Effects of polyvalent immunoglobulins in patients with recurrent pregnancy loss and antibodies to the choriocarcinoma cell line JEG-3.

OBJECTIVE: The benefit of polyvalent immunoglobulins (IVIG) for patients with recurrent pregnancy loss (RPL) is controversially discussed. Anti-trophoblast antibodies as an expression of immune pathology have been associated with RPL. We investigated whether the antibody activity against the choriocarcinoma cell line JEG-3 in RPL patients is influenced in vitro by IVIG. STUDY DESIGN: Sera of 110 unexplained RPL patients with positive anti-JEG-3 antibodies were coincubated with IVIG in different concentrations (10mg/ml, 20mg/ml, 40mg/ml). Coincubation with human albumin in identical concentrations served as control. Anti-JEG-3 reactivity was measured by using flow cytometry in comparisons with two in-house standards antibody probes of low and high reactivity as described before. Anti-JEG-3 reactivity above the 95% confidence interval of controls was defined as positive. RESULTS: Incubating RPL sera with 10mg/ml IVIG significantly decreased anti-JEG-3 activity (p<0.001). Increasing IVIG concentration to 40mg/ml resulted in a slightly additionally reduction (p=0.42). In contrast, coincubation with albumin in identically concentrations did not affect anti-JEG-3 activity (p>0.40). CONCLUSION: Coincubation with IVIG in vitro leads to a significant suppression of anti-JEG-3 activity in the sera of RPL patients.

Increased chromosome 16 disomy rates in human spermatozoa and recurrent spontaneous abortions.

OBJECTIVE: To investigate if unexplained recurrent spontaneous abortions (RSA) are associated with increased rates of aneuploidy in spermatozoa of RSA partners ("RSA-men"). DESIGN: Case-control study. SETTING: Academic research center. PATIENT(S): Patients enrolled at the Hormone and Fertility Center and controls at the Department of Urology (LMU-Munich). INTERVENTION(S): Sperm samples of 11 partners of unexplained RSA cases evaluated for elevated diploidy and disomy levels of chromosomes 1-22, X, and Y by multicolor sperm fluorescence in situ hybridization (FISH). MAIN OUTCOME MEASURE(S): Aneuploidy rates obtained in RSA-men compared with controls from the literature and internally; an increase of the aneuploidy rate was considered statistically significant, when it differed ≥ 2 standard deviations from the mean baseline level in controls. RESULT(S): Our sperm FISH data on RSA men showed increased disomy rates for at least three chromosomes in more than 60% of patients but no statistically significant increase of the overall mean sperm disomy or diploidy rate. In particular, meiotic errors involving chromosome 16 contributed to increased sperm disomy in more than 60% of our patients. CONCLUSION(S): These data suggest that among paternal meiotic errors nondisjunction of chromosome 16 might have similar relative influence on fetal aneuploidy compared with maternal chromosome 16 disomy.

Effective ovarian stimulation in a patient with resistant ovary syndrome and antigonadotrophin antibodies.

PROBLEM: We report on a successful ovarian stimulation and pregnancy in a patient with 'resistant ovary syndrome' (ROS) and antigonadotrophin antibodies. ROS is characterized by high endogenous gonadotrophins, low estradiol, normal ovarian antral follicle counts and normal antimuellerian hormone values. METHOD OF STUDY: After cyclical hormone treatment, downregulation with GnRH analogue and ICSI procedure followed. Granulosa cells were treated with LH, FSH or hMG and expression of receptors for FSH, LH, oestrogen receptor beta (ERb) and progesterone receptor A (PR-A) was determined. Serum of the patient was analysed for antibodies directed against hMG. RESULTS: After fertilization of ten metaphase II oocytes and transfer of two blastocysts, a singleton pregnancy was established. Stimulation of granulosa cells with FSH, LH and hMG upregulated ERb and PR-A. Dot blot analysis showed strong reactivity with hMG but not with recFSH. CONCLUSION: This patient with normal expression of gonadotrophin receptors showed antibodies directed to hMG but not to recFSH.

Thyroid hormone receptor (TR)alpha and TRbeta expression in breast cancer.

