Cholesteryl Ester Transfer Research Articles

Association between plasma HDL-cholesterol concentration and Taq1B CETP gene polymorphism in non-insulin-dependent diabetes mellitus

The effects of CETP gene Taq1B polymorphism on plasma lipoproteins were investigated in 176 patients with non-insulin-dependent diabetes mellitus. The distribution of CETP genotypes was similar to that previously described in the general population. A significant association was found between CETP genotype and both CETP and HDL cholesterol (HDL-c) concentrations. B1B1 had the highest CETP and the lowest HDL-c whereas B2B2 had the lowest CETP and the highest HDL-c. However, HDL-c was not correlated with CETP concentration, even when genetic groups were separately considered. By multivariate analysis, the determinants of HDL were body mass index, triglycerides concentration, net mass CE transfer, and CETP genotype. No association was found between CETP genetic groups and HDL or LDL size distribution. In contrast, net mass CET was positively and HDL and LDL sizes were negatively correlated with plasma triglyceride concentration. Overall, our work demonstrates that, in a population of diabetic patients where lipoprotein-related parameters vary over a large range, the association of CETP gene polymorphism with HDL-c is independent of plasma CETP concentration.—Bernard, S., P. Moulin, L. Lagrost, S. Picard, M. Elchebly, G. Ponsin, F. Chapuis, and F. Berthezène. Association between plasma HDL-cholesterol concentration and Taq1B CETP gene polymorphism in non-insulin-dependent diabetes mellitus. J. Lipid Res. 1998. 39: 59–65.

Suppression of lipid transfer inhibitor protein activity by oleate. A novel mechanism of cholesteryl ester transfer protein regulation by plasma free fatty acids.

Cholesteryl ester transfer protein (CETP) mediates the interlipoprotein exchange of cholesteryl ester (CE) and triglyceride. A second plasma protein, lipid transfer inhibitor protein (LTIP), binds to lipoproteins and inhibits CETP activity by displacing CETP from the lipoprotein surface. Since free fatty acids (FFAs) enhance the binding of CETP to lipoproteins, we have examined the possible role of FFAs in modulating LTIP activity. Partially purified CETP, LTIP, and lipoproteins were incubated with 0 to 30 mumol/L sodium oleate, and the transfer of CE between a labeled donor lipoprotein and a given acceptor lipoprotein was measured. Without LTIP, oleate stimulated CETP-mediated CE transfer between VLDL, LDL, and HDL up to threefold. This stimulation was unique in both magnitude and oleate concentration dependence for each donor-acceptor lipoprotein pair. In contrast to CETP activity, in transfer reactions involving LDL or VLDL as donor, LTIP activity was suppressed (> 80%) by 10 to 15 mumol/L oleate. LTIP activity in transfer reactions with HDL as donor was less sensitive. Similar results to these were observed when lipid transfer reactions were measured in the total lipoprotein fraction isolated from FFA-enriched plasma. The FFA content of lipoproteins was strongly influenced by the concentration of FFA in plasma; lipoprotein FFA levels sufficient to suppress LTIP activity by 50% to 100% were achieved in plasma containing 0.8 to 1.0 mmol/L FFA. We conclude that LTIP may be functionally inactive during periods of transient elevations of plasma FFA levels, such as during postprandial lipemia or overnight fasting, or chronically suppressed in disease states in which plasma FFA levels are increased. The suppression of LTIP activity by FFA allows for maximum CETP-mediated lipid transfer between all lipoproteins, including lipid transfer reactions involving LDL that are normally preferentially suppressed by LTIP.

4.P.349 Effect of alcohol-induced changes in plasma lipoproteins and lipids on cholesteryl ester transfer

4.P.349 Effect of alcohol-induced changes in plasma lipoproteins and lipids on cholesteryl ester transfer

3.P.2 The effect of vitamin E on lipoprotein oxidation and cholesteryl ester transfer

3.P.2 The effect of vitamin E on lipoprotein oxidation and cholesteryl ester transfer

Lipid transfer inhibitor protein activity deficiency in normolipidemic uremic patients on continuous ambulatory peritoneal dialysis.

