Changes in the lipid component of dried doughnut mixtures during storage

Changes in various indices of lipid components of dried doughnut mixtures during their joint or separate 3-month storage were studied. Methyl oleate oxidation model was used to determine the prooxidant properties of the initial mixture. The antioxidant activity, inhibition of oxidation, and inhibitory properties of lipids were shown to be enhanced during storage. The composition of phospholipids varied considerably due to changes in the degree of unsaturation of lipids in the mixture during storage. Joint storage of components resulted in greater stability and better preservation of lipid components (as compared to their separate storage).     

Free and bound fatty acid oxidation products in archaeological ceramic vessels

While oxidation products of unsaturated fatty acids, for example dicarboxylic acids (hereafter diacids), must form during the use of unglazed ceramic vessels for the processing of animal and plant products, such components have never been observed during studies of absorbed lipids. Their absence from the extractable lipid fraction is presumed to be the result of their loss from potsherds through groundwater leaching. Lipid oxidation products including short-chain dicarboxylic acids, ω-hydroxy acids and longer-chain hydroxy and dihydroxy acids have now been observed as components probably covalently bound into solvent insoluble residues of potsherds recovered from waterlogged deposits. These components were only revealed following alkaline treatment of the insoluble residues. A similar mixture of diacids was observed in high abundance in the free lipid fraction of vessels recovered from an exceptionally arid deposit where groundwater leaching would never have occurred. These results confirm the formation of oxidation and probable polymerization products of unsaturated fatty acids during vessel use and burial.     

Effect of lecithin on liver microsomal lipid peroxidation]

The effect of exogeneous (egg) lecithin on peroxidation of microsomal lipids was studied with the view of elucidating the role of various components of lipid substrate in the overall oxidation rate of the lipids. The following processes were studied a) NADPH-dependent microsomal lipid peroxidation in the presence of lecithin; b) ascorbate-dependent microsomal lipid peroxidation in the presence of lecithin; c) oxidation of lipid mixture, isolated from the microsomes, and that of lecithin in the presence of the Fe2+ + ascorbate system; 4) oxidation of lecithin induced by the Fe2+ + ascorbate system. It was found that in the presence of exogeneous lecithin the oxidation of microsomal lipids in inhibited, which is probably due to the peculiarities of lecithin oxidation. It was shown that the specific rate of lecithin oxidation is decreased with an increase in lecithin concentration. Possible mechanisms of lecithin effect on microsomal lipid peroxidation are discussed.     

The effect of the chain length distribution of free fatty acids on the mixing properties of stratum corneum model membranes

The stratum corneum (SC) plays a fundamental role in the barrier function of the skin. The SC consists of corneocytes embedded in a lipid matrix. The main lipid classes in the lipid matrix are ceramides (CERs), cholesterol (CHOL) and free fatty acids (FFAs). The aim of this study was to examine the effect of the chain length of FFAs on the thermotropic phase behavior and mixing properties of SC lipids. Fourier transform infrared spectroscopy and Raman imaging spectroscopy were used to study the mixing properties using either protonated or deuterated FFAs. We selected SC model lipid mixtures containing only a single CER, CHOL and either a single FFA or a mixture of FFAs mimicking the FFA SC composition. The single CER consists of a sphingoid base with 18 carbon atoms and an acyl chain with a chain length of 24 carbon atoms. When using lignoceric acid (24 carbon atoms) or a mixture of FFAs, the CER and FFAs participated in mixed crystals, but hydration of the mixtures induced a slight phase separation between CER and FFA. The mixed crystalline structures did not phase separate during storage even up to a time period of 3 months. When using palmitic acid (16 carbon atoms), a slight phase separation was observed between FFA and CER. This phase separation was clearly enhanced during hydration and storage. In conclusion, the thermotropic phase behavior and the mixing properties of the SC lipid mixtures were shown to strongly depend on the chain length and chain length distribution of FFAs, while hydration enhanced the phase separation.     

