Lipid composition and endogenous respiration of pig heart mitochondria

Lipid composition and endogenous respiration of pig heart mitochondria were studied in parallel, since the level of endogenous respiration affects the oxidation of added substrates and therefore the regulation of oxidative phosphorylation; mitochondrial lipids can interfere either as substrates or as partner in the energy conservation mechanism. O₂ uptake kinetics were measured in presence of different additives: ATP, ADP, NAD⁺ and hexokinase + glucose. The lipid composition of pig heart mitochondria was determined by chromatographic and spectrophotometric methods. Total lipids were 90% phospholipids; the main phosphatides were cardiolipin, phosphatidyl choline and phosphatidyl ethanolamine; the two latter were rich in plasmalogens. The main nonpolar lipids were triglycerides and free fatty acids. The fatty acid composition of total lipids, phospholipids, free fatty acids and triglycerides was determined by gas liquid chromatography. Mitochondrial lipids were characterized by a high content of unsaturation. Part of this work is included in “Thèse de Doctorat de Spècialitè en Biochimie” de J. Comte, Lyon, June 26, 1970.     

Lipid and fatty acid composition of the endogenous energy sources of striped bass (Morone saxatilis) eggs

The unique physiological flexibility of the early life stages of striped bass is attributed to the calorierich endogenous energy sources of the striped bass egg. Eggs of different aged striped bass from geographically separate populations were examined for lipid and fatty acid compositions and were found to be basically similar. Yolk components of the eggs contained significantly less total lipid than oil globules, were more diverse in lipid class composition and consisted mostly of polar lipids. Oil globules were entirely lipid material consisting predominantly of steryl/wax esters. Fatty acid compositions of yolk and oil globules differed according to their respective lipid compositions. The functional significance of these lipids is discussed in relation to the ecological context of the early life stages.     

Lipid composition and protoplast-forming capicity ofStreptomyces antibioticus

The lipid and fatty acid composition of a strain ofStreptomyces antibioticus has been studied as a function of culture age and glycine concentration in the growth medium. Under all conditions, the main polar lipids were phosphatidylethanolamine, cardiolipin and phosphomannoinositides in order of decreasing abundance; no ornithinolipids were detected. Acylglucoses and menaquinones were found among the nonpolar lipids. The main fatty acids present were anteiso 15∶0 and anteiso 17∶0. The lipid composition of the cells varied with the age of the culture, but no uniform pattern of variation was found in the cultures grown on different amounts of glycine. Among the cells harvested at the end of the exponential phase of growth, those grown on 2% glycine give the highest yield of protoplast formation. These cells were found to contain low amounts of nonpolar lipids and of phosphatidylethanolamine, and high proportions of anteiso fatty acids. We propose that the membrane bilayer of these cells, because of its peculiar lipid composition, is particularly stable and fluid. As a consequence, the yield and stability of derived protoplasts should be increased.     

Lipid composition ofbacillus megaterium spores and spore membranes

Bacillus megaterium QM B1551 spore lipids were extracted by an improved technique, and the phospholipid and fatty acid compositions were determined. Phospholipids accounted for 65% of the total fatty acids; the neutral lipid fraction contained 15% and the remaining fatty acids were in the interphase, aqueous phase and pellet from the lipid extraction. Each phospholipid had similar fatty acid compositions as did the delipidated pellet. However, the aqueous phase and, to some extent, the interphase had unique fatty acid compositions. Also, fatty acids were found acylated to proteins, which was observed by electrophoresis of delipidated proteins from spores grown in [1-¹⁴C]palmitate. Therefore, spores contain unique non-phosphatide fatty acid components that can now be analyzed.     

