Plasma lipoproteins in dairy cows with naturally occurring severe fatty liver: Evidence of alteration in the distribution of apo A-I-containing lipoproteins

The relationships between fatty liver in dairy cows and reduced levels of plasma lipoproteins, and particularly of low density lipoproteins (LDL), has been previously described. Since electrophoretic heterogeneity of ultracentrifugally isolated LDL (d, 1.006–1.063 g/ml) has been found, the exact nature of this reduction in cows with fatty liver was investigated. Lipoproteins from control and severely afflicted animals were isolated by ultracentrifugation and affinity chromatography on heparin-Sepharose CL 6 B. Gradient gel electrophoresis of lipoproteins on 4–30% gels and an immunolocalization study of apoprotein A-I (apo A-I) showed that control animals have two subpopulations of apo A-I-containing particles with a mean radius of 6.52 and 5.05 nm. In the fatty liver cows, the former was clearly shifted toward smaller particles. We concluded that the depressed level and compositional modifications of LDL in severe fatty liver cows result from a decrease in the oversized apo A-I-containing lipoproteins which can be isolated in the LDL density range. This could stem from the decreased supply of triglyceride-rich lipoprotein surface components for the production of these lipoproteins. The modifications can be plausibly explained by a reduced synthesis or secretion of very low density lipoproteins (VLDL) by the liver.     

Effect of chronic glucagon administration on lipoprotein composition in normally fed,fasted and cholesterol-fed rats

Male adult Wistar rats received daily (at 9 a.m. and 5 p.m.) 10 μg of zinc-protamine glucagon by subcutaneous injection for 8 days. Plasma cholesterol levels were decreased by 36% in fed rats, 33% in cholesterol-fed rats and by 55% in fasted rats. Lipoproteins were separated into 22 fractions by ultracentrifugation using a density gradient. Glucagon administration decreased the cholesterol content in all lipoproteins except low density lipoprotein (LDL₁) (1.006–1.040) and very low density lipoprotein (VLDL) from cholesterol-fed rats. The main decrease (−57 to −81%) was observed in 1.050–1.100 g/mL lipoproteins (LDL₂ and HDL₂), which contained a large amount of apo E, while HDL₃ cholesterol was not affected. Triacylglycerol levels were decreased only in chylomicrons and VLDL (−70%) of fed and cholesterol-fed rats, while plasma and lipoprotein triacylglycerol levels were not changed in fasted rats treated with glucagon. In normally fed rats glucagon administration increased by 42% the fractional catabolic rate of [¹²⁵I]HDL₂ while the absolute catabolic rate appeared to be unchanged. Glucagon seems to be a potent hypolipidemic agent affecting mainly the apo E-rich lipoproteins. Its chronic administration limits lipoprotein accumulation which occurs upon cholesterol feeding.     

Plasma high density lipoprotein subgroup distribution in rats fed diets with varying amounts of sucrose and sunflower oil

The effect of varying the dietary sunflower oil/sucrose (SO/SU) ratio on rat plasma lipid concentration and lipoprotein distribution was studied. Four groups of 10 rats were fed for 4 weeks diets with varying SO/SU ratios. Lipoprotein components were then estimated in whole plasma and after cumulative density ultracentrifugation. Whole plasma triacylglycerol (TG), total cholesterol (TC) and free cholesterol (FC) decreased with increasing SO/SU ratio; the CE/FC ratio increased, because CE remained virtually unaltered. Plasma TG-lowering was due to a decrease in VLDL and LDL-TG. Protein, CE and FC in d=1.063–1.100 g/ml (HDL_2b) and d=1.100–1.125 g/ml (HDL_2a) lipoproteins decreased upon increasing the SO/SU ratio. In contrast, in d=1.125–1.200 g/ml (HDL₃) lipoproteins, there was a concomitant increase in these components. Although increasing the SO/SU ratio effected more protein and CE transportation in HDL₃ and less in HDL₂, the total amount of these components in high density lipoproteins (d=1.063–1.200 g/ml) remained constant. Apo A-I and apo C-III decreased in HDL₂ but increased in HDL₃ upon increasing the SO/SU ratio. Also, HDL₂ apo E, and the apo C-II/apo C-III and small apo B/large apo B ratios in VLDL and LDL were lowered by increasing the SO/SU ratio. The hepatic VLDL-TG output during isolated liver perfusion was lowest in rats fed the diet with the highest SO/SU ratio. In perfusate, like in plasma, the VLDL and LDL apo C-II/apo C-III ratio, as well as the small apo B/large apo B ratio, decreased upon increasing the dietary SO/SU ratio. The results indicate that there can be appreciable diet-dependent variations in plasma HDL subgroup distribution in spite of unchanged total HDL levels.     

