) were defined in women with and without histories of recurrent pregnancy loss (RPL). Using cDNA microarray analysis, ZIP modulations of specific genes associated with T-cell activation signaling were defined. Alterations of defined components were confirmed at the protein level using chromatographically purified ZIP from normal pregnancies versus analogous material from women experiencing RPL. Based on microarray analyses, ZIP from normal pregnancies induced an increase (> or =2-fold) in the expression of 19 genes and a decrease (> or =2-fold) in 15 genes, when incubated with cultured T-cells. In contrast, when T-cells were incubated with analogous material from RPL or non-pregnant controls, no significant differences were observed in the expression of these genes. At the protein level, ZIP from normal pregnancies induced decreases in CD3-zeta (2.36-fold), JAK3 (2.41-fold), STAT5 (1.85-fold), and NF-kappaB (4.24-fold) and a 2.05-fold increase in SOCS2 (all at p<0.001 compared to RPL and non-pregnant controls). The suppressive effects of Zip can lead to the failure of T-cell production of Th1 cytokines, such as IL-2. The 14 kDa circulating ZIP from normal pregnancies suppressed components within the JAK/STAT pathway and induced suppressors of cytokine stimulation, SOCS2.