Skin surface lipids of the mole Scalopus aquaticus

Skin surface lipids of the mole Scalopus aquaticus were found to consist principally of squalene (70%), wax esters (15%), and sterol esters (5%), together with small amounts of triglycerides, free fatty acids, free fatty alcohols, and free sterols. Analysis of the fatty acids occurring free and as wax esters and sterol esters showed these to consist of approximately equal amounts of saturated and monounsaturated compounds. The saturated fatty acids consisted predominantly of odd-carbon anteiso and even-carbon straight-chain compounds, with minor amounts of even-carbon iso-branched chains. The unsaturated fatty acids had double bond positions that would have been produced by delta 9-desaturation of C14, C16 and C18 straight chain saturated precursors. Both the free and the esterified fatty alcohols had chain structures corresponding with those of the fatty acids but of somewhat greater average chain length. Discovery of a major proportion of squalene in the sebum of this animal extends the number of non-human species that have this characteristic to four, all of which inhabit a damp environment, suggesting that squalene conveys some biological advantage under these conditions.     

Lipids of chicken epidermis

The lipids from chicken epidermis were analyzed by a combination of quantitative thin-layer and gas-liquid chromatography and by chemical and spectroscopic methods. The lipid groups present included wax diesters (34%), triglycerides (32%), sterols (11%), phospholipids (11%), nonphosphorus-containing sphingolipids (3%), beta-D-glucosylsterols (3%), 6-O-acyl-beta-D-glucosylsterols (2%), steryl esters (1%), cholesteryl sulfate (1%), and free fatty acids (1%). The major phospholipids were phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin, and the sphingolipids included ceramides, glucosylceramides, O-acylceramides, and O-acylglucosylceramides. Glucosylsterols and acylglucosylsterols have not been found in mammalian skin, and may be relevant to the evolutionary history of the epidermal water barrier. The wax diesters contained mainly 16-, 18-, and 20-carbon saturated fatty acids esterified to 20- through 24-carbon threo and erythro 2,3-diols, while the chicken epidermal triglycerides contained some very long-chain (26-40 carbon) saturated fatty acids. These wax diesters and unusual triglycerides may be of significance in human health.     

Skin surface lipids of the mink

Skin surface lipids from mink (Mustela vison) were collected in acetone and analyzed by thin-layer chromatography and gas chromatography. The principal components were wax monoesters (92%), cholesteryl esters (5%), free fatty acids (1%), fatty alcohols (1%) and cholesterol (1%). The fatty acids and alcohols contained in these lipids were composed principally of homologous series of straight chained omega 7-unsaturated structures (C16-C24), accompanied by lesser proportions of homologous series of saturated (C14-C22) and omega 9-unsaturated (C18-C22) structures.     

Oxidative degradation of model lipids representative for main paper pulp lipophilic extractives by the laccase–mediator system

Different model lipids-alkanes, fatty alcohols, fatty acids, resin acids, free sterols, sterol esters, and triglycerides-were treated with Pycnoporus cinnabarinus laccase in the presence of 1-hydroxybenzotriazole as mediator, and the products were analyzed by gas chromatography. The laccase alone decreased the concentration of some unsaturated lipids. However, the most extensive lipid modification was obtained with the laccase-mediator system. Unsaturated lipids were largely oxidized and the dominant products detected were epoxy and hydroxy fatty acids from fatty acids and free and esterified 7-ketosterols and steroid ketones from sterols and sterol esters. The former compounds suggested unsaturated lipid attack via the corresponding hydroperoxides. The enzymatic reaction on sterol esters largely depended on the nature of the fatty acyl moiety, i.e., oxidation of saturated fatty acid esters started at the sterol moiety, whereas the initial attack of unsaturated fatty acid esters was produced on the fatty acid double bonds. In contrast, saturated lipids were not modified, although some of them decreased when the laccase-mediator reactions were carried out in the presence of unsaturated lipids suggesting participation of lipid peroxidation radicals. These results are discussed in the context of enzymatic control of pitch to explain the removal of lipid mixtures during laccase-mediator treatment of different pulp types.     

