matured MII or IVM-MII oocytes were collected and processed rapidly. RNA was extracted and amplified using a 2-round in vitro transcription procedure, then labeled and hybridized to Applied Biosystems Human Genome Survey Microarrays. This array interrogates 29,098 genes. Up-or down-regulated genes were classified according to the molecular function, biological process, or pathway in which they participate. Statistical analysis was employed to compare each class of genes between samples.RESULTS: The in vitro transcription protocol generated an average of 12 mg of RNA per sample. GV, in vivo matured-MII and IVM-MII oocytes expressed 14,188, 16,402 and 18,299 genes, respectively. There was extensive overlap in the genes expressed by each group, but also some significant differences. Samples showing the least similarity in expression profile were GV and in vivo MII oocytes. IVM-MIIs also displayed significant differences in expression when compared to in vivo MII oocytes, indicating that the process of in vitro maturation does not perfectly mimic that achieved in vivo. A disproportionate number of genes for storage proteins displayed down-regulation in IVM-MII oocytes (relative to in vivo MII). A reduction in expression was also seen for genes involved in processes of homeostasis. Significant up-regulation of gene expression for the ubiquitin proteasome pathway was observed in both IVM-MII and GV samples.CONCLUSIONS: This study provided a detailed insight into gene expression of human oocytes, assessing virtually every gene in the genome. GV oocytes were highly active for processes related to transcription, RNA processing and early stages of protein synthesis, whereas MII oocytes were active for pathways involved in processing of mature proteins. The results suggest that although IVM-MII oocytes closely resemble in vivo-MII oocytes for pathways related to nuclear maturity, a number of those associated with cytoplasmic functions continue to be expressed in a GV-like manner. Additionally, IVM-MII oocytes have significant differences in the expression of genes related to cellular storage and homeostasis. The pathways abnormally expressed during IVM highlight specific deficiencies in culture media and point the way to further optimization of IVM protocols.

This case report represents one of the few documented cases of parthenote embryo retrieval from an IVF patient with a history of ovarian teratomas. A 29-year-old woman presented at our centre with a history of primary infertility for 6 years due to male factor. She had undergone left oophorectomy 4 years before due to an ovarian teratoma. An ultrasound scan performed during basal evaluation revealed two complex images in the right ovary suggesting teratomas, measuring 2.5 x 2.4 and 1.7 x 1.3 cm. A significant extent of sonographically normal ovarian parenchyma was present, and the patient underwent the long leuprolide acetate protocol of ovarian stimulation with recombinant FSH for an IVF-ICSI cycle. She had 13 metaphase II (MII), four metaphase I (MI), two germinal vesicle (GV) oocytes and one 4-cell embryo retrieved. Eight out of nine injected oocytes were fertilized normally while one was unfertilized. Embryo transfer was carried out 72 h after retrieval. The 4-cell (parthenote) embryo recovered at oocyte retrieval continued to cleave in culture, developing into a 7-cell embryo by the next day. The embryo was morphologically normal, presenting an evident nucleus in each blastomere. Fluorescent in situ hybridization (FISH) returned two signals for the X chromosome in each blastomere that was analysed. Of the eight normally fertilized embryos, three were transferred, resulting in a normal singleton pregnancy and the birth of a healthy baby.

Objectives: To assess survival and long term arch patency rates in a consecutive group of children after extended arch repair for coarctation of the aorta. Methods: Review of 191 consecutive children (154 (81%) under 1 year of age) operated on between 1990 and 2002 by a single surgeon using extended arch reconstructive techniques. For assessment of survival patients were divided into three groups: 1, coarctation alone, n = 104; 2, coarctation and ventricular septal defect, n = 38; and 3, coarctation in association with complex intracardiac anomalies, n = 49. A prospective and systematic clinical and echocardiographic evaluation of the aortic arch was undertaken. Results: Median time to follow up was 4.2 years (range 1-10.6 years). Overall actuarial survival was 92%, 88%, and 88% at two, five, and 10 years. Mortality was significantly higher in those patients with complex intracardiac anatomy. Arch obstruction recurred in seven of 165 (4.2%) patients: four of 139 (2.9%) term and three of 10 (30%) premature infants (p , 0.001). Conclusions: Survival after extended arch reconstruction for coarctation is excellent. At long follow up recurrent arch obstruction is rare, with prematurity the only risk factor.

Intracytoplasmic sperm injection (ICSI) into mouse oocytes involves a very low survival rate. This study was designed to determine why ICSI frequently fails in mice. Metaphase II oocytes were obtained from superovulated 4-6 week old F1 hybrid mice. Spermatozoa were retrieved from the epididymis of 12-14 week old F1 hybrid mice. The spiked microinjection pipette used to inject a spermatozoon into the ooplasm had outer and inner diameters of 10 and 8 microns respectively. The oocytes used in the first part of the study were not activated (group 1). Some oocytes were incubated with calcium ionophore for 5 min (group 2). The injected oocytes were evaluated 6, 20, 48 and 72 h after injection. A total of 143 eggs in each group underwent ICSI. In group 1, sperm heads escaped into the perivitelline space. In all, 63 (47%) of the remaining oocytes were damaged during the injection or had degenerated by the first evaluation. The survival rate was 53%, but fertilization did not occur. In group 2, 31 oocytes (22%) were damaged during microinjection or soon degenerated. Two oocytes underwent accidental subzonal insemination. Six oocytes were fertilized (4.2%) among the 78% of survivors. After injection, the sperm tail was found in the cytoplasm (27 and 31% in groups 1 and 2 respectively), the perivitelline space (45% in both groups) or protruding through the zona pellucida (28 and 23% respectively). More oocytes degenerated when the tail remained in the cytoplasm, i.e. 78% in group 1 and 36% in group 2.

Tay-Sachs disease is a lysosomal storage disease, which in its most severe form leads inexorably to death during infancy. We have developed a method for preimplantation diagnosis, using polymerase chain reaction (PCR) technology, by which the two most frequent mutations occurring in this disease can be amplified simultaneously. We have tested this method on single blastomeres and have compared four lysis methods: (i) boiling in water at 94 degrees C for 15 or (ii) 30 min, and (iii) incubation in an alkaline lysis buffer for 30 min at 94 degrees C or (iv) at 65 degrees C for 10 min. The amplification percentages were 21, 67, 71 and 91% respectively.