Lipid lowering effects of high linoleate and high alpha-linolenate diets in rats and mice. Consequence of long-term feedings.

Diets high in linoleate (safflower oil) or high in alpha-linolenate (perilla oil) were fed to rats for 11 months, and the effects of the diets on plasma and tissue lipids were compared. The plasma levels of total cholesterol (Cho), phospholipids (PL) and triacylglycerol (TG) were significantly lower in the high alpha-linolenate group than in the high linoleate group, the differences being more than 30% in the levels of total Cho and TG. The diets had differential effects on the lipid contents of major tissues: the TG level in muscle was higher but both the TG level in depot fat and the PL level in muscle were lower in the high alpha-linolenate group than in the high linoleate group. In order to clarify whether or not the hypolipidemic effect of the high alpha-linolenate diet was due to changes in the distribution of lipids among tissues, whole body lipids were estimated in mice fed these diets for 5 months. The whole body Cho content was significantly lower, by 28%, in the high alpha-linolenate group compared with the high linoleate group, but the total lipid content, PL and neutral lipids were similar between the groups. Our results indicate that the high alpha-linolenate diet has a more potent cholesterol lowering effect in plasma and body tissue than the high linoleate diet; interestingly, whole body TG levels are similar but tissue distributions of TG are different between the two dietary groups.     

Identification of novel n-4 series polyunsaturated fatty acids in a deep-sea clam, Calyptogena phaseoliformis

Novel fatty acids originated from the cold-seep clam Calyptogena phaseoliformis, collected from hydrothermal vents in the Japan Trench at a depth of 6354-6367 m, were determined by using gas chromatography-mass spectrometry analysis of the 4,4-dimethyloxazoline derivatives. The major fatty acids present in the C. phaseoliformis lipids belong to the n-4 family non-methylene interrupted polyunsaturated fatty acids (NMI-PUFA): 20:3n-4,7,15, 20:4n-1,4,7,15, and 21:3n-4,7,16, with significant levels of 20:2n-7,15 and 21:2n-7,16 as non-methylene interrupted n-7 dienes. Compared with the lipids of shallow-water clam Mactra chinensis, which contains photosynthetic n-3 PUFA, such as docosahexaenoic acid and icosapentaenoic acid, C. phaseoliformis might have an intrinsic mechanism in vivo so as to maintain the fluidity of the high-melting fatty acids in the membrane lipids by exogenous n-4 family PUFA adaptation as substitutes. Such special kinds of fatty acids are assimilated by the symbiotic chemosynthetic bacteria, which use geothermal energy and minerals from the cold-seep vents. Its unique fatty acid composition corresponding to the novel n-4 family NMI-PUFA markedly differs from those of the reported lipid compositions of other marine animals, which depend on the marine grazing food chain originating from phytoplankton. Thus, the present findings confirm a perfect and closed novel food chain in the cold-seep bivalve and in its symbionts, which is independent from the photosynthetic food chain.     

Dietary,but not topical,alpha-linolenic acid suppresses UVB-induced skin injury in hairless mice when compared with linoleic acids

Peroxidizability of fatty acids in the air is roughly proportional to the number of double bonds, but in vivo peroxidation proceeds in a more complex manner. Here, we compared the effects of dietary and topically applied oils enriched with linoleic acid (LA, 18:2n-6) or alpha-linolenic acid (ALA, 18:3n-3) on UV-induced skin injury in a strain of hairless mice. The UVB-induced erythema score was significantly lower in mice with topically applied creams containing LA and ALA than in mice with the basal cream; no significant increase in the score was detected in the ALA group compared with the LA group. However, dietary ALA inhibited the increase in erythema score after UVB irradiation compared with LA. The peroxidizability index of the skin total lipids was significantly higher, but UVB-induced prostaglandin E2 (PGE2) production was significantly lower in the group fed an ALA-rich diet compared with the group fed an LA-rich diet. The levels of thiobarbituric acid-reactive substances, estimated in the presence of butylated hydroxytoluene in the assay mixture, were not affected by UVB treatment or by the dietary fatty acids, but the severity of the skin lesion was associated with PGE2 levels. These results indicate that the type of fatty acids, n-6 or n-3, is critical for the suppression of UVB-induced skin lesion when the skin fatty acids are modified by dietary manipulation. Anti-inflammatory activity of dietary flaxseed oil with relatively high ALA and low LA contents was demonstrated in UVB-irradiated hairless mice.     