UNLABELLED: There is evidence that breast cancer patients suffer from thyroid disorders. However, the relation between thyroid receptor (TR) expression and breast cancer remains unknown so far. Therefore, the aim of this study was an immunohistochemical analysis of TR expression in breast cancer patients. MATERIALS AND METHODS: The expression of the combined antibody TRalpha1 and 2 and TRalpha1 or 2 alone as well as the expression of combined TRbeta1 and 2 and TRbeta1 or 2 alone was investigated with specific monoclonal or polyclonal antibodies in 82 patients. All patients presented with a first diagnosis of sporadic breast cancer. The ABC method was used for staining and staining intensities were analyzed using the IRS score. RESULTS: Both TRalpha and TRbeta were expressed in the nuclei of breast cancer cells. In 24% (28/78) of the slides TRalpha1 and 2 IRS was positive. Immunopositivity for TRalpha1 was found in 55/78 slides, for TRalpha 2 in 54/79 slides (71 and 68%, respectively). The expression of TRbeta1 and 2 showed a positive detection in 33/77 (43%) of the slides, for TRbeta1 it was 43/79 (54%), for TRbeta2 60/76 (79%). Significant correlations of the expression of TRs - especially TRalpha2 - were found with further prognostic histopathological parameters such as tumor size, axillary lymph node involvement, grading and hormone receptor status. Multivariate analysis showed a trend for TRalpha2 as an independent predictor of disease-free and overall survival. DISCUSSION: Our results revealed specific alterations in the expression of TRs - especially of TRalpha2 - in breast cancer patients, suggesting it as a marker with possible prognostic validity.

FSH prevents depletion of the resting follicle pool by promoting follicular number and morphology in fresh and cryopreserved primate ovarian tissues following xenografting.

BACKGROUND: Cryopreservation and transplantation of ovarian tissue is one option for re-establishing ovarian function, but optimal conditions for graft sustainment and follicular survival are still considered experimental. The present study aims to analyze the effect of FSH treatment on the resting follicle pool in fresh and cryopreserved primate ovarian tissues following xenografting. METHODS: Ovarian tissues from adult marmosets were grafted freshly or following cryopreservation to ovarectomized nude mice treated with FSH 25 IU twice daily post transplantation or left untreated as controls. Grafts were retrieved 2 or 4 weeks after transplantation to evaluate the number and morphological appearance of follicles. RESULTS: Early start of FSH treatment within 1 week following transplantation partly prevents primordial follicle loss in fresh and frozen-thawed tissues, whereas after a 3 weeks time interval this effect is present only in fresh tissues. A similar positive effect of early, but not later FSH treatment on primary follicles is seen in fresh tissues compared to only marginal effects in frozen-thawed tissues. The percentage of morphologically normal follicles is generally increased in FSH treated tissues, whereas the percentage of primary follicles over all primordial and primary follicles is increased by FSH only in freshly-grafted tissues. CONCLUSIONS: FSH treatment alleviates depletion of the resting follicle pool and promotes normal follicular morphology both in freshly and frozen-thawed grafted tissues. In previously cryopreserved tissues, applying to most of the tissues intended for clinical use in fertility preservation attempts, its positive effect on primordial follicle numbers and potential graft sustainment is dependent on an early start of treatment within one week of transplantation.

Subtle progesterone rise on the day of human chorionic gonadotropin administration is associated with lower live birth rates in women undergoing assisted reproductive technology: a retrospective study with 2,555 fresh embryo transfers.

OBJECTIVE: To evaluate the association between serum P levels on the day of hCG administration and pregnancy outcome in women undergoing controlled ovarian hyperstimulation, prevention of premature ovulation by GnRH analogues, and fresh ET after 5 days of embryo culture. DESIGN: Retrospective, observational, cohort study. SETTING: Private IVF center. PATIENT(S): A total of 2,555 women undergoing fresh ET on day 5 in 2,062 GnRH agonist and 493 GnRH antagonist cycles. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Live birth rate. RESULT(S): Live birth rate in cycles with GnRH agonists was significantly lower in women with P levels ≥ 2.0 ng/mL (17.4%) on the day of hCG administration as compared with women with P levels <1.5 ng/mL (24.6%) and 1.5-1.99 ng/mL (26.7%). No such significant differences in live birth rates in cycles with GnRH antagonist could be observed. CONCLUSION(S): A rise of serum P levels ≥ 2.0 ng/mL on the day of hCG administration is associated with impaired early embryo implantation and reduced live birth rate in cycles with GnRH agonists after day-5 fresh ET.

