Distribution of cholesteryl esters and other lipid in subcellular fractions of the adrenal gland of the pig

Total lipids from whole pig adrenal glands as well as from their mitochondria, microsomes, liposomes, and cell sap were extracted and fractionated first into neutral lipids and phospholipids. The highest percentage of neutral lipids was found in the cell sap, and the lowest in the microsomal fraction. Neutral lipids were subfractionated into cholesteryl esters, free cholesterol, triglycerides, and free fatty acids. Cholesteryl esters were distributed throughout the liposomes. Free fatty acids represented a substantial part of cell sap lipids, but were present also in the mitochondria, microsomes, and liposomes. Fatty acids of all fractions were analyzed by gas liquid chromatography. Free fatty acids and cholesteryl ester fatty acids from all cellular fractions were similar in composition and were characterized by considerable quantities of linoleic and arachidonic acid. Triglycerides were characterized by an increased percentage of palmitic and a low content of arachidonic acid. Phosphatidyl choline, phosphatidyl ethanolamine, diphosphatidyl glycerol, and sphingomyelin plus phosphatidyl inositol were isolated from the lipids by preparative thin layer chromatography, and their fatty acids analyzed by gas liquid chromatography. Phosphatidyl choline and phosphatidyl ethanolamine from mitochondria, microsomes, and cell sap were very similar in respect of their fatty acid composition. Sphingomyelin plus phosphatidyl inositol was characterized by a high content of C22:2omega6. Diphosphatidyl glycerol was present in mitochondria and in the cell sap.     

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.     

The structures of the principal glycerolipids of pig liver

The lipid composition of pig liver has been determined. The principal glycerolipids, i.e., triglycerides, phosphatdyl choline, phosphatidyl ethanolamine and phosphatidyl inositol, were isolated and the positional distribution of fatty acids in each determined by stereospecific analysis procedures. Previous results for the triglycerides were confirmed, while the phospholipids were similar in structure to those found in most other animal livers. The triglycerides were separated into simpler molecular species by combinations of silver nitrate thin layer chromatography and high temperature gas liquid chromatography, but the proportions found did not agree well with those calculated assuming a 1-random, 2-random, 3-random arrangement. The phospholipids were hydrolyzed with phospholipase C and converted to diglyceride acetates that were fractionated into simpler molecular species by the same procedures as were used with triglycerides. Highly specific fatty acid combinations were found in molecular species, and these specificities were very similar to those reported in similar lipids from the livers of such disparate species as the rat and chicken.     

Quantitative and qualitative analyses of isolated lipid droplets from interstitial cells in renal papillae from various species

The lipid droplets of renal papillae homogenates from four different species were obtained by ultracentrifugation. Ca. 80–98% of the lipids (triglycerides, phospholipids, free fatty acids, and cholesterol esters) consist of triglycerides. The triglycerides were fractionated by argentation thin layer chromatography and each fraction characterized by gas liquid chromatography. No fraction contained any unique triglyceride. The fatty acid composition of the total triglycerides, as analyzed by gas liquid chromatography and ozonolysis, differed markedly from the fatty acid composition of the corresponding plasma triglycerides. The papillary triglycerides were characterized by higher concentrations of stearic acid, arachidic acid, and polyunsaturated acids with 20 or more carbon atoms. Particularly interesting was the presence in the lipid droplets of docosa-7,10,13,16-tetraenoic acid. This acid has been shown to be a major component in the cholesterol ester fraction of rat and canine adrenal lipids. In the papillary triglycerides, this acid accounted for 7%, 15%, and more than 20% of the total fatty acids in the dog, rat, and rabbit, respectively. The pig differs from these three species in having only ca. 1% of this acid. These observations suggest that the interstitial cells produce these triglycerides. This production could occur either by a transacylation from phospholipids and cholesterol esters and by a de novo synthesis from locally produced fatty acids. The possibility that the triglyceride production may be involved in a control of the prostaglandin production of the renal medulla is discussed.     

Structural comparison between triglycerides and phospholipids from pig kidney

The compositional specificity of the major diacyl phosphatides, plasmalogens and triglycerides of pig kidney has been determined. The triglycerides have been shown to be esterified predominantly with saturated fatty acids in the 2 position while the phosphatides have predominantly unsaturated fatty acids in this position. Such complete “inversion” of the structure of triglycerides and phospholipids is not normally seen in mammalian tissue and suggests that the synthesis of structural polar lipids involves other steps than the reaction of a diglyceride with CDP-ethanolamine or CDP-choline. This marked difference in the fatty acid distribution of the polar lipids and triglycerides of the pig kidney may render this tissue especially suitable for studies on the turnover and synthesis of structural lipids.     

