Fabien Milliat

Plasma lipoprotein distribution in the turkey (Meleagris gallopavo)

The plasma lipoprotein profile has been determined in fasted 7-week-old male turkeys. Lipoprotein classes were subfractionated by density gradient ultracentrifugation. According to phospholipid concentration over the density gradient, an initial peak was visible in the usual LDL density range, whereas two peaks were detected in that of HDL. As density increased, the lipid composition of particles showed an increase in cholesteryl esters and decrease in triglycerides. VLDL were recovered in the first fraction (d<1.013) on the top of the gradient and IDL in fractions 2-5 (d=1.013-1.028 g/ml). The LDL and HDL populations in the density range 1.028-1.090 (fractions 6-12) differ from that found in the other bird species analyzed under the same experimental conditions. LDL predominated in fractions 6-8 with mostly beta-motility and apoB100 as the major protein component. HDL predominated in fractions 10-12 (d=1.055-1.090 g/ml) and corresponded to the first HDL peak (HDL-(A)), with mostly alpha-mobility and apoA-I as the major protein component. Both LDL- and HDL-like particle populations were present in fractions 6-12, making the separation between the two classes of lipoproteins difficult. The second peak in the HDL density range (HDL-(B), d=1.076-1.146 g/ml) contained only HDL-type particles above d=1.090 g/ml. This points out the specificity of the lipoprotein distribution in the turkey that is unique among animals. The density limit at d=1.048 g/ml is a good compromise for the separation of LDL from HDL; however, the presence of HDL-like particles in the LDL density range, and the existence of two, and even three HDL subclasses should be taken into account in the design of further metabolic studies.

Intralipid 10%: Physicochemical Characterization

OBJECTIVES Parenteral fat emulsions contain two populations of particles: artificial chylomicrons rich in triacylglycerols (TAG), and liposomes (bilayer of phospholipids [PL] enveloping an aqueous phase). Centrifugation permits isolating the liposomes in the infranatant called mesophase. The aim of the present work was to better characterize this mesophase chemically and to view the particles it contains by electron microscopy. METHODS Electron microscopy (Philips 410) was performed after cryofracture on native 10% Intralipid, mesophase (centrifugation for 1 h at 27 000 g), and a liposome-enriched fraction (ring of density 1.010-1.030 g/l obtained after centrifuging mesophase in a KBr density gradient at 100 000 g for 24 h). The TAG and protein content of the mesophase was analyzed and the proteins partially characterized by immunodetection (Western-blot). RESULTS This electron microscope study of 10% Intralipid gives evidence for the coexistence of artificial chylomicrons (mean diameter, 260 nm) and liposomes (43 nm), the latter being smaller than expected and containing 8% w/w TAG after purification. The solubilization of TAG in PL bilayers (reported to be < or = 3.1% w/w) might have been increased in parenteral emulsions by the manufacturing process or/and the high TAG/PL ratio. Minute amounts of proteins have also been detected and partially characterized using a specific antibody raised against the human 7 kDa Anionic Polypeptide Factor (APF), known to strongly interact with PL in bile. CONCLUSIONS This work has shown that the size (mean diameter, 43 nm) of the liposomes present in 10% Intralipid is smaller than that usually assumed. Traces of hydrophobic proteins in the emulsion may account for certain allergic reactions sometimes observed in infused patients.

Overexpression of SR-BI in hamsters treated with a novel ACAT inhibitor (F12511).

The effect of a novel acyl-coenzyme A: cholesterol acyltransferase (ACAT) inhibitor on cholesterol metabolism was studied in hamsters. Oral administration of F12511 (10 mg/kg/d) for 4 weeks produced a decrease in dietary cholesterol absorption (-18%) and in the liver concentration of esterified cholesterol (-75%), as compared with control values in untreated hamsters. While the hepatic expression of LDLr was unchanged by the treatment, that of SR-BI was increased (+142%), which suggests that the hepatic expression of SR-BI could be upregulated by a depletion of the cholesterol stores, due to ACAT inhibition. This SR-BI overexpression, however, did not induce a fall in plasma HDL-cholesterol concentration, in contrast with previous reports in transgenic mice overexpressing SR-BI at a higher extent.

Effects of insulin deficiency on lipoproteins and their hepatic receptors in Rico rats.

The present study was designed to examine the effect of streptozotocin (STZ)-induced diabetes on the plasma lipoprotein profile and hepatic expression of the LDL receptor and HDL binding protein (HB2) in hypercholesterolemic Rico rats. The plasma level of HDL1 (density range 1.040-1.063), which is particularly high in this rat strain, decreased (-25%) 28 d after STZ administration (50 mg/kg). In contrast, the treatment increased (+54%) the plasma concentration of HDL2 (density range 1.063-1.210). These variations in the lipoprotein concentrations were associated with inverse changes in the hepatic protein levels of the LDL receptor (+118%) and HB2 (-46%). These results suggest that the hepatic expression of HB2, a putative HDL receptor, can influence the plasma level of apo Al-rich HDL as has already been shown for the LDL receptor for apo B/E containing lipoproteins.