Hypertension does not stimulate the development of hypercholesterolemia or fatty liver induced by a high cholesterol/cholate diet in rats

A high cholesterol/cholate diet induced hypercholesterolemia and fatty liver in both spontaneously hypertensive rats (SHR) and normotensive control rats (WKY). However, in contrast to previous concepts, the levels of cholesterol ester, triacylglycerol and phosphatidylcholine in plasma as well as triacylglycerol in liver were higher in WKY than in SHR fed a normal diet. The high cholesterol/cholate diet elevated the levels of plasma cholesterol, plasma cholesterol ester and hepatic triacylglycerol, and the extent of elevation was significantly higher in WKY than in SHR. Increases both in monoene/saturated ratios, an indication of elevated delta 9-desaturase activity, and in linoleate/arachidonate ratios, a possible indication of impaired desaturation-elongation activity, were observed in hepatic and plasma lipids of both strains fed the high cholesterol/cholate diet. The increases in monoene/saturated ratios were similar in both strains, but the increases in the linoleate/arachidonate ratios were higher for the plasma cholesterol esters of WKY than of SHR. The n-6/n-3 ratios of plasma and hepatic lipids were higher in WKY than in SHR throughout the experiments. These diet-induced changes observed in hepatic and plasma lipids were not reflected in the aortic lipids. Thus, hypertension per se does not promote the development of hyperlipemia and fatty liver induced by a high cholesterol/cholate diet. Our results also suggest that the metabolism of polyenoic fatty acids is different between SHR and WKY.     

Cis-trans isomerization of polyunsaturated fatty acid residues in phospholipids catalyzed by thiyl radicals.

Phospholipids containing trans-unsaturated fatty acid residues are the major products of the thiyl radical attack on L-alpha-phosphatidylcholine from soybean lecithin in homogeneous solution or in liposomes (LUVET). Thiyl radicals act as the catalyst for the cis-trans isomerization, and the number of catalytic cycles depends on the reaction conditions. The presence of approximately 0.2 mM oxygen does not influence the reaction outcome but accelerates the efficiency of cis-trans isomerization in homogeneous solution. Under these conditions, the PUFA peroxidation is found to be unimportant. A detailed study of the isomerization of methyl linoleate including product studies indicates the formation of a small amount of conjugated dienes that act as inhibitors. Indeed, all-trans-retinol substantially retarded the isomerization process.     

Fatty acid composition and volatile compounds of caviar from farmed white sturgeon (Acipenser transmontanus)

The present study was conducted to characterize caviar obtained from farmed white sturgeons (Acipenser transmontanus) subjected to different dietary treatments. Twenty caviar samples from fish fed two experimental diets containing different dietary lipid sources have been analysed for chemical composition, fatty acids and flavour volatile compounds. Fatty acid make up of caviar was only minimally influenced by dietary fatty acid composition. Irrespective of dietary treatments, palmitic acid (16:0) and oleic acid (OA, 18:1 n-9) were the most abundant fatty acid followed by docosahexaenoic acid (DHA, 22:6 n-3) and eicopentaenoic (EPA, 20:5 n-3). Thirty-three volatile compounds were isolated using simultaneous distillation-extraction (SDE) and identified by GC-MS. The largest group of volatiles were represented by aldehydes with 20 compounds, representing the 60% of the total volatiles. n-Alkanals, 2-alkenals and 2,4-alkadienals are largely the main responsible for a wide range of flavours in caviar from farmed white surgeon.     

