Plasma cholesteryl ester transfer protein and lipoprotein levels during treatment of growth hormone-deficient adult humans

The incidence of atherosclerosis is increased in growth hormone (GH) deficient-individuals. Nonetheless, the antiatherogenic benefits of GH replacement therapy remain uncertain. In this study the effect of human recombinant growth hormone (hrGH) replacement therapy administered to GH-deficient adults on the plasma cholesteryl ester transfer protein (CETP) concentration and activity was analyzed. These findings were related to changes in the concentrations of the plasma lipoproteins. The hrGH was administered for 12 mon to human GH-deficient patients (n=13; 8 men, 5 women). During the study plasma lipoproteins were separated by ultracentrifugation, and plasma cholesterol esterification rate (CER), endogenous CETP activity, and CETP concentration were measured. GH replacement therapy transiently (at 3 mon) lowered plasma concentration of CETP and low density lipoprotein-cholesterol (LDL-C) and raised total triglycerides. Furthermore, hrGH permanently increased both the plasma lipoprotein(a) [Lp(a)] concentration, which is known as atherogenic, and the proportion of cholesteryl ester in the high density lipoprotein₂ (HDL₂) particles, which is potentially atheroprotective. The simultaneous decrease of the plasma CETP and LDL-C concentrations elicited by hrGH indicated a close relationship between LDL metabolism and the regulation of the CETP gene expression. Endogenous CETP activity and the CER were not modified because these parameters are regulated in opposite ways by plasma levels of triglycerides; that is, CER increased and CETP decreased.     

Purification and properties of aortic cholesteryl ester hydrolase

The enzyme(s) present in acetonedried powder of rat and rabbit aortas, which catalyzes the synthesis and hydrolysis of cholesteryl ester, was purified partially by acid precipitation, acetone fractionation, 0-(diethylaminoethyl) cellulose chromatography, and Sephadex G-100 filtration. The synthetic activity was purified by 120-fold (rat) and 140-fold (rabbit). Purification of hydrolytic activity was 90-fold (rat) and 103-fold (rabbit). Cholesteryl ester hydrolase activity was separated from nonspecific, esterase by column chromatography. Both synthetic and the hydrolytic activities are apprently the functions of one enzyme. The mol wt of the enzyme was estimated to be 140,000 dalton as determined by Sephadex G-200 gel filtration. The extracts of the acetone-dired powders of aortas of both species contained an inhibitor of synthetic activity. The inhibitor was nondialyzable and was precipitated at pH 5.7 Both activities were found to be fairly nonspecific with regard to sterol and fatty acids. With oleic acid, the relative rates of sterol ester synthesis were: cholesterol, 100; cholestanol, 94; desmosterol, 35; corprostanol, 24; ergosterol, 20; and β-sitosterol, 19. Epicholesterol was not esterified. Oleic acid was most active in cholesteryl ester synthesis, the relative rates being: oleic>linoleic>arachidonic>palmitic>stearic>butyric. The rate of hydrolysis was maximum with cholesteryl linoleate followed by oleate, linolenate, palmitate, stearate and laurate in decreasing order.     

Association between cholesteryl ester transfer protein Taq1B polymorphism with lipid levels in primary hyperlipidemic patients

Primary hyperlipidemia, characterized by hypertriacylglycerolemia and/or hypercholesterolemia, is considered to be one of the most important risk factors for atherosclerosis and coronary heart disease. This study was performed by using polymerase chain reaction and restriction fragment length polymorphism analysis. We measured lipids and cholesteryl ester transfer protein (CETP) activity in primary hyperlipidemic and normolipidemic subjects, with and without Taq1B polymorphism. Genotype distribution and allelic frequencies of polymorphism were determined and compared in both groups. Our results showed that plasma CETP activity was significantly higher in primary hyperlipidemia than in controls (p = 0.001). Plasma lipids were also remarkably increased in primary hyperlipidemic subjects. In both patient and control groups, individuals with B₁B₁ and B₁B₂ genotypes had higher plasma CETP activity, lower total cholesterol, lower high‐density lipoprotein cholesterol and higher triacylglycerol than those with the B₂B₂ genotype. The values of low‐density lipoprotein cholesterol were significantly increased in primary hyperlipidemic patients with the B₂B₂ genotype. The genotype and allelic frequencies for this polymorphism differed significantly between primary hyperlipidemic patients and controls (p = 0.022 and p = 0.039, respectively). Taq1B polymorphism of the CETP gene was associated with changes in lipid profile and plasma CETP activity in the selected population.     

