Three singleton deliveries with healthy children from one couple after cryo-tESE and ICSI.

UNLABELLED: We report on a couple who delivered three healthy babies in three deliveries after cryo-TESE combined with ICSI. The male patient suffers from congenital bilateral absence of the vas deferens (CBAVD). METHODS: Three testicular sperm extraction (TESE) operations were performed in the male accompanied by six stimulated ICSI cycles in the female patient. Altogether, 59 oocytes were retrieved. Fifty-one oocytes (86%) were in metaphase II and 38 fertilized regularly (75%). Sixteen embryos, in the 3-6 cell stage, were transferred to the uterus. RESULTS: The first, fifth and sixth embryo transfers of fresh embryos led to intact intrauterine singleton pregnancies. The pregnancy and implantation rates with fresh embryos were 50% and 20%, respectively. CONCLUSIONS: TESE or microscopic epididymal sperm aspiration in patients with CBAVD in combination with a healthy female partner is likely to yield very good results in ICSI/ET. As azoospermia can be caused by cystic fibrosis and cystic fibrous transmembrane conductance regulator gene mutation range varies dramatically in patients of different ethnic groups.

In vivo formation of a human beta-globin locus control region core element requires binding sites for multiple factors including GATA-1, NF-E2, erythroid Kruppel-like factor, and Sp1.

The active elements of the beta-globin locus control region (LCR) are located within domains of unique chromatin structure. These nuclease hypersensitive sites (HSs) are characterized by high DNase I sensitivity, erythroid specificity, similar nucleosomal structure, and evolutionarily conserved clusters of cis-acting elements that are required for the formation and function of the core elements. To determine the requirements for HS core formation in the setting of nuclear chromatin, we constructed a series of artificial HS cores containing binding sites for GATA-1, NF-E2, and Sp1. In contrast to the results of previous in vitro experiments, we found that when constructs were stably integrated in mouse erythroleukemia cells the binding sites for NF-E2, GATA-1, or Sp1 alone or in any combination were unable to form core HS structures. We subsequently identified two new cis-acting elements from the LCR HS4 core that, when combined with the NF-E2, Sp1, and tandem inverted GATA elements, result in core structure formation. Both new cis-acting elements bind Sp1, and one binds erythroid Kruppel-like factor (EKLF). We conclude that in vivo beta-globin LCR HS core formation is more complex than previously thought and that several factors are required for this process to occur.

Mullerian inhibiting substance inhibits ovarian cell growth through an Rb-independent mechanism.

Müllerian inhibiting substance (MIS), a transforming growth factor-beta family member, causes regression of the Müllerian duct in male embryos. MIS overexpression in transgenic mice ablates the ovary, and MIS inhibits the growth of ovarian cancer cell lines in vitro, suggesting a key role for this hormone in postnatal development of the ovary. This report describes a mechanism for MIS-mediated growth inhibition in both a human epithelial ovarian cancer cell line and a cell line derived from normal ovarian surface epithelium, which is the origin of human epithelial ovarian cancers. MIS-treated cells accumulated in the G(1) phase of the cell cycle and subsequently underwent apoptosis. MIS up-regulated the cyclin-dependent kinase inhibitor p16 through an MIS type II receptor-mediated mechanism and inhibited growth in the absence of detectable or inactive Rb protein. Prolonged treatment with MIS down-regulated the Rb-related protein p130 and increased the Rb family-regulated transcription factor E2F1, overexpression of which inhibited growth. These findings demonstrate that p16 is required for MIS-mediated growth inhibition in ovarian epithelial cells and tumor cells and suggest that up-regulation of E2F1 also plays a role in this process.