High density lipoproteins from bovine plasma and follicular fluid do not possess a high affinity for glycosaminoglycans

Possible interactions between glycosaminoglycans and high density lipoproteins (HDL) in plasma and follicular fluid were examined. Total lipoproteins (d<1.21 g/ml) were obtained from plasma of five Holstein cows by ultracentrifugation and fractionated by gel filtration. Every other fraction from the HDL peak or fractions corresponding to the base and ascending portion of the HDL peak were composited and applied to a heparin-Sepharose affinity chromatography column. Elution profiles from both composites showed a peak that did not bind to the column that contained HDL devoid of apolipoprotein-E as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and silver staining and immunoblot analysis. Elution of lipoproteins from the ascending portion of the HDL peak resulted in a second minor peak eluting at 0.35 M NaCl, which was low density lipoprotein (LDL) contamination. Lipoproteins (d<1.21g/ml) isolated from follicular fluid obtained from small, medium or large follicles also were subjected to heparin-Sepharose affinity chromatography. Two peaks were observed, one corresponding to the lipoprotein that did not bind to the column, the other eluted at 0.5M NaCl and accounted for less than 2% of the protein applied. The second peak did not contain apolipoprotein-E or LDL. Bovine follicular fluid glycosaminoglycans (GAG) were isolated and subjected to HDL-Sepharose affinity chromatography. Less than 2% of the total GAG bound to the HDL column. Therefore, HDL in bovine specimens did not interact appreciably with heparin or GAG isolated from follicular fluid.     

The influence of protein and carbohydrate type on serum and liver lipids and lipoprotein cholesterol in rabbits

The non-lipid portions of semi-synthetic diets appear to be important determinants of hypercholesterolemia and atherosclerosis in the rabbit. Serum and liver lipid concentrations were determined in rabbits which had been pair-fed various protein (casein or soy protein isolate) and carbohydrate (sucrose or dextrose) sources as part of low fat, low cholesterol, semi-synthetic diets. It was verified that caseincontaining diets render rabbits hypercholesterolemic, while soy protein caused a degree of hypocholesterolemia. Additionally, sucrose, when fed in conjunction with casein, appears to augment this hypercholesterolemic effect. The distribution of total cholesterol among lipoprotein subclasses was increased in both the intermediate density lipoprotein (IDL) (1.006–1.019 g/ml) and low density lipoprotein (LDL) (1.019–1.063 g/ml) fractions and decreased in the high density lipoprotein (HDL) (1.063–1.21 g/ml) fraction when casein is fed. Soy protein feeding caused relatively more cholesterol to appear only in the IDL fraction when compared with commercial chow fed rabbits. Reasons for these differences may involve the saturation or suppression of endogenous lipoprotein hepatic receptors.     