不对称催化中的手性配体和底物之间的相互作用

讨论了手性配体和底物之间的相互作用及其在对映选择催化中的应用。     

Regulation by long-chain fatty acids of the expression of cholesteryl ester transfer protein in HepG2 cells

Cholesteryl ester transfer protein (CETP) is an important determinant of lipoprotein function, especially high density lipoprotein (HDL) metabolism, and contributes to the regulation of plasma HDL levels. Since saturated and polyunsaturated fatty acids (FA) appear to influence the CETP activity differently, we decided to investigate the effects of FA on the expression of CETP mRNA in HepG2 cells using an RNA blot hybridization analysis. Long-chain FA (>18 carbons) at a 0.5 mM concentration were added to the medium and incubated with cells for 48 h at 37°C under 5% CO₂. After treatment with 0.5 mM arachidonic (AA), eicosapentaenoic (EPA), and docosahexaenoic acid (DHA), the levels of CETP mRNA were less than 50% of the control levels (AA, P=0.0005; EPA, P<0.01; DHA, P<0.0001), with a corresponding significant decrease in the CETP mass. These results suggest that FA regulate the gene expression of CETP in HepG2 and this effect is dependent upon the degree of unsaturation of the acyl carbon chain in FA.     

The activity and properties of a hepatic acid cholesteryl ester hydrolase obtained from rats of different age groups

The activity of lysosomal acid cholesteryl ester hydrolase (acid CEH, EC 3.1.1.13) in rat liver was determined at 3, 5, 7, 10 and 20 wk following birth. The levels of acid CEH activity showed a marked decrease as rats grew older, whereas those of other lysosomal marker enzymes, such as acid phosphatase, β-glucuronidase and cathepsin B and D, showed only a slight decrease. On the other hand, acid CEH activity was detected in all subcellular fractions obtained from rat liver, but the enzyme activity in these fractions did not show the age-related decrease observed in the lysosomal fraction. The results presented here suggest that the marked alteration of lysosomal acid CEH activity that accompanies aging may be related to its possible involvement in the regulation of cholesterol concentration in rat liver.     

A micro‐method for lipoprotein cholesterol profiles: Impact of CETP in KKAy mice

The aim of the present study was to assess cholesterol‐containing lipoprotein profiles in minute serum samples. The lipoprotein profiles of KKA^y and transgenic KKA^y‐CETP mice and of other species were determined. The transgenic KKA^y‐CETP mice express the simian enzyme cholesteryl ester transfer protein (CETP). The serum profile of the cholesterol‐containing high‐density (HDL), low‐density (LDL) and very‐low‐density lipoproteins (VLDL) was monitored on a Superose 6 column using fast protein liquid chromatography. Serum from several mouse and rat strains, rabbit, hamster, pig and man was included for comparative and method validation purposes. The chromatograms showed that the transgenic KKA^y‐CETP mice had significantly decreased relative levels of HDL vs. VLDL and LDL cholesterol (p <0.001). Introduction of the CETP gene shifted the serum profile of the cholesterol‐containing lipoproteins of the KKA^y‐CETP mice closer to the human profile in a dose‐dependent manner, thus making these mice an interesting model for man. The described lipoprotein separation technology offers promising and reliable opportunities for studies of blood lipoprotein profiles with minute serum samples, in both animals and man.     

Processing and properties of co-injected resin transfer molded vinyl ester and phenolic composites

Vacuum Assisted Resin Transfer Molding type processes have been proven to be cost effective manufacturing techniques for large composite structures. However, their use has been limited to a single resin system. A large variety of composite structures require multiple resins to serve different purposes while being integrated into a single structure. Co-Injection Resin Transfer Molding (CIRTM) is a new manufacturing process, developed at the University of Delaware's Center for Composite Materials in collaboration with the U.S. Army Research Laboratory, that enables the user to manufacture multi-layer hybrid composite parts in a single processing step (1). In this paper, CIRTM is used to manufacture a dual layered structure consisting of a vinyl ester layer for structural integrity and a phenolic layer for fire, smoke, and toxicity protection. The two resins are simultaneously injected into a mold filled with a stationary fiber bed and are co-cured. Resin separation is maintained by a 0.0254 mm (0.001 in) thick polysulfone film sandwiched between two layers of 0.165 mm (0.0065 in) thick adhesive. A Differential Scanning Calorimeter (DSC) is used to select the optimum cure cycle for all of the materials. Mechanical testing is used to evaluate the performance of the interphase formed between dissimilar materials. Short beam shear (SBS) is used to evaluate the overall quality of the part produced. Double cantilever beam (DCB) is used to quantify the fracture toughness of the interphase, and the wedge test is used to evaluate the durability of the interphase. Experimental results show that co-injected, co-cured materials offer properties equivalent, or in some cases, superior, to those provided by single injection resin composites. This case is used to develop and present a methodology that can be followed to co-inject different resins.     