Cuticular lipids of insects. VI. Cuticular lipids of the grasshoppers Melanoplus sanguinipes and Melanoplus packardii

The cuticular lipids of the grasshoppers Melanoplus sanguinipes and Melanoplus packardii contain 60 and 68% alkanes and 28 and 18% secondary alcohol wax esters, respectively, with lesser amounts of normal and sterol wax esters, triglycerides, alcohols, sterols, and free fatty acids. All the hydrocarbons are saturated, and four types of alkanes are present: n-alkanes, 3-methylalkanes, internally branched monomethylalkanes, and internally branched dimethylalkanes. The principal n-alkanes in both insects are C(29) and C(27), with a range from C(21) to C(33). Trace amounts of 3-methylalkanes of 28, 30, and 32 total carbons are present. The principal internally branched monomethylalkanes are C(32) and C(34), whereas the main dimethylalkane contains 35 carbons. The n-alkanes do not correspond in chain length to the secondary alcohols. The primary alcohols range from C(22) to C(32) in both insects, with C(24) and C(26) predominating. The fatty acids in the triglyceride and free fatty acid fractions range from C(12) to C(24) in M. sanguinipes and from C(12) to C(18) in M. packardii.     

Separation and analysis of steryl and wax esters from Dicranum elongatum

Abstract Complete separation of the steryl and wax esters in the subarctic moss Dicranum elongatum was achieved on MgO thin-layer plates without any notable alteration of the acyl and alkyl moieties of the esters. Gas chromatography-mass spectrometry of the hydrolyzed fraction showed that the sterols (campesterol, stigmasterol, sitosterol, cycloartenol, 24-methylene cycloartanol and an unidentified sterol) were primarily esterified with unsaturated fatty acids 18:2 ω 6, 18:3 ω 3 and 20:4 ω 6. In contrast, the wax alcohols (l-octadecanol, phytol and geranylgeraniol) were mainly esterified with saturated fatty acids with 16:0, 18:0 and 20:0 as major components. No great differences were found in the fatty acid pattern of the steryl esters between different portions of the shoot. Slight differences, however, were found in the proportions of ω 3 and ω 6 fatty acids. In the wax esters a clear decrease was found in the proportions of 18:0 and 20:0 acids with increased shoot age accompanied by a slight increase in the proportions of 14:0, 20:4 ω 6 and phytenic acid.     

Cholesterol and oxysterol metabolism and subcellular distribution in macrophage foam cells. Accumulation of oxidized esters in lysosomes

Cholesterol- and cholesteryl ester-rich macrophage foam cells, characteristic of atherosclerotic lesions, are often generated in vitro using oxidized low density lipoprotein (OxLDL). However, relatively little is known of the nature and extent of sterol deposition in these cells or of its relationship to the foam cells formed in atherosclerotic lesions. The purpose of this study was to examine the content and cellular processing of sterols in OxLDL-loaded macrophages, and to compare this with macrophages loaded with acetylated LDL (AcLDL; cholesteryl ester-loaded cells containing no oxidized lipids) or 7-ketocholesterol-enriched acetylated LDL (7KCAcLDL; cholesteryl ester-loaded cells selectively supplemented with 7-ketocholesterol (7KC), the major oxysterol present in OxLDL). Both cholesterol and 7KC and their esters were measured in macrophages after uptake of these modified lipoproteins. Oxysterols comprised up to 50% of total sterol content of OxLDL-loaded cells. Unesterified 7KC and cholesterol partitioned into cell membranes, with no evidence of retention of either free sterol within lysosomes. The cells also contained cytosolic, ACAT-derived, cholesteryl and 7-ketocholesteryl esters. The proportion of free cholesterol and 7KC esterified by ACAT was 10-fold less in OxLDL-loaded cells than in AcLDL or 7KCAcLDL-loaded cells. This poor esterification rate in OxLDL-loaded cells was partly caused by fatty acid limitation. OxLDL-loaded macrophages also contained large (approximately 40-50% total cell sterol content) pools of oxidized esters, containing cholesterol or 7KC esterified to oxidized fatty acids. These were insensitive to ACAT inhibition, very stable and located in lysosomes, indicating resistance to lysosomal esterases. Macrophages loaded with OxLDL do not accumulate free sterols in their lysosomal compartment, but do accumulate lysosomal deposits of OxLDL-derived cholesterol and 7-ketocholesterol esterified to oxidized fatty acids. The presence of similar deposits in lesion foam cells would represent a pool of sterols that is particularly resistant to removal.     