Characterization of plasma lipoproteins in swine with different propensities for obesity

Yorkshire (lean) and Ossabaw (obese) swine ca. one year of age were used to characterize the quantity and composition of plasma lipoproteins in animals with markedly different adiposity. While lean swine weighed more (175 vs 88 kg for obese), they had less backfat than obese swine (2.64 vs 5.97 cm; P<0.05). Fasting plasma triacylglycerol (Tg) and cholesterol (CH) levels were elevated in obese swine. Swine plasma lipoproteins were fractionated into very low density lipoprotein (VLDL; d<1.006), low density lipoprotein₁ (LDL₁; d=1.019–1.063), low density lipoprotein₂ (LDL₂; d=1.063–1.09), and high density lipoprotein (HDL; d=1.09–1.21) by density ultracentrifugation. Obese VLDL-Tg, CH and protein (Pr) were elevated more than 2-fold. VLDL from obese swine were 2-fold larger than VLDL from lean swine. No alterations in LDL₁ or LDL₂ composition were observed. HDL-Tg, CH, Pr and phospholipid levels were significantly higher in obese swine. Plasma and VLDL-Tg levels were highly correlated with backfat thickness (r=0.67 and r=0.73, respectively). These was a positive correlation between adiposity and HDL-CH as well as VLDL-Tg and HDL-CH. These data indicate that (a) there are marked alterations in swine plasma lipoprotein composition between lean and obese swine; (b) that swine plasma lipoprotein levels may be useful parameters in estimating body composition; and (c) that HDL-CH is positively correlated with adiposity in swine. Department of Dairy and Animal Science, College of Agriculture. Nutrition Program, College of Human Development.     

Experimental nephrotic syndrome in the rat induced by puromycin aminonucleoside: Hepatic synthesis of lipoproteins and apolipoproteins

Hepatic synthesis of lipoproteins and apolipoproteins was investigated in male Wistar rats with severe nephrotic syndrome induced by puromycin aminonucleoside by incubating liver slices with a mixture of¹⁴C-amino acids. Labeled lipoproteins were separated by preparative ultracentrifugation from the incubation medium after the addition of carrier plasma. The incorporation of¹⁴C-amino acids into very low density lipoproteins (VLDL) (1.006 g/ml), low density lipoproteins (LDL) (1.006–1.063 g/ml) and high density lipoproteins (HDL) (1.063–1.210 g/ml) was increased in nephrotic liver 6.1-, 5.7- and 5.0-fold, respectively. The measurement of radioactivity associated to apolipoproteins isolated by SDS-PAGE documented an increased incorporation into apolipoprotein E (apoE) of nephrotic VLDL (33.1% vs 20% of the total radioactivity incorporated into VLDL apoproteins) and a markedly increased incorporation into apolipoprotein A-I (apoA-I) of nephrotic HDL (44.3% vs 16.3% of the total radioactivity incorporated into HDL apoproteins). In nephrotic liver, the total incorporation of amino acids into apolipoproteins (apoVLDL+apoLDL+apoHDL) was increased 12.6 times for apoA-I, 6.4 times for apoB, 5.0 times for apoE, 4.2 times for apoC+apoA-II and 2.5 times for apoA-IV. We suggest that, in nephrotic liver: (a) the synthesis of VLDL, LDL and HDL is increased, and (b) the total synthesis of apoA-I is selectively increased when compared to that of the other apolipoproteins. Preliminary reports of this work were presented at the Annual Meeting of the European Society for the Study of the Liver (Düsseldorf, September 13–15, 1979); at the 5th International Symposium on Atherosclerosis (Houston, November 6–9, 1979) and at the Annual Meeting of the Italian Society for the Study of the Liver (Rome, December 14–15, 1979).     