Cholesterol metabolism in the rat lactating mammary gland: The role of cholesteryl ester hydrolase

An acid cholesteryl ester hydrolase activity associated with a fraction containing mitochondria and lysosomes from rat lactating mammary glands was found to have a pH optimum of 5.0. Its sedimentation pattern was closely related to that of the lysosomal enzyme markers acid phosphatase and β-glucuronidase, suggesting that the activity is associated with the lysosomes. The enzyme was strongly inhibited by Cu²⁺, but was inhibited little by other divalent metal ions. Acid cholesteryl ester hydrolase activity was almost completely abolished byp-hydroxymercuribenzoate, but this effect was reversed in the presence of an equimolar concentration of reduced glutathione (GSH), indicating that the enzyme requires free sulfhydryl groups for activity. These properties are similar to those of acid, lysosomal cholesteryl ester hydrolases found in other tissues. Acid cholesteryl ester hydrolase activity was 8–14 fold higher in mammary tissue from lactating as compared to virgin rats. Neutral cholesteryl ester hydrolase activities associated with the microsomal and cytosolic subcellular fractions were also increased in lactating glands, but to a lesser extent. In addition, a 2-fold increase in the activities of both the acid and microsomal neutral enzymes was seen during the first few days of lactation, while the cytosolic neutral activity remained constant. These results suggest that mammary gland cholesteryl ester hydrolases have a role in the regulation of cholesterol metabolism in mammary cells, and in the provision of cholesterol for secretion into milk.     

The inhibition of neutral cholesteryl ester hydrolase by a cytosolic protein factor in female rat liver: The influence of varying hormonal and nutritional conditions on the inhibitory activity

A cytosolic protein, that is inhibitory to neutral cholesteryl ester hydrolase, has been investigated in the livers of female rats using microsomes isolated from the mammary gland of lactating rats as an enzyme source. To facilitate comparisons, inhibitory activity is expressed in terms of the amount (μg) of cytosolic protein required to reduce esterase activity by 50% and is compared to the hepatic content of both cholesterol and cholesteryl esters. The experiments revealed a sexual difference in the level of inhibitory activity, with the livers of both suckling and mature male animals containing less of the material than the corresponding females. Alterations in the physiological status of the females, such as pregnancy and lactation, led to a decrease in the activity of the protein. This was reversed by blocking lactation with a combination of an antiserum to rat growth hormone and the anti-prolactin drug, bromcoriptine, but not by premature weaning of the animals. Food withdrawal for 24 hr also had the effect of increasing inhibitory activity. In general the cholesteryl ester content of the livers correlated with the level of inhibitory activity. Thus the activity of the cytosolic inhibitor of neutral cholesteryl ester hydrolase responded to changes in both the hormonal and the nutritional status of the female animal. It is suggested that the presence of the greater cholesteryl ester hydrolase inhibitory activity in the female liver may help to explain the lower risk of coronary heart disease in premenopausal females by facilitating increased hepatic storage of the sterol in the form of the ester.     

Esterification of cholesterol and hydrolysis of cholesteryl ester in alcohol induced fatty liver of rats

Repeated oral administrations of ethanol to rats induced accumulation of cholesteryl ester, as well as triglyceride in the livers. The contents of free cholesterol and phospholipid in the livers were not changed significantly in the present experiment in which ethanol ingestions were repeated four times. Although in vitro esterification of cholesterol by particle fractions of the alcoholic fatty liver was not affected, hydrolysis of cholesteryl palmitate by the supernatant fraction of the liver homogenate was reduced when compared with those of the control group which was given water or isocaloric glucose. The results of in vitro esterification of cholesterol and hydrolysis of cholesteryl palmitate in the liver of the rats which were ingested with glucose were larger than those of the control rats which were given water.     