气相色谱全分析海水和淡水养殖南美白对虾组织中脂肪酸组成与含量

采用超声提取-毛细管气相色谱法分析测定了海水养殖和淡水养殖的南美白对虾的肌肉和肝胰腺(中肠腺)中脂肪酸的组成与含量,以用于对虾繁殖与营养的研究。采用Folch 液超声提取对虾组织中的脂质,脂质经氢氧化钾-三氟化硼法甲酯化后上机分析。采用BPX70(60 m×0.25 mm i.d.)石英毛细管柱分离,火焰离子化检测器检测。以线性程序升温等当碳长度(ECLT)值结合标准品对照的方法对17种脂肪酸定性,用面积归一化法定量。结果表明,两种对虾的脂肪酸组成均以不饱和脂肪酸为主,而且不饱和多烯酸的含量明显高于不饱和单烯酸。两种对虾的肌肉脂肪酸中高度不饱和脂肪酸,如二十碳五烯酸(EPA)与二十二碳六烯酸(DHA)的总含量均较高,达20%以上;肝胰腺中EPA和DHA的总含量较低,为11%～17%。淡水虾中n-6族不饱和脂肪酸含量比海水虾高,海水虾中n-3族与n-6族脂肪酸含量之比值比淡水虾高。     

Production of aliphatic aldehydes on peroxidation of various types of lipids.

IN vitro peroxidation by air, or xanthine-xanthine oxidase (xanthine-XOD) was performed to estimate the production of aliphatic aldehydes from free polyunsaturated fatty acids (PUFA), triglycerides, phospholipids and rat liver microsomes and mitochondria. The aldehyde contents in peroxidized lipids were determined by liquid chromatography and fluorescence detection. In both peroxidation, pentanal, (E)-4-hydroxy-2-nonenal (4-HN), and hexanal were produced from omega-6 PUFA rich lipids and propanal was markedly enhanced by increasing the degree of fatty acid unsaturation. The ratios of 4-HN to hexanal production in xanthine-XOD peroxidation of the omega-6 PUFA rich lipids, and rat liver microsomes and mitochondria were much higher than those in air peroxidation. The ratios (4-HN/hexanal) obtained in microsomes and mitochondria by xanthine-XOD were similar to those in rat liver observed in vitamin E deficient studies. The determination of these aldehydes may be useful to estimate the kinds of fatty acids peroxidized and investigate in vivo lipid peroxidation mechanism.     

硒化合物对不饱和脂肪酸过氧自由基的作用

本文用电子自旋共振谱技术研究了硒代二乙酸和硒代二丙酸对不饱和脂肪酸过氧自由基的作用。结果表明,硒化合物清除不饱和度为2和2以上的脂肪酸的过氧自由基,但促进仅含一个双键的不饱和脂肪酸的过氧自由基的生成。     

WAVELENGTH DEPENDENCE OF PHOTOINDUCED PEROXIDATION AND CYTOTOXICITY OF HUMAN LOW DENSITY LIPOPROTEINS

The relative abilities of UV-A, B and C radiations to initiate lipid peroxidation and apolipoprotein (apo) B modification of human purified low density lipoproteins have been compared. Ultraviolet-B and C (at 310 and 254 nm, respectively) exhibited similar efficacy as shown by the increase in lipid peroxidation markers (conjugated dienes, thiobarbituric acid reactive substances and fluorescent lipid soluble products) and in oxysterols, as well as by the decrease of the contents of natural antioxidants (tocopherols and carotenes) and in polyunsaturated fatty acids. In contrast, UV-A (at 360 nm) was found poorly effective and only at very high radiation intensities. Under all the conditions used, apoB was not affected by the UV radiations as shown by the stability of amino acid composition (except tryptophan level) and of trinitrobenzenesulfonic acid reactive amino group content. Similarly, the low density lipoprotein size was not altered. By comparison, low density lipoproteins oxidized by transition metal presented strong alterations of apoB and major changes of the apparent low density lipoprotein size. Finally, low density lipoproteins irradiated by UV-B. or C exhibited a much higher cytotoxicity on cultured cells than those irradiated by UV-A. Under the conditions used in this paper, the cytotoxic effect of the irradiated low density lipoproteins was positively correlated with their content in lipid peroxidation products and inversely correlated with their tocopherol content.     

