AP500B Sigma-AldrichGoat Anti-Mouse IgM µ chain Antibody, biotin conjugate, Species Adsorbed

Diabetes accelerates atherosclerosis through undefined molecular mechanisms. Hyperglycemia induces formation of advanced glycation end product (AGE)-modified low-density lipoprotein (LDL). Anti-AGE-LDL autoantibodies favor atherosclerosis (AS) progression in humans, while anti oxidized LDL immunization inhibits AS in hypercholesterolemic, non-diabetic mice. We here investigated if AGE-LDL immunization protects against AS in diabetic mice.After diabetes induction with streptozotocin and high fat diet, both low density lipoprotein receptor (LDLR)-/- and apoE female mice were randomized to: AGE-LDL immunization with aluminum hydroxide (Alum) adjuvant; Alum alone; or PBS.AGE-LDL immunization: significantly reduced AS; induced specific plasma IgM and IgG antibodies; upregulated splenic Th2, Treg and IL-10 levels, without altering Th1 or Th17 cells; and increased serum high density lipoprotein(HDL) while numerically lowering HbA1c levels.Subcutaneous immunization with AGE-LDL significantly inhibits atherosclerosis progression in hyperlipidemic diabetic mice possibly through activation of specific humoral and cell mediated immune responses and metabolic control improvement.

The intracellular uptake of nanoparticles (NPs) is an important process for molecular and cellular labeling, drug/gene delivery and medical imaging. The vast majority of investigations into NP uptake have been conducted using confocal imaging that is limited to observation of a small number of cells. Such data may not yield quantitative information about the cell population due to the tiny sample size and the potential heterogeneity. Flow cytometry is the technique of choice for studying cell populations with single cell resolution. Unfortunately, classic flow cytometry detects fluorescence from whole cells and does not shed light on subcellular dynamics. In this report, we demonstrate the use of microfluidics-based total internal reflection fluorescence flow cytometry (TIRF-FC) for examining initial quantum dot (QD) entry into cells and the associated subcellular movement at the single cell level with a rate of ?200 cells s(-1). Our cytometric tool allows extraction of quantitative data from a large cell population and reveals details about the QD transport in the periphery of the cell membrane (?100 nm deep into the cytosol). Our data indicate that the fluorescence density at the membrane vicinity decreases after initial QD dosage due to the decline in the density of QDs in the evanescent field and the transport into the cytosol is very rapid.