Thermophilic fungi: III. The lipids ofHumicola grisea var.thermoidea

The lipids of the thermophilic fungusHumicola grisea var.thermoidea were qualitatively and quantitatively determined. The polar lipids consisted of 38.4–42.3% of the total lipids. The relative per cent phospholipids based upon the total phospholipids were as follows: phosphatidyl choline, 32.3–33.7%; phosphatidic acid, 24.5–31.7%; phosphatidyl ethanolamine, 15.8–20.9%; phosphatidyl inositol, 12.5–13.0%; phosphatidyl serine, 2.3–5.4%; and diphosphatidyl glycerol, 3.9–4.0%. The relatively high concentration of phosphatidic acid may be characteristic of fungi grown at elevated temperatures. Several sterol glycosides (3.1–6.0%) were present in the polar lipids. The neutral lipids consist of triglycerides, 28.6–36.0%; free fatty acids, 5.3–13.5%; sterols, 11.4–13.9%; sterol esters, 1.8–3.0%; and diglycerides, 2.2–3.4%. The sterols and derivatives comprise an unusually large fraction of the total lipids (16.3–22.9%) suggesting a role in thermostability.     

Lipid metabolism of the yellow clam,Mesodesma mactroides: I. Composition of the lipids

The lipid composition of the yellow clam,Mesodesma mactroides, that lives in the northern beaches of the Buenos Aires province of Argentina was studied. The main nonpolar lipids are triglycerides and alkoxyglycerides. Phosphatidyl choline, phosphatidyl ethanolamine, and phosphatidyl serine are the main phospholipids. The predominant fatty acids are 16∶0, 16∶1ω7, 18∶0, 18∶1ω9, 20∶5ω3, and 22∶6ω3. The are mainly provided by the clam's food and stored in the hepatopancreas. The content of polyunsaturated acids increases in summer together with an increase in nonpolar lipids and is correlative with an increase in phytoplankton in the sea water. Sexual maturity modifies the lipid composition of gametes.     

The lipid composition of microsomal preparations from lactating bovine mammary tissue

The microsomes isolated from lactating bovine mammary tissue contained 4.3 mg lipid per milligram nitrogen. Phospholipids comprised 83% of the lipids. The neutral lipids were composed of triglycerides (20–30%), diglycerides (5–10%), free fatty acids (15–30%, cholesterol (35–40% and cholesterol esters (10–12%, respectively. Phosphatidylcholine was the predominant phospholipid component (>50%), and the remainder consisted of phosphatidylethanolamine (21–13%), phosphatidylserine (4–6%), phosphatidylinositol (8%), sphingomyelin (9%) and lysophosphatidylcholine (2%) respectively. The composition of the microsomal phospholipids was similar to that of isolated mammary cells and tissue homogenates but quite different from milk and fat globule membrane phospholipids. The triglycerides contained short chain fatty acids but their relative concentrations were lower than in milk triglycerides. The various lipid fractions had a variable proportion of saturated fatty acids, i.e., triglycerides (47.7%), diglycerides (86.7%), free fatty acids (70.6%), phosphatidylcholine (50.6%), phosphatidylethanolamine (50.8%), phosphatidylserine (35.3%), phosphatidylinositol (40.5%) and sphingomyelin (82.3%), respectively. The molecular distribution of fatty acids in the microsomal triglycerides and phosphatidylcholine was similar to that occurring in milk, i.e., the short chain and unsaturated fatty acids were concentrated in the primary positions (sn1 andsn3) of the triglycerides, and the unsaturated acids were preferentially located in positionsn2 of the phosphatidylcholine. The compositional data indicate that mammary microsomes are not the direct source of the phospholipids of the milk fat globule.     

Phospholipid Composition of <Emphasis Type="Italic">Jatropha curcus</Emphasis> Seed Lipids

Jatropha curcus L. oil has emerged as one of the most important raw materials for biodiesel production. However, no detailed study has been reported on characterizing the lipid constituents of jatropha oil. The present study revealed that the total oil content of jatropha seeds was 32% with a composition of 97.6% neutral lipids, 0.95% glycolipids and 1.45% phospholipids. The fatty acid composition of total lipids, neutral lipids, phospholipids and glycolipids was also determined and found to contain oleic acid (18:1) and linoleic acids (18:2) as major fatty acids. The phospholipids fraction was further characterized and quantified and found to contain phosphatidyl choline (PC) 60.5%, phosphatidyl inositol (PI) 24% and phosphatidyl ethanolamine (PE) 15.5%. The fatty acid composition and the positional distribution of the fatty acids of individual phospholipids were also reported.     