We previously demonstrated that lipid transfer inhibitor protein (LTIP) is a potent modifier of lipid transfer protein (LTP) function in vitro. Based on these studies, we proposed that LTIP activity is an important determinant of lipoprotein size and composition, which leads to a stimulation of reverse cholesterol transport. To further evaluate this hypothesis, we have studied a normolipidemic, uremic patient population undergoing continuous ambulatory peritoneal dialysis (CAPD) that is deficient in LTIP activity (< 18% of control). LDL from CAPD plasma was triglyceride enriched; the diameters of both CAPD LDL and HDL were increased and CAPD HDL was dominated by the largest subfraction, HDL2b. In CAPD patients, the plasma cholesterol esterification rate was only 61% of control; this decrease was due mainly to the poor reactivity of CAPD lipoproteins. CAPD lipoprotein-deficient plasma promoted twofold greater transfer of radiolabeled cholesteryl ester (CE) between standard lipoproteins than control, although LTP itself was increased only 39%. This twofold increase was not equally expressed among individual lipoprotein classes; CE transfers involving LDL were increased 2.4-fold, whereas those not involving LDL were increased only 50%. In whole plasma, CE net mass transfer to VLDL was slightly increased in CAPD plasma; relative to their CE content, control HDL contributed twofold more CE mass to VLDL than control LDL, but in CAPD plasma this preferential transfer of CE from HDL was absent. Collectively, the aberrations in CAPD lipoprotein composition and metabolism are consistent with the hypothesized role of LTIP. The data further support the role of LTIP in modulating the participation of HDL in CE mass transfers to VLDL. This is the first report of LTIP activity deficiency in humans.

Lack of Effect of Vitamin E on Cholesteryl Ester Transfer and Lipoprotein Composition in Cholesterol-fed Rabbits

The concentration and activity of cholesteryl ester transfer protein (CETP) is increased in plasma in hypercholesterolemic humans and in experimental animals fed cholesterol. While the concentration of lipoproteins appears to be the major determinant of CETP activity, we have found previously that dietary measures and pharmacologic agents that alter their lipid composition reduce the activity of CETP in plasma (CET). Since vitamin E is lipophilic and is incorporated into lipoproteins, we have examined the question of whether it too attenuates CET in cholesterol-fed New Zealand White rabbits prior to and 14 weeks after treatment with differing doses (5, 15, 30, 45 mg/kg) of vitamin E. Plasma triglycerides (TG), cholesterol (TC) and phospholipids (Lys, Sph, Lec, PI, PE) all increased significantly to a comparable degree in the rabbits fed cholesterol compared to those fed chow (p < 0.05; p < 0.01); the levels achieved were similar in the vitamin E-treated and untreated groups. As was observed with plasma lipids, cholesteryl ester transfer (CET) was accelerated to the same degree in each of the cholesterol-fed groups independent of whether they received vitamin E compared to chow-fed controls (p < 0.01) and the distribution of cholesterol in apo-B containing lipoproteins (VLDL, IDL, and LDL) was similar in the vitamin E-treated and untreated groups. These findings indicate that vitamin E has no discernible effect on CET when cholesterol levels are markedly elevated.

Modification of the N-terminal cysteine of plasma cholesteryl ester transfer protein selectively inhibits triglyceride transfer activity

An invariant cysteine residue is found at the N-terminus of cholesteryl ester transfer protein (CETP) isolated from plasma of humans, rabbits and cynomolgus monkeys. We previously reported the expression of recombinant rabbit cholesteryl ester transfer protein in yeast (Kotake et al., J. Lipid Res. 1996; 37: 599–605). The recombinant CETP secreted into the medium contains an altered N-terminal sequence but was fully capable of facilitating both cholesteryl ester (CE) and triglyceride (TG) transfer between lipoproteins. We investigated the importance of the conserved N-terminal cysteine of plasma CETP in the lipid transfer activity by chemical modification of the free sulfhydryl groups of the recombinant CETP and CETP from human and rabbit plasma. The unmodified forms of these CETPs had similar specific activities of CE and TG transfer. Neither 5,5′-dithiobis-(2-nitrobenzoate) nor N-ethyl maleimide altered the lipid transfer activity. In contrast, p-chloromercuriphenyl sulfonate selectively inhibited the TG transfer activity of both human and rabbit plasma CETP. The TG and CE transfer activities of the recombinant CETP, which lacks the N-terminal cysteine residue, was not affected. These results demonstrate that the N-terminal cysteine residue of both human and rabbit plasma CETP is free and is likely to be involved in the construction of a critical part of the active site of CETP that can determine the selectivity of the lipid molecule for the transfer reaction.