Antitrophoblast antibodies are associated with recurrent miscarriages.

OBJECTIVE: To investigate whether antitrophoblast antibodies are associated with unexplained recurrent miscarriages, we used choriocarcinoma cells JEG-3, since these cells are negative for class I and II antigens, but they do express HLA-G, resembling an antigen expression of endovascular and interstitial trophoblasts. DESIGN: Case-control study. SETTING: Academic research center. PATIENT(S): One hundred ninety-four patients with two or more consecutive, idiopathic recurrent miscarriages (RM; <20 weeks of gestation) were compared with 110 controls with normal pregnancies and without pregnancy complications. INTERVENTION(S): Anti-JEG-3 reactivities were measured by using flow cytometry and comparisons with two in-house standards antibody samples of low and high reactivity. MAIN OUTCOME MEASURE(S): Anti-JEG-3 reactivities above the 95% confidence interval of controls were defined as positive. RESULT(S): Sera of RM patients reacted significantly stronger with JEG-3 cells than that of controls. In addition, RM patients significantly more often had positive anti-JEG-3 reactivities (17.5%) than controls 5%. This difference was markedly increased with a subgroup of 80 RM patients who had three or more miscarriages, as 27 of these women (34%) were anti-JEG-3 positive. CONCLUSION(S): Antitrophoblast antibodies show significantly more mean channel shift reactivities, and positive reactivities are significantly more prevalent in RM patients as compared with controls. Such antibodies may be involved in mechanisms affecting pregnancies.

Divergent effects of the 677C>T mutation of the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene on ovarian responsiveness and anti-Müllerian hormone concentrations.

OBJECTIVE: To investigate the influence of the 5,10-methylenetetrahydrofolate reductase (MTHFR) 677C>T mutation on serum anti-Müllerian hormone (AMH) concentrations and on the numbers of oocytes retrieved (NOR) following controlled ovarian hyperstimulation (COH). DESIGN: Prospective cohort study. SETTING: University-based infertility clinic. PATIENT(S): Two hundred and seventy women undergoing COH for IVF with or without intracytoplasmic sperm injection. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): AMH levels were determined from blood samples collected after 10 days of GnRH superagonist treatment and before COH. The MTHFR 677C>T genotype was characterized by a TaqMan 5' nuclease assay. RESULT(S): AMH serum concentrations correlated significantly with the NOR in all individuals studied. Average (±SD) AMH levels of TT carriers (2.85±2.23 ng/mL) were significantly higher than those of homozygous CC (1.91±1.59 ng/mL) or heterozygous CT individuals (2.23±1.74 ng/mL). When evaluated by multiple regression analysis, AMH had a significant positive effect on NOR, whereas age and MTHFR 677TT genotype had significant negative effects. CONCLUSION(S): The MTHFR 677TT genotype is associated with higher serum AMH concentrations and has a negative effect on NOR. This apparent paradox might be resolved in light of recent findings describing a negative feedback function of AMH in the coordination of follicle development.

Menopausal status in breast cancer patients with past chemotherapy determines long-term hypoactive sexual desire disorder.

INTRODUCTION: Chemotherapy and endocrine treatment in young breast cancer patients are frequently associated with abrupt menopause. Little is known about the long-term prevalence of hypoactive sexual desire disorder (HSDD) in these patients. AIMS: To examine the effects of adjuvant endocrine therapy on sexual desire in premenopausal patients with breast cancer and past chemotherapy. METHODS: A controlled, cross-sectional study enrolled 47 women with breast cancer or benign breast disease at a tertiary care center. A standardized questionnaire (Sexual Interest and Desire Inventory-Female; SIDI-F) on HSDD was utilized. Serum concentrations for estradiol were measured by a specific assay. MAIN OUTCOME MEASURES: The SIDI-F interview was applied in 35 women with breast cancer (mean age: 42.3 years) with eventual adjuvant endocrine therapy, 2-8 years after chemotherapy, and 13 women with benign breast tumors (mean age: 39.8 years), 2-5 years after diagnosis. RESULTS: Mean SIDI-F scores were similar in the breast cancer group (32.9) and the benign breast disease group (34.0). Subgroup analysis revealed no statistical differences in the mean SIDI-F scores with respect to the actual endocrine therapy. However, in breast cancer patients with menopause induced by chemotherapy or gonadotropin-releasing hormone (GnRH) agonists, the SIDI-F scores were significantly lower (30.7) compared to breast cancer patients with menorrhea (40.4). In breast cancer patients, amenorrhea was associated with significantly lower estradiol levels compared to menorrhea (24 pg/mL vs. 91 pg/mL; P = 0.02). CONCLUSIONS: Cancer treatment that leads to long-term ovarian failure in breast cancer patients has a negative impact on sexual desire. Patients with menopause induced by chemotherapy or GnRH agonists show significantly reduced sexual desire as compared to menstruating patients with past chemotherapy.