Distribution of antithrombin III and glucosylceramide in human plasma lipoproteins and lipoprotein deficient plasma

We have investigated the distribution of antithrombin-III and glucosylceramide (Glc-Cer) in human plasma, plasma lipoproteins and lipoprotein-deficient plasma. Antithrom bin III activity was measured employing immunochemical and biological assays. Glc-Cer was quantified by gas liquid chromatography (GLC). Whole plasma contained 145 μg antithrombin III/ml plasma, all of which was associated with the lipoprotein-deficient plasma (d>1.25 g/ml). Whereas, most if not all the plasma GlcCer was associated with plasma low density lipoproteins (LDL) (d-1.022–1.055 g/ml) and high density lipoproteins (HDL) (d-1.063–1.25). GlcCer was not found in the lipoprotein-deficient plasma. We conclude that GlcCer on lipoproteins does not contribute to antithrombin III activity. Moreover, the absence of GlcCer in lipoprotein-deficient plasma does not impair antithrombin-III activity.     

Plasma,apolipoprotein A-I and A-II levels in hyperlipidemia

Some of the component moieties of high denisty lipoproteins (HDL) were analyzed in normal subjects and in patients with hyperlipidemia. Apoproteins A-I and A-II were quantified by radioimmunoassay, HDL cholesterol and triglycerides were assessed on heparin-MnCl₂ supernates of fasting plasmas. We found that HDL is enriched in triglycerides in all forms of hyperlipidemia, while the proportion of ApoA-II is unaltered and the proportion of ApoA-I is decreased. Thus, the composition of HDL is altered in hypertriglyceridemia. The molecular associations of ApoA-I and ApoA-II in plasma were also examined by assaying the apoprotein contents of plasma fractions prepared by ultracentrifugation and by gel filtration column chromatography. The ApoA-I contents of d<1.063 fraction increased in hyperlipidemia from <0.5% to ∼2%, but the ApoA-I contents of the d>1.21 fraction remained at <12% of total in plasmas with triglyceride levels <1500 mg/dl. d>1.21 ApoA-I rose to 23% in one plasma with a triglyceride level of >1700 mg/dl. On column chromatography, ApoA-I eluted with the lipoproteins and also in a fraction whose molecular weight (MW) appreared to be ∼50,000 daltons. The proportion of plasma ApoA-I which eluted in the 50,000 MW peak was positively correlated with plasma triglyceride levels, but at triglyceride levels of <1500 mg/dl, <20% of ApoA-I was in the 50,000 MW peak. Between levels of ∼2000 and 12,000 mg/dl, the percentage “50,000 M.W. ApoA-I” was 20–25%. The ApoA-II contents of d<1.063 fractions were also increased in hyperlipidemia, but >95% of ApoA-II was found in the HDL fractions in both normal and hyperlipidemic plasma both by column chromatography and ultracentrifugation. Thus, the molecular association of ApoA-I appears to be altered in hyperlipidemia.     

Isolation of human serum low-density lipoproteins with the aid of an immune-specific adsorber

Human serum lipoproteins containing B-protein have been isolated using an immunoadsorber. Bromoacetyl cellulose was combined with pure antibodies to low density lipoprotein (LDL) and an immunoadsorber of high capacity was obtained. With 1 g of this immunoadsorber all LDL and very low density lipoprotein (VLDL) from 30 ml pooled human serum were adsorbed and then eluted with glycine-HCl buffer pH 3.2 at 0 C. The isolated lipoproteins were investigated by electrophoresis, immunodiffusion and ultracentrifugation, and found to be identical to LDL+VLDL isolated by ultracentrifugation.     

Interaction of plasma high density lipoprotein HDL2b (d 1.063–1.100 g/ml) with single-bilayer liposomes of dimyristoylphosphatidylcholine