Evidence of membrane lipid oxidation of spray-dried Lactobacillus bulgaricus during storage

P. TEIXEIRA, H. CASTRO AND R. KIRBY. 1996. Membrane fatty acids of Lactobacillus bulgaricus were analysed by gas-liquid chromatography before and after spray drying. The ratio unsaturated/saturated fatty acids decreased following spray drying, indicating the formation of lesions in cellular lipid-containing structures. The same method was used to analyse membrane lipids of Lact. bulgaricus during storage. Similarly the ratio of unsaturated/saturated fatty acids in dried cells decreased further during storage in air, presenting evidence of lipid oxidation after prolonged storage. The mechanisms of cell death during storage in the dried state are still unknown, but from these results and those presented in the literature, it seems evident that lipid oxidation and survival during storage may be related.     

Changes in the Lipid Composition of Thalassiosira pseudonana during Its Life Cycle

The lipid and fatty acid (FA) compositions of a marine diatom alga Thalassiosira pseudonana grown in culture were investigated. The relative content of separate lipid classes and their FA composition varied during of the life cycle. During the periods of active cell division and resting cell production, the proportion of polar lipids, as the structural components of cell membranes, increased. Changes in the proportion of lipid classes resulted in shifts in the FA composition of total lipids. It is suggested that the structural components of photosynthetic and cells membranes accumulate in the resting cells. Thereby, a rapid cell growth and an extensive development of the species under favorable environmental conditions is provided.     

The membrane lipids of Halobacterium halobium

The lipid content of the cell membrane of Halobacterium halobium increased from about 15% to 21% during exponential growth of the organism. Total lipid phosphorus more than doubled during the growth cycle. The mixture of membrane lipids from stationary-phase organisms was similar to lipid mixtures from whole cells of other halobacteria inasmuch as 80% of the lipid phosphorus occurred in a diether analogue of phosphatidylglycerophosphate and an additional 7.5% occurred in the ether analogue of phosphatidylglycerol. The lipid mixture was more complex than those reported for other halophils, however, 12 components being recognized in the acetone-insoluble fraction and 17 in the acetone-soluble fraction. There were major changes in the proportions of some minor components of the acetone-insoluble fraction during a growth cycle. Three nitrogenous lipids were recognized in the acetone-insoluble fraction, but all were present in relatively low proportion. One, which was not a phospholipid, contained a bound peptide. Of the 17 acetonesoluble compounds, 15 were pigments. The major carotenoids were alpha- and beta-bacteriorubrin. The carotenoid pigments occurred at maximal concentration after 6-7 days' growth.     

Analysis and optimization of the cationic lipid component of a lipid/peptide vector formulation for enhanced transfection in vitro and in vivo

We have previously described a lipopolyplex formulation comprising a mixture of a cationic peptide with an integrin-targeting motif (K16GACRRETAWACG) and Lipofectin, a liposome consisting of DOTMA and DOPE in a 1:1 ratio. The high transfection efficiency of the mixture involved a synergistic interaction between the lipid/peptide components. The aim of this study was to substitute the lipid component of the lipopolyplex to optimize transfection further and to seek information on the structure-activity relationship of the lipids in the lipopolyplex. Symmetrical cationic lipids with diether linkages that varied in alkyl chain length were formulated into liposomes and then incorporated into a lipopolyplex by mixing with an integrin-targeting peptide and plasmid DNA. Luciferase transfections were performed of airway epithelial cells and fibroblasts in vitro and murine lung airways in vivo. The biophysical properties of lipid structures and liposome formulations and their potential effects on bilayer membrane fluidity were determined by differential scanning calorimetry and calcein-release assays. Shortening the alkyl tail from C18 to C16 or C14 enhanced lipopolyplex and lipoplex transfection in vitro but with differing effects. The addition of DOPE enhanced transfection when formulated into liposomes with saturated lipids but was more variable in its effects with unsaturated lipids. A substantial improvement in transfection efficacy was seen in murine lung transfection with unsaturated lipids with 16 carbon alkyl tails. The optimal liposome components of lipopolyplex and lipoplex vary and represent a likely compromise between their differing structural and functional requirements for complex formation and endosomal membrane destabilization.     

