vider of these samples is covered as defined in the HIPPA Act of 1996 as is providing them to us with a limited data set of protected health information. Blood was collected in vacuum tubes containing sodium heparin. The tubes were centrifuged at 2000 g67 minutes, and the plasma was then removed and aliquoted for storage at 280uC. All samples were obtained in the course of IRB-approved studies, following the documentation of informed consent in accordance with university policy at both Loma Linda University and the University of California at Irvine. Human Survivin Immunoassay Whole plasma samples were subjected to a commercially available human Survivin Immunoassay using the manufacturer’s instructions in order to quantitate Survivin concentrations. Exosome Isolation Plasma microvesicles were isolated as previously described, with minor modifications. Thawed, cryopreserved plasma was centrifuged for 30 min at 5006 g, 45 min at 12,0006 g and 18 h at 110,0006 g. Pellets were resuspended in a large volume of PBS, filtered through a 0.22-mm filter and centrifuged at 110,0006 g for 1 h. Microvesicle pellets were washed once in a large volume of PBS, centrifuged at 110,0006 g for 1 h and re-suspended in 50200 ml of PBS. The amount of 110,0006g pellet proteins recovered was measured using the BCA protein assay kit. Exosomes were used as fresh preparations for immunoblotting or were conserved at 280uC for later use. For serum samples, the commercially available ExoQuick was MedChemExpress GFT-505 employed as described by the vendor. Prostate Cancer Exosomes Contain Survivin Western Blot Analysis For Western blot analysis, cells or exosomal preparations were lysed using lysis buffer, 1% NP40, 0.25% 
DOC, 150 mM NaCl2, 1 mM PMSF, 10 mg/ml Aprotinin/ leupeptin/pepstatin, 20 mM NaF, 0.2 mM EGTA, 1 mM EDTA, H2O). For protein concentrations the BCA assay was used. Proteins from exosomes were separated using 12% BisTris polyacrylamide gels, transferred onto polyvinylidene difluoride membranes and probed using the following antibodies: mouse monoclonal anti-LAMP1, and rabbit polyclonal anti-Survivin. Secondary antibodies were goat anti-rabbit and goat anti-mouse immunoglobulin. Immunoreactive bands were detected using the Odyssey Imaging System and quantified using ImageQuant software. Statistical Analysis Multiple comparisons among different groups were calculated by using Multiple Analysis of Variance. Student t-test was used to evaluate the significance of changes between control groups and experimental groups. Probability values P,0.05 were considered statistically significant. doi:10.1371/journal.pone.0046737.t005 at 4uC followed by centrifugation at 15006g for 30 minutes. After centrifugation the exosomes appear as a beige or white pellet at the bottom of the vessel which is then reconstituted with 500 mL of dH2O. ~~ Hypercholesterolemia and low high density lipoprotein cholesterol contribute to coronary heart disease but little is known about their direct effects on myocardial function. Based on echocardiographic data, an early cardiomyopathy characterized by systolic and diastolic dysfunction was described in patients with primary hypercholesterolemia without evidence of coronary heart disease. In patients with a first myocardial infarction, postinfarct left ventricular ejection fraction is adversely influenced by elevated non-HDL cholesterol levels and lower HDL cholesterol levels irrespective of the severity of coronary atherosclerosis. Low HDL and raised non-HDL c