Incubation of a major subfraction, HDL_2b (d 1.063–1.100 g/ml), of human plasma high density lipoproteins, HDL (d 1.063–1.21 g/ml), with single-bilayer liposomes of dimyristoylphosphatidylcholine (DMPC) resulted in uptake of DMPC by the HDL_2b and dissociation of lipid-free apolipoprotein A-I (apoA-I). In the presence of excess DMPC, the dissociated apoA-I was also incorporated with DMPC into discoidal complexes. Preliminary studies with model apoA-I-DMPC complexes indicated that they also can interact with native HDL_2b with the resultant transfer of their DMPC to HDL_2b and the concomitant release of their apoA-I. After interaction of HDL_2b with DMPC liposomes, the DMPC-enriched HDL_2b product showed a lower hydrated density and a larger particle size than the control HDL_2b. The molecular properties of the lipoprotein product suggest that stabilization of the apoA-I-depleted HDL_2b probably occurred via substitution of DMPC for the apoA-I at the HDL_2b surface rather than by fusion of the apoA-I-depleted HDL_2b. The above interactions of HDL_2b with single-bilayer liposomes and discoidal complexes indicate pathways of phospholipid transfer relevant to the possible role of HDL in the metabolism of lipoprotein surface components in vivo.     

Glycosphingolipids of human plasma lipoproteins

The content and structure of glycosphingolipids (GSL) in human plasma lipoproteins were studies. The quantitative distribution of the neutral GSL(Glc-Cer, Gal-Glc-Cer, Gal-Gal-Glc-Cer, and GalNAc-Gal-Gal-Glc-Cer) and the principal ganglioside (AcNeu-Gal-Glc-Cer) within the different lipoprotein classes was similar to that of whole plasma. The total amounts (μmol glucose/100 ml plasma) of GSL in the plasma lipoproteins of three normal subjects were VLDL (very low density lipoproteins) (trace to 0.46), LDL (low density lipoproteins) (1.08–1.48), HDL₂ (high density lipoproteins₂) (0.62–0.85), and HDL₃ (high density lipoproteins₃) (trace to 0.28). In subjects with Lp(a) lipoproteins, HDL₂ rather than HDL₃ contained most of the GSL in HDL. When the data were corrected for differences in the plasma concentrations of the lipoproteins, the total amounts of GSL(nmol glucose/mg lipoprotein cholesterol) were VLDL(trace to 21.20), LDL(11.70–15.36), HDL₂(8.50–9.10), and HDL₃(3.12). No GSL were detected in lipoprotein deficient plasma. Mass spectrometry of the trimethylsilyl derivatives of the GSL in LDL showed major fragment ions characteristic of their individual structural components. The elevated plasma levels of the GSL(2–18 fold), in a homozygote for familial hypercholesterolemia, resided in LDL which contained an absolute increase (per mg lipoprotein cholesterol) of GSL. Most, if not all, of the plasma GSL are associated with plasma lipoproteins and may have an important role in their biological functions.     

Alpha- and gamma-tocopherol levels in lipoproteins fractionated by affinity chromatography

Alpha- and gamma-tocopherol levels of nine women were measured in whole serum and in serum lipoproteins separated by heparin affinity chromatography. Alphatocopherol levels (mean±SD) in whole serum, low density plus very low density lipoproteins and high density lipoproteins were 10.8±2.7, 6.4±1.6 and 4.6±1.4 (μg/ml), respectively. Corresponding values (μg/ml) for gamma-tocopherol were 1.2±0.5, 0.7±0.3 and 0.6±0.2. Recoveries of serum alpha- and gamma-tocopherol from the heparin columns were 102±5% and 105±7%, respectively. Serum alpha-tocopherol was linearly correlated with components of high density lipoprotein (apolipoproteins, high density lipoprotein cholesterol), but not with serum total lipids or indices of low density lipoprotein, even though high density lipoprotein carried less than half of the serum alpha-tocopherol. However, serum gamma-tocopherol was highly correlated with indices of serum lipids, such as serum cholesterol (r=0.92, p=0.005). The coefficient for the correlation of low density lipoprotein (+very low density lipoprotein) tocopherol with high density lipoprotein tocopherol was r=0.66 (p=0.06) for alpha-tocopherol and r=0.84 (p=0.004) for gamma-tocopherol. These differences in the relationships of the two tocopherols to lipids and lipoproteins support the view that when the two tocopherols are present at normal dietary levels, gamma-tocopherol partitions between lipoproteins based on their relative lipid content, but a portion of the alpha-tocopherol in high density lipoprotein is specifically bound.     