Identification by gas chromatography and mass spectrometry of lipids from the rat Harderian gland

Lipids from rat Harderian glands were extracted with ethyl acetate, hydrolysed with base and examined by gas chromatography (GC) and gas chromatography—mass spectrometry (GC—MS) as trimethylsilyl (TMS), [²H₉]TMS, methyl ester—TMS, picolinyl, nicotinate and nicotinylidene derivatives. The latter three derivatives were used to reveal the structures of the alkyl chains of fatty acids, alcohols and glycerol ethers, respectively. Forty-eight compounds were identified, representing about 97% of the total extracted lipids as measured by GC peak areas. The major constituents were fatty acids with chain lengths from 12 to 22 carbon atoms (mainly C₁₈ and C₂₀) and fatty alcohols (C₁₆ to C₂₆) derived from wax esters. Most of these acids and alcohols were unsaturated in the ω-7 position and were accompanied by smaller amounts of the saturated and ω-5 monounsaturated analogues. Glycerol ethers were also identified for the first time in this secretion; the ether chains contained from 14 to 19 carbon atoms (mainly 16) and were straight-chain saturated, unsaturated (ω-5 and ω-7) and branched (iso). The only sterol found was cholesterol amounting to 1.24% of the total extract.     

The lipid biochemistry of calanoid copepods

Calanus species, particularly those in high latitudes, can accumulate large oil reserves consisting predominantly of wax esters. These wax esters consist predominantly of 16:0, 20:1 (n–9) and 22:1 (n–11) fatty alcohols, mainly formed de novo by the animals from non-lipid dietary precursors, esterified with various fatty acids that are often polyunsaturated fatty acids and largely of dietary, phytoplanktonic origin. Wax ester formation is maximal in copepodite stages IV and V. The lipids are elaborated not primarily for buoyancy regulation but as a source of metabolic energy during overwintering, particularly for reproduction. Large quantities of wax esters are utilised for gonadal development when stage V copepodites mature to females. Development of stage V copepodites to males is not accompanied by wax ester utilisation but males consume large amounts of these lipids in physical activity during reproduction. The role of wax esters in the life history of calanoids is illustrated with particular reference to a comparison of Calanus finmarchicus and Metridia longa in Balsfjord, northern Norway.     

Lipid and sterol composition of the pollen of the west african oilpalm, Elaeis guineensis

The lipid classes, fatty acid methyl esters and the sterols of oilpalm pollen were analysed. The neutral lipid fraction consisted of triglycerides, esterified and free sterols and trace amounts of hydrocarbons. Monogalactosyl and digalactosyl diglycerides, phosphatidyl choline, phosphatidyl inositol and phosphatidyl ethanolamine represented the polar lipids. The major fatty acids were linoleic, palmitic and linolenic acids together with small to trace amounts of oleic, stearic, arachidic, myristic, lauric, palmitoleic and margaric acids. Unsaturated fatty acids predominated over saturated ones in the ratio of 3:2. The 4-desmethyl sterols were the major phytosterols in the free form but they constituted a lower proportion of the sterols in the esterified state. 28-Isofucosterol was isolated and characterized as the principal sterol.     

Aliphatic Chains of Esterified Lipids in Isolated Eyespots of Euglena gracilis var. bacillaris

Isolated eyespot granules of Euglena gracilis Klebs var. bacillaris Pringsheim contained approximately 6% lipids (based on protein). Separation of the lipid extracts by thin layer chromatography revealed four major fractions: wax esters, triacylglycerols, free fatty acids, and phospholipids. Methanolysis of each fraction yielded between 27 and 29 different fatty acids ranging from 12:0 to 22:6. Acetates of the fatty alcohols of the wax fraction consisted of 11:0 to 18:0 carbon chains, with 14:0 being the major component; unsaturated alcohols were not detected.     