Effect of dietary omega-3 eicosapentaenoic acid supplements on cholesteryl ester transfer from HDL in cholesterol-fed rabbits

We investigated the reactivities of cholesteryl ester transfer protein (CETP) in Japanese white rabbits fed either a low-cholesterol diet containing 0.1% cholesterol (Control group) or a diet containing 0.1% cholesterol plus 17.5% ω-3 eicosapentaenoic acid ( ω-3 20:5, EPA) of 4.5% (w/w) total lipid (EPA group) for 6 weeks. The plasma total and LDL cholesterol levels and aortic cholesterol content were all significantly higher in the EPA group than in the control group. The aortic cholesterol content significantly correlated with LDL cholesterol ( r=0.81). HDL cholesterol levels tended to be lower in the EPA group compared with control group, which was not statistically significant. The plasma VLDL cholesterol levels did not differ significantly between the groups. In addition, no significant differences were observed in the plasma CETP activity or lecithin:cholesterol acyltransferase (LCAT) activity between the groups. However, the cholesteryl ester (CE) mass transfer from fractionated HDL in the EPA group to excess VLDL and/or LDL as acceptors by purified CETP increased significantly compared with the control group, even if the acceptors were fractionated from either the EPA or the control group. Fatty acid analyses of CE showed that the ω-3 18:3, 20:4 or ω-3 20:5 fatty acid acyl groups in CE of HDL were significantly more transferred to apo B-containing lipoproteins compared with the 14:0,16:0, 18:0, 18:1 or 18:2 fatty acid acyl groups in CE of HDL during the incubation period. The amount of CE in HDL containing ω-3 18:3 and ω-3 20:5 fatty acid acyl groups was greater, while the amount of CE containing 18:2 fatty acid acyl groups was smaller in the EPA group than in the control group. These results show that although CETP itself did not change, the transfer of CE in HDL to apo B-containing lipoproteins by CETP increased in the rabbits fed a diet containing EPA as the HDL is modified by the diet, which may partly explain why atherogenicity was thus found to progress in the rabbits fed a cholesterol plus EPA diet. © 1997 Elsevier Science B.V. All rights reserved.

Indirect energy transfer of Ce 3+ → Eu 2+ in CaAl 12O 19 phosphor

Photoluminescence characteristics and the energy transfer between Ce 3+ and Eu 2+ in CaAl 12O 19 host lattice have been investigated. A series of concentrations of Ce 3+ ion with a fixed Eu 2+ concentration in doubly doped CaAl 12O 19 : Ce 3+Eu 2+ have been studied. The indirect energy transfer has been observed between Ce 3+ and Eu 2+ through the Ce 3+O Me complex in CaAl 12O 19 host lattice. The two nonradiative relaxation processes from the excited levels of Ce 3+O Me complex; one, to the lower emitting level of Ce 3+O Me complex themselves and the other, to the Eu 2+ site are competing processes.

Systematic approach to links between separations in capillary electrophoresis and liquid chromatography IV. Application of binding constant-retention factor relationship to the separation of 2-, 3- and 4-methylbenzoate anions using β-cyclodextrin as selector

Quantitative links between binding constants determined in CE and retention factors in HPLC for 1:1 analyte-selector complexes, developed in a previous paper and applied to separations of cationic tioconazole enantiomers with β-cyclodextrin (β-CD), have been extended to the separation of anionic positional isomers. In this paper we apply our unified theory to the separation of anionic methylbenzoate anions (MB −, again using β-CD as selector. There is only partial resolution for all of the three isomeric methylbenzoates without β-CD present, either in the CE background electrolyte or immobilised in the HPLC experiments. Using the same buffer as in CE (ammonium phosphaae pH 7.0), and calculating the effective concenyration of selector from the amount of β-CD divided by the volume of the mobile phase in the column, HPLC is shown to give effective binding constants, K′, which are in excellent agreement with binding constants, K, determined from the variation of CE mobility with β-CD concentration. K′=11,41,87 M′ and K=13.0, 45.5, 85.3 M −1 for 2-, 3-, 4-MB − respectively. This demonstrates the quality of our unified treatment of separations based on binding equilibria, and suggests the potential of method development in CE and information transfer to HPLC or vice versa. Quantitative determination of K in CE also generates mobilities of free analyte and analyte-selector complexes, which after correction for non-ideality give radii and diffusion coefficient of the MB − and MB −: β-CD ions. This in turn allows a quantitative treatment of variance contributions from diffusion to CE peak widths. Good agreement is found between calculated and observed peak widths when these are measured with the fastest possible data acquisition parameters.

