Release of sICAM-1 in Oocytes and In Vitro Fertilized Human Embryos

Background: During the last years, several studies have reported the significant relationship between the production of soluble HLA-G molecules (sHLA-G) by 48–72 hours early embryos and an increased implantation rate in IVF protocols. As consequence, the detection of HLA-G modulation was suggested as a marker to identify the best embryos to be transferred. On the opposite, no suitable markers are available for the oocyte selection. Methodology/Principal Findings: The major finding of the present paper is that the release of ICAM-1 might be predictive of oocyte maturation. The results obtained are confirmed using three independent methodologies, such as ELISA, Bio-Plex assay and Western blotting. The sICAM-1 release is very high in immature oocytes, decrease in mature oocytes and become even lower in in vitro fertilized embryos. No significant differences were observed in the levels of sICAM-1 release between immature oocytes with different morphological characteristics. On the contrary, when the mature oocytes were subdivided accordingly to morphological criteria, the mean sICAM-I levels in grade 1 oocytes were significantly decreased when compared to grade 2 and 3 oocytes. Conclusions/Significance: The reduction of the number of fertilized oocytes and transferred embryos represents the main target of assisted reproductive medicine. We propose sICAM-1 as a biochemical marker for oocyte maturation and grading

Molecular Mechanisms Generating and Stabilizing Terminal 22q13 Deletions in 44 Subjects with Phelan/McDermid Syndrome

In this study, we used deletions at 22q13, which represent a substantial source of human pathology (Phelan/McDermid syndrome), as a model for investigating the molecular mechanisms of terminal deletions that are currently poorly understood. We characterized at the molecular level the genomic rearrangement in 44 unrelated patients with 22q13 monosomy resulting from simple terminal deletions (72%), ring chromosomes (14%), and unbalanced translocations (7%). We also discovered interstitial deletions between 17–74 kb in 9% of the patients. Haploinsufficiency of the SHANK3 gene, confirmed in all rearrangements, is very likely the cause of the major neurological features associated with PMS. SHANK3 mutations can also result in language and/or social interaction disabilities. We determined the breakpoint junctions in 29 cases, providing a realistic snapshot of the variety of mechanisms driving non-recurrent deletion and repair at chromosome ends. De novo telomere synthesis and telomere capture are used to repair terminal deletions; non-homologous end-joining or microhomology-mediated break-induced replication is probably involved in ring 22 formation and translocations; non-homologous end-joining and fork stalling and template switching prevail in cases with interstitial 22q13.3. For the first time, we also demonstrated that distinct stabilizing events of the same terminal deletion can occur in different early embryonic cells, proving that terminal deletions can be repaired by multistep healing events and supporting the recent hypothesis that rare pathogenic germline rearrangements may have mitotic origin. Finally, the progressive clinical deterioration observed throughout the longitudinal medical history of three subjects over forty years supports the hypothesis of a role for SHANK3 haploinsufficiency in neurological deterioration, in addition to its involvement in the neurobehavioral phenotype of PMS

704-3 Relation of Neurohormonal Activation to Functional Class in Patients with Primary or Precapillary Secondary Pulmonary Hypertension

Intense neurohormonal activation has been demonstrated in patients with primary or precapillary secondary pulmonary hypertension (PH) but the relation of neurohormons to functional impairment is not well known. Plasma levels of atrial natriuretic peptide (ANP), aldosterone (ALD), renin activity (PRA), epinephrine (PE), norepinephrine (PNE) and endothelin (ET) were assessed from the antecubital vein in 12 patients with primary PH, 7 patients with precapillary secondary PH (2 connective tissue disease, 2 chronic thromboembolic, 3 closed atrial septal defect) and 10 control subjects. Twelve patients were in NYHA functional class II (PH-II) and 7 in class III/IV (PH-III/IV). Mean PA pressure (PAP), cardiac index (CI), pulmonary vascular resistance (PVR) and right atrial pressure were assessed by heart catheterization:PAPCIPVRRAP(mmHg)(l/minlm2)(RU)(mmHg)PH-II53 ± 132.4 ± 0.412 ± 33 ± 2PH-III/IV61 ± 92.0 ± 0518 ± 611 ±5P0.170.070.0090.0001Neurohormons plasma levels in control subjects (C), PH-II and PH-III/IV patients were as follows:ANPALDPRAPEPNEET(pg/ml)(pg/ml)(ng/ml/h)(pg/ml)(pg/ml)(pg/ml)C58 ± 18110 ± 650.7 ± 0.433 ± 19220 ± 1011.7 ± 0.3PH-II167 ± 96*144 ± 10215 ± 2.0298 ± 263*420 ± 294*3.6 ± 1.4*PH-III/IV276 ± 153*†209 ± 2703.2 ± 3.9*462 ± 524*820 ± 693*†8.4 ± 3.9*°*p < 0.05 vs C° p < 0.001 vs PH-II †P < 0.09 vs PH-IIConclusionsNeurohormonal activation in primary or precapillary secondary PH is detectable also in patients without overt clinical and hemodynamic signs of right heart failure (PH-II). Neurohormonal activation seems to be progressive and is more severe in functional class III/IV patients. ET shows the best statistically significant relation with functional impairment