A comparison of the composition of epicuticular wax from red raspberry (Rubus idaeus L.) and hawthorn (Crataegus monogyna Jacq.) flowers

Epicuticular waxes have been characterised from the flowers of raspberry and hawthorn, on both of which adult raspberry beetles (Byturus tomentosus) can feed. The flower wax from both species had similar alkane profiles and also contained long-chain alcohols, aldehydes and fatty acids. The range of the carbon numbers detected for these classes of compounds was broadly similar in both but the relative amounts of each differed between species. Raspberry flower wax also contained fatty acid methyl esters, a group of compounds that has rarely been detected in plant epicuticular waxes, however, these were not observed in hawthorn flower wax. Long-chain alcohol-fatty acid esters with carbon numbers ranging from C36 to C48 were also detected in both plant species. However, an examination of their constituent acids indicated that in hawthorn the esters based on the C16 fatty acid predominated, whilst in raspberry flower wax, esters based on the C20 fatty acid were most abundant. Both species also contained pentacyclic triterpenoids, which accounted for, on average, over 16 and 48% of the total wax extracted from raspberry and hawthorn flowers respectively. In the former, ursolic and oleanolic acids accounted for over 90% of the pentacyclic triterpenes, whilst hawthorn flower wax, in addition to containing these acids, also contained high relative concentrations of both free and esterified alpha- and beta-amyrins.     

Waxes and lipids associated with the external waxy structures of nymphs and pupae of the giant whitefly, Aleurodicus dugesii

The nymphs and pupae of the giant whitefly, Aleurodicus dugesii, produce large quantities of external lipids, both as waxy particles and as waxy filaments. The nymphs and pupae extrude filaments from two dorsal rows of five pores each. Filaments can attain lengths of 5-8 cm. The external lipids of nymphs and pupae consist largely of long-chain aldehydes, alcohols, acetate esters and wax esters. Hydrocarbons are minor components. Soon after hatching, the nymph produced an unidentified waxy fringe extruded laterally from its margin. After molting to the second instar, long, hollow, waxy filaments were produced by the immature stages. The major lipid class associated with the filaments was saturated wax esters (89%), mainly C44, C46 and C60. Associated with formation of the filaments were waxy particles in the shape of curls, which peeled off of the extruding filaments. Similar but more tubular-shaped curls were also produced by numerous lateral pores so that, eventually, the curls completely camouflaged the nymph. The major lipid class of the curls was wax esters (50%), mainly C44 and C46. The cuticular surface lipids of the nymphs were mainly long-chain aldehydes (43%) and wax esters (27%). Unsaturated fatty acid moieties constituted 2 and 19% of the wax esters of curls and nymph cuticular surface lipids, respectively. The major lipid classes of pupae and of their palisade were long-chain aldehydes and alcohols. No unsaturated wax esters were detected in the filaments, but 30% of pupal and 21% of palisade surface wax esters were unsaturated in their fatty acid moieties, 16:1, 18:1 and 20:1.     

Lipid composition of the extracellular matrix of Botrytis cinerea germlings

Six simple lipid classes (mono-, di- and tri-acylglycerols, free fatty acids, free fatty alcohols and wax esters) were identified by TLC in the extracellular matrix of Botrytis cinerea germlings and the molecular components of each class were characterized using GC-MS. The relative amounts of fatty acids and fatty alcohols within each lipid class were determined by GC-FID. Over all the lipid classes, the most abundant saturated fatty acids were palmitic (ca. 30%) and stearic acid (ca. 22%). Palmitoleic and oleic acids made up ca. 21% and 24% (respectively) of the free fatty acids, while erucic (ca. 4.1%) and linoleic (ca. 3.6%) acids were the most abundant unsaturated fatty acids in the acylglycerides. The acylglycerides also contained almost 35% long chain fatty acids (C20:0 to C28:0). Six fatty acids were identified which had odd-numbered carbon chain lengths (C15:0, C17:0, C19:0, C21:0, C23:0 and C25:0). Of these, pentacosanoic acid made up almost 14% of the fatty acids in the acylglycerides. Three methyl-branched chain fatty acids, namely isopalmitic, isoheptadecanoic and anteisopalmitic, were identified in the ECM, all in small amounts. Of the fatty alcohols identified, only palmityl and stearyl alcohols were found in the free form (ca. 57% and 43%, respectively) but arachidyl alcohol (ca. 47%) and 1-octacosanol (ca. 30%) were the most abundant fatty alcohols found in the wax ester fraction.     

