Interaction of discoidal complexes of dimyristoyl phosphatidylcholine-cholesterol-apolipoprotein A-I with human plasma high density lipoprotein HDL3

The interaction of human plasma high density lipoproteins (HDL₃) with discoidal complexes of apolipoprotein A-I (apoA-I) and dimyristoyl phosphatidylcholine (DMPC) containing 0, 10, 20 or 30 mol % cholesterol was investigated. Discoidal complexes containing various amounts of cholesterol were prepared by incubating apoA-I and DMPC-cholesterol liposomes for 12 hr at 25 C; the protein-lipid complexes were isolated by gel filtration chromatography on Bio-Gel A15m. Increasing the cholesterol content from 0 to 30 mol % caused a decrease in the fluidity of the discoidal complexes as determined by fluorescence polarization with 1,6-diphenyl-1,3,5-hexatriene; a reduced phase-transition amplitude; a decrease in the ratio of apoA-I to DMPC; and an increase in the width of the discoidal complexes as determined by electron microscopy after negative staining. Incubation of the apoA-I-lipid complexes with HDL₃ resulted in a complete breakdown of the discoidal structures and a transfer of DMPC and cholesterol to HDL₃. As a result of lipid transfer, there was an increase in the size of HDL₃. These in vitro results may be of significance as they relate to the interconversion of HDL subfractions during lipoprotein-lipase-induced lipolysis of triglyceride-rich lipoproteins.     

The composition and metabolism of high density lipoprotein subfractions

The composition and metabolism of high density lipoprotein (HDL) subfractions were investigated in seven nomal individuals. Mean HDL₂ (d, 1.063–1.125 g/ml) composition (by weight) was 43% protein, 28% phospholipid, 23% cholesterol, and 6% triglyceride, and mean HDL₃ (d, 1.125–1.21 g/ml) composition was 58% protein, 22% phospholipid, 14% cholesterol, and 5% triglyceride. The mean apoA-I; apoA-II weight ratio was 4.75 for HDL₂ and 3.65 for HDL₃.HDL₂ protein was proportionally slightly richer in C apolipoproteins and higher molecular weight constituents (including apoE) than HDL₃. Kinetic studies utilizing radiolabeled HDL_A (d, 1.09–1.21 g/ml), HDL₂, and HDL₃ demonstrated rapid exchange of apoA-I and apoA-II radioactivity among HDL subfractions, similar fractional rates of catabolism of apoA-I and apoA-II within HDL, and similar radioactivity decay within HDL subfractions. Mean plasma residence time was 5.74 days for radiolabeled HDL₂ and 5.70 days for radiolabeled HDL₃. Differences in HDL protein mass among individuals were largely due to alterations in catabolism, and in general both HDL₂ and HDL₃ were catabolized via a plasma and a nonplasma pathway. Data from simultaneous radiolabeled very low density lipoprotein and HDL studies in 2 individuals are consistent with the concept that apoC-II and apoC-III are catabolized at a different rate than are apoA-I and apoA-II within the HDL density range.     

The role of high density lipoprotein apolipoprotein CII in triglyceride metabolism