LIPID PEROXIDATION AND OTHER MEMBRANE DAMAGE PRODUCED IN Escherichia coli K1060 BY NEAR-UV RADIATION AND DEUTERIUM OXIDE

Abstract— With accumulating evidence that the membrane is an important site for near-UV inactivating events, we have investigated the effects of near-UV radiation on the unsaturated fatty acid auxotroph E. coli K1060 following incorporation into the membrane phospholipids of fatty acids with varying degrees of unsaturation. Sensitivity, lipid peroxidation and membrane damage, as determined by ⁸⁶Rb⁺ leakage, have been found to increase with increasing unsaturation in log-phase cells. Similar experiments carried out in D₂O also show an increase in sensitivity, lipid peroxidation and membrane damage, indicating that singlet oxygen may be responsible for such near-UV-radiation-induced damage     

Polyunsaturated but not conjugated linoleic acid supplementation of leukemic U937 cells can act as an amplification factor for photofrin-mediated photodynamic therapy

Polyunsaturated fatty acids located in leukemia cell membranes are excellent targets for peroxidation. They can significantly enhance the effectiveness of Photofrin-mediated photodynamic therapy (PDT)-induced cell killing. In this study, the peroxidizability of conjugated fatty acid isomers (9c,11t-linoleic acid and 9c,11c-linoleic acid) and polyunsaturated fatty acids (PUFAs; linoleic acid, gamma-linolenic acid and arachidonic acid) with 2,2'-azo-bis(2-amidinpropane)dihydrochloride, soybean lipoxygenase and photomediated peroxidation are compared with each other. Peroxidation was determined using different methods: by means of gas chromatography to estimate the fatty acid (FA) consumption, by photometry for the level of FA peroxides or phospholipid peroxides and by definition of the content of malondialdehyde for thiobarbituric acid reactive substances (TBARS). The results suggest that the generation of oxidation products from individual FAs indicate a different formation rate of oxidation products. Radical FA peroxides were produced most by polyunsaturated arachidonic acid, followed by linoleic acid and gamma-linolenic acid, whereas conjugated FA isomers did not generate peroxides. Accordingly, the levels of lipid peroxides and TBARS were substantially increased after incorporation and oxidation of polyunsaturated FAs into U937 cells and could significantly enhance the effectiveness of Photofrin-PDT-induced cytotoxicity. The results showed that PUFA, but not conjugated FA supplementation of U937 cells, can act as a PDT amplification factor.     

STUDY OF THE EFFECTS OF ULTRAVIOLET LIGHT ON BIOMEMBRANES—IV. THE EFFECT OF OXYGEN ON UV-INDUCED HEMOLYSIS AND LIPID PHOTOPEROXIDATION IN RAT ERYTHROCYTES AND LIPOSOMES

Abstract— Hemolysis induced by irradiation with ultraviolet (UV) light at 254 nm showed a pronounced oxygen effect: under irradiation in vacuum, the rate of hemolysis was decreased by an order of magnitude. Irradiation at 254 nm in air but not under vacuum caused the peroxidation of erythrocyte membrane lipids. These results suggest that membrane lipid photoperoxidation is one of the causative factors of UV hemolysis. Irradiation at different wavelengths showed that UV-induced lipid photoperoxidation in erythrocyte membranes developed while the antioxidant α-tocopherol was directly photooxidized. It is shown that the process of lipid photolysis in erythrocyte membranes involves sensitization, possibly by protoporphyrin, whose presence in liposomes accelerates the photoperoxidation at 254 and 365 nm of unsaturated fatty acid residues in lecithin. Possible mechanisms of photochemical damage to erythrocyte membranes are discussed.     

Electron-transfer oxidation properties of unsaturated fatty acids and mechanistic insight into lipoxygenases