Mesoporous solid acid catalysts: relationship between amine TPD data and catalytic activities

The concentrations of surface acid sites on Al³⁺- and H⁺-exchanged AlMCM-41 catalysts have been measured by temperature-programmed desorption (TPD) of a series of primary, secondary and tertiary amines, and by adsorption of ammonia. Surface acid site concentrations have been compared with cation exchange capacities (CECs) and catalytic activities in the alkylation of toluene with benzyl alcohol. The results show that the catalytic activities of the two ion-exchanged forms of the AlMCM-41 are similar. This is reflected in the acid site concentrations measured with tri-n-butylamine, and to a lesser extent tri-i-butylamine, which are similar for both ion-exchanged forms of the catalyst, and close to the CEC. Acid site concentrations measured with smaller amines and ammonia are similar to those based on tributylamine TPD for H⁺-exchanged AlMCM-41, but the Al³⁺ form of the catalyst appears to chemisorb significantly more of the smaller amines and ammonia, resulting in higher than expected acid site concentrations when these adsorbates are used. Larger amines than the tributylamines show very low chemisorption capacities on both forms of AlMCM-41, and appear to access only a small proportion of the surface acid sites. The primary amines tested show poor separation of chemisorbed and physisorbed amine in TPD experiments, making the determination of acid site concentration difficult. The results illustrate the importance of choosing an appropriate amine in using TPD to measure catalyst surface acidities.     

Correlation between electrophoretic clearcoats properties and electrochemical characteristics of noble substrates

The use of transparent organic coatings as protection for noble metal layers has a wide application especially in the costume jewellery and glass frames industry. The role of a clearcoat is to protect the noble substrate from scratches, tarnishing, and changes in gloss due to the interaction with the environment and human body. In this study electrodeposited organic coatings have been considered for this purpose. The application of clearcoats by electrophoresis usually assures a good adhesion to the substrate and the formation of a compact and protective layer. Electrophoresis has been developed especially under cathodic deposition and was mainly optimized for corrosion protection of active metals, but in this work both anodic and cathodic e-coats have been applied and analyzed. Since the substrates were noble metals, the surface could show a different reactivity during the application of the deposition voltage. The relationship between electrochemical behaviour of the substrate and the final properties of the clear coatings was investigated with electrochemical tests such as polarization curves, and electrochemical impedance spectroscopy. The correlation between deposition parameters and coatings properties as thickness and presence of defects was discussed according to the electrochemical results.     

Cholesterol metabolism in human monocyte-derived macrophages: Stimulation of cholesteryl ester formation and cholesterol excretion by serum lipoproteins

The role of lipoproteins and serum in the formation and accumulation of cholesteryl esters in human monocyte-derived macrophages (HMD macrophages) was investigated; studies were also carried out with IC21 cells (a cell line derived from mouse peritoneal macrophages). Following preincubation of HMD macrophages with lipoprotein-depleted serum (LPDS), both native and acetylated low density lipoprotein (LDL and AcLDL, respectively) stimulated the formation of cholesteryl esters with a resultant increase in cellular cholesteryl ester content. Cholesteryl ester formation and accumulation was also stimulated in macrophages exposed continuously to 25-hydroxycholesterol. However, the stimulation of cholesterol esterification by either lipoproteins or 25-hydroxycholesterol was not inhibited by progesterone in HMD macrophages, but was in the IC21 cells. Cholesterol efflux and the hydrolysis of cellular cholesterol ester, promoted by serum components, were studied in HMD macrophages preloaded with cholesteryl ester by incubation with 25-hydroxy cholesterol. Replacement of the medium with one devoid of 25-hydroxycholesterol resulted within 24 hr in at least a 30% decrease in the cholesteryl ester content of the HMD macrophages; replacement with a medium high in cholesterol acceptor content (LPDS or high density lipoprotein) and incubation for three days led to the most marked decreases in cellular cholesterol content. Thus, hydrolysis of the cholesteryl esters by HMD macrophages was not dependent on the presence of cholesterol acceptors in the medium, but cellular cholesterol content was.     