The purpose of these studies was (a) to examine the relationship between total plasma triglycerides (TG) and the amount of apolipoprotein CII (apo CII) in triglyceride rich lipoproteins (TRL), and (b) to determine whether TRL could be enriched with apo CII in vitro. In 13 patients with primary endogenous hypertriglyceridemia, (log₁₀) total plasma TG correlated inversely with the amount of apo CII per unit very low density lipoprotein (VLDL) protein (r=−0.76;p<0.005) and VLDL TG (r=−0.75; p<0.005). The potency of VLDL to activate milk lipoprotein lipase (LPL) in hydrolyzing triolein was studied in vitro. LPL activator potency per unit VLDL protein or VLDL TG correlated inversely with (log₁₀) total plasma TG (r=−0.86 and r=−0.76, respectively; p<0.005). LPL activator potency per nM VLDL apo CII also correlated inversely with (log₁₀) total plasma TG (r=−0.49; p<0.01). In seven patients with familial type V hyperlipoproteinemia, the average amount of apo CII in TRL protein was subnormal (5.86±0.62% vs 10.0±0.51% in normal subjects). The higher the (log₁₀) total plasma TG, the lower was the apo CII content in TRL protein (r=−0.93; p<0.01). To determine the factors governing the distribution of apo CII between lipoproteins and whether TRL could be enriched with apo CII, five approaches were undertaken: (a)¹²⁵I apo CII was added to mixtures of VLDL and HDL. The amount of labelled apo CII in VLDL was proportional to the ratio of VLDL to HDL. (b) TRL from four patients with familial type V hyperlipoproteinemia was incubated with high density lipoprotein (HDL) from a normal subject. An increase in the TRL/HDL ratio was associated with transfer of apo CII from HDL to TRL and a reciprocal transfer of non-apo CII protein from TRL to HDL. Net apo CII enrichment of TRL protein was possible below a HDL/TRL protein ratio of ca. 6 under the experimental conditions. (c) A fixed amount of normal plasma feed of TRL was incubated with different amounts of TRL from two patients with familial type V hyperlipoproteinemia. The amount of apo CII that transferred from normal TRL free plasma to the patient’s TRL was proportional to the amount of TRL in the mixture. (d) A doubling and tripling in the amount of apo CII in TRL was found when apo CII was added directly to TRL from a normal subject and TRL from a patient with familial type V hyperlipoproteinemia, respectively. (e) When apo CII was added directly to normal plasma and plasma from a patient with primary type IV hyperlipoproteinemia, the peptide was taken up mainly by VLDL and HDL, indicating enrichment of these fractions. The distribution of the added apo CII in each lipoprotein fraction resembled the distribution in the native plasma. TRL was isolated after addition of apo CII to plasma from two patients with familial types IV and V, respectively. Enrichment of TRL with apo CII was associated with an approximate 1.5-fold increase in the LPL activator potency per unit TRL protein. These studies suggest that firstly, the amount of apo CII in TRL is inversely related to the severity of hypertriglyceridemia. Secondly, the distribution of apo CII between TRL and HDL is governed by the mass ratios of these two lipoprotein classes. Thirdly, plasma TRL and HDL have a reserve binding capacity of apo CII and fourthly, it is possible to enrich these lipoproteins with this functionally important peptide. Whether net enrichment of TRL with apo CII and also an increase in its biological activity to activate LPL in vitro is related to increased in vivo catabolic rate requires to be determined.     

Intercorrelations among plasma high density lipoprotein,obesity and triglycerides in a normal population

The interrelationships among fatness measures, plasma triglycerides and high density lipoproteins (HDL) were examined in 131 normal adult subjects: 38 men aged 27–46, 40 men aged 47–66, 29 women aged 27–46 and 24 women aged 47–66. None of the women were taking estrogens or oral contraceptive medication. The HDL concentration was subdivided into HDL_2b, HDL_2a and HDL₃ by a computerized fitting of the total schlieren pattern to reference schlieren patterns. Anthropometric measures employed included skinfolds at 3 sites, 2 weight/height indices and 2 girth measurements. A high correlation was found among the various fatness measures. These measures were negatively correlated with total HDL, reflecting the negative correlation between fatness measures and HDL₂ (as the sum of HDL_2a and_2b). Fatness measures showed no relationship to HDL₃. There was also an inverse correlation between triglyceride concentration and HDL₂. No particular fatness measure was better than any other for demonstrating the inverse correlation with HDL but multiple correlations using all of the measures of obesity improved the correlations. Partial correlations controlling for fatness did not reduce any of the significant correlations between triglycerides and HDL₂ to insignificance. The weak correlation between fatness and triglycerides was reduced to insignificance when controlled for HDL₂. Presented (in part) at the Annual Meeting of the Oil Chemists' Society in St. Louis, MO, May 1978.     

Interaction of human plasma high density lipoprotein HDL2 with synthetic saturated phosphatidylcholines

The interaction of human plasma high density lipoprotein HDL₂ (d 1.063–1.125 g/ml) with sonicated dispersions of synthetic saturated phosphatidylcholines, dipalmitoyl- (diC₁₆PC), dimyristoyl- (diC₁₄PC), didodecanoyl- (diC₁₂PC), didecanoyl- (diC₁₀PC), and dioctanoyl- (diC₈PC) L-alpha phosphatidylcholine, was investigated. Incubation (4.5 hr, 37 C) of HDL₂ with diC₁₄PC, diC₁₂PC, diC₁₀PC and diC₈PC followed by gradient gel electrophoresis or preparative ultracentrifugation resulted in a redistribution of apolipoprotein A-I (apoA-I). The extent of redistribution depended on the molar ratio of the phospholipid to HDL₂ in the incubation mixture. Redistributed apoA-I occurred as lipid-free apoA-I and/or as complexes of apoA-I with phosphatidylcholine. Increasing the length of time of ultracentrifugation of the interaction mixtures did not increase the extent of redistribution. No redistribution of apoA-I was detected following incubation and gradient gel electrophoresis or preparative ultracentrifugation of mixtures of HDL₂ with dispersions of diC₁₆PC. Presented in part at the Joint Meeting of the American Oil Chemists' Society and the Japan Oil Chemists' Society, 1979.     