Rate constants of photoinduced electron-transfer oxidation of unsaturated fatty acids with a series of singlet excited states of oxidants in acetonitrile at 298 K were examined and the resulting electron-transfer rate constants (k(et)) were evaluated in light of the free energy relationship of electron transfer to determine the one-electron oxidation potentials (E(ox)) of unsaturated fatty acids and the intrinsic barrier of electron transfer. The k(et) values of linoleic acid with a series of oxidants are the same as the corresponding k(et) values of methyl linoleate, linolenic acid, and arachidonic acid, leading to the same E(ox) value of linoleic acid, methyl linoleate, linolenic acid, and arachidonic acid (1.76 V vs SCE), which is significantly lower than that of oleic acid (2.03 V vs SCE) as indicated by the smaller k(et) values of oleic acid than those of other unsaturated fatty acids. The radical cation of linoleic acid produced in photoinduced electron transfer from linoleic acid to the singlet excited state of 10-methylacridinium ion as well as that of 9,10-dicyanoanthracene was detected by laser flash photolysis experiments. The apparent rate constant of deprotonation of the radical cation of linoleic acid was determined as 8.1 x 10(3) s(-1). In the presence of oxygen, the addition of oxygen to the deprotonated radical produces the peroxyl radical, which has successfully been detected by ESR. No thermal electron transfer or proton-coupled electron transfer has occurred from linoleic acid to a strong one-electron oxidant, Ru(bpy)3(3+) (bpy = 2,2'-bipyridine) or Fe(bpy)3(3+). The present results on the electron-transfer and proton-transfer properties of unsaturated fatty acids provide valuable mechanistic insight into lipoxygenases to clarify the proton-coupled electron-transfer process in the catalytic function.     

Gas chromatographic profiles of plasma total lipids as indicators of dietary history. Correlation with fat intake based on 24-h dietary recall

Fasting plasma total lipid profiles were determined by high-temperature gas chromatography on a total of 1246 free living urban subjects, ages 20-59 years, from the Toronto-McMaster Lipid Research Clinic Population Study. Quantitative estimates of the major molecular species, lipid classes and lipid class ratios were correlated with a total of twelve dietary lipid components, including total saturated and unsaturated fats. oleic and linoleic acids, and cholesterol, to give appropriate Spearman coefficients (rS) and tests of significance (P) for groups of 775 males and 471 females. The intake of the various nutrients was derived from a 24-h dietary recall. The most significant correlations varying from rs +/- 0.1-0.4 and P less than 0.0001-0.0005 were between the intake of total fat, individual saturated and unsaturated fats, and the ratios of C50/C54 triacylglycerols and the C34/C36 phosphatidylcholines, which reflected the nature and quantity of the dietary fat consumed. Increases in dietary cholesterol and saturated fat produced small increases in plasma cholesterol and saturated triacylglycerols, while unsaturated dietary fat produced small decreases in saturated and increases in unsaturated plasma triacylglycerols. These changes in the plasma lipid parameters are consistent with those observed previously in much more limited dietary experiments with accurately known composition of ingested fats. It is, therefore, concluded that direct gas chromatographic profiling of plasma total lipids provides a simple and rapid method of verifying the overall correctness of the dietary recall.     

High-light-induced changes on photosynthesis, pigments, sugars, lipids and antioxidant enzymes in freshwater (Nostoc spongiaeforme) and marine (Phormidium corium) cyanobacteria

We studied the effects of high-light exposure (500 micromol m(-2) s(-1) of photosynthetic active radiation) on the cyanobacteria Nostoc spongiaeforme Agardh, a fresh-water alga, and Phormidium corium Agardh (Gomont), a marine alga, with respect to photosynthesis, pigments, sugar content, lipid peroxidation, fatty acids composition, antioxidant enzymes activity and DNA. It was seen that the ratio of variable fluorescence (Fv) to maximum fluorescence (Fm), which is indicative of photosynthetic efficiency, decreased because of the light treatment. The damage to photosynthesis occurred in the antenna system and the photosynthetic II reaction center. Photobleaching of photosynthetic pigments was also observed. High-light treatment also resulted in decreased sugar content, which was probably due to the effect on photosynthesis. Peroxidation of membrane lipids, indicating oxidative damage to lipids and a high level of unsaturation in the cell membrane, was also observed. The activity of antioxidant enzyme superoxide dismutase and ascorbate peroxidase was increased, probably as a result of oxidative damage observed in the form of lipid peroxidation. Quantitative decreases in phospholipid and glycolipid levels were also observed. The level of unsaturated fatty acids in total lipids and glycolipids remained unchanged in both species; however, the level of saturated fatty acids decreased, which slightly changed the ratio in favor of unsaturated fatty acids. Degradation of DNA was also observed in both species. There was a transient plateau 2-4 h after exposure to high-light treatment in the Fv/Fm ratio and in levels of phycobilisome pigments, sugars and antioxidant enzymes after an initial decrease 1 h after the treatment. These findings may indicate a period of partial adaptation to high light that is due to the efficiency of protective processes operational in the two species, which subsequently failed after a longer exposure duration of 4-6 h.     