Lipid composition of normal and hypertrophic bovine thyroids

The phospholipid content of bovine thyroid tissue amounts to 70% of total lipid. Triglycerides and cholesterol are the main neutral lipids. Only trace amounts of free fatty acid and esterified cholesterol are found, while two not yet identified components also are present. The distribution of lipid phosphorus in the different phospholipid classes is as follows: phosphatidyl choline, 43.0%; phosphatidyl ethanolamine, 25.2%; phosphatidyl serine, 5.6%; phosphatidyl inositol, 6.5%; sphingomyelin, 14.0%; cardiolipin, 2.8%; lysophosphatidyl choline, <1%; and phosphatidic acid, <1%. The phosphatidyl ethanolamines are rich in plasmalogens. The fatty acid patterns in the different lipid classes are reported. The essential differences between normal and hypertrophic bovine thyroid tissue are higher water content and lower triglyceride and sphingomyelin values for hypertrophic tissue.     

Lipid composition and biosynthesis of human omental tissue

Two groups of five males each were selected for total lipid analyses of their omental tissue. One of these groups had been subjected to a severe caloric restriction and had undergone total weight reduction of about 20%. The other group served as control. Both of these groups of patients required elective surgical procedures during which it was possible to obtain small samples of omental tissue, adipose pad, and/or mesenteric tissue. Total lipid analyses were performed on all of the materials. A distinctive positional distribution of the acyl groups was maintained in the triglycerides of omental tissue for all the patients regardless of dietary state. Patients under caloric restriction showed a reduction in their total triglyceride content, a reduction in their content of unsaturated fatty acyl groups, and a relative increase in phospholipid content. The de novo lipid biosynthetic capacity of omental tissues, as determined by 1-[¹⁴C]-acetate incorporation, showed an inverse proportionality to the lipid content of the samples. Omental tissue is biosynthetically a very versatile material capable of yielding rapidly many types of fatty acids. This ability, among others, could account for the usefulness of omental tissue as a supporting base in many types of restorative surgery.     

Lipid composition of beef and human pituitary glands

The lipid composition of beef and human pituitary was determined by chromatographic and spectrophotometric methods. Beef pituitary lipid contained about 25% nonpolar lipids and 75% phospholipids whereas nonpolar lipids made up approximately 60% of the total in human pituitaries. The main nonpolar (i.e., low polarity) lipids in human pituitary were triglycerides, cholesterol, free fatty acids and an unidentified component in the triglyceride fraction. Cholesterol was the major nonpolar lipid component in freshly collected beef anterior and posterior pituitary, but the amount of free fatty acids appeared to increase during storage. Preliminary investigation of the unknown nonpolar lipid in human pituitaries suggested that it was an unsaturated hydroxy compound with no carbonyl functions. Thin layer chromatography indicated that it was also present in smaller amounts in freshly collected beef pituitaries. The main phospholipids of beef anterior, posterior and human pituitary were phosphatidyl ethanolamine, phosphatidyl choline, phosphatidyl inositol, phosphatidyl serine and sphingomyelin. The fatty acid composition of total nonpolar lipids, free fatty acids, total phospholipids, phosphatidyl ethanolamine and phosphatidyl choline of beef anterior and posterior pituitary was determined by gas liquid chromatography. Mixtures of saturated and unsaturated fatty acids ranging from C₁₂ to C₂₂ were present; the main fatty acids were palmitic, stearic, oleic, linoleic and arachidonic.     

Lipid composition of liver mitochondria and microsomes in hyperthyroid rats

Triiodothyronine-induced alteration of the lipid pattern in rat-liver mitochondria and microsomes has been investigated. In mitochondria, a 25% total cholesterol decrease and a 14% phospholipid increase have been detected. In these hyperthyroid rat liver organelles, a strong decrease in the total cholesterol/phospholipid molar ratio occurs. On the contrary, in microsomes from the same animals, a decrease of about 23% has been measured for both total cholesterol and phospholipids; hence, in this fraction, the total cholesterol/phospholipid molar ratio is unaffected by hyperthyroidism. The liver mitochondrial phospholipid composition, unlike the microsomal composition, is altered significantly in hyperthyroid rats; a 7.4% phosphatidylcholine decrease is accompanied by a similar additive percentage increase of both phosphatidylethanolamine and cardiolipin. In regard to total phospholipid fatty acid composition in liver microsomes from hyperthyroid rats, no variation has been observed compared with the control rats, whereas in mitochondria from the same animals, a meaningful linoleic acid decrease with a similar arachidonic acid increase has been found. In addition to fatty acid alteration, the separated mitochondrial phospholipid classes also exhibit some increase in stearic acid. Among phospholipids, cardiolipin changes the most of the esterified fatty acids in hyperthyroid rat liver. In this compound, a strong increase in the percentage of both palmitic and stearic acid and a 32.4% decrease of linoleic acid have been found.     

