Vincenzo D'Orio

Impact of the presence of a clinical pharmacist in university hospital wards on the elderly or polymedicated patients care

IntroductionThis paper uses data from My Health @ Age (2008-2010), an EU, Northern Periphery funded project. Its aim is to offer the rapidly ageing population in the northern periphery regions of E ...Age and ageing related humour expressed in birthday cards has been suggested to support negative attitudes towards aged people. Little attention has been paid to how retired people themselves exper ...It is well known that reduced ability to perform activities of daily living (ADL) and declining mobility are both related to advanced age. This relation is also valid for low quality-of-life. It is ...The HPA-1 polymorphism of alphaIIbbeta3 arises from a Leu to Pro exchange at residue 33 of the beta3 subunit resulting in HPA-1a (Leu33) or HPA-1b (Pro33). We have documented that patients with coronary artery disease who are carriers of HPA-1b experience their myocardial infarction 5.2 years earlier than HPA-1a/1a patients (JTH 2005; 3: 1522). Based on these observations, it has been postulated that HPA-1b is a prothrombotic variant of alphaIIbbeta3. To explore the molecular nature of this phenotype, we have now generated a model overexpressing fluorescent proteins fused with alphaIIbbeta3 in transfected HEK293 cells. The yellow (YFP) and the cyan fluorescent protein (CFP) were cloned to the C-termini of the beta3 and alphaIIb subunits prior to transfection of HEK293 cells, subsequently expressing the fusion proteins of both HPA-1 isoforms. Using flow cytometry, Western blotting and specific antibodies directed against alphaIIb or beta3, we identified 12 HPA-1a and 11 HPA-1b positive clones expressing equal amounts of fluorescent fusion proteins, i.e. a 140 kD alphaIIb-CFP and a 113 kD beta3-YFP. Functional integrity of both integrin variants and proper membrane insertion were documented by intact activation through G protein-coupled receptors with organic acid, PMA-induced activation of protein kinase C and by specific binding of Alexa647 fibrinogen to alphaIIbbeta3. In the presencence of pertussis toxin or abciximab, activation or ligand binding of alphaIIbbeta3 were completely (> 98%) inhibited in both isoforms. Analysis of Src, a tyrosine kinase associated with alphaIIbbeta3, revealed that activation of the phosphotyrosine motif at residue 418 was higher in adherent HPA-1b than HPA-1a cells (P < 0.01). Upon activation of alphaIIbbeta3, analysis by FRET showed equal kinetics with signal disappearance of 40 5% in both HPA-1 variants. We therefore conclude that the postulated prothrombotic phenotype of HPA-1b (Pro33) is related to increased outside-in signaling rather than to allosteric changes of the C-terminal cytoplasmic tails of alphaIIbbeta3. Disclosure of interest: none declared.