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Effect of intrajejunal acidity on lipid digestion and aqueous solubilisation of bile acids and lipids in health, using a new simple method of lipase inactivation.

We have investigated whether acid-mediated bile acid precipitation, pancreatic enzyme inactivation, and fatty acid partitioning occur in health when intraluminal pH falls below 5. In order to assess lipolysis and aqueous solubilisation of lipid, we first developed a new technique for inactivating lipase in jejunal aspirate (acid inactivation), and showed it to be more effective and simpler than the established technique (heat inactivation). We then studied 14 healthy subjects, aspirating jejunal content for three hours after a liquid meal, and pooling according to pH. Eighteen per cent of the total aspirate was collected at pH less than 5 compared with 56% at pH greater than 6 (p less than 0.01). Forty eight per cent of the bile acids were precipitated at pH less than 5 compared with 18% at pH greater than 6 (p less than 0.01), leading to a reduction in aqueous phase bile acid concentration at low pH (2.1 mmol/l at pH less than 5 vs 5.8 mmol/l at pH greater than 6, p less than 0.01). Lipase activity was reduced at low pH (133 IU/l at pH less than 5 vs 182 IU/l at pH greater than 6, p less than 0.01), leading to reduced lipolysis at low pH (14% at pH less than 5 vs 32% at pH greater than 6, p less than 0.01). Aqueous phase lipid concentration was reduced at low pH (3.5 mmol/l at pH less than 5 vs 12.5 mmol/l at pH greater than 6, p less than 0.01). This reduction was less dependent on bile acid precipitation than on lipase inactivation and fatty acid partitioning. We conclude that intraluminal acidity influences aqueous solubilisation of bile acids and lipid in health.

Adult↗

Cellular lipid metabolism and the role of lipids in progressive renal disease.

Dyslipidemia contributes to the rate of progression of atherosclerosis and chronic kidney disease. Also, chronic kidney disease leads to the development of secondary abnormalities in lipid metabolism that contribute to increased cardiovascular morbidity and mortality. This review presents the mechanisms that underlie this risk. The mechanisms of normal cellular lipid metabolism and the abnormalities that develop in association with inflammation are reviewed. There is a special emphasis on foam cells in the kidney and on lipid-mediated changes in intrinsic kidney cells that lead to glomerulosclerosis and interstitial fibrosis. Correlates to studies performed in whole animals and humans are included.

Animals↗

Effects of dietary phytosterols on liver lipids and lipid metabolism enzymes.

The effects of dietary phytosterols on lipid metabolism have been assessed through determination of liver lipids (sterols and fatty acids) and lipid metabolism enzymes (acetyl-CoA carboxylase, malic enzyme, glucose-6-phosphate dehydrogenase) in rats fed 12 or 24 mg cholesterol a day and 0-96 mg phytosterols. The results indicate that, provided the dietary phytosterol to cholesterol ratio is at least 1 and in the presence of a dietary cholesterol excess, phytosterols do exert a regulatory role through decreases of both acetyl-CoA carboxylase and malic enzyme activities. A ratio of 2 enhances this effect. At the same time, liver fatty acids and cholesterol contents significantly decrease.

Acetyl-CoA Carboxylase↗

Unexpectedly low incorporation of isotopic acetate into lipids of Ehrlich ascites tumor cells cultured in lipid-poor medium.

The present communication reports data on the lipid biosynthesis of Ehrlich ascites tumor cells grown in culture media supplemented with modified sera. Whereas the metabolisms of [14C]pyruvate and [14C]mevalonate are identical in all media tested, the incorporation of [14C] acetate is higher in medium with dialyzed serum than in medium with delipidized serum; it is suppressed in the absence of all lipids in culture medium. Cellular integrity is not impaired in modified media. The results indicate that acetyl-CoA synthetase of Ehrlich ascites tumor cells is not regulated by exogenous lipids as is known to be the case in nonmalignant cells.

Acetate-CoA Ligase↗

Relationships of nutrient intake to lipids and lipoproteins in 1234 white children. The Lipid Research Clinics Prevalence Study.