Autophagy is required for lipid homeostasis during dark-induced senescence

Autophagy is an evolutionarily conserved mechanism that mediates the degradation of cytoplasmic components in eukaryotic cells. In plants, autophagy has been extensively associated with the recycling of proteins during carbon-starvation conditions. Even though lipids constitute a significant energy reserve, our understanding of the function of autophagy in the management of cell lipid reserves and components remains fragmented. To further investigate the significance of autophagy in lipid metabolism, we performed an extensive lipidomic characterization of Arabidopsis (Arabidopsis thaliana) autophagy mutants (atg) subjected to dark-induced senescence conditions. Our results revealed an altered lipid profile in atg mutants, suggesting that autophagy affects the homeostasis of multiple lipid components under dark-induced senescence. The acute degradation of chloroplast lipids coupled with the differential accumulation of triacylglycerols (TAGs) and plastoglobuli indicates an alternative metabolic reprogramming toward lipid storage in atg mutants. The imbalance of lipid metabolism compromises the production of cytosolic lipid droplets and the regulation of peroxisomal lipid oxidation pathways in atg mutants.

Autophagy is required for the mobilization of membrane lipid components and lipid droplet dynamics during extended darkness in Arabidopsis.     

Analysis and Optimization of the Cationic Lipid Component of a Lipid/Peptide Vector Formulation for Enhanced Transfection In Vitro and In Vivo

We have previously described a lipopolyplex formulation comprising a mixture of a cationic peptide with an integrin-targeting motif (K₁₆GACRRETAWACG) and Lipofectin^®, a liposome consisting of DOTMA and DOPE in a 1:1 ratio. The high transfection efficiency of the mixture involved a synergistic interaction between the lipid/peptide components. The aim of this study was to substitute the lipid component of the lipopolyplex to optimize transfection further and to seek information on the structure-activity relationship of the lipids in the lipopolyplex. Symmetrical cationic lipids with diether linkages that varied in alkyl chain length were formulated into liposomes and then incorporated into a lipopolyplex by mixing with an integrin-targeting peptide and plasmid DNA. Luciferase transfections were performed of airway epithelial cells and fibroblasts in vitro and murine lung airways in vivo. The biophysical properties of lipid structures and liposome formulations and their potential effects on bilayer membrane fluidity were determined by differential scanning calorimetry and calcein-release assays. Shortening the alkyl tail from C18 to C16 or C14 enhanced lipopolyplex and lipoplex transfection in vitro but with differing effects. The addition of DOPE enhanced transfection when formulated into liposomes with saturated lipids but was more variable in its effects with unsaturated lipids. A substantial improvement in transfection efficacy was seen in murine lung transfection with unsaturated lipids with 16 carbon alkyl tails. The optimal liposome components of lipopolyplex and lipoplex vary and represent a likely compromise between their differing structural and functional requirements for complex formation and endosomal membrane destabilization.     

Antioxidant properties of essential oils: autoxidation of essential oils from laurel and fennel and effects of mixing with essential oil from coriander

Changes in the composition of essential oils from the seeds of laurel (Laurus nobilis L.) and fennel (Foeniculum vulgare Mill., var. dulce Thelling) and their mixture with essential oil from coriander were studied by capillary gas-liquid chromatography during storage in the dark and in light. Under these conditions, essential oil of laurel retained its composition for 12 months. Essential oil of fennel was rapidly oxidized in light. However, the rate of its oxidation in the dark was lower. The major component of essential oil of fennel, transanethol, had a lower antioxidant activity than essential oil of coriander. The mixture of essential oils from laurel and coriander possessed antioxidant properties and strongly inhibited the oxidation of components of the fennel oil.     

Effect of hydrophobicity of yeast cell envelope on the rate of autooxidation of methyloleate

The relationships between the antioxidant and antiperoxide properties and the lipid composition of yeast cell envelope prior to, and after the reaction with a liquid culture medium were studied. Correlations between the hydrophobicity of envelopes, their lipid composition and the parameters of the kinetic curves of the oxidation of model methyloleate in the presence of lipids were established. It was found that, irrespective of the general content of lipids in yeast cell envelope, preparations with high antiperoxide activity of lipids had a high hydrophobicity and sorbed lipophilic prooxidants from medium, whereas preparations with low antiperoxide activity were less hydrophobic and adsorbed predominantly lipophilic inhibitors. It was found that the most comprehensive information on the physicochemical properties of lipids adsorbed from medium is provided by an analysis of kinetic curves of oxidation in toto.     