Subfractionation and characterization of native and incubation enlarged human plasma high density lipoprotein particles by high performance gel filtration

Incubation of human plasma in vitro at 37 C results in an increase of the mean particle size of the high density lipoproteins (HDL) accompanied by an almost complete disappearance of the original particles present prior to incubation. A rapid high performance gel filtration technique has been developed in order to study the chemical composition of subfractions of native and incubation enlarged HDL particles as a function of particle size. Subfractionation of HDL isolated by preparative ultracentrifugation from 3 normal human plasmas incubaed in vitro at 0 and 37 C for 24 hr have been performed using a 150 cm long TSK-G 3000 SW column. The separation time was less than 65 min. The curves obtained at high performance gel filtration of HDL, by monitoring the effluents from the column at 280 nm, agreed well both in positions of peak maxima and relative peak intensities with the particle distribution patterns observed at polyacrylamide gradient gel electrophoresis of the corresponding HDL preparations run in parallel. The different HDL particle subfractions of the effluents from the gel filtration column have been characterized by quantification of free and esterified cholesterol, total phospholipids and apolipoprotein A-I and A-II. The incubation enlarged HDL particles, subfractionated by the high performance gel filtration technique, were found to have a composition which differed from that of native HDL particles of corresponding size. Incubation enlarged HDL had a generally higher and almost constant relative cholesteryl ester content over the whole particle range compared to native HDL in which a continuous increase in relative cholesteryl ester content could be observed when going from large to small particles. The molar ratio of phospholipids to free cholesterol was higher in small native HDL particles than in the corresponding large ones. The relation between apolipoprotein A-I and A-II remained nearly constant between small and large HDL particles in each subfractionation experiment. The results demonstrate that the high performance gel filtration technique is a rapid and reproducible means for studying the composition of subfractions of HDL particle populations.     

Effect of dietary palm oil and its fractions on rat plasma and high density lipoprotein lipids

Male Sprague Dawley rats were fed semipurified diets containing 20% fat for 15 weeks. The dietary fats were corn oil, soybean oil, palm oil, palm olein and palm stearin. No differences in the body and organ weights of rats fed the various diets were evident. Plasma cholesterol levels of rats fed soybean oil were significantly lower than those of rats fed corn oil, palm oil, palm olein or palm stearin. Significant differences between the plasma cholesterol content of rats fed corn oil and rats fed the three palm oils were not evident. HDL cholesterol was raised in rats fed the three palm oil diets compared to the rats fed either corn oil or soybean oil. The cholesterol-phospholipid molar ratio of rat platelets was not influenced by the dietary fat type. The formation of 6-keto-PGF_1α was significantly enhanced in palm oil-fed rats compared to all other dietary treatments. Fatty acid compositional changes in the plasma cholesterol esters and plasma triglycerides were diet regulated with significant differences between rats fed the polyunsaturated corn and soybean oil compared to the three palm oils.     