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.     

Comparison of the clearances of serum chylomicron triglycerides enriched with eicosapentaenoic acid or oleic acid

Rat mesenteric lymph chylomicrons containing triglycerides enriched with either [¹⁴C]oleic acid (OA) or [¹⁴C]-eicosapentaenoic acid (EPA) were prepared by ultracentrifugation of lymph samples collected for 6 hr after a single duodenal infusion of an emulsion containing either fatty acid. These chylomicrons were injected into the jugular vein of recipient rats and, at various time intervals, blood was drawn and serum was assayed for radioactivity. In separate animals, serum lipoprotein fractions were separated by ultracentrifugation, and the redistribution of labeled fatty acid among circulating lipoproteins was determined by liquid scintillation spectrometry. When the early disappearance rates (10 min) of either total serum radioactivity or specifically the chylomicron fraction were compared, there were no differences between the groups receiving OA-or EPA-enriched chylomicrons. However, disappearance rates of EPA-enriched chylomicrons were slower than those of OA-enriched chylomicrons from 25 to 90 min. The small but significant differences in the disappearance rates for the longer time periods cannot be ascertained without further studies. At 5 min after injection of either type of chylomicron, the d<1.006 g/ml lipoprotein fraction of serum chylomicrons and very low density lipoproteins contained almost 90% of the original radioactivity. By 240 min, when less than 2% of the radioactivity remained, this radioactivity in the d<1.006 g/ml fraction was 43–46%, with concomitant increases in the low and high density lipoprotein fractions and in the lipoprotein-free serum. Deceased.     

Induction of long-lasting hypercholesterolemia in the rat fed a cystine-enriched diet

The influence of dietary excess (5%) of L-cystine on rat plasma lipoproteins was examined. After only one week of cystine feeding, an increase in the plasma cholesterol level and a decrease in triglyceride levels were observed. The increase in cholesterol level became greater when the duration of cystine-enriched diet increased until eight weeks (+131% after eight weeks), but no further increase occurred between 8 and 20 weeks. This change was essentially due to the progressive increase in cholesterol levels in high density lipoproteins (HDL) and in lipoproteins isolated between 1.040 and 1.063 g/ml, i.e., certain low density lipoproteins (HDL₂), and containing mainly apoE-rich lipoproteins (HDL₁). The decrease in plasma triglycerides resulted from that of chylomicrons and very low density lipoproteins (VLDL). The effects observed after four or eight weeks of cystine feeding were maintained for eight weeks after replacing the cystine diet by the standard diet. Ingestion of the standard diet containing either cholestyramine (2%) or probucol (0.25%) following eight weeks of cystine feeding significantly decreased plasma cholesterol levels. It is concluded that cystine-fed rats are a useful tool of investigation for understanding mechanisms leading to increased plasma cholesterol level and for hypocholesterolemic drug trials.     

Subfractionation of Sf 4–105, Sf 4–20 and high density lipoproteins

Subfractionation of the total low density S_f 4–10⁵, the low density S_f 4–20 and high density plasma (or serum) lipoproteins has been accomplished using a cumulative flotation rate procedure. Fractionation employs nonlinear salt gradients and high performance swinging bucket rotors. Subfractionation of the total low density lipoproteins with minimal contamination allows and extremely accurate lipoprotein mass measurement of S_f > 400, total very low density lipoproteins and low density lipoproteins (LDL) by elemented CHN analysis. Physical and chemical data on LDL and high density lipoprotein (HDL) subfractions are in general agreement with earlier data. Lower molecular weight data are obtained for HDL subfractions than reported earlier; however this may be the result of the different fractionation procedures used. Presented in part at AOCS Meeting, New Orleans, April 1970.     

