Antimüllerian Hormone as a Tool to Predict the Age at Menopause.

This study aimed to assess an eligible cut-off value of anti-Müllerian hormone (AMH) to detect ovarian senescence in a group of premenopausal Greek women to evaluate the possible link between AMH-values and the severity of climacteric symptoms during a follow-up of 24 months. This study included 180 women (group A, 96 women of late reproductive stage/early perimenopause; group B, 84 women in late perimenopause). We measured AMH blood levels and assessed climacteric symptoms using the Greene scale. Log-AMH is inversely associated with postmenopausal status. The AMH cut-off of 0.012 ng/mL predicts the postmenopausal status with a sensitivity of 24.2% and specificity of 30.5%. The postmenopausal stage associated with age (OR = 1.320, 95%CI: 1.084-1.320) and AMH (values ≥ vs. <0.012 ng/mL, OR = 0.225, 95%CI: 0.098-0.529, p-value < 0.001). Moreover, the severity of vasomotor symptoms (VMS) was only associated inversely with AMH (b-coefficient = -0.272, p-value = 0.027). In conclusion, AMH levels measured in the late premenopausal period are inversely associated with the time to ovarian senescence. In contrast, AMH levels measured in the perimenopausal period are inversely associated only with the severity of VMS. Therefore, a cut-off of 0.012 ng/mL predicts menopause with low sensitivity and specificity, making it challenging to use in a clinical setting.

Searching HPV genome for methylation sites involved in molecular progression to cervical precancer.

Background: Human Papilloma Virus has been considered as the main cause for cervical cancer. In this study we investigated epigenetic changes and especially methylation of specific sites of HPV genome. The main goal was to correlate methylation status with histological grade as well as to determine its accuracy in predicting the disease severity by establishing optimum methylation cutoffs. Methods: In total, sections from 145 cases genotyped as HPV16 were obtained from formalin- fixed, paraffin-embedded tissue of cervical biopsies, conization or hysterectomy specimens. Highly accurate pyrosequencing of bisulfite converted DNA, was used to quantify the methylation percentages of UTR promoter, enhancer and 5' UTR, E6 CpGs 494, 502, 506 and E7 CpGs 765, 780, 790. The samples were separated in different groupings based on the histological outcome. Statistical analysis was performed by SAS 9.4 for Windows and methylation cutoffs were identified by MATLAB programming language. Results: The most important methylation sites were at the enhancer and especially UTR 7535 and 7553 sites. Specifically for CIN3+ (i.e. HSIL or SCC) discrimination, a balanced sensitivity vs. specificity (68.1%, 66.2% respectively) with positive predictive value (PPV) and negative predictive value (NPV) (66.2%, 68.2% respectively) was achieved for UTR 7535 methylation of 6.1% cutoff with overall accuracy 67.1%, while for UTR 7553 a sensitivity 60.9%, specificity 69.0%, PPV=65.6%, NPV=64.5% and overall accuracy=65.0% at threshold 10.1% was observed. Conclusion: Viral HPV16 genome was found methylated in NF-1 binding sites of UTR in cases with high grade disease. Methylation percentages of E6 and E7 CpG sites were elevated at the cancer group.

New algorithm for OHSS prevention.

Ovarian hyperstimulation syndrome (OHSS) still remains a life-threatening complication of in vitro fertilization treatment (IVF), keeping patients and especially those, who previously experienced OHSS, from attempting infertility treatment and childbearing. The recent implementation of four new modalities: the GnRH antagonist protocol, GnRH agonist (GnRHa) triggering of ovulation, blastocyst transfer and embryo/oocyte vitrification, renders feasible the elimination of OHSS in connection with ovarian hyperstimulation for IVF treatment. The proposed current algorithm is based on the number of follicles developed after ovarian stimulation, setting a cut-off level at the development of 18 or more follicles. Further, fulfilling this criterion, the algorithm is based on four decision-making points: the final day of patient work-up, the day of triggering final oocyte maturation, day-1 post oocyte pick-up (OPU) and day-5 post OPU. If the physician decides to administer hCG for final oocyte maturation regardless the type of analogue used, he has the option on day-1 to either freeze all embryos or to proceed to day-5. On this day, based on the clinical condition of the patient, a decision should be made to either transfer a single blastocyst or to vitrify all blastocysts available. However, this strategy will not guarantee an OHSS free luteal phase especially if a pregnancy occurs. If the physician decides to trigger ovulation with GnRHa, feasible only with the antagonist protocol, embryos can be cultured until day-5. On this day a transfer can be performed with no risk of OHSS and spare blastocysts may be vitrified. Alternatively, on day-1 or day-2 post OPU, all embryos could be frozen. Hopefully, in a near future, GnRHa triggering and vitrification of oocytes will become everyday practice. Only the combined use of a GnRH antagonist protocol with GnRHa triggering and subsequent single blastocyst transfer or embryo/oocyte freezing will completely abolish the risk of OHSS after ovarian hyperstimulation.