Effect of hydrophobicity of yeast cell envelopes on the rate of methyl oleate autooxidation

The relationships between the antioxidant and antiperoxide properties and the lipid composition of yeast cell envelopes before and after interaction with a liquid culture medium was studied. Correlations between the hydrophobicity of cell envelopes, their lipid composition, and parameters of kinetic oxidation curves of model methyl oleate in the presence of lipids were found. Irrespective of the total lipid content in yeast cell envelopes, the preparations with a high antiperoxide activity of lipids exhibited a higher hydrophobicity and adsorbed lipophilic prooxidants from the medium more readily, whereas the preparations with a low antiperoxide activity were less hydrophobic and adsorbed predominantly lipophilic inhibitors. It was found that analysis of kinetic oxidation curves in toto provides the most comprehensive information on the physicochemical properties of lipids adsorbed from medium.     

A Calorimetric Study of the Changes in Lipids during Seed Storage under Dry Conditions

Changes in the lipid constituents of seeds are believed to be associated with losses of seed viability during storage. The physical properties of lipids in seeds of genetically similar backgrounds but varying ages were compared using differential scanning calorimetry. The technique measures the temperature and energy associated with lipid-melting transitions and is noninvasive. Although differences in the temperature at which lipids melted were sometimes observed among deteriorated and fresh seeds, the direction of the change was inconsistent among species tested. For all species tested except tomato, there was a decrease in the energy associated with the lipid melt in deteriorated samples, and the change occurred at a similar rate as the loss of seed vigor. The data suggest that there are changes in the lipid components of seeds that are associated with seed deterioration and that these changes can be measured using differential scanning calorimetry.     

Changes in adrenocortical lipid fluidity of hyperfunctioning human adrenals

Lipid droplets, the storage places of cholesterol in adrenocortical cells, exhibit a relatively uniform appearance studied by the electron microscope but they are heterogeneous in respect of their optical polarizing properties. Optical birefringency was studied in cryosections of normal and hyperfunctioning adrenal cortex by a polarizing microscope, equipped with a cold/hot stage working in the temperature range from −40 to 40°C. The majority of lipid droplets in normal adrenal cortex were optically anisotropic in each cortical zone at room temperature (22°C) indicating a long-range molecular order of the lipid components. The lipids of the zona glomerulosa, in the cases of Conn's and Bartter's syndromes, became anisotropic when the temperature was lowered below ambient. The birefringency of the lipids of the zona fasciculata in the case of Cushing's disease was observed at temperatures below −10°C indicating ordered packing of the components of lipid droplets at this temperature. Thus the lipids were more fluid in the hyperfunctioning, hormone-producing cells—this may represent an optimal precondition for their mobilization and processing by the hydrolyzing enzyme system. The changes in fluidity of the intracellular lipids can be attributed to different functional states in the adrenal cortex. Study of the thermotropic phase transitions of the lipid droplets by polarizing microscopy may be a useful additional method for the diagnosis of some adrenocortical diseases.     

Liposomes composed of unsaturated lipids for membrane modification of human erythrocytes

Previous studies have shown that certain saturated lipids protect red blood cells (RBCs) during hypothermic storage but provide little protection during freezing or freeze-drying, whereas various unsaturated lipids destabilize RBCs during hypothermic storage but protect during freezing and freeze-drying. The protective effect of liposomes has been attributed to membrane modifications. We have previously shown that cholesterol exchange and lipid transfer between liposomes composed of saturated lipids and RBCs critically depends on the length of the lipid acyl chains. In this study the effect of unsaturated lipids with differences in their number of unsaturated bonds (18:0/18:1, 18:1/18:1, 18:2/18:2) on RBC membrane properties has been studied. RBCs were incubated in the presence of liposomes and both the liposomal and RBC fraction were analyzed by Fourier transform infrared spectroscopy (FTIR) after incubation. The liposomes caused an increase in RBC membrane conformational disorder at suprazero temperatures. The fluidizing effect of the liposomes on the RBC membranes, however, was found to be similar for the different lipids irrespective of their unsaturation level. The gel to liquid crystalline phase transition temperature of the liposomes increased after incubation with RBCs. RBC membrane fluidity increased linearly during the first 8 hours of incubation in the presence of liposomes. The increase in RBC membrane fluidity was found to be temperature dependent and displayed Arrhenius behaviour between 20 and 40°C, with an activation energy of 88 kJ mol?1. Taken together, liposomes composed of unsaturated lipids increase RBC membrane conformational disorder, which could explain their cryoprotective action.     