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 effect of dietary fats on the plasma lipid composition of sheep

This study reports on the plasma lipid compositions of sheep fed either a control diet (C), a control diet supplemented with tallow (A) or polyunsaturated fatty acid (B) that had been protected against hydrolysis and hydrogenation in the rumen, or a control diet supplemented with maize oil (D). Diet B considerably increased the 18∶2 content of all the major plasma lipid fractions. Although the feeding of diet D also resulted in an increase in the 18∶2 contents within the cholesteryl ester, unesterified fatty acid, and phospholipid fractions the increases were considerably less than those observed with diet B; the levels of 18∶2 within the triglyceride fraction remained similar to that for the sheep which received the control diet. The effect of feeding diet A was confined solely to the triglyceride fraction where the concentrations of 16∶0 and 18∶1 were increased. The lipoproteins of the plasma were separated into very low density lipoproteins (d<1.006), low density lipoproteins (1.006<d<1.063), and high density lipoproteins (1.063<d<1.21), and the distribution of the major lipids between these lipoprotein fractions was investigated. Diet B increased considerably the proportion of triglyceride found in association with the very low density fraction and the concentrations of 18∶2 within all the lipoprotein fractions; these increases in the concentrations of 18∶2 were not confined to any particular lipid fraction of the lipoproteins. In contrast, the increases in the concentrations of 18∶2 produced as a result of feeding diet D were confined to the low and high density lipoproteins. The effect of feeding diet A was confined to fatty acid changes within the triglycerides of the low and very low density lipoproteins.     

Pulmonary surfactant: Distribution of lipids,proteins and surface activity in ultracentrifugation of rabbit pulmonary washing and derived fractions

Lavage from normal adult rabbit lung and two known derived fractions, Fraction T and Fraction S, were subjected to either differential ultracentrifugation in 1.090 g/ml KBr or sucrose density gradient ultracentrifugation; the surface activity of the lipid extract of selected fractions was measured. In differential ultracentrifugation, the three starting materials yielded a pellicle containing > 85% of the phospholipid with <1% protein. In sucrose density gradient ultracentrifugation: pulmonary washing, containing about equal weights of phospholipid and protein (60% albumin, 20% sIgA, 10% IgG, 10% minor proteins), produced one single band at density 1.040, containing virtually one single protein, namely >80% of the total sIgA (protein T) and up to 60% of the total phospholipid, whereas all the albumin and IgG were found at very low densities, 1.010 and 1.025, respectively; Fraction T, having nearly equal weights of one signle protein, sIgA, and phospholipid, produced two contiguous bands at densities 1.059 and 1.078, totalling >85% of its phospholipid and <25% of its protein, the balance of which was found free of phospholipid at densities 1.020 to 1.050; comprising >80% of the phospholipid and <20% of the protein of pulmonary washing, Fraction S yielded two small bands at densities 1.028 and 1.044 and a major band at d=1.059. In surface activity measurements: when the total lipid extract of the bands from the sucrose density gradient ultracentrifugation was spread as a film, in spite of similarly high dipalmitoyl lecithin contents (about 70% palmitoyl lecithin contents (about 70% palmitoyl residue), the lipid of the band of Fraction T and that of the high density band of Fraction S were very active (γ _min=0); whereas the lipid of the band of pulmonary washing and that of the lowest density band of Fraction S were not active,γ _min being 18 dyne/cm and 21 dyne/cm, respectively. This wokk brings forth three major conclusions. First, under conditions which are used to isolate serum lipoproteins, no lipoprotein was obtained from either of the three surfactant fractions and most of the lipid was found virtually free of protein. Second, the isopicnic equilibrium of a given ultracentrifugation fraction varied with the molecular structure of its constituents and could not be accounted for by the latter’s average densities; instead, major roles must be player by particle geometry and their water contents. Third, although the various lipid samples contained the same quantities of palmitoyl residues (70%), the surface activities varied with the physical state of the lipid, method of assay, and some other undefined factors.