The effect of nasal oxytocin on pregnancy rates following intrauterine insemination: double-blind, randomized, clinical pilot study.

PURPOSE: We tested the hypothesis that the application of intranasal oxytocin (8 IU) following intrauterine insemination (IUI) would increase pregnancy rates, without causing major side effects. METHODS: Randomized, double-blind, placebo-controlled pilot study: eighty-six couples with idiopathic infertility, polycystic ovary syndrome and/or male sub-fertility treated with 132 homologous IUI cycles with nasal application of placebo or 8 IU oxytocin following IUI. RESULTS: In 132 IUI cycles of 86 women, 17 pregnancies were achieved, accounting for a pregnancy rate of 12.9% (17/132) per IUI cycle. The pregnancy rates were 13.4% (9/67) per IUI cycle in the placebo group, and 12.3% (8/65) per IUI cycle in the oxytocin group, the difference not being statistically significant. No relevant side effects were observed in both groups. CONCLUSIONS: Intranasal application of 8 IU oxytocin has no major side effects but at the same time did not affect pregnancy rates after IUI in our population. This study does not exclude that a larger patient group, a different time interval between oxytocin application and IUI, higher or multiple oxytocin applications or a different mode of application would have achieved different effects on pregnancy rates.

Predictive value of early serum beta-hCG levels after single blastocyst transfer.

OBJECTIVE: Analysis of serum beta-human chorionic gonadotropin (beta-hCG) levels on standardized days after oocyte retrieval (OR) following single blastocyst transfer to predict pregnancy outcome after assisted reproductive technology. DESIGN: Retrospective study. SETTING: Private fertility center. POPULATION: A total of 230 women with an intact intrauterine pregnancy 21 days after OR and a vital term delivery (n = 191) or a miscarriage (< 10+0 gestational weeks) (n = 39) in the course of the pregnancy following single blastocyst transfer between 1999 and 2008. METHODS: Blood was sampled 11, 14 and partly 21 days after OR. The serum concentration of total beta-hCG was measured by an automated quantitative immunoassay. Receiver operating characteristic curves were generated to calculate sensitivity and specificity. MAIN OUTCOME MEASURES: A total of 509 beta-hCG measurements. RESULTS: The mean beta-hCG concentrations on days 11, 14 and 21 after OR were significantly lower in the group of patients with a miscarriage compared to the group with a term delivery (p = 0.02, < 0.001 and < 0.001 respectively). With the cut-off values of 15 mIU/mL, 80 mIU/mL and 1500 mIU/mL on days 11, 14 and 21, the positive predictive value was 89%, 93% and 92%. The negative predictive value was 28%, 40% and 54%. CONCLUSIONS: The predictive value of a single beta-hCG measurement for pregnancy outcome after single blastocyst transfer increases between days 11 and 21 after OR. A single beta-hCG measurement on day 14 may balance the accuracy of prediction and the necessity of an early reassuring test.

Effects of progesterone and its antagonist mifepristone on progesterone receptor a expression in human umbilical vein endothelial cells.

Effects of female steroid hormones on endothelial cells are gaining increased importance due to several studies on the effects of hormonal treatment on cardiovascular risk. Recent data argue for an improvement of endothelium-derived relaxation and impaired vascular contraction by estradiol, whereas progesterone and testosterone might entail contrary effects. So far, gestagenic influence on endothelial cell physiology is poorly understood. Human umbilical vein endothelial cells (HUVECs) exposed to the female sex hormones estradiol and progesterone show expression of estrogen receptor-beta (ERbeta) and progesterone receptor A (PR-A), and are negative for ERalpha and PR-B. The aim of this study was to analyze the expression and stimulation of PR-A and -B in HUVECs after stimulation with progesterone and PR antagonists that are commercially available. PR-B expression or upregulation was abrogated after application of progesterone or antagonists to HUVECs. Expression of PR-A could be significantly upregulated with progesterone and mifepristone. Unexpectedly, stimulation with the progesterone antagonist RU486 (mifepristone) was accomplished by an upregulation of PR-A expression in our study. We conclude that gestagenic effects on HUVECs independent of modulators are mediated via the PR-A.