Lipid composition and erucic acid in rat liver cells in culture

Erucic acid (Δ¹³-docosenoic acid) was added to fetal calf serum, then fed to rat liver epithelial cells in culture, and uptake measured at intervals over 24 hr. During the first 6 hr. of incubation, uptake of the docosenoic acid was 21 nmoles/hr/mg protein in 7-day cells, and 15 mmoles/hr/mg protein in 14-day cells. Of¹⁴C-labeled erucic acid taken up by the cells in 24 hr, radioactivity measurements showed 60% of the total lipid¹⁴C activity derived from [1-¹⁴C] 22∶1 in neutral lipid (NL) and 40% in phospholipid (PL); whereas 55% of lipid¹⁴C activity was in NL and 45% in PL when the substrate was [14-¹⁴C] 22∶1. Within the NL fraction, 75% of¹⁴C activity derived from [1-¹⁴C] 22∶1 was in triglyceride (TG) and 11% in cholesterol (CHL), while 79% was in TG and 6.5% in CHL when the substrate was [14-¹⁴C] 22∶1. Triglycerides and cholesteryl esters accumulated in the cells during incubation with erucic acid. Among phospholipids separated by thin layer chromatography, 75% of¹⁴C activity was in lecithin (PC), 10% in phosphatidylethanolamine (PE), 5% in sphingomyelin (SPH), and 1% or less in cardiolipin (DPG). The highest specific activity (SA) was in PC, followed by SPH and PE. Incubation with erucic acid altered fatty acid composition of PC, PE, and SPH, although amounts of phospholipids were unaffected. Gas liquid chromatography analyses detected 18% erucic acid in PC, 2% in PE, and 4–5% in SPH.     

Alkali-silica reaction and pore solution composition in mortars in sea water

The promotion of expansion of mortars containing a reactive aggregate in 1N NaCl solution at 38 °C was attributed to a rise of OH⁻ ion concentration in the pore solution in the mortars. However, it is ambiguous whether the promotion of expansion of mortars in sea water at a room temperature can be explained in the same way as in NaCl solution at an elevated temperature. This study aims at pursuing the expansion behavior of mortars containing a reactive aggregate relating it to their pore solution composition and the extent of alkalisilica reaction occurring within reactive grains. The alkali-silica reaction in mortars in sea water and 0.51N NaCl solution at 20 °C appears to progress differently from that in mortars in 1N NaCl solution at an elevated temperature of 38 °C. The promotion of expansion of mortars in sea water at 20 °C was found to be responsible for an effect of Cl⁻ ions on the alkali-silica reaction at early stages of immersion. Only when OH⁻ ion concentration in the pore solution was relatively high, NaCl and sea water could accelerate the alkali-silica reaction in mortars at 20 °C.     