Lipid Composition of Garlic

Neutral, glyco- and phospholipids of garlic were resolved into their component fractions by thin layer chromatography. Neutral lipids contained considerable quantities of monoglycerides (18.5%), diglycerides (14.2%), sterols (16.3%) and triglycerides (41.5%) respectively. The phospholipid fraction was rich in phosphatidyl choline (23.5%), phosphatidyl ethanolamine (17.9%), lysophosphatidyl choline (11.8%) and lysophosphatidyl ethanolamine (8.2%). Digalactosyl diglyceride (10.1%), sterol glycoside (15.6%), cerebrosides (8.1%), acylsterol glycoside (38.6%) and monogalactosyl diglyceride (22.5%) were the major components of the glycolipids of garlic. Lauric, myristic, palmitic and linoleic acids constituted the major fatty acids of monoglycerides, diglycerides and free fatty acid fractions whereas palmitic, linoleic and linolenic acids were the major fatty acids of triglycerides. Palmitic and linoleic acids were the major fatty acids of garlic phospholipids. Except the acylsterol glycoside fraction glycolipids were rich in lauric, palmitic, linoleic and linolenic acids; palmitic acid was the only major fatty acid of acylsterol glycosides.     

The lipid components of white potato tubers (Solanum tuberosum)

Four Canadian varieties of potatoes were examined for their lipid composition. Lipids, extracted with chloroformmethanol, were shown by TLC and column chromatography to consist of 16.5% neutral lipids, 45.5% phospholipids and 38.1% glycolipids. Among the phospholipids and glycolipids, phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl inositol, the galactolipids and the sterol glucosides were the major lipids. The predominant fatty acids were palmitic (19.5%), linoleic (44.8%) and linolenic (30.4%, in Kennebec). Analyses of the fatty acids of stored potatoes showed a marked decrease in linoleic acid and an increase in linolenic acid, in the Irish Cobbler and Sebago potatoes. β-sitosterol comprised 85.0% of total sterols. Nearly half of the carotenoids was lutein (xanthophyll), the others being α-carotene, β-carotene, an unidentified pigment and lutein epoxide. Contribution No. 101 of the Food Research Institute, Canada Department of Agriculture.     

Lipid composition ofNeurospora crassa

The lipids ofNeurospora crassa, isolated in pure form from freeze-dried mycelium, were found to contain squalene, sterol esters, triglycerides, free fatty acids, geranylgeraniol, free sterols, carotenoids, cardiolipin, phosphatidyl ethanolamine, phosphatidyl choline, phosphatidyl serine, and phosphatidic acid. The above compounds were isolated in pure form by column and thin layer chromatography and were characterized by infrared spectroscopy and chromatographic mobilities. Fatty acid moieties were characterized by gas liquid chromatographic retention times of their methyl esters relative to those of authentic standards. The fatty acid composition of the triglycerides was found to be similar to that of phosphatidic acid, cardiolipin, and lecithin.     

Lipids of some thermophilic fungi

Total lipid content in the thermophilic fungi—Thermoascus aurantiacus, Humicola lanuginosa, Malbranchea pulchella var.sulfurea, andAbsidia ramosa—varied from 5.3 to 19.1% of mycelial dry weight. The neutral and polar lipid fractions accounted for 56.4 to 80.2% and 19.8 to 43.6%, respectively. All the fungi contained monoglycerides, diglycerides, triglycerides, free fatty acids, and sterols in variable amounts. Sterol ester was detected only inA. ramosa. Phosphatide composition was: phosphatidyl choline (15.9–47%), phosphatidyl ethanolamine (23.4–67%), phosphatidyl serine (9.3–17.6%), and phosphatidyl inositol (1.9–11.9%). Diphosphatidyl glycerol occurred in considerable quantity only inH. lanuginosa andM. pulchella var.sulfurea. Phosphatidic acid, detected as a minor component only inM. pulchella var.sulfurea andA. ramosa, does not appear to be a characteristic phosphatide of thermophilic fungi as suggested earlier. The 16∶0, 16∶1, 18∶0, 18∶1, and 18∶2 acids were the main fatty acid components. In addition,A. ramosa contained 18∶3 acid. Total lipids contained an average of 0.93 double bonds per mole of fatty acids, and neutral lipids tend to be more unsaturated than phospholipids.     

