With GT, BSS, SSD, parallel controls (drawn simultaneously to patients), and controls drawn at the time of analysis. All values are mean ?standard deviation. *Always PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/8086425 those cases blood specimens were only collected from fresh (not parallel) controls. Abbreviations: ADP Adenosine diphosphate, BSS Bernard Soulier Syndrome, GT Glanzmann Thrombasthenia, TRAP thrombin receptor activating peptide, PMA phorbol myristate acetate, SSD platelet secretion and signal transduction defects.S chez-Guiu 5-Fluoro-3-nitropyridin-2(1H)-one et al. Orphanet Journal of Rare Diseases (2014) 9:Page 7 ofsite mutation in the IIb gene inherited from the mother- were paternal in origin and cosegregated in the same allele across three generations [31]. In our patient, however, throughout investigation of other family members (father, mother, brother) did not detect the two missense mutations in any of the individuals, and DNA profiles generated from blood from the patient and parents revealed double paternal/maternal contribution at different loci in the propositus, confirming the maternal and parental relationship. To define the structural impact of the new missense mutation p.Tyr190Cys, identified in two of our patients (patients GT-4 and GT-11), the role of the amino acid PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/9282946 substitutions in 3 was analyzed in silico. The three dimensional diagram of the Cys190 suggested a disruption of the hydrogen bond created in between Tyr190 and Tyr115. This change would also generate new hydrogen bond with Gly189, predicting a destabilizing effect on the fold (data not shown).Bernard Soulier syndromeThis syndrome was diagnosed in eight individuals (2 men and 6 women with a median age of 40 years) (Tables 1 and 5). In these patients, platelet count was consistently less than 20 ?109 L-1 by commercial cell counters, with 1-(3-Aminopiperidin-1-yl)ethan-1-one hydrochloride persistent findings of giant platelets in the peripheral blood smear. Patients had slightly diminished aggregation responses to TRAP compared to their parallel controls, but RIPA was absent or severely defective in all patients (Table 4). Flow cytometry revealed that the GPIb and GPIX expression was severely reduced (<10 compared to controls). MFI of other platelet GPs were normal or increased due to the increased size of the cells. Further studies using Western-blot analysis revealed an absent or severely reduced GPIb band in all patients (data not shown). More than half of the patients (BSS-2, BSS-5, BSS-6, BSS-7 and BSS-8) had been splenectomized without having any benefit, as they had been previously misdiagnosedwith primary immune thrombocytopenia (ITP). The severity grade, measured by BS for epistaxis and menorrhagia was 2 3 in 100 of patients. Other less prevalent locations of moderate-severe blood loss were cutaneous bleeding (50 of patients), and following procedures of dental extraction, surgery, or delivery (57 , 71 , and 75 of patients, respectively). Mild bleeding symptoms were reported at other sites (bleeding from minor wounds, oral cavity, or gastrointestinal tract). The overall BS in BSS patients was 11.8 (Table 1). Disease-causing homozygous mutations were found in 7 out of 8 patients -five lacking a history of known family consanguinity-, while in the remaining patient a mutation was i.