Showing papers in "Lipids in 1983"

Fatty acids in plasma and red cell membranes in normal humans

Abstract: A detailed study was made of the fatty acid composition of plasma triglycerides, free fatty acids, phospholipids, red cell total phospholipids, phosphatidylcholine and phosphatidylethanolamine in 32 normal males and 18 normal females. No sex differences could be detected. There were substantial differences in the compositions of the various fractions and long-chain polyunsaturated fatty acids were particularly important in the red cells.

Detection and determination of lipase (acylglycerol hydrolase) activity from various sources

Abstract: Methods for the detection and determination of lipases (acylglycerol hydrolases) and preparation of assays are reviewed including substrates, conditions and screening. Some newer methods for the determination of lipase activity are discussed. Several of these are: (a) titrimetry, (b) colorimetry of Cu soaps of free fatty acids (FFA), (c) colorimetry of chromophores in the acyl chain of FFA or in glycerol, (d) radioassay, (e) gas liquid chromatography, (f) enzymatic treatment of FFA and measurement of the resulting products, and (g) direct immunological determination of the lipase. Examples and sensitivities are given and advantages and disadvantages are described.

Inhibition of cholesterol and fatty acid biosynthesis in liver enzymes and chicken hepatocytes by polar fractions of garlic

Abstract: Different concentrations of polar fractions, methanol-soluble (MESF), or water-soluble (WASF), of 1–8% equivalent to fresh garlic paste were added to yellow corn-soybean based diets and fed to 5-week-old male broiler chickens for 3 weeks to measure the inhibition of hepatic β-hydroxy-β-methylglutaryl coenzyme A (HMG-CoA) reductase, cholesterol 7α-hydroxylase (7α-hydroxy) and fatty acid synthetase (FAS). Dose-related decreases in the activities of these enzymes were obtained. Decreases in serum total cholesterol and in low density lipoprotein (LDL) levels were also observed. There was no effect on the level of cholesterol in high density lipoprotein (HDL). The most effective dose for these decreases was found 0.54% (MESF) and 1.2% (WASF) equivalent to 6% of the fresh garlic. The inhibition of HMG-CoA reductase and FAS by 25–300 μm of MESF or WASF for 15 min was tested in vitro, in male and female chicken hepatocytes. Inhibitions of activity were dose-dependent and the degree of inhibition increased with duration of incubation (150 μg of MESF or WASF 5 to 60 min). Dietary supplementation of odorless WASF of garlic was found to be very effective in lowering the total and LDL cholesterol levels compared to control chickens.

Bovine pulmonary surfactant: chemical composition and physical properties.

Abstract: Bovine pulmonary surfactant was obtained by endotracheal lavage of lungs from newly slaughtered cows followed by differential centrifugation. Lipid extracts of bovine surfactant contained 3% neutral lipid, mainly as cholesterol and diacylglycerol and 97% phospholipid. Phosphatidylcholine (79%) and phosphatidylglycerol (11%) accounted for most of the phospholipids with smaller amounts of phosphatidylethanolamine, phosphatidylinositol, lyso-bis-phosphatidic acid and sphingomyelin. Fatty acid analysis revealed high levels of palmitate in phosphatidylcholine and to a lesser extent phosphatidylglycerol, but not in the other diacylphospholipids. Phosphatidylcholine was 53% disaturated and phosphatidylglycerol was 23% disaturated. Monoenoic species accounted for the major proportion of the remaining lipid. The protein content was 10% as estimated by the Lowry procedure and 5% when determined by amino acid analysis. Extraction with chloroform/methanol removed ca. 90% of the protein but had no effect on the surfactant properties as evaluated by a pulsating bubble technique.

A comparison of the oleaginous yeast,Candida curvata, grown on different carbon sources in continuous and batch culture

Abstract: The oleaginous yeast,Candida curvata D, was grown in both batch and continuous culture on 5 different carbon sources to compare the efficiency of fat production from the various substrates. Maximum lipid accumulation occurred in batch culture with xylose as the carbon source on nitrogenlimited medium reaching a level of 49% (w/w) of the biomass, but this was reduced to 37% at the optimum dilution rate (D=0.05/hr) in a chemostat. Both the highest biomass and lipid yields were attained in continuous culture with lactose as the sole carbon source at a dilution rate of D=0.04/hr, giving an efficiency of substrate conversion of 60 g of biomass and 18.6 g lipid per 100 g lactose utilized. The relative proportions of the major fatty acids (16∶0, 18∶0, 18∶1, 18∶2) in the lipid were found to vary considerably in batch culture and in continuous culture under carbon-limited conditions. However, on nitrogen-limited media in the chemostat, the fatty acid composition remained relatively constant over the whole range of dilution rates employed. Lipid from xylose-grown cells contained the greatest percentage of stearic acid (18∶0) 15% and the lowest linoleic acid (18∶2) 4%, whereas lipid from ethanol-grown cells contained elevated levels of oleic acid (18∶1) 51% and decreased palmitic acid (16∶0) 25%.