High-density lipoproteins from probucol-treated patients have increased capacity to promote cholesterol efflux from mouse peritoneal macrophages loaded with acetylated low-density lipoproteins.

To elucidate the anti-atherogenic effect of probucol, high-density lipoprotein (HDL) was isolated from probucol-treated patients (n = 14) and compared with that from control subjects (n = 12). The HDL obtained from probucol-treated patients was low in cholesteryl ester (CE) in comparison with that from control subjects (21.3 +/- 3.9 mol per cent vs. 27.6 +/- 3.2 mol% of total lipids. P < 0.001), and the peak diameters of patients' HDL were significantly smaller than those of control subjects on 4-30% non-denaturing polyacrylamide gradient gel electrophoresis (10.6 +/- 0.6 nm vs. 12.1 +/- 0.4 nm, P < 0.001). These data may be explained by the increased cholesteryl ester transfer protein (CETP) activities of probucol-treated patients (129 +/- 12% of control subjects, P < 0.001). The in vitro ability of HDL to remove CE from lipid-laden macrophages induced by incubation with acetylated low-density lipoprotein (Ac-LDL) was studied. The small and CE-poor HDL obtained from probucol-treated patients had a greater capacity to promote CE efflux from macrophages than did control HDL (59.8 +/- 6.9% vs. 44.2 +/- 5.4%, P < 0.01). Furthermore, the ability of HDL to promote cholesterol efflux correlated negatively with the CE content and particle diameter of HDL (r = -0.561 and r = -0.583 respectively; P < 0.01). When the inhibitory effect of HDL on the incorporation of [14C]-oleate into cellular cholesteryl ester was compared, the HDL from patients and control subjects inhibited CE formation to a similar extent. The enhanced ability of probucol-treated patients' HDL may, therefore, be involved in the acceleration of hydrolysis of the CE pool in macrophages. Taken together, we conclude that CETP plays a crucial role in making HDL more active in its anti-atherogenic function by reducing CE and making HDL smaller, and that probucol may enhance reverse cholesterol transport by activating CE transfer in vivo. The current study demonstrated, for the first time, that HDL modified by enhanced CETP activity in vivo is potentially anti-atherogenic.

Effects of multiple daily insulin injections and intraperitoneal insulin therapy on cholesteryl ester transfer and lipoprotein lipase activities in NIDDM

Effects of multiple daily insulin injections and intraperitoneal insulin therapy on cholesteryl ester transfer and lipoprotein lipase activities in NIDDM

Transfer of dehydroepiandrosterone- and pregnenolone-fatty acid esters between human lipoproteins.

Dehydroepiandrosterone (DHEA)- and pregnenolone (PREG)-fatty acid esters (FA) are formed in plasma high density lipoproteins (HDL), whereas they accumulate in very low density lipoproteins (VLDL), low density lipoproteins (LDL), and HDL. We have hypothesized that these lipoidal steroids could be transferred from HDL to VLDL and LDL by the cholesteryl ester (CE) transfer protein (CETP), which mediates CE transfer activity in human plasma. In this study, we further investigated this hypothesis. Lipoproteins and lipoprotein-deficient plasma (LPDP) were purified and analyzed by Western blots. LPDP was rich in CETP, in contrast to lipoprotein preparations, which contained very low amounts. Using these preparations in in vitro transfer assays, CE transfer from radiolabeled steroid ester-HDL to VLDL or LDL was only observed in the presence of LPDP. In contrast, time- and temperature-dependent transfer of DHEA-FA and PREG-FA were observed in the absence of LPDP. The addition of LPDP had no effect on the DHEA-FA transfer rate, whereas the PREG-FA transfer rate was increased. Moreover, in the absence of LPDP, no decrease in the transfer levels of DHEA-FA and PREG-FA was observed after the removal of CETP from lipoprotein preparations by immunoaffinity column chromatography. The PREG-FA transfer activity of LPDP was studied using the anti-CETP monoclonal antibody TP-2, which is known to block the CE transfer activity of CETP. In the presence of LPDP, this antibody led to a dose-dependent decrease in CE transfer activity, whereas PREG-FA transfer activity was unaffected. In conclusion, we have shown that DHEA-FA and PREG-FA are transferred from HDL to VLDL and LDL by a CETP-independent mechanism. There is a major difference in the transport of lipoidal steroids by human lipoproteins compared to that of CE.