Inhibition of lipid oxidation by serum and albumin

The influence of serum and albumin on enzymatic and non-enzymatic lipid oxidation was investigated. Intensity of oxidation was measured as the amount of oxygen consumed by the sample and by quantitation of malonaldehyde formed during breakdown of lipid peroxides. Non-enzymatic lipid oxidation was stimulated by ascorbic acid or ferrous ions and enzymatic by NADPH-dependent oxidase, 15-lipoxygenase and 12-lipoxygenase. Albumin inhibits lipid oxidation only when pure fatty acid (arachidonic or linoleic) is the substrate for this oxidation. Serum was a stronger inhibitor than an equivalent amount of albumin and it also inhibited oxidation of a mixture of lipids from liver microsomes. It is concluded that serum contains two antioxidant factors: albumin which binds fatty acids and probably another factor which is a true antioxidant.     

Changes in the physico-chemical characteristics of mouse liver lipids after administration of a mixture of unsaturated fatty acids

The effects of a mixture of unsaturated fatty acids (UFA) on changes in the lipid oxidation substrate, a system of natural antioxidants and the functioning of a system controlling lipid peroxidation were studied. It was shown that arachidonate induces the incorporation of fatty acids into individual lipid fractions and the dilution of lipids by UFA esters, as a result of which the relative amount of natural antioxidants diminishes. Under these conditions, the oxidative capacity of lipids decreases, which manifests itself in a reduction of the rate of natural antioxidants utilization and an increase in their number. It was shown that purposeful modification of the lipid substrate does not interfere with the interaction between individual parameters of the system controlling lipid peroxidation.     

A lipid-phase separation model of low-temperature damage to biological membranes

An hypothesis is proposed to explain the damage caused to biological membranes exposed to low temperatures. The thesis rests on the general observation that the lipid components of most membranes are heterogeneous and undergo phase transitions from gel-phase lamellae to liquid-crystalline lamellae and some to a non-lamellar, hexagonal-II phase over a wide range of temperatures. As a consequence of these phase transitions the lateral distribution of the lipids characteristic of the growth temperature is disturbed and redistribution takes place on the basis of the temperature at which phase transitions occur. When membranes are cooled, first the non-lamellar forming lipids pass through a transition to a fluid lamellar phase and are miscible with bilayer-forming lipids into which they diffuse. On further cooling the high-melting-point lipids begin to crystallize and separate into a lamellar gel phase, in the process excluding the low-melting point lipids and intrinsic proteins. The lipids in these remaining regions form a gel phase at the lowest temperature. It is suggested that, because the non-lamellar lipids tend to undergo a liquid-crystalline to gel-phase transition at higher temperatures than lamellar-forming lipids, these will tend to phase separate into a gel phase domain rich in these lipids. Damage results when the membrane is reheated, whereupon the hexagonal-II-forming lipids give rise to non-lamellar structures. These probably take the form of inverted micelles sandwiched within the lipid bilayer and they completely destroy the permeability barrier properties of the membrane. The model is consistent with the phase behavior of membrane lipids and the action of cryoprotective agents in modifying lipid phase properties.     

Triacylglycerol lipases of the yeast

All eukaryotes including the yeast contain a lipid storage compartment which is named lipid particle, lipid droplet or oil body. Lipids accumulating in this subcellular fraction serve as a depot of energy and building blocks for membrane lipid synthesis. In the yeast, the major storage lipids are triacylglycerols (TGs) and steryl esters (SEs). An important step in the life cycle of these non-polar lipids is their mobilization from their site of storage and channeling of their degradation components to the appropriate metabolic pathways. A key step in this mobilization process is hydrolysis of TG and SE which is accomplished by lipases and hydrolases. In this review, we describe our recent knowledge of TG lipases from the yeast based on biochemical, molecular biological and cell biological information. We report about recent findings addressing the versatile role of TG lipases in lipid metabolism, and discuss non-polar lipid homeostasis and its newly discovered links to various cell biological processes in the yeast.