The influence of AMH on IVF success.

PURPOSE: Anti-Müllerian hormone has a regulative function in the activation of folliculogenesis and an influence on atresia rate. It is considered a marker for the ovarian reserve. We know that a relationship exists between AMH levels and oocyte retrieval numbers, antral follicle count, pregnancy rates and birth rates. The role of AMH as an efficient prognostic factor in determining the probability of pregnancy has been largely discussed in the literature. The aim of this study is to determine the role age and AMH levels play in success rates of IVF/ICSI therapies. To date, the sample group we examined was one of the biggest ever included in a single study of the subject. METHODS: All patients who underwent an IVF/ICSI treatment with FSH stimulation in the Wiesbaden Kinderwunschzentrum between 2003 and 2010, were no older than 44 years old, and had an evaluation of serum AMH levels before treatment were included in this study. In total, 1287 patients were analysed retrospectively. Statistical analysis was performed with SPSS. RESULTS: Females' mean age was 34.89, ranging from 21 to 44 years. The patients underwent between 1 and 11 IVF cycles. Younger women had significantly higher AMH levels (p = 0.001). Patients with higher AMH levels had significantly lower break-off rates (p < 0.0005) and a significantly higher number of oocytes retrieved (p < 0.0005). Higher levels of AMH corresponded to higher pregnancy rates (p = 0.017). AMH levels do not influence pregnancy rates in younger patients (<36 years). CONCLUSIONS: AMH is a useful parameter that should be measured before performing an IVF/ICSI treatment. In younger patients, AMH levels do not predict pregnancy outcomes. In patients older than 36 years, AMH can be used as a prognostic factor. Even when a woman's AMH levels are too low to be detected, she still an acceptable chance of becoming pregnant.

Broad DNA methylation changes of spermatogenesis, inflammation and immune response-related genes in a subgroup of sperm samples for assisted reproduction.

Aberrant sperm DNA methylation patterns, mainly in imprinted genes, have been associated with male subfertility and oligospermia. Here, we performed a genome-wide methylation analysis in sperm samples representing a wide range of semen parameters. Sperm DNA samples of 38 males attending a fertility centre were analysed with Illumina HumanMethylation27 BeadChips, which quantify methylation of >27 000 CpG sites in cis-regulatory regions of almost 15 000 genes. In an unsupervised analysis of methylation of all analysed sites, the patient samples clustered into a major and a minor group. The major group clustered with samples from normozoospermic healthy volunteers and, thus, may more closely resemble the normal situation. When correlating the clusters with semen and clinical parameters, the sperm counts were significantly different between groups with the minor group exhibiting sperm counts in the low normal range. A linear model identified almost 3000 CpGs with significant methylation differences between groups. Functional analysis revealed a broad gain of methylation in spermatogenesis-related genes and a loss of methylation in inflammation- and immune response-related genes. Quantitative bisulfite pyrosequencing validated differential methylation in three of five significant candidate genes on the array. Collectively, we identified a subgroup of sperm samples for assisted reproduction with sperm counts in the low normal range and broad methylation changes (affecting approximately 10% of analysed CpG sites) in specific pathways, most importantly spermatogenesis-related genes. We propose that epigenetic analysis can supplement traditional semen parameters and has the potential to provide new insights into the aetiology of male subfertility.

Success Rate of Inseminations Dependent on Maternal Age? An Analysis of 4246 Insemination Cycles.

Intrauterine insemination (IUI) has latterly become less important in reproductive medicine. The aim of this retrospective analysis was to identify and evaluate the success rates of repeated insemination cycles in women of different ages. All women who underwent intrauterine insemination in the Wiesbaden Fertility Clinic between 1998 and 2010 were included in the analysis. Additional inclusion criteria were: not more than 45 years old, previous FSH stimulation and slight to moderate subfertility of the male partner. A total of 4246 insemination cycles in 1612 patients were included in the analysis. The average number of IUI cycles per patient was 2.24 (1-14). Patient age ranged from 19 to 45 years (mean: 33.9 years). Logistic regression analysis showed a drop in pregnancy rates with increasing age (p = 0.000). However, for the first three cycles the pregnancy rates for women aged 40 and 41 did not differ from those of women aged between 35 and 39 years. Overall pregnancy rates were stable in women up to the age of 40, even after several insemination cycles (7.5 and 10 %). Insemination is therefore still an effective procedure in selected patients. Stable pregnancy rates were recorded even after more than 3 cycles. After 3 cycles, the success rates for women aged 40 and 41 did not differ from those of women below the age of 40.

Methylation status of imprinted genes and repetitive elements in sperm DNA from infertile males.

Stochastic, environmentally and/or genetically induced disturbances in the genome-wide epigenetic reprogramming processes during male germ-cell development may contribute to male infertility. To test this hypothesis, we have studied the methylation levels of 2 paternally (H19 and GTL2) and 5 maternally methylated (LIT1, MEST, NESPAS, PEG3, and SNRPN) imprinted genes, as well as of ALU and LINE1 repetitive elements in 141 sperm samples, which were used for assisted reproductive technologies (ART), including 106 couples with strictly male-factor or combined male and female infertility and 28 couples with strictly female-factor infertility. Aberrant methylation imprints showed a significant association with abnormal semen parameters, but did not seem to influence ART outcome. Repeat methylation also differed significantly between sperm samples from infertile and presumably fertile males. However, in contrast to imprinted genes, ALU methylation had a significant impact on pregnancy and live-birth rate in couples with male-factor or combined infertility. ALU methylation was significantly higher in sperm samples leading to pregnancy and live-birth than in those that did not. Sperm samples leading to abortions showed significantly lower ALU methylation levels than those leading to the birth of a baby.