Lipid composition of beef brain,beef liver,and the sea anemone: Two approaches to quantitative fractionation of complex lipid mixtures

Two new schemes for fractionation of complex lipid mixtures are presented. Their use for the study of lipids of beef brain, beef liver, and the sea anemone are described. Apparatus and techniques for working in an inert atmosphere, evaporation of solutions in the cold under nitrogen, use of infrared spectroscopy for examination of lipids and their hydrolysis products, preparation and clution of diethylaminoethyl (DEAE) cellulose and silicic acid-silicate columns and general column combinations that can be used to fractionate complex lipid mixtures are considered in detail. The first scheme, employing DEAE cellulose columns followed by thin layer and paper chromatographic examination of the fractions, was applied to liver lipids. The many components, some of them new lipids not previously detected, are clearly seen with this technique but are not seen when paper or thin layer chromatography alone or silicic acid chromatography are used. The second scheme employing DEAE for initial fractionation, followed by complete separation on silicic acid and silicic acid-silicate columns, was applied to lipids of the sea anemone and beef brain. Typical lecithin and phosphatidyl ethanolamine were isolated, but sphingomyelin was not found. A new sphingolipid, ceramide aminoethylphosphonate, with a free amino group and a direct carbon to phosphorus bond was isolated and characterized. The methods used for quantitative isolation, the infrared spectra, and the amounts of cholesterol, ceramide, cerebroside, galactosylglyceride, sulfatide, sphingomyelin, lecithin, phosphatidyl ethanolamine, phosphatidyl serine, phosphatidyl inositol, triphosphoinositide, phosphatidic acid, cardiolipin, and ganglioside of beef brain are presented. Finally, the types of lipid-nonlipid interactions disclosed by column chromatography and their potential application to biological problems are discussed.     

Lipid composition of glucose-stimulated pancreatic islets and insulin-secreting tumor cells

The effect of glucose stimulation (25 mM for 5 min) on the phospholipid and neutral lipid composition of isolated pancreatic islets was studied to find out whether there is a change in the mass of potential lipid mediators or modulators of insulin secretion. For comparison, the lipid compositions of homogenates and subcellular fractions from RINm5F insulin-secreting tumor cells and of glucose-stimulated streptozotocin/nicotinamide-induced islet cell tumors were analyzed. After separation of the lipid extract into a neutral and an acidic fraction by anion-exchange chromatography, lipids were separated by high-performance thin-layer chromatography and quantitated byin situ densitometry of the cupric sulfate-charred bands. In glucose-stimulated islets, the molar percentages of phosphatidic acid (PA) and of phosphatidylinositol were significantly increased (3.1 vs. 4.7 mol% and 8.6 vs. 11.8 mol%), while those of all other phospholipids and neutral lipids, including 1,2-diacylglycerol, were not significantly changed. In stimulated islet cell tumors, an increase of PA was visible in the microsomal fraction, and there was an increase of lysophosphatidylcholine in the mitochondrial fraction. However, in both tumoral tissues, particularly in RINm5F cells, the lipid distribution pattern showed abnormalities which can be regarded as a loss of differentiation and which limit the usefulness of these tissues for the study of the physiological regulation of lipid metabolism during glucose stimulation. In conclusion, the data are in accordance with a role of PA early in stimulus-secretion coupling. The well-known stimulation of phospholipid synthesis in pancreatic islets during glucose-induced insulin secretion does not result in an increase in the total phospholipid mass. The results were taken in part from the M.D. thesis of A.M.     

Lipid and fatty acid composition of testes of quaking mice

Testes of quaking mice (sterile mutants) and of controls were analyzed for major lipid classes and fatty acid composition. Of the main lipid classes, only cholesterol esters differed significantly in concentration between the two groups (1.01 for quakers vs 0.69 mg/g wet wt of tissue for controls). The concentration of triglycerides was 4.5–5.0, that of total phosphatides 18–19, and that of free cholesterol 1.9–2.0 mg/g for mutants and controls. The concentrations of phosphatidyl ethanolamine and of sphingomyelin were both lower in quaking than in normal mice, but only the change in the former was statistically significant. Phosphatidyl choline was the major phosphatide (43–45% of total phosphatides) followed by phosphatidyl ethanolamine (24–26%) and sphingomyelin, phosphatidyl serine, and phosphatidyl inositol (all ca. 7% of total phosphatides). Minor differences between the mutants and controls were observed in concentrations of fatty acids of major lipid classes. The mutants, sterile because of faulty spermatid differentiation, had normal quantities of 22∶6 w3 and 22∶5 w6. These data are consistent with the hypothesis that the 22-carbon polyenes are associated with the formation of spermatids, rather than with their final differentiation into spermatozoa.     