Occurrence of steryl and wax esters in Dicranum elongatum

The steryl and wax esters of the frozen subarctic moss Dicranum elongatum Schleich contained fatty acids 39.8 mg per gram dry green tissue. The content decreased with increasing age of the moss shoots, but no great changes were found in the fatty acid pattern of the esters. The major part of the steryl and wax ester fraction of the green shoots was made up of esterified sterols (85%), and the rest (15%) of esterified aliphatic alcohols. No great changes were found in their relative proportions with increased age of the shoots. Some changes were evident in the pattern of individual esterified sterols, however. The proportion of cycloartenol was lower in the older parts than in the green part, and the proportion of campesterol, stigmasterol, and β-sitosterol were lower in the green part. The major esterified aliphatic alcohols were 1-octadecanol, phytol and geranylgeraniol. The proportion of geranylgeraniol was highest in the green part and that of phytol in the older parts. The main alcohol of the surface lipids was 1-octadecanol.     

Lipid storage compounds in marine bacteria

Forty psychrophile or psychrotrophic crude-oil-utilizing marine bacteria were investigated for their ability to accumulate lipid storage compounds in the cytoplasm during cultivation under nitrogen-limiting conditions. Most of them (73%) were able to accumulate specialized lipids like polyhydroxyalkanoic acids (PHA) while other lipids such as wax esters occurred in two isolates. Accumulation of PHA occurred predominantly at low temperatures (4–20 °C) as demonstrated for three isolates. Electron microscopy revealed polyphosphate inclusions occurring in two isolates in addition to PHA. Cells of the isolate Acinetobacter sp. 211 were able to synthesize and accumulate lipid inclusions during growth on acetate, ethanol, olive oil, hexadecanol and heptadecane. The composition of the lipid inclusions depended on the compounds provided as carbon source. Wax esters and acylglycerols occurred mainly during the cultivation on olive oil; in contrast, wax esters and free alcohols occurred during cultivation on hexadecanol. Total fatty acids in cells of the Acinetobacter sp. 211 amounted to 25% of the cellular dry weight in olive-oil-grown cells. Palmitic acid was the main fatty acid in the lipids when the cells were cultivated on acetate or ethanol (44% and 32% of total fatty acids respectively). In contrast, fatty acids occurring in the lipids during cultivation on hexadecanol, heptadecane or olive oil were related to the carbon source. The fatty acids present in the accumulated lipids consisted predominantly of saturated and unsaturated straight-chain fatty acids with a chain length ranging from 12 to 18 carbon atoms. Analysis of the lipid-granule-associated proteins in cells of Acinetobacter sp. 211 revealed a protein of 39 kDa as the predominant protein species. Received: 2 July 1996 / Received revision: 3 September 1996 / Accepted: 28 September 1996     

Cuticular hydrocarbons and wax esters of the ectoparasitoid Habrobracon hebetor: Ontogenetic,reproductive, and nutritional effects