Alterations of high density lipoprotein subclasses in obese subjects

The object of this study was to investigate the characteristics of lipid metabolism in obese subjects, with particular emphasis on the alteration of HDL subclass contents and distributions. A population of 581 Chinese individuals was divided into four groups (25 underweight subjects, 288 of desirable weight, 187 overweight, and 45 obese) according to body mass index (BMI). Apoprotein A-I (apoA-I) contents of plasma HDL subclasses were determined by 2-D gel electrophoresis associated with an immunodetection method. The concentrations of TG and the apoA-I content of pre-α₁-HDL were significantly higher (P<0.01 and P<0.01, respectively), but the levels of HDL cholesterol, and the apoA-I contents of HDL_2a and HDL_2b were significantly lower (P<0.01, P<0.05, and P<0.01, respectively) in obese subjects than in subjects having a desirable weight. Moreover, with the elevation of BMI, small-sized pre-α₁-HDL increased gradually and significantly, whereas large-sized HDL_2b decreased gradually and significantly. Meanwhile, the variations in HDL subclass distribution were more obvious with the elevation of TG levels in obese as well as overweight subjects. In addition, Pearson correlation analysis revealed that BMI and TG levels were positively correlated with pre-α₁-HDL but negatively correlated with HDL_2b. Multiple regression analysis also showed that TG concentrations were associated independently and positively with high pre-α₁-HDL and independently and negatively with low HDL_2b in obese and overweight subjects. The HDL particle size was smaller in obese and overweight subjects. The shift to smaller size was more obvious with the elevation of BMI and TG, especially TG levels. These observations, in turn, indicated that HDL maturation might be abnormal, and reverse cholesterol transport might be impaired. The first two authors contributed equally to this study.     

Evidence for the lipoprotein heterogeneity of human plasma high density lipoproteins isolated by three different procedures

Quantitative electroimmunoassays of apolipoproteins in ultracentrifugally isolated high density lipoproteins (HDL) of normolipidemic subjects showed that A-I and A-II are the major (80–85% of total HDL protein) and B, C-III, E, D and F are the minor protein constituents of this density class. A comparison between the apolipoprotein composition of ultracentrifugally isolated HDL and heparin-Mn⁺⁺ supernates showed no significant difference in the levels of A-I and C-III. However, the concentration of ApoE in the heparin-Mn⁺⁺ supernates was almost twice as high as that in the ultracentrifugally isolated HDL; ApoB was only detectable in trace amounts in the heparin-Mn⁺⁺ supernates. To establish whether these apolipoproteins are parts of a single macromolecular complex or form separate, discrete lipoprotein particles, the high density lipoproteins were isolated by three different procedures including ultracentrifugation, heparin-Mn⁺⁺ precipitation and agarose column chromatography. The double diffusion analyses of each of these HDL preparations with antisera to A-I, A-II, ApoB, C-III, ApoD, ApoE, and ApoF showed nonidentity reactions between each possible combination of these antisera. The only exception was a reaction of partial identity between antisera to A-I and A-II polypeptides indicating the occurrence of two types of lipoprotein particles, a major one (LP-A) containing both polypeptides and a minor one (LP-A-I) containing A-I as the sole protein constituent. These findings indicate that high density lipoproteins, regardless of the manner of isolation, do not consist of a single macromolecular complex, but represent a mixture of several, discrete lipoprotein families. Presented at the AOCS Meeting, St. Louis, May 1978.     

Normalization of high density lipoprotein in fish eye disease plasma by purified normal human lecithin: Cholesterol acyltransferase

Plasma from a patient with fish eye disease has been enriched with autologous high density lipoproteins (HDL) and supplemented with highly purified normal human plasma lecithin:cholesterol acyltransferase (LCAT). Incubation of such plasma at 37 C in vitro resulted in normalization of its low HDL cholesteryl ester percentage, from 23% to 79%, associated with a two-fold increase in both the cholesteryl ester and triglyceride contents of the HDL fraction, as compared to incubation experiments with absent or heat-inactivated purified normal LCAT. The normalization of the HDL cholesteryl ester percentage induced by incubation with purified normal LCAT also was accompanied by an increase in the size of the original fish eye disease HDL particles, which had a mean mass of 115 kd, to HDL particle populations with mean particle masses ranging from 130–220 kd, depending on the concentration of purified LCAT in the incubate. Both HDL cholesterol esterification and particle enlargement were abolished completely by the LCAT inhibitor DTNB and by heat inactivation of the purified normal LCAT. The results give further evidence that fish eye disease is an α-LCAT deficiency.     