Using the geographically and socioeconomically varied collaborative Lipid Research Clinics Prevalence Study data, this report focuses upon relationships between dietary intake and plasma lipids and lipoproteins in 1234 white children, 661 boys and 573 girls, aged 6-19 years who were sampled in a random recall (Visit 2) from large populations in six Lipid Research Clinics. Using multiple regression analysis, we found that in 6- to 12-year-old boys the dietary polyunsaturated-to-saturated fat ratio was inversely associated with plasma total and low density lipoprotein cholesterol and dietary cholesterol was positively associated with plasma high density lipoprotein cholesterol. In 13- to 19-year-old boys, high density lipoprotein cholesterol and the ratio of high density lipoprotein cholesterol to total cholesterol were inversely related to sucrose intake. In 6- to 12-year-old girls, plasma triglycerides were positively related to dietary sucrose. Using analysis of covariance in children having the lowest, middle, and top decile nutrient intakes, we found that higher carbohydrate intakes were associated with lower plasma total cholesterol in boys. The highest polyunsaturated fat intake (in 6- to 12-year-old girls) was associated with the lowest plasma cholesterol and very low density lipoprotein cholesterol. The highest cholesterol intake (in 6- to 12-year-old boys) was associated with the highest high density lipoprotein cholesterol levels. In girls, the highest sucrose intakes were associated with the highest plasma triglyceride levels. We conclude that the weak, but statistically significant, associations that we observed were meaningful relative to etiologies of nutrient-lipoprotein interrelationships, and should be useful in forming new hypotheses for focused metabolic ward studies.

Adolescent↗

Effect of exogenous lipid on lipid synthesis by bone and bone cell cultures.

Newborn rat calvaria and isolated calvaria cells are capable of de novo lipid synthesis when grown in the presence or absence of exogenous lipid sources. Synthesis decreases when exogenous lipids are supplied. Several cholesterol precursors were found in these tissues and the presence of dihydrocholesterol was established for the first time.

Acetates↗

Serum lipids, lipoproteins, and lipid metabolizing enzymes in identical twins discordant for obesity.

Obesity is associated with adverse changes in plasma lipoprotein metabolism, but it is not known completely how this association is modified by genetic factors. We assessed the contribution of obesity to serum lipid and lipoprotein levels and lipid metabolizing enzyme activities by examining 23 identical twin pairs (9 male, 14 female) who had, on the average, an 18-kg intrapair difference in BW. Compared with lean co-twins, obese co-twins had approximately 20% higher low-density lipoprotein (LDL) cholesterol (P < 0.01), 20% lower high-density lipoprotein2 cholesterol (P = 0.010), and 90% (men) or 35% (women) higher (P < or = 0.06) total, very-low-density lipoprotein and LDL triglycerides. The pairs were divided into subgroups by the gender-specific median value of abdominal visceral fat (AVF) area in the obese co-twin and by apolipoprotein E 4 phenotype. The intrapair differences in serum cholesterol fractions were similar in twin pairs with high or low AVF, whereas only high AVF pairs showed significant differences in triglyceride fractions. The greatest intrapair differences in total, very-low-density lipoprotein and LDL triglycerides were observed in apolipoprotein E 4-positive pairs expressing high AVF. Compared with lean co-twins, lecithin cholesterol acyltransferase activity was 18% higher (P < 0.001) and hepatic lipase activity was 38% higher (P = 0.016) in obese co-twins with high AVF. When genetic factors are identical, obesity is associated with an atherogenic lipid profile, especially in subjects with high visceral fat accumulation.

Apolipoprotein E4↗

Lipids in blood-brain barrier models in vitro I: Thin-layer chromatography and high-performance liquid chromatography for the analysis of lipid classes and long-chain polyunsaturated fatty acids.

The objectives of this study were to optimize a sensitive high-performance liquid chromatography (HPLC) method for fatty acid (FA) analysis for the quantification of polyunsaturated FAs (PUFAs) in cell lipid extracts and to analyze the lipid and FA patterns of three cell lines used in blood-brain barrier (BBB) models: RBE4, ECV304, and C6. Thin-layer chromatographic analysis revealed differences in the phosphatidylcholine-phosphatidylethanolamine (PC:PE) ratios and the triglyceride (TG) content. The PC:PE ratio was <1 for RBE4 cells but >1 for ECV304 and C6 cells. ECV304 cells displayed up to 9% TG depending on culture time, whereas the other cell lines contained about 1% TG. The percentages of docosahexaenoic acid were 9.4 +/- 1.7% of the unsaturated FAs in RBE4 cells (n = 5; 4 d in culture; 9.9% after 10 d), 8.1 +/- 2.0% in ECV304 cells (n = 11; 10 to 14 d), and 6.7 +/- 0.6% in C6 cells (n = 6; 10 to 14 d) and were close to the published values for rat brain microvascular endothelium. The percentage of arachidonic acid (C20:4) was about half that in vivo. ECV304 cells contained the highest fraction of C20:4, 17.8 +/- 2.2%; RBE4 cells contained 11.6 +/- 2.4%; and C6 cells 15.8 +/- 1.9%. It is concluded that a sensitive HPLC method for FAs is now optimized for the analysis of long-chain PUFAs. The results provide a useful framework for studies on the effects of lipid modulation and give reference information for the development of further BBB models.