Photosensitizing action of furocoumarins on membrane components and consequent intracellular events

The photodamage induced in membrane components by furocoumarins is reviewed. The oxygen-dependent photoreactions between furocoumarins and cell membrane constituents lead mainly to lipid peroxidation and the formation of cross-linking in ghost proteins, whereas the oxygen-independent photoreactions lead essentially to a C4 cycloaddition between the furocoumarin and the unsaturated fatty acids. In the latter, cycloadducts are formed between the 3,4 double bond of the furocoumarin and the olefinic double bond of the unsaturated fatty acid. The stereochemical structures of these cycloadducts and the reaction mechanism of the cycloaddition are discussed. Finally, the modulation of several membrane systems by furocoumarins and the consequent intracellular events are reviewed.     

Photohemolysis Sensitized by the Furocoumarin Derivative Alloimperatorin and its Hydroperoxide Photooxidation Product

The dark and photosensitized effects of alloimperatorin methyl ether 1 (hereafter simply alloimperatorin) and its photooxygenation product alloimperatorin hydroperoxide 2 were investigated on human erythrocytes. The results reveal that the furocoumarin 1 photosensitizes efficiently the hemolysis of erythrocytes. The rate of photohemolysis increases on raising the temperature of the postirradiated incubation from 4°C to 37°C. Thermal activation of the photohemolysis and inhibition by 2,6‐di‐tert‐butyl‐p‐cresol (BHT) suggest that the furocoumarin 1 photosensitizes lipid peroxidation, increasing permeability in the erythrocyte membrane. The hydroperoxide 2 induces dark and photosensitized hemolysis more efficiently than the furocoumarin 1. The rate of hemolysis induced by 2 increases with the incubation temperature and decreases in the presence of tert‐butanol and BHT. The hydroperoxide 2 photosensitizes the formation of lipid peroxidation products as shown by the reaction with thiobarbituric acid. This process is diminished by BHT. Our data imply that the photohemolysis sensitized by the furocoumarin 1 is caused by the in situ‐formed photooxygenation product 2. Such hydroperoxides are potent hemolytic agents in the dark and especially on photosensitization.     

Gas chromatography and silver-ion high-performance liquid chromatography analysis of conjugated linoleic acid isomers in free fatty acid form using sulphuric acid in methanol as catalyst

This study used GC and silver-ion HPLC to examine the effects of temperature and time on methylation of individual and mixtures of conjugated linoleic acid (CLA) isomers in free fatty acid form using sulphuric acid as catalyst. In the conditions tested (temperatures between 20 and 50 degrees C and times between 10 and 60min) methylation was complete while avoiding isomerization of conjugated dienes and the formation of artefacts that could interfere with chromatographic determinations. An analytical method using solvent extraction of the lipids followed by selective elution of the free fatty acids from aminopropyl bonded phase columns and methylation with H(2)SO(4) in mild conditions was then applied to determine the CLA isomers in free fatty acid form in rumen fluid, and the results were evaluated.     

UVB-INDUCED PHOTOPEROXIDATION OF LIPIDS OF HUMAN LOW and HIGH DENSITY LIPOPROTEINS. A POSSIBLE ROLE OF TRYPTOPHAN RESIDUES

Ultraviolet radiation of the UVB region readily destroy tryptophan (Trp) residues of low (LDL) and high (HDL) density lipoproteins. The photooxidation of tryptophan residues is accompanied by the peroxidation of low and high density lipoproteins unsaturated fatty acids, as measured by the thiobarbituric acid assay. Moreover, low and high density lipoproteins are natural carriers of vitamin E and carotenoids. These two antioxidants are also rapidly bleached by UVB. The UVA radiation promotes neither tryptophan residue destruction nor lipid photoperoxidation. The redox cycling Cu2+ ions considerably increase lipid photoperoxidation. The synergistic action of photo and auto (Cu2(+)-induced) peroxidation induces marked post-irradiation modifications of apolipoproteins as illustrated by the degradation of most tryptophan residues after overnight incubation in the dark of pre-irradiated samples.