A prospective randomized study comparing gonadotropin-releasing hormone agonists or gonadotropin-releasing hormone antagonists in couples with unexplained infertility and/or mild oligozoospermia.

OBJECTIVE: To compare the efficacy of GnRH antagonist vs. GnRH agonist administration for controlled ovarian hyperstimulation (COH) in assisted reproduction. DESIGN: A prospective, randomized trial. SETTING: Clinical research unit at a tertiary care medical center. PATIENT(S): Sixty-five patients with unexplained infertility or mild male subfertility undergoing COH for IUI. INTERVENTION(S): Twenty-nine women (group A) were randomized to receive 600 microg of busereline acetate per day starting in the midluteal phase of the cycle (long protocol), whereas 36 women (group B) were treated with 0.25 mg/d of the GnRH antagonist Cetrorelix starting from day 6 of the cycle. The starting dose of recombinant FSH was 150 IU in women of both groups. Insemination was performed 34 hours after hCG injection. MAIN OUTCOME MEASURE(S): Clinical and successful ongoing pregnancy rate (PR), measurements of serum FSH, LH, E2, and P, number of recruited follicles, duration of stimulation period, and amount of gonadotropins used. RESULT(S): Women in group A required significantly more days of treatment (median: 12.0 vs. 9.0) and significantly more total units of recombinant FSH (median 1,800 vs. 1,550) as compared with the corresponding values of the antagonist group (group B). Serum FSH, LH, E2, and P were significantly higher on the antagonist group on days 2 and 6 of stimulation. However, these differences regress until the day of hCG administration. CONCLUSION(S): The GnRH antagonists have facilitated short and simple treatment, and are particularly attractive for administration in women undergoing COH, achieving comparable PR with the long protocol regimen.

Uterine and embryonic trophoblast CRH promotes implantation and maintenance of early pregnancy.

Epithelial cells of human endometrium and differentiated endometrial stromal cells express the corticotropin-releasing hormone (CRH) gene. CRH is also produced by the human placental cytotrophoblast. Endometrial and placental CRH is under the endocrine control of gonadal steroids as well as under an autocrine/paracrine regulation by prostanoids and interleukins. Human endometrium, myometrium and placenta also express the relevant receptors. Invasive trophoblasts promote apoptosis of activated Fas-expressing human T lymphocytes, an effect potentiated by CRH and inhibited by the CRH type 1 antagonist, antalarmin. Female rats treated with antalarmin during the first 6 days of gestation had a dose-dependent decrease of implantation sites and live embryos, and significantly decreased endometrial FasL expression. Our data suggest important physiological roles of endometrial and placental CRH in the regulation of decidualization, blastocyst implantation, and early maternal tolerance.

Corticotropin-releasing hormone (CRH) and immunotolerance of the fetus.

The hypothalamic neuropeptide corticotropin-releasing hormone (CRH) is produced by several tissues of the female reproductive system, including the endometrial glands and decidualized stroma, as well as the trophoblast, syncytiotrophoblast, and placental decidua. CRH is also secreted at inflammatory sites and possesses potent pro-inflammatory properties influencing both innate and acquired immune processes. Recent experimental findings show that uterine CRH participates in local immune phenomena associated with early pregnancy, such as differentiation of endometrial stroma to decidua and protection of the fetus from the maternal immune system. CRH induces the expression of apoptotic Fas ligand (FasL) on invasive extravillous trophoblast and maternal decidual cells at the fetal-maternal interface. Furthermore, CRH increases the apoptosis of activated T lymphocytes through FasL induction, participating in the processes of both implantation and early pregnancy tolerance.

Cervical regeneration after diathermy excision of cervical intraepithelial neoplasia as assessed by transvaginal sonography.

OBJECTIVE: To evaluate regeneration in cervical craters following large loop excision of the transformation zone (LLETZ) and to investigate possible differential healing patterns depending on the cone's size. STUDY DESIGN: A prospective study of 100 nulliparous women who underwent LLETZ. They underwent transvaginal scanning estimation of the cervical craters (diameter, depth) immediately post-operatively and at 3, 6 and 12 months. The crater dimensions of the women with the 25 largest cones were compared to those of the women with the 25 smallest cones in each of the above points of time. RESULTS: The mean crater size of all women at 12 months was significantly smaller from the crater size immediately post-operatively. Although, there was a statistically significant difference in mean crater dimensions between the two quartile groups immediately post-operatively, no difference was found at 6 and 12 months. CONCLUSION: There is a healing process of the cervical crater, which is almost completed by the sixth post-treatment month. The defect remaining in the cervix is similar whether a large or small excision was performed.