Inhibition of cholesterol ester transfer protein by CGS 25159 and changes in lipoproteins in hamsters

As a result of screening, several isoflavans were identified to be antagonists of cholesterol ester transfer protein (CETP) activity. The present study evaluates CGS 25159, a synthetic isoflavan, as a putative inhibitor of CETP activity of human and hamster plasma. Determined by [3H]CE transfer from HDL to VLDL+LDL fraction or by fluorescent-CE transfer assay, CGS 25159 inhibited CETP in both human plasma bottom fraction (d=1.21 g/ml) and in plasma from Golden Syrian Hamsters with an IC50<10 μM. The compound also inhibited (IC50≈15 μM) the reciprocal transfer of triglycerides in the incubated whole plasma from normal and hyperlipidemic hamsters. When orally administered to normolipidemic hamsters, CGS 25159 (10 mg/kg, 4 days) reduced plasma transfer activity by 35–60%. Treatment with CGS 25159 (10 and 30 mg/kg, p.o.) resulted in dose dependent and time dependent changes in CETP activity. After two weeks of treatment at 10 mg/kg, the changes in VLDL+LDL cholesterol, total triglycerides and HDL cholesterol were −22±4.6*, −23±7.5 and +10±2.8%, respectively. The corresponding changes at 30 mg/kg were −28±5.5*, −38±6.8* and +29±4.4*%, (*, P, 0.05; mean±S.E.M., n=6). A single spin gradient density ultracentrifugation of plasma lipoproteins from treated animals showed an increase in HDL cholesterol and a redistribution to larger HDL particles. These data support the contention that pharmacological down regulation of CETP activity could result in favorable changes in lipoprotein profile.

Physical and kinetic characterization of recombinant human cholesteryl ester transfer protein.

Cholesteryl ester transfer protein (CETP) mediates the exchange of triglycerides (TGs), cholesteryl esters (CEs) and phospholipids (PLs) between lipoproteins in the plasma. In order to better understand the lipid transfer process, we have used recombinant human CETP expressed in cultured mammalian cells, purified to homogeneity by immunoaffinity chromatography. Purified recombinant CETP had a weight-average relative molecular mass (MW) of 69561, determined by sedimentation equilibrium, and a specific absorption coefficient of 0.83 litre.g-1.cm-1. The corresponding hydrodynamic diameter (Dh) of the protein, determined by dynamic light scattering, was 14 nm, which is nearly twice the expected value for a spheroidal protein of this molecular mass. These data suggest that CETP has a non-spheroidal shape in solution. The secondary structure of CETP was estimated by CD to contain 32% alpha-helix, 35% beta-sheet, 17% turn and 16% random coil. Like the natural protein from plasma, the recombinant protein consisted of several glycoforms that could be only partially deglycosylated using N-glycosidase F. Organic extraction of CETP followed by TLC showed that CE, unesterified cholesterol (UC), PL, TG and fatty acids (FA) were associated with the pure protein. Quantitative analyses verified that each mol of CETP contained 1.0 mol of cholesterol, 0.5 mol of TG and 1.3 mol of PL. CETP mediated the transfer of CE, TG, PL, and UC between lipoproteins, or between protein-free liposomes. In dual-label transfer experiments, the transfer rates for CE or TG from HDL to LDL were found to be proportional to the initial concentrations of the respective ligands in the donor HDL particles. Kinetic analysis of CE transfer was consistent with a carrier mechanism, having a Km of 700 nM for LDL particles and of 2000 nM for HDL particles, and a kcat of 2 s-1. The Km values were thus in the low range of the normal physiological concentration for each substrate. The carrier mechanism was verified independently for CE, TG, PL and UC in 'half-reaction' experiments.