On the phospholipids ofCulex pipiens fatigans

The lipids of different developmental stages ofCulex pipiens fatigans, vector of bancroftian filariasis, have been investigated. The phospholipid composition of the developmental stages and of the subcellular fractions of fourth instar larvae of the insects were analyzed. The composition of fatty acids and their positional distribution have also been examined in the major phospholipids of the larvae. The insect eggs contained higher amounts of lipids than larvae suggesting that they were utilized during embryogenesis. Phosphatidyl ethanolamine (PE) and phosphatidyl choline (PC) comprised over 75% of the insect phospholipids. Of these, PE was present in the greatest amounts during all stages of growth and in the subcellular fractions of larvae. An ethanolamine containing sphingolipid was found as a component of the phospholipids of the insects. About 50% of the lipids of the larvae were localized in the cell debris and nuclei fraction which also contained most of the lysolipids of the insects. As in other Diptera 16∶0, 16∶1 and 18∶1 were the major fatty acids present in the insect lipids of which the fatty acid found in greatest amounts was 16∶1. Similar to the phospholipids of animal species, saturated fatty acids were predominantly linked to the 1 position of the major phospholipids of the insects while the unsaturated fatty acids were in higher amounts at the 2 position.     

Quantitative effects of dietary polyunsaturated fats on the composition of fatty acids in rat tissues

A method combining data on fatty acid composition into subsets is used to illustrate general relative competitive selectivities in the metabolic and transport events that maintain fatty acid compositions in tissue lipids and to minimize differences among tissues or species in the amount of individual fatty acids. Fatty acid compositions of triglycerides and phospholipids in several tissues of the rat were maintained with simple relationships between the exogenous n−3 and n−6 dietary polyunsaturated fatty acids and the endogenous n−7 and n−9 types of fatty acid. The general pattern of fatty acids in triglycerides was similar for liver, plasma and adipose tissue, averaging about 30% as saturated acids, 67% as 16- and 18-carbon unsaturated acids and only about 2% as 20- and 22-carbon highly unsaturated acids. The tissues maintained a linear relationship between the amount of 18-carbon polyunsaturated fatty acids in the diet and in the tissue triglycerides, with the proportionality constant for 18∶3n−3 being 60% of that for 18∶2n−6. The total phospholipids of liver, plasma and red blood cells maintained about 45% of the fatty acids in the form of saturated fatty acids and 20–30% as 20- and 22-carbon highly unsaturated fatty acids irrespective of very different proportions of n−3, n−6 and n−9 types of fatty acids. In all three tissues, the 20-carbon highly unsaturated fatty acids of the n−3, n−6 and n−9 type were maintained in a competitive hyperbolic relationship with apparent EC₅₀ values for dietary 18∶2n−6 and 18∶3n−3 near 0.1% of dietary calories. The consistent quantitative relationships described in this study illustrate an underlying principle of competition among fatty acids for a limited number of esterification sites. This approach may be useful in predicting the influence of diet upon tissue levels of the substrates and antagonists of eicosanoid biosynthesis.     

A comparative study of the lipids of globule membrane and fat core and of the milk serum of cows

Nine samples of fresh raw cow's milk were separated into fat globules and milk serum by centrifugation. After destabilization by freezing and thawing, the milk fat globules were resolved into membranes and fat cores. The lipid composition of these structures was compared to that of the surrounding milk serum. Of the total milk fat, 95–98% was in the fat cores, 0.5–1% in the globule membranes and the rest (1.5–4%) in the milk serum. The fat cores contained 88–93% triglyceride, 5.2–9.8% diglyceride, 1.5–7.3% free fatty acid and 0.2–0.4% cholesterol, but no phospholipid. The lipids of the membrane contained 21–44% phospholipid, made up of about equal proportions of phosphatidyl ethanolamine, phosphatidyl choline, and sphingomyelin. The other lipids of the membrane (56–79%) consisted of 83–88% triglyceride, 5.1–10.7% diglyceride, 1–5.1% free fatty acid and 0.4–1.9% cholesterol. The milk serum contained 30–45% phospholipid divided about equally among phosphatidyl ethanolamine, phosphatidyl choline and sphingomyelin. The rest (55–70%) of the milk serum lipids was made up of 71–83% triglycerides, 4.3–10.1% diglycerides, 8.7–15.7% free fatty acids, and 1.2–8.4% cholesterol. Corresponding phospholipid classes of milk serum and globule membranes had identical fatty acid compositions. The triglycerides and diglycerides of the globule membranes possessed increased proportions of palmitic and stearic acids in comparison to the glycerides of the fat cores. Taken in part from a PhD thesis submitted by T. C. Huang to Queen's University, Kingston, Canada in April, 1965. Presented in part at the 47th Canadian Chemical Conference and Exhibition held in Kingston, Canada, June 1–3, 1964.     