Animals↗

Distinguishing individual lipid headgroup mobility and phase transitions in raft-forming lipid mixtures with 31P MAS NMR.

A model membrane system composed of egg sphingomyelin (SM), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and cholesterol was studied with static and magic angle spinning (31)P NMR spectroscopy. This model membrane system is of significant biological relevance since it is known to form lipid rafts. (31)P NMR under magic angle spinning conditions resolves the SM and DOPC headgroup resonances allowing for extraction of the (31)P NMR parameters for the individual lipid components. The isotropic chemical shift, chemical shift anisotropy, and asymmetry parameter can be extracted from the spinning side band manifold of the individual components that form liquid-ordered and liquid-disordered domains. The magnitude of the (31)P chemical shift anisotropy and the line width is used to determine headgroup mobility and monitor the gel-to-gel and gel-to-liquid crystalline phase transitions of SM as a function of temperature in these mixtures. Spin-spin relaxation measurements are in agreement with the line width results, reflecting mobility differences and some heterogeneities. It will be shown that the presence of DOPC and/or cholesterol greatly impacts the headgroup mobility of SM both above and below the liquid crystalline phase transition temperature, whereas DOPC displays only minor variations in these lipid mixtures.

Cholesterol↗

Ceramide-domain formation and collapse in lipid rafts: membrane reorganization by an apoptotic lipid.

The effect of physiologically relevant ceramide concentrations (< or = 4 mol %) in raft model membranes with a lipid composition resembling that of cell membranes, i.e., composed of different molar ratios of an unsaturated glycerophospholipid, sphingomyelin, and cholesterol (Chol) along a liquid-disordered-liquid-ordered tie line was explored. The application of a fluorescence multiprobe and multiparameter approach, together with multiple fluorescence resonance energy transfer (FRET) pairs, in the well-characterized palmitoyl-oleoyl-phosphocholine (POPC)/palmitoyl-sphingomyelin (PSM)/Chol ternary mixture, revealed that low palmitoyl-ceramide (PCer) concentrations strongly changed both the biophysical properties and lipid lateral organization of the ternary mixtures in the low-to-intermediate Chol/PSM-, small raft size range (<25 mol % Chol). For these mixtures, PCer recruited up to three PSM molecules for the formation of very small ( approximately 4 nm) and highly ordered gel domains, which became surrounded by rafts (liquid-ordered phase) when Chol/PSM content increased. However, the size of these rafts did not change, showing that PCer did not induce the formation of large platforms or the coalescence of small rafts. In the high Chol/PSM-, large raft domains range (>33 mol % Chol), Chol completely abolished the effect of PCer by competing for PSM association. Lipid rafts govern the biophysical properties and lateral organization in these last mixtures.

Absorption↗

Relationship of blood thromboxane-B2 (TxB2) with lipid peroxides and effect of vitamin E and placebo supplementation on TxB2 and lipid peroxide levels in type 1 diabetic patients.

OBJECTIVE: To study the effect of vitamin E supplementation on platelet hyperaggregability in type 1 diabetic patients. RESEARCH DESIGN AND METHODS: Written informed consent according to the Institutional Review Board on Human Experimentation guidelines was obtained from diabetic patients (n = 29) and their age-matched normal siblings (n = 21) to participate in this study. Diabetic patients were supplemented with DL-alpha-tocopherol (vitamin E) capsule (orally, 100 IU/day) or placebo for 3 months in a double-blind clinical trial. Alternate diabetic patients were assigned to vitamin E or placebo during regular visits to the clinic. Fasting blood was collected from each diabetic patient before the start and after the vitamin E or placebo supplementation. Platelet aggregability was assessed by competitive enzyme-linked immunosorbent assay of the blood TxB2 (a stable thromboxane metabolite). Plasma vitamin E and MDA (malondialdehyde, a product of lipid peroxidation) was assessed by high-performance liquid chromatography. Data were analyzed statistically on 12 diabetic patients on vitamin E and 12 on placebo supplementation. RESULTS: Diabetic patients (n = 29) had 62% higher (P < 0.05) levels of TxB2 and 15% higher levels (P < 0.05) of MDA in comparison to normal subjects (n = 21). Plasma TxB2 levels had a significant correlation with MDA levels (r = 0.45, P < 0.02) but not with the HbA1 (r = -0.08), glucose (r = -0.13), duration of diabetes (r = -0.04), or age (r = 0.12) of diabetic patients. Vitamin E supplementation lowered MDA levels by 30% (P < 0.04), TxB2 levels by 51% (P < 0.03), and triglyceride levels by 22% (P < 0.04) in diabetic patients. There were no differences in these parameters before versus after placebo supplementation. CONCLUSIONS: The elevated blood level of TxB2 (hyperaggregability of platelets) is significantly related to the level of lipid peroxidation products (oxidative stress) in type 1 diabetic patients. Supplementation of modest doses of vitamin E (100 IU/day) significantly lowers blood TxB2 and lipid peroxidation products levels in type 1 diabetic patients.