Role of Lp A-I and Lp A-I/A-II in cholesteryl ester transfer protein-mediated neutral lipid transfer. Studies in normal subjects and in hypertriglyceridemic patients before and after fenofibrate therapy.

The two major subclasses of HDL contain apo A-I only (Lp A-I) or both apo A-I and apo A-II (Lp A-I/A-II). We have carried out experiments to quantify the participation of Lp A-I and Lp A-I/A-II in the neutral lipid transfer reaction in normal and hypertriglyceridemic subjects. Thirteen hypertriglyceridemic subjects were studied before and after fenofibrate therapy. Fenofibrate treatment resulted in decreases in total cholesterol, triglycerides (TG), and VLDL cholesterol of 19%, 48%, and 70%, respectively, and a 28% increase in HDL cholesterol, with no significant change in the proportion of Lp A-I and Lp A-I/A-II particles. The abundance of cholesteryl ester transfer protein (CETP) mRNA in peripheral adipose tissue decreased with treatment in four of five patients studied; however, no change occurred in plasma CETP mass. Using an isotopic transfer assay, we demonstrated that both Lp A-I and Lp A-I/A-II participated in the CE transfer reaction, with no change after fenofibrate therapy. This finding suggests that the marked increase in HDL cholesterol during fenofibrate therapy is due to normalization of plasma TG and hence decreased opportunity for mass transfer of lipid between HDL and TG-rich proteins in vivo. In this population of hypertriglyceridemic subjects, CETP was distributed in both the Lp A-I and Lp A-I/A-II subfractions of HDL, with preferential association with the smaller Lp A-I poor. In contrast, in nine normal subjects studied, negligible amounts of CETP were associated with Lp A-I/A-II. Nonetheless, the Lp A-I/A-II fraction of HDL contributed significantly to total CE mass transfer in normolipidemic plasma. Lp A-I/A-II is an efficient donor for CE transfer to TG-rich lipoproteins, and its low affinity for CETP may in fact facilitate neutral lipid transfer either by a shuttle mechanism or by formation of a ternary complex.

An inhibitor of the microsomal triglyceride transfer protein inhibits apoB secretion from HepG2 cells.

The microsomal triglyceride (TG) transfer protein (MTP) is a heterodimeric lipid transfer protein that catalyzes the transport of triglyceride, cholesteryl ester, and phosphatidylcholine between membranes. Previous studies showing that the proximal cause of abetalipoproteinemia is an absence of MTP indicate that MTP function is required for the assembly of the apolipoprotein B (apoB) containing plasma lipoproteins, i.e., very low density lipoproteins and chylomicrons. However, the precise role of MTP in lipoprotein assembly is not known. In this study, the role of MTP in lipoprotein assembly is investigated using an inhibitor of MTP-mediated lipid transport, 2-[1-(3, 3-diphenylpropyl)-4-piperidinyl]-2,3-dihydro-1H-isoindol-1-o ne (BMS-200150). The similarity of the IC50 for inhibition of bovine MTP-mediated TG transfer (0.6 microM) to the Kd for binding of BMS-200150 to bovine MTP (1.3 microM) strongly supports that the inhibition of TG transfer is the result of a direct effect of the compound on MTP. BMS-200150 also inhibits the transfer of phosphatidylcholine, however to a lesser extent (30% at a concentration that almost completely inhibits TG and cholesteryl ester transfer). When BMS-200150 is added to cultured HepG2 cells, a human liver-derived cell line that secretes apoB containing lipoproteins, it inhibits apoB secretion in a concentration dependent manner. These results support the hypothesis that transport of lipid, and in particular, the transport of neutral lipid by MTP, plays a critical role in the assembly of apoB containing lipoproteins.