The evidence for the antiatherogenicity of high density lipoprotein in man

It has long been recognized that patients with clinical coronary heart disease (CHD) have, on average, higher concentrations of plasma very low density and low density lipoproteins than do healthy subjects. The same, studies clearly demonstrated that coronary victims tend also to have low plasma concentrations of high density lipoprotein (HDL). It is only recently, however, that the possible significance of this second observation has been examined. Direct evidence for an inverse relationship between HDL cholesterol concentration and the prevalence of clinical CHD, independent of other plasma lipoproteins, has been provided by the Honolulu Heart and Cooperative Lipoprotein Phenotyping Studies. The Tromsø Heart and Framingham Studies subsequently demonstrated that this relationship precedes the clinical manifestation of coronary disease. More recently, angiographic studies have confirmed that the severity of existing coronary atherosclerosis is inversely related to HDL cholesterol concentration. Other investigations have shown that coronary victims also have low mean concentrations of apolipoproteins AI and AII (the major protein components of HDL), although the reduction of apoAI concentration may be less marked than that of HDL cholesterol, and preliminary findings from Tromsø have suggested that apolipoprotein AI may be less powerful than HDL cholesterol as a predictor of CHD. Such observations have supported the proposal that HDL may exert a protective effect against coronary atherosclerosis. Final confirmation (or otherwise) of this hypothesis, however, must await the results of carefully controlled animal experiments and of regression studies in patients with angiographically defined atherosclerosis.     

The distribution of serum high density lipoprotein subfractions in non-human primates

The ultracentrifugal flotation patterns in 1.2 g/ml solvent and ultracentrifugal gradient distribution of high density lipoproteins (HDL) from the primates-human, apes and monkeys-were determined, with emphasis on the gorilla species of apes and rhesus monkeys. Diets for non-human primates were commercial chow, which is low in cholesterol. Molecular weights and protein, cholesterol, phospholipid and triglyceride compositions of various density fractions were determined on human, gorilla and rhesus HDL. The HDL₂/HDL₃ ratio was determined from the two peaks observed upon flotation in high salt in the analytical ultracentrifuge. The HDL₂ of all three species of apes-gorillas (Gorilla gorilla), chimpanzees (Pan troglodytes) and orangutans (Pongo pygmaeus)—was always greater than HDL₃, while that of all six species of Old World monkeys-Rhesus (Macaca mulatta), sooty mangabeys (Cercocebus atys), cynomolgus (Macaca fascicularis), stumptails, (Macaca arctoides) patas (Erythrocebus patas) and African greens (Cercopithecus aethiops)—was less. In addition, the HDL₃ concentration in five gorillas was about 15 mg/dl as cholesterol while the HDL₂ concentration was 92 mg/dl, much lower and higher, respectively, than humans. HDL₂ of gorillas was similar in density and molecular weight to that of humans. The distribution of densities in gorilla HDL was predominantly in HDL₂, while rhesus HDL usually, but not always, was unimodal, having a density distribution similar in heterogeneity to human HDL₃, but somewhat less dense (peaking at 1.109 vs 1.129 g/ml). The molecular weight of rhesus HDL was about the same as human HDL₃ in all three density subfractions and at the peak density. Likewise, the chemical compositions were similar for the subfractions 1.10–1.125 and>1.125 g/ml for rhesus HDL and human HDL₃. Consequently most but not all chow-fed rhesus HDL was very similar to human HDL₃, but lighter in density. A preliminary report of this study was given at the American Society for Biological Chemists Meeting in New Oreleans in April 1982.     

Effects of sex on formation and properties of plasma very low density lipoprotein in vivo

The concentration and composition of the very low density lipoprotein (VLDL) lipids and the behavior of the VLDL in a density gradient in the zonal ultracentrifuge were examined in plasma obtained from normal fed male and female rats before and after intravenous injection of Triton WR-1339. Concentration of lipids in plasma VLDL of female rats was about half that of male animals. Following injection with Triton WR-1339, the concentration of VLDL lipids was higher in female rats (triacylglycerol) or similar (phospholipid, cholesterol, and cholesteryl esters) in both sexes. Female rats secreted much more VLDL triacylglycerol into the plasma compartment than did the male animals under the same experimental conditions. No differences were observed in lipid composition of the VLDL or in the position of the VLDL in the zonal rotor after ultracentrifugation in a density gradient of the lipoprotein from plasma of normal male and female rats before treatment with the detergent. However, after treatment with Triton, a higher proportion of the VLDL particles isolated from plasma of female rats displayed a more rapid rate-zonal flotation in the ultracentrifuge than did the VLDL produced by the male. The VLDL secreted by female rats contained fewer moles of phospholipid and free sterol per mol triacylglycerol than did the VLDL secreted by male animals under identical experimental conditions. The molar ratio of free cholesterol: cholesteryl ester in the VLDL secreted after treatment with Triton increased in both male and female rats. Simultaneously, the content of arachidonic acid in phospholipid of VLDL increased with a concomitant decrease in cholesteryl ester. These changes in fatty acid composition suggest that the inhibitory effect of Triton on lecithin-cholesterol acyl transferase activity affects the exchange of lipids between VLDL and high density lipoprotein. It can be concluded from the data reported here that sex influences the concentration of plasma lipids in vivo and the output and properties of the VLDL. Presented in part at the 59th annual meeting of the Federation of American Societies for Experimental Biology, Atlantic City, NJ, April 1975 (1). Recipient of a Career Development Award from the U.S. Public Health Service, No. 1-K4-HL-70329.     