Total,polar and neutral lipids ofRhizopus arrhizus fischer

The changes in total lipid content, neutral and polar lipids, total fatty acids, and free fatty acids were investigated over a 4 day period in the zygomycete,Rhizopus arrhizus Fischer. The highest concentration of lipids occurred at the 72 hr period. The degree of unsaturation in the total fatty acid fraction increased during the growth period, whereas the degree of unsaturation decreased in the free fatty acid fraction during the same time period. The ratios of neutral to polar lipids over the 4 day period were: 0.75, 0.22, 1.94 and 0.94. The major components of polar lipids were phosphatidyl ethanolamine, lecithin, lysolecithin, and fatty acids. The fatty acids in the mono- and diglycerides were predominately saturated (67–96%). The fatty acids in the triglycerides shifted from a predominately unaturated (69%, 24 hr) to a more saturated pattern (62%, 96 hr).     

Lipid labeling with32P-orthophosphate and14C-acetate in marine copepods

Freshly collectedCalanus pacificus were maintained in sea water containing 25 μCi/ml [³²P]orthophosphate or 1 μCi/ml [¹⁴C]acetate at 10 C for 24 hr. The animals took up label from the environment and incorporated it into various lipid fractions. After incubation with [¹⁴C]acetate the order of specific activity of the different lipid classes was: phospholipids > free fatty acids > wax esters > triglycerides. Argentation thin layer chromatography of the fatty acid methyl esters showed that ca. 50% of the activity was in saturated fatty acids and 34% in polyunsaturated acids. When the animals were exposed to [³²P]orthophosphate, lysophosphatidyl choline became most heavily labeled, followed by lysophosphatidyl ethanolamine, sphingomyelin, phosphatidyl ethanolamine, and phosphatidyl choline. Comparison of the data obtained with those available for decapods and mammals revealed striking similarities between these phylogenetically distant groups. It is believed that labeling the lipids of marine and freshwater planktonic crustaceans in this way will provide much information about the metabolism of lipids in these organisms.     

In vitro and in vivo effects of exogenous lipids on the enzymatic hydrolysis of rat bile phospholipids

The addition of total phospholipids, phosphatidylcholines, triglycerides, cholesterol or glycerol to incubation media containing rat pancreatic juice and bile labeled with [9,10³H₂] oleic acid (90% of the radioactivity present as phospholipids) had no effect on the hydrolysis of bile endogenous phospholipids. The introduction of 2 or 10 mg of phosphatidylcholines and 0.5 ml of bile (≈ 1.5 mg of phospholipids)into the rat upper duodenum decreased the rate of absorption of rative bile phospholipids. It was not followed by an increase of free fatty acids released from biliary phospholipids in the intestinal lumen. The introduction of bile (0.5 ml) and small amounts of triolein (1.4–3.5 mg) into the duodenum had little effect on the rate of hydrolysis and absorption of native bile phospholipids, but caused a reduced absorption of the free fatty acids released or those coming from initial nonphosphorus biliary lipids. The introduction of bile (0.5 ml) and large amounts of triolein (30 mg) into the duodenum increased the rates of hydrolysis and absorption of endogenous bile phospholipids. These observations suggest that luminal lipid components can modify the organization of luminal micelles and, consequently, the action of the pancreatic phospholipase A₂ and the absorption of bile lipids.     

Effects of lupin oil on carp lipids during chilling 1. Changes in lipid class composition

The effect of lupin (Lupinus termis) oil on the muscle lipids of carp (Cyprinus carpio) during chilling was studied. During chilling, total lipids decreased whereas triglycerides remained almost constant. Neither the behavior of total lipids nor that of triglycerides during chilling was affected by the composition of the dietary lipids. The proportions of ?free”? fatty acids increased and the proportions of phospholipids decreased during chilling. These changes were markedly affected by the composition of the diet.