Differing Effects of Pancreas-kidney Transplantation With Systemic Versus Portal Venous Drainage on Cholesteryl Ester Transfer in IDDM Subjects

Cholesteryl ester transfer (CET) is accelerated in patients with IDDM treated with conventional (subcutaneous) insulin therapy (CIT) and a number of other disorders associated with premature cardiovascular disease. We have shown that in IDDM this disturbance is closely linked to iatrogenic hyperinsulinemia (HI), because it was reversed when insulin was administered by the intraportal (i.p.) route. In this study, we sought to determine whether HI after successful pancreas-kidney transplantation (PKT) has the same adverse effect on CET. CET was measured by both mass and isotopic assays and compared in two groups of euglycemic non-insulin-requiring IDDM PKT patients with either systemically draining allografts and persistent HI or grafts with portal vein anastomoses that were normoinsulinemic (PK-P). A third group of eight nondiabetic kidney transplant (KT) patients receiving the same immunosuppressive drugs served as control subjects. CET in pancreas-kidney transplantation subjects with systemic venous drainage (PK-S) was increased (P < 0.001) to the same level we have reported previously in IDDM patients receiving CIT and was significantly higher (P < 0.001) than in those subjects with PK-P. CET in the PK-P group did not differ from that of the KT control patients. CET is affected by variations in systemic insulin levels in pancreas transplant patients with allografts that have differing venous drainage. Because high systemic insulin levels are linked to the activation of (ET, euglycemic HI IDDM pancreas allograft recipients may continue to be at high risk for macrovascular complications.

Plasma cholesteryl ester synthesis, cholesteryl ester transfer protein concentration and activity in hypercholesterolemic women: effects of the degree of saturation of dietary fatty acids in the fasting and postprandial states

Hypercholesterolemic women ( n = 19) sequentially maintained on a long-term saturated (SAT) or a polyunsaturated (PUFA) fatty acid-rich diet, respectively, were studied in the fasting state and after a meal rich in SAT or PUFA. When apo B-containing lipoprotein was excluded from plasma the in vitro HDL- 14C-cholesterol esterification rate was identical for the saturated (SAT) and polyunsaturated (PUFA) fatty acid diets, and did not increase during the postprandial period. Rates of transfer of 14C-cholesteryl ester to apo B-containing lipoproteins from HDL were also similar for both diets in the fasting state and increased to the same extent in the postprandial period in parallel with the rise in plasma triglycerides. When transfer data were related to the plasma concentration of apo B, the gain of cholesteryl ester by the triglyceride-containing particles (VLDL + LDL) also increased in the postprandial period to a similar extent for both diets. Cholesteryl ester transfer protein (CETP) concentration measured by radioimmunoassay was similar during both experimental diets, although greater in the postprandial period for the PUFA diet. The rate limiting factor for CETP-mediated transfer of HDL-derived cholesteryl ester (CE) was the plasma triglyceride concentration, that is, the content of triglycerides per lipoprotein particle and the quantity of TG-containing particles (VLDL + LDL). In contrast, the fatty acid composition of these particles had less effect on CETP-mediated CE transfer.

Plasma cholesteryl ester transfer activity is modulated by the phase transition of the lipoprotein core

Previous studies have shown that lipid transfer protein (LTP) activity is strongly temperature dependent, demonstrating a dramatic rise in activity near 37 degrees C. We have investigated the origin of this rapid rise in LTP activity. LTP-mediated transfers of radiolabeled cholesteryl ester (CE) from LDL to HDL, HDL to LDL, LDL to biotin-LDL, HDL to biotin-HDL, and between liposomes were determined as a function of assay temperature. Only assays containing LDL demonstrated this rapid rise in CE transfer activity. In contrast, TG transfer was almost linear with assay temperature. As human LDL CE undergoes a thermal phase transition near 37 degrees C, we investigated whether the rapid rise in CE transfer was dependent on this transition. Monkey LDL were isolated from animals consuming diets containing cholesterol and enriched in saturated, monounsaturated, or polyunsaturated fatty acids. With these LDL as substrate, the CE transfer between 21 degrees and 49 degrees C could be described by two straight lines, the intersection of which defined the inflection temperature. Among eight LDL samples, the inflection temperature was highly correlated with the CE phase transition determined by differential scanning calorimetry (r2 = 0.86). Both calorimetry and CE transfer activity inflection values were correlated with the saturated + monoene/polyene ratio of the LDL cholesteryl esters (r2 = 0.733 and 0.612, respectively). For LDL with inflection temperatures below 37 degrees C, CE transfer activity at 37 degrees C increased 10-14% for each 1 degree C decrease in the inflection temperature. We conclude that LTP activity is markedly affected by the physical state of the core CE. Diets rich in saturated fatty acids may result in LDL that are poor LTP substrates, which may hinder LTP's ability to promote normal lipoprotein remodeling.