Child↗

Influence of dietary palm oil supplementation on serum lipid metabolites, carcass characteristics, and lipid composition of carcass tissues of growing ram and ewe lambs.

The objective of this research was to determine the influence of dietary palm oil supplementation on carcass characteristics and lipid composition of tissues from growing lambs. Twenty-eight Suffolk x Hampshire lambs were weaned at 60 d of age (average 36 kg BW) and assigned to a 2 x 2 factorial arrangement consisting of diet (control [NPO] or 10.6% added dietary palm oil [PO]) and sex (ram vs ewe). The NPO diet (77% forage and 23% concentrate) contained 11.2% CP and 2.66 Mcal of ME/kg. Palm oil replaced molasses in the PO diet. Lambs were individually given ad libitum access to feed for 60 d to a final BW average of 50.1 kg. Lipid composition of the longissimus muscle and corresponding subcutaneous (s.c.) adipose tissue was determined by gas-liquid chromatography (GLC). Lambs fed PO were fatter (P less than .01) than lambs fed NPO (.77 vs .56 cm, s.c. fat). Diet had no effect on cholesterol content of lean tissue; however, feeding PO increased the saturated fatty acids of lean tissue. The s.c. fat from lambs fed PO had less (P less than .01) cholesterol (64.79 vs 89.67 mg/100 g) and more saturated fatty acids than that from lambs fed NPO. Ewes were fatter (P less than .01) than rams, yet they had less cholesterol content in the s.c. adipose tissue (68.71 vs 85.74 mg/100 g). High amounts of dietary palm oil fed to growing lambs caused changes in fatty acid deposition and cholesterol metabolism and may be a useful investigative tool to study lipid metabolism in growing ruminants.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue↗

Kinetic modelling of in vitro lipid peroxidation experiments--'low level' validation of a model of in vivo lipid peroxidation.

Kinetic modelling overcomes some of the drawbacks of purely intuitive thinking in integrating information accumulated on chemical reactions involved in oxidative stress. However, it is important to assess if current knowledge about the reactions that mediate lipid peroxidation already allows satisfactory modelling of this process in near-to-physiological conditions. In this paper, a set of increasingly complex in vitro experiments on antioxidants (alpha-tocopherol and ascorbate) and lipid peroxidation in heterogeneous systems is simulated. Quantitative to semiquantitative agreement is found between experimental and simulation results. In addition, this theoretical analysis provided useful insights, suggested new hypotheses and experiments and pointed out relevant aspects needing further research. The results encourage and serve as partial validation for the formulation of relatively detailed mathematical models of in vivo lipid peroxidation. Some important aspects of the formulation and analysis of such models are discussed.

Antioxidants↗

Lipid-laden macrophages in tracheal aspirates of newborn infants receiving intravenous lipid infusions: a cytologic study.

Seventy-four neonates with respiratory distress syndrome had cytologic study of tracheal aspirates. Thirteen, all of whom were receiving intravenous lipid and nothing orally had abundant macrophages with foamy cytoplasm, positive for intracytoplasmic lipid by Sudan black stain, in the tracheal aspiration specimens. Cytologic surveillance of tracheal aspiration specimens may be a useful method of monitoring infants receiving intravenous lipid infusions.

Fat Emulsions, Intravenous↗

Mammary lipid metabolism and milk fatty acid secretion in alpine goats fed vegetable lipids.