Lipid composition and amino acid transport in a nystatin-resistant mutant ofAspergillus niger

A nystatin-resistant mutant ofAspergillus niger has been isolated and used as a model system to study the effect of altered sterol levels on lipid composition, transport behavior and physical properties of membrane lipids. There is a decrease in the sterol to phospholipid ratio in the mutant compared to the wild type. Although there is no qualitative change in phospholipid composition, the mutant contains a higher amount of phosphatidylcholine and a lower amount of phosphatidylethanolamine compared to the wild type. The most significant change is the elevated level of linoleic acid in the mutant, concomitant with a decreased level of oleic acid. These adaptive changes to nystatin resistance are manifested in the altered thermotropic behavior of membrane lipids as studied by the steady-state fluorescence polarization technique. These changes are also associated with altered membrane permeability as evidenced by the change in V_max values for uptake of some amino acids in the mutant compared to the wild type.     

Lipid and fatty acid characterization and metabolism in the sea anemonePhymactis clematis (Dana)

Neutral lipid, phospholipids and fatty acids of the sea anemonePhymactis clematis from the south-west Atlantic were characterized and quantified in spring and autumn. Neutral lipids predominated over phospholipids in both seasons. Triacylglycerol and diacylglycerol ethers were the major lipids. In spring, an increase of esterified sterols was noted. The major fatty acids found were 22∶5ω3, 20∶5ω3 and 16∶0. The sea anemones were also incubated in vivo with either [1-¹⁴C]linoleate or [1-¹⁴C] α-linolenate for 2 hr. Isotope incorporation into lipids and their transformations into higher fatty acids were examined. Both precursors were incorporated into the lipids, mainly in triacylglycerols and mono-acylglycerols, while α-linolenate was also incorporated into phospholipids. The radioactive linoleate was elongated to 20∶2, 22∶2 and 24∶2 fatty acids, but not desaturated to 18∶3ω6. α-Linolenate was desaturated by Δ6 desaturase to 18∶4ω3. The specificity of Δ6-desaturase is discussed.     

Lipid composition of subcellular membranes from larvae and prepupae ofDrosophila melanogaster

Subcellular membranes were analyzed for their lipid composition and protein content at two developmental points representing the third instar wandering larvae and prepupal stages ofDrosophila. At both stages, phosphatidylethanolamine (PE) and phosphatidylcholine (PC) were the major constituents with phosphatidylinositol (PI), phosphatidylserine (PS), diphosphatidylglycerol (DPG) and phosphatidic acid (PA) being relatively minor components. In total homogenates and in the nuclear-enriched fraction there was no significant difference in the phospholipid composition of the wandering larvae and prepupae. In mitochondria only a significant increase in the minor component PS was observed in the prepupae. In lysosomal membranes on the other hand, the relative abundance of the major components PE and PC increased in the prepupae although the molar ratios of the two lipids remained almost constant. The fatty acid composition of the phospholipids remained virtually unchanged in all of the fractions examined, including the lysosomes, and there was no evidence of lipid peroxidation. With regard to cellular degeneration and the involvement of lysosomes, we conclude that mechanisms other than gross modification of the lipid and/or lipid/protein ratio of their membranes are involved in the liberation of the acid phosphatase contents.     