Effects of diet and high density lipoprotein subfractions on the removal of cellular cholesterol

The effects of isocaloric substitutions of dietary polyunsaturated and saturated fat on the composition and function of plasma high density lipoproteins (HDLs) were studied in 3 normal subjects who were fed saturate-rich and polyunsaturate-rich diet programs. Compared to the saturated diets (P/S=0.4), polyunsaturated fat diets (P/S=4 or 2) reduced both plasma cholesterol and triglyceride levels. In 2 of the subjects, HDL cholesterol concentrations increased with polyunsaturated fat caused a reduction in HDL fatty acyl content of oleate and an increase in linoleate. To determine whether the altered composition affected the removal of cell membrane cholesterol, HDL and their subfractions, HDL₂ and HDL₃, which were isolated from each of the diets, were incubated with Ehrlich ascites cells in vitro. The cells were prelabeled with [³H] cholesterol, and the release of labeled cholesterol from the cells into the medium containing the various HDL fractions was determined. HDL, irrespective of the type of dietary fat, caused a release of [³H] cholesterol from the cells into the medium. The amount of [³H] cholesterol recovered in the medium was dependent on the absolute concentration of HDL cholesterol added to the cells and was independent of the type of diet. These results indicate that HDL facilitates the removal of cholesterol from cells, but that the amount and rate of removal are independent of the changes in HDL composition that can be obtained by dietary perturbations.     

The effect of exercise on plasma high density lipoproteins

The influence of vigorous activity in man on plasma lipids and lipoproteins is reviewed, with particular emphasis on high density lipoproteins. Both cross sectional and longitudinal (or training) studies have been reported, many of them of less than ideal design. Nonetheless, a consistent pattern emerges in which increased exercise levels lead to lower plasma concentrations of triglycerides and very low density lipoproteins, and of low density lipoproteins. High density lipoprotein levels increase. Sometimes, but not uniformly, plasma total cholesterol level falls as the result of these changes. The increase in plasma high density lipoprotein appears to be the result largely of an increase in the less dense HDL₂ subfraction. Plasma apolipoprotein A-I levels (but not apo-A-II levels) seem to increase concomitantly. The precise biochemical mechanism responsible for these changes has not been elucidated; but the recent finding of increased lipoprotein lipase activity in adipose tissue and muscle of endurance runners suggests that increased lipolytic rate of trigly ceride-rich lipoproteins may be an initial step in a sequence of events leading to higher plasma levels of HDL₂.     

Notching characteristics of un-doped amorphous silicon in high density plasma etching

Notching in un-doped amorphous silicon gate etching, a profile distortion phenomenon, has been studied and analyzed with low pressure, high density plasmas using HBr/Cl₂/O₂ chemistries. In contrast with the notching phenomenon observed in doped polysilicon etching, a notch was evolved at the outer sidewall foot of the edge line, where the outer sidewall of the edge line was exposed to a microscopic open area. A considerable recess was observed at the upper corner of a trench etched silicon substrate in photoresist-masked gate etching, while an oxynitride-masked gate structure showed little silicon recess at the same region. The subsequent accumulation of ions caused by deflected ions on the oxide surface adjacent to the outermost gate line leads to the formation of a local electric field near the outermost gate line, intensifying the ion trajectory distortion toward the sidewall foot of the outermost gate line.     

Heat transfer between wafer and electrode in a high density plasma etcher

Heat transfer between a wafer and electrode has been studied in a planar-type inductively coupled plasma reactor in terms of temperatures of wafer, chamber wall and electrode. A substantial increase in the wafer temperature was attributed mainly to bombardment of incident ions onto the wafer surface. The decrease in the wafer temperature at a higher pressure was attributed to the decrease in plasma density and a resistance to heat transfer in a micro gap formed between the wafer and the electrode. Compared to the case of no rf-chuck power applied, the wafer temperature when the electrode was biased with 13.56 MHz if power showed a greater increase mainly due to increased ion bombardment. Since the electrode having a water-cooled-backside geometry gains heat from the bulk plasma, it may lead to fast etch rates of hard materials whose etch products are less volatile at low temperatures, but not be good for photoresist materials.     