Fourteen Alpine goats at midlactation were fed a diet of hay and concentrate (55:45), without (control) or with formaldehyde-treated linseed (FLS) or oleic sunflower oil (OSO) at 11.2 or 3.5% of dry matter intake, respectively, in a 3 x 3 Latin Square design with three 3-wk periods. Milk yield was lower in goats fed FLS than control or OSO (2.13 vs. 2.32 kg/d). Milk fat content was higher with FLS or OSO than control (40.8 vs. 33.8 g/kg). Formaldehyde-treated linseed and OSO caused a significant decrease (23 and 18%, respectively) of C10 to C17 fatty acids secretion compared with control. The secretion of cis-9 C18:1 and cis-9, trans-11 C18:2 were increased 1.44- and 1.54-fold for FLS and 1.78- and 1.36-fold for OSO, compared with control. The C18:3 (n-3) secretion was increased 2.61-fold with FLS compared with control. Milk cis-9 C14:1/C14:0, cis-9 C16:1/C16:0, and cis-9 C18:1/C18:0 ratios decreased with the supplemented diets compared with control. Mammary stearoyl-CoA desaturase mRNA and activity were decreased by the lipid supplements, whereas no significant change was observed for acetyl-CoA carboxylase and fatty acid synthase. The activities of glucose-6-phosphate dehydrogenase, malic enzyme, and glycerol-3-phosphate dehydrogenase were not affected by the lipid supplements. Mammary lipoprotein lipase mRNA increased with OSO, whereas lipoprotein lipase activity tended to decrease with FLS compared with control. Milk lipoprotein lipase activity sharply decreased with lipid supplement (by 59 and 71%, for FLS and OSO, respectively). The changes in milk fatty acid profile due to FLS and OSO supplements were partly related to changes in the levels of mammary enzyme activities or mRNA.

Animal Feed↗

Short communication: hydroperoxides in circulating lipids from dairy cows: implications for bioactivity of endogenous-oxidized lipids.

This study was conducted to investigate the potential for increased oxidative stress of high- vs. average-producing dairy cows. Two experiments were performed using 11 and 13 Holstein cows (53 +/- 2 d postpartum). Lipohydroperoxides (LHP) were determined in serum lipids (experiment 1) and low-density lipoprotein (experiment 2) via oxidation of ferrous to ferric ions through LHP using thiocyanate as chromogen. In experiment 1, differing milk yield and milk energy output corresponded to different concentrations of LHP. In experiment 2, analysis of regression resulted in a significant relationship between milk yield and LHP. Phospholipids isolated from lipids with 6.5 microM of LHP evoked in monocytic cells a transient increase in superoxide formation, indicating inflammatory potential. The results show that high milk productivity can associate with oxidative stress indicated by oxidative modifications of circulating lipids and their changed bioactivity.

Animals↗

Changes in platelet aggregation and lipid metabolism in rats given dietary lipids containing different n-3 polyunsaturated fatty acids.

We compared the effects of different n-3 polyunsaturated fatty acids (PUFA) on platelet aggregation and lipid metabolism in rats. alpha-Linolenic acid (ALA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were used as n-3 PUFA sources. The rats were fed diets containing 10% lipids (polyunsaturated/saturated fatty acid (P/S) ratio = 1.0; n-3/n-6 = 0.02 for the control group, 0.2 for the test groups) for two weeks. The platelet counts, platelet aggregation, and production of thromboxane A2 (TXA2), plasma total cholesterol (TC) and triacylglycerols (TG) were not different between the ALA group and the control group, but showed a decreasing tendency for the EPA group and significant decreases for the DHA group. The production of prostacyclin in the aorta was significantly decreased in all of the n-3 PUFA groups when compared with that in the control group. Liver TC and TG concentrations were significantly decreased in the DHA group when compared with those in the control group. Based on the above, it is assumed that the physiological action exerted by n-3 PUFA differs by type and that DHA is a more effective n-3 PUFA, both for suppressing platelet aggregation and for modulating lipid metabolism in the plasma and liver of rats.

Animals↗

Studies on lipolytic system of human placenta, blood lipids and lipid mobilizing activity in pregnant women and newborns.

Lipolytic activity of human placenta and lipid mobilizing activity (LMA), free fatty acids (FFA) as well as esterified fatty acid (EFA) levels in serum of pregnant women in different periods of pregnancy and in newborn's serum were studied. Lipolytic activity of human placenta is low in the first trimester of pregnancy and increases in samples examined after delivery. The levels of FFA and EFA in blood of pregnant women are significantly increased in the second and third trimester of pregnancy. The newborn's blood lipid level is significantly lower than that in the mother's blood. Lipid mobilizing activity in serum of pregnant women increases especially in the third trimester of pregnancy and is also high in newborn's blood. This may indicate that fatty acids released from newborn's adipose tissue are metabolized as a source of energy.

Adipose Tissue↗