Hydrocarbon and wax ester components of cuticular lipids of the braconid parasitoid Habrobracon hebetor Say reared at 25 degrees C on larvae of a pyralid moth have been identified by GC-MS and analyzed with respect to adult age, mating status, and diet. The hydrocarbons range in carbon number from C(21) to C(45) and consist of a homologous series of n-alkanes, 11-, 13-, and 15-methyl alkanes, 13,17-dimethyl alkanes, and Z-5, Z-7, and Z-9-alkenes. The wax esters found in the cuticular lipid fraction are a series of homologous compounds with the acid portion being short chain, unbranched, even carbon number acids from C(8) to C(20) (predominately C(8) to C(16)). The alcohol portions of the esters are secondary alcohols with carbon number from C(22) to C(25) (predominately C(23) and C(25)) with the hydroxyl function located at C(6), C(7), C(8), and C(9). Gender, age, and nutritional states were significant factors for variation in several of the individual esters, but mating status did not affect wax ester composition. Ontogenetic examinations indicated that prepupal, and early pupal cuticular lipids contain only hydrocarbons. Low levels of wax esters are detectable in late stage pupae, and somewhat greater quantities of wax esters are present on newly eclosed adults. When pharate adults emerge from the cocoon, however, their cuticular lipids consist of approximately equal amounts of hydrocarbons and wax esters, and 6d post emergence from the cocoon, wax esters are the predominant lipid component.     

A Comparison of the Composition of Wax Ester Molecular Species of Different Coral Groups (Subclasses Hexacorallia and Octocorallia)

Wax esters, which are esters of fatty alcohols and fatty acids (FAs), are one of the main classes of reserve lipids in all coral species. The chemical structures and the content of wax ester molecular species were determined for the first time in nine coral species from three taxonomic groups: symbiotic reef-building corals, (Hexacorallia subclasses), symbiotic soft coral alcyonarians, and asymbiotic soft coral gorgonians (Octocorallia subclasses) collected in the South China Sea (Vietnam). Our comparison of these groups showed that the absence of symbiotic microalgae (zooxanthellae) and the exoskeleton affects the profile of molecular species of wax esters considerably. The main components of wax esters of all corals were cetyl palmitate (16:0-16:0) and other saturated wax esters containing 30, 34, and 36 carbon atoms. The content of unsaturated molecular species 6:0–16:1, 16:0–18:1, and 16:0–20:1 in wax esters of symbiotic soft corals (alcyonarians) was greater than that in wax esters of reef-building corals. In contrast to symbiotic coral species, wax esters of asymbiotic soft corals, namely azooxanthellate gorgonians, contained a considerable amount of long-chain molecular species (C₃₇-C₄₁) with an odd number of carbon atoms. The presence of such molecular species indicates that asymbiotic gorgonians may use bacterial FAs in biosynthesis of their own wax esters. This observation confirms our hypothesis that bacterial community is important for maintaining the energy balance of azooxanthellate corals.     

Biosynthesis of wax esters in tissues of Sinapis alba L. seeds

The biosynthesis of wax esters has been investigated in maturing seeds of Sinapis alba. Exogenous long-chain alcohols are incorporated exclusively into alkyl moieties of wax esters. Oxidation of the long-chain alcohols is not detected. Exogenous fatty acids are incorporated into acyl moieties of wax esters to a low extent. A reduction of fatty acids to alcohols is not observed. Synthesis of wax esters is localized exclusively in the testa; both outer and inner integument are equally active in wax ester biosynthesis. The biosynthesis of wax esters is specific with regard to both chain length and degree of unsaturation of long-chain alcohols. Exogenous and endogenous sterols are not esterified.     

Quantitative analysis of fatty acid precursors in marine samples: direct conversion of wax ester alcohols and dimethylacetals to FAMEs

To apply fatty acid analyses to the study of foraging ecology and diet determination, all compounds that may be deposited as fatty acids in a predator must be quantified in the prey. These compounds include the usual fatty acids in acyl lipids, but also the alcohols of wax esters and the vinyl ethers of plasmalogens. In routine fatty acid analysis, samples are extracted and transesterified (methylated), resulting in the formation of fatty acid methyl esters (FAMEs); however, fatty alcohols and dimethylacetals (DMAs) are also generated if wax esters or plasmalogens are present. Here, we present a new method using a modified Jones' reagent to oxidize these alcohols and DMAs to free fatty acids (FFAs). These FFAs are then easily methylated and quantitatively recombined with FAMEs from the same sample. This generates a fatty acid signature of prey that is equivalent to that which the predator has available for deposition upon digestion of that prey. This method is validated with alcohol and DMA standards. Its application to typical marine samples is also presented, demonstrating the change in effective fatty acid signature after inclusion of fatty acids derived from wax esters and plasmalogens.