Lipid profile and fatty acid composition of two silurid fish eggs

The lipid content and the composition pattern of the lipid class including fatty acid composition in the eggs of two different Indian silurid cat fishes Ompok pabda and Wallagu attu have been examined. The lipid content of O. pabda and W. attu (on dry basis) are about 14.7% and 17.8% respectively. The major lipid classes are phospolipid (PL) and triacylglycerol (TAG). The O. pabda egg lipid contains more PL while the W. attu egg lipid contains more TAG. Phosphatidylcholine (PC) constitutes the major phospholipid followed by phosphatidylinositol (PI). PI represents in about 31.7% and 21.3% of total PC in O. pabda and W. attu respectively while phosphatidylethanolamine (PE) (about 28.0%) is significantly higher in the egg of W. attu than O. pabda (9.6%). Cholesterol content in egg of O. pabda is also higher (about 9.6%) than W. attu (4.1%). The lipids are rich in polyunsaturated fatty acids (PUFAs) and they are mainly concentrated in the respective PL fractions. Among PUFAs the arachidonic acid (20:4 n-6 AA) is present at about 9.3% in both egg PL. Eicosapentaenoic acid (20:5 n-3 EPA) is significantly lower in egg lipids of both W. attu (1.8%) and O. pabda (3.2%), whereas docosahexaenoic acid (22:6 n-3 DHA) is predominantly higher (14.6% and 18.1% in W. attu and O. pabda respectively) in their PL fractions.     

Lipid composition of cultured endothelial cells in relation to their growth

Human endothelial cells in culture were examined in different growth conditions. The human endothelial cell line, EA.hy 926 cell line, was used and cells were studied either in exponential growth phase, at confluence, or growth-arrested by serum deprivation. Phospholipids were separated and analyzed by high-performance thin-layer chromatography, and their fatty acids were quantified by gas-liquid chromatography. No significant differences in the phospholipid distributions were found between exponentially growing and confluent endothelial cells in which phosphatidylcholine (PC) represented the major phospholipid. In comparison, serum-deprived cells exhibited higher proportions of sphingomyelin and lower content of PC. We also found that among the total lipids, cholesterol level for dividing endothelial cells was lower than for cells growth-arrested either by serum deprivation or by contact inhibition at confluence. The global fatty acid distribution was not affected by the growth conditions. Thus, oleate (18∶1n−9 and 18∶1n-7), palmitate (C_16∶0), and stearate (C_18∶0) were the main components of endothelial cell membranes. However, the fatty acid distributions obtained from each phospholipid species differed with the growth status. Altogether, the data indicated that subtle modulations of endothelial cell metabolism appear upon cell growth. The resulting membrane-dependent cellular functions such as cholesterol transport and receptor activities can be expected to be relevant for lipid trafficking within the vessel wall in vitro and in vivo.     

Lipid composition and de novo lipid biosynthesis of human palmar fat in Dupuytren's disease

Seventy-two surgically obtained Dupuytren's disease palmar-fat (DDPF) specimens and 18 location-matched specimens from patients not suffering from this disease (controls) were studied for their total lipid composition and de novo lipogenic activity. Incubation of "DDPF" with 1-[14C]acetate in oxygen produced [14C]palmitate and [14C]stearate in approximately equal yields as those obtained from "controls." No [14C]octanate was formed in any of the palmar-fat preparations. The lipids and fatty acid analysis revealed differences: (a) DDPF specimens were richer in free fatty acids, methyl esters of fatty acids and free-cholesterol than specimens of controls. (b) DDPF specimens contained less phospholipids. (c) DDPF specimens showed a significantly higher content of octanoate and other short-chain fatty acids than specimens of controls. The above findings are not incompatible with the results expected if some mild hypoxia occurred in DDPF; this has been suggested in the statistical correlations observed for this disease and alcoholism with liver involvement.     

Lipid composition in tomato skin supercritical fluid extracts with high lycopene content

The use of supercritical fluid extraction is proposed for obtaining stable extracts with high added value from natural and economical sources. Lipid composition, namely, of FFA, TAG, and FAME, in tomato skin extracts with high lycopene content was determined. Separation of different classes of lipids was achieved from tomato extracts using TLC followed by transesterification and GC, and lycopene and other carotenoids were analyzed by HPLC with a photodiode array detector. In lycopene extracts obtained using supercritical fluids, no FFA were found and polyunsatureted TAG represented only 9.2% of the total TAG content.