The relationship between immunosuppressive activity and immunoregulatory cytokines in seminal plasma: influence of sperm autoimmunity and seminal leukocytes.

While the contributions of prostasomes, polyamines and prostaglandins to the immunosuppressive activity (ISA) of human seminal plasma have been well-characterised, the contribution of immunoregulatory cytokines found in seminal plasma has received relatively little attention. Semen samples were collected from adult men displaying normospermic parameters, sperm antibodies or substantially elevated seminal leukocytes. Samples were processed through ultracentrifugation and dialysis (<3500Da) to remove prostasomes, polyamines and prostaglandins, and then assayed for ISA by an in vitro T lymphocyte inhibition assay, as well as by specific immunoassays for transforming growth factor beta(1) (TGFbeta(1)), interleukin-10 (IL-10), activin A and the activin-binding protein, follistatin. Seminal plasma from all groups retained substantial ISA following processing. Compared with normospermic men, this 'large' molecular weight ISA fraction was significantly increased in a subset of men with sperm antibodies, but was not altered in the group with elevated leukocytes. There was no relationship between ISA and any cytokine examined, and only TGFbeta(1) was present at levels sufficient to contribute to ISA. Inhibition with a TGFbeta-specific antibody reduced ISA in seminal plasma by approximately 50%. Across all patients, TGFbeta(1) levels were positively correlated with sperm numbers in the ejaculate and with activin A, but not with follistatin or IL-10. Activin A and IL-10 also displayed a positive relationship, and elevated leukocytes was associated with a significant elevation of IL-10 and activin A, but not TGFbeta(1). It is concluded that 'large' molecular weight molecules, the most important of which appears to be TGFbeta(1), make a significant contribution to immunosuppression by human seminal plasma.

Pregnancy complications, obstetric risks, and neonatal outcome in singleton and twin pregnancies after GIFT and IVF.

PURPOSE: In vitro fertilization (IVF) and to a lower extent gamete intra-fallopian transfer (GIFT) have become routine infertility treatments in industrialized countries. Our purpose is to compare the obstetric and neonatal characteristics of singleton and twin pregnancies after GIFT and IVF with those conceived spontaneously. METHODS: This case-control study was conducted in a tertiary care medical center. The 322 singleton and 78 twin pregnancies after GIFT or IVF from 1991 through 1996 were evaluated and compared with each other, and with a control group that conceived spontaneously and matched for parity, maternal and gestational age. Statistical significance of differences was assessed by chi(2) test or two-tailed Fisher exact test. Continuous variables were compared by the paired t-test. RESULTS: Pregnancy-induced hypertension (PIH) and vaginal bleeding were significantly more frequent maternal complications in the GIFT/IVF singleton groups compared to controls. In twin pregnancies the rate of cesarean sections, vaginal bleeding and preterm labor were more common after GIFT/IVF but did not reach statistical significance. Assisted reproduction was associated with low birth weight only in twin pregnancies when controlled for confounding variables, however perinatal outcome was comparable. There was no significant difference in the outcome measures between GIFT and IVF pregnancies. CONCLUSION: After controlling for parity, maternal and gestational age, singleton pregnancies conceived by GIFT/IVF are at increased obstetrical risk, however the perinatal outcome is comparable despite a lower average birth weight.

The contributions of deficient androgen action in spermatogenic disorders.

The contributions of deficient androgen actions in spermatogenic disorders causing idiopathic male infertility are reviewed. The physiological role of androgens in spermatogenesis, the mechanism of actions of testosterone and the clinical implication of androgen deficiency are explained. The role of mutations in the androgen receptor (AR) in idiopathic infertility and, in particular, the contribution of expanded cytosine-adenine-guanine (CAG) repeats in exon 1 of the AR gene to the occurrence of male idiopathic infertility is highlighted. Possible future aspects of treatment for such patients are discussed.