Structural modifications of rat serum high density lipoprotein by pancreatic phospholipase A2

An important and unusual aspect of the high density lipoprotein (HDL) in the rat is its tendency to undergo marked alterations in structure in response to physiological perturbations. In this study, the role of the surface lipids for maintenance of HDL integrity were investigated. Hydrolysis by pancreatic phospholipase A₂ of the phospholipids of rat HDL in the presence of the d>1.21 g/ml fraction of rat serum results in an increase in the particle diameter and an uptake of apo-E and apo A-IV from the lipoprotein-free fraction; augmentation of the albumin concentration in the incubation mixture intensified the observed changes, probably due to enhancement of the compositional changes brought about by phospholipase treatment. Phospholipase A₂ treatment of the d<1.21 g/ml fraction of rat serum produces only minor changes in the properties of the isolated HDL. These data suggest that changes in apoprotein content reflect an uptake of A-IV and E by the rat HDL, rather than a net loss of apo A-I. Likewise, titration of the action of pancreatic phospholipase A₂ on HDL apoprotein composition showed that initially a modest increase in apo A-IV content occurred, but with more extensive phospholipolysis there was a considerably greater increase in the apo-E content of the particle. The data suggest that hydrolysis of phospholipids such as occurs physiologically through the action of lecithin: cholesterol acyl transferase and hepatic lipase may alter the HDL structure independently from changes effected in the neutral lipid core.     

A new relationship between total/high density lipoprotein cholesterol and polyunsaturated fatty acids

Dietary and plasma fatty acids have been linked to total cholesterol but not to the ratio of total/high-density lipoprotein cholesterol (TC/HDLC). To evaluate the relationship between dietary and plasma levels of polyunsaturated fatty acids (PUFA) and TC/HDLC, we analyzed cross-sectional and longitudinal data using 519 plasma samples (50% men, 50% women) from subjects participating in the Framingham Heart Study and results from a study feeding diets rich in either n-6 linoleic acid or n-3 α-linolenic acid with or without fish oil supplements (n-3 derivatives). Values of TC/HDLC are inversely related to the percent of plasma PUFA when both variables are measured at the same time in different subjects,R=0.82,P<0.000001. PUFA in phospholipids increase in response to increased dietary intake of different PUFA, either n-3 or n-6 or fish oils. There was a highly significant inverse relationship between TC/HDLC and the percent of PUFA in phospholipids,R=0.97,P<0.001. The relationship was similar regardless of the source and type of dietary fatty acids. A similar relationship existed when only the baseline points were considered. When plasma PUFA % increases, either in response to a diet high in PUFA or across different subjects, TC/HDLC ratios decline. Evaluation of plasma fatty acid profiles and increased balanced dietary intake of PUFA to bring fatty acid profiles of subjects with low PUFA plasma levels closer to the profile of a healthy reference group is an effective approach to reduce high TC/HDLC. Reductions of more than 50% in TC/HDLC appear feasible with dietary modification alone. Further research into fatty acid metabolic activity may determine the biochemical basis of common dysplipidemias.     

Separation of the apoprotein components of human very low density lipoproteins by ion-paired,reversed-phase high performance liquid chromatography

A number of crude apolipoprotein samples isolated from human very low density lipoproteins (VLDL) were analyzed by reversed phase high performance liquid chromatography. The mobile phase consisted of a 1% solution of the polar ion-pairing reagent triethylammonium phosphate. A slow, nonlinear gradient of acetonitrile (37–42%) was used to elute the apolipoproteins. The order of elution was as follows: apolipoprotein C_x, apolipoprotein C-I, apolipoprotein C-III₂, apolipoprotein C-III₁, apolipoprotein C-III₀ and apolipoprotein C-II. This order is consistent with the known polarity of the proteins, i.e., the most nonpolar, apolipoprotein C-II, was the last to be eluted, whereas apolipoprotein C-I, with the lowest nonpolar surface area eluted first. The recovery of the individual apolipoproteins was 80–95% and the individual peaks were characterized by amino acid analysis, UV absorption spectra and chromatography of pure protein standards.