Relationship between the network morphology and properties of commercial vinyl ester resins

Two commercial vinyl ester resins, Derakane 411‐350 (resin D) and Derakane 411‐350 Momentum (resin M), were characterized. Despite the large quantity of publications in the literature about vinyl ester resins, few experimental results have been reported for resin M. The effect of the styrene content on the mechanical properties and morphological structure was studied. An increase in the styrene content produced a network with a low storage modulus in the rubber state and a glass‐transition temperature. The apparent average molecular weight between crosslink points and glass‐transition temperature were slightly higher for resin D than for resin M. The Fourier transform infrared spectra and molecular weight distributions were similar. However, resin M was tougher than resin D, and this may have been due to the closer structure in the fully cured state. Atomic force microscopy was performed for each cured resin and confirmed the difference in the nanostructures. The main reason for the differences in the developed structures was the use of an accelerator, which influenced the final morphology. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 100: 3895–3903, 2006     

Purification and characterization of a 28 kDa cytosolic inhibitor of cholesteryl ester hydrolases in rat testis

A 28 kDa inhibitory protein was purified from ratestis cytosol by sequential 40–65% ammonium sulfate precipitation, cation exchange chromatography, anion exchange chromatography, and preparative SDS-polyacrylamide gel electrophoresis. The heat-stable, trypsin-labile protein exhibited nonenzymatic, concentration-dependent inhibition of testicular and pancreatic cholesteryl ester hydrolases at all stages of putification. Copurifying at each stage was a 26.5 kDa protein which comprised 25% of the mass of the two proteins. Polyclonal antibodies raised to either or both 28 kDa and 26.5 kDa proteins by direct injection of excised electrophoretic bands cross-reacted with both proteins on western blots, immunoprecipitated both proteins, and neutralized inhibitory activity. Amino acid compositions of the individual proteins electroeluted from SDS-polyacrylamide gels were different from those of other surface-active proteins of similar molecular weights. Both proteins exhibited identical pl of 4.8 on chromatofocusing columns and two-dimensional gel electrophoresis. Although the subcellular distribution of the 28 kDa protein is unknown, its testicular cytosolic concentration, calculated from the purified protein mass, was 8×10⁻⁹ mols/L, which probably underestimates the actual concentration by an order of magnitude. This is greater than the minimum concentration required forin vitro inhibition (10⁻⁹ mols/L), consistent with a physiological role for this protein.     

Quantitative relationship between microstructure and mechanical properties in Nd: YAG transparent ceramics

In this work, stereology and fractals were applied to identify the quantitative relation between stereology parameters, fractal dimension, and mechanical properties of Nd: YAG transparent ceramics sintered at 1750 °C for 8–50 h. Mechanical properties and microstructure of the samples were investigated by using universal testing machine, micro-hardness tester, and scanning electron microscopy (SEM), respectively. When the ceramics were sintered at 1750 °C for 50 h, the compressive strength, flexural strength, and Vickers hardness reached 381.6 ± 5.2 MPa, 275.0 ± 5.5 MPa, and 1330.4 ± 18.5 MPa, respectively. Besides, the fracture toughness of ceramic samples was calculated by Vickers hardness. Micrographs of the sample surface and frequency distribution of crystal grains were analyzed by using metallographic image analyzer software. Findings suggest that compressive strength, flexural strength, and Vickers hardness linearly increase upon an increase in equivalent sphere diameter (D_3S). However, compressive strength, flexural strength, and Vickers hardness decrease as a function of specific surface area per unit volume of the grains (S_V) and discrete grains (S_VP) and mean free distance (λ). Perimeter and area of crystal grains were obtained by using Image-Pro Plus image analysis software. The relationship between the fractal dimension of grain boundary and mechanical properties was analyzed based on the area-perimeter (small-island) method. When the grain boundary fractal dimension is close to 1.0, the geometry of ceramic grains tends to be regular, and mechanical properties of ceramic samples increases.     

The relationship between mechanical properties and structure in rolled polypropylene

The relationship between mechanical properties and fine structure has been studied in polypropylene rolled both unidirectionally and biaxially (cross rolled). In unidirectionally rolled samples, a complex dependence with cold work is observed with a substantial change being observed at about 50 percent cold work. At 70 percent cold work, the yield strength and tensile strength increase substantially in the roll direction as compared with the starting billet but decrease only slightly in the transverse direction. Above 50 percent cold work, Young's modulus increases rapidly in the roll direction with a smaller increase in the transverse direction. The elongation to freak decreases in the roll direction but increases in the transverse direction. A striking feature is the large increase in ductility due to a small amount of cold work (ca., 10-20 percent). Analogous property changes are observed for cross rolled samples although no significant variation with direction in the sheet was found. The complex property changes are accompanied by complex changes in the molecular orientation as observed by wide angle X-ray pole figures and by changes in the morphology as observed with small angle x-ray scattering. These changes are interpreted in terms of a model incorporating tilting both of lamellae and of chain stems within the lamellae at early stages of rolling followed by breakup of lamellae and molecular rearrangement at later stages.