How do polyunsaturated fatty acids lower plasma cholesterol levels

Abstract: For 30 years it has been known that linoleic acid can lower elevated cholesterol levels. Large increases in linoleic acid have been widely recommended as a way of reducing the risk of cardiovascular disease. Such recommendations have resulted in major dietary shifts in some countries, including the USA. Yet the precise characteristics of the linoleic acid molecule which confer on it cholesterol-lowering properties are unknown. gamma-Linolenic acid, the first essential fatty acid metabolite of linoleic acid, has been found to have cholesterol-lowering actions ca. 170 times greater than the parent molecule, suggesting that linoleic acid must be converted to gamma-linolenic acid to exert its desirable effects on cholesterol metabolism. Aging, sex, diabetes mellitus, alcohol, catecholamines and trans fatty acids and saturated fats can all modulate the delta-6-desaturase enzyme which converts linoleic acid to gamma-linolenic acid. This provides a possible unifying explanation for the actions of these known risk factors for cardiovascular disease.

Determination of lipase specificity

Abstract: Specificity of lipases is controlled by the molecular properties of the enzyme, structure of the substrate and factors affecting binding of the enzyme to the substrate. Types of specificity are as follows. I. Substrate: (a) different rates of lipolysis of TG, DG, and MG by the same enzyme; (b) separate enzymes from the same source for TG, DG and MG. II. Positional: (a) primary esters; (b) secondary esters; and (c) all three esters or nonspecific hydrolysis. III. Fatty acid, preference for similar fatty acids. IV. Stereospecificity: faster hydrolysis of one primary sn ester as compared to the other. V. Combinations of I-IV. Lipases with these specificities are: Ia, pancreatic; Ib, postheparin plasma. IIa, pancreatic; IIb, Geotrichum candidum, for fatty acids with cis-9-unsaturation, and IIc, Candida cylindracea. III, G. candidum for unsaturates. IV. sn-1, postheparin plasma and sn-3 human and rat lingual lipases. V. Rat lingual lipase. Methods for determination involve digestion of natural fats of known structure and synthetic acylglycerols followed by analysis of the lipolysis products. All of the types of specificity have been detected with use of synthetic acylglycerols. Detection of stereospecificity requires enantiomeric acylglycerols which are difficult to synthesize, so other methods have been developed. These involve the generation of 1,2-(2,3)DG and resolution of the enantiomers. Trioleoylglycerol or racemic TG can be used as substrates. If the lipase is stereospecific, then either the sn-1,2- or 2,3-enantiomer will predominate. The relative amounts of the enantiomers can be determined by measurement of specific rotation, and nuclear magnetic resonance spectra. The DG can also be separated by conversion to phospholipids and hydrolysis with phospholipases A-2 or C. Applications of these procedures are discussed and data on the specificity of various lipases presented.

Effects of purified eicosapentaenoic acid on arachidonic acid metabolism in cultured murine aortic smooth muscle cells, vessel walls and platelets

Abstract: The effects of highly purified eicosapentaenoic acid (97% pure) on the arachidonic acid cascade in isolated murine vascular cells and platelets were studied. The incorporation of eicosapentaenoic acid was not as active as that of arachidonic acid in platelets. The ratio of incorporation of eicosapentaenoic acid to arachidonic acid into platelet phospholipids was about 0.7. Analysis of the phospholipid fractions of platelets after labeling with 14C-eicosapentaenoic acid and 14C-arachidonic acid revealed that the incorporation of 14C-eicosapentaenoic acid into the fraction is significantly less than that of 14C-arachidonic acid, while the incorporation of both fatty acids into other phospholipid fractions was almost the same. On the other hand, no significant difference between either fatty acid in incorporation rate, kinetics or distribution in cellular phospholipids was found in cultured aortic smooth muscle cells. Following treatment with eicosapentaenoic acid, cells produced less prostacyclin from endogenous arachidonic acid than did control cells. This was not due to the decrease in fatty acid cyclooxygenase activity, but rather, due to the decrease in arachidonic acid content in cellular phospholipids. In addition, eicosapentaenoic acid was neither converted to prostaglandin I3 by the vascular cells nor to thromboxane A3 by platelets. Furthermore, similar results were also obtained by in vivo experiments in which rats were fed with eicosapentaenoic acid enriched diet.

Systematic protocol for the accumulation of fatty acid data from multiple tissue samples: Tissue handling, lipid extraction and class separation, and capillary gas chromatographic analysis

Abstract: A systematic procedure was developed for detailed fatty acid profiling of both neutral and polar lipid fractions isolated from hundreds of related bovine muscle and adipose tissue samples. A regimen was established for a nonbiased handling of tissue samples, which included their handling in a predetermined random order. Lipid class separation was accomplished concomitantly during the extraction of the tissues by a selective dry column method, which allowed a detailed analysis of minor but important polyunsaturated fatty acids associated with the polar fraction. Neutral lipids were derivatized to fatty acid methyl esters (FAME) by a literature procedure. However, to protect against lysis of plasmalogens in the polar fraction, a modified nonacidic esterification procedure was developed. FAME profiles were obtained on a program-mable high resolution capillary gas chromatograph (GC). Run programs for unattended GC operation and data storage are described. By this overall procedure, the quantitation and peak identification were obtained for major and minor fatty acid constituents from bovine tissue in a manner that prepares for valid statistical interpretation of the resulting data.

Different fatty chain compositions of alkenylacyl, alkylacyl and diacyl phospholipids in rabbit alveolar macrophages: High amounts of arachidonic acid in ether phospholipids

Abstract: High levels of ether phospholipids were found in rabbit alveolar macrophages. Choline phosphoglycerides (CPG) contained a significant amount of alkylacyl compound (32.5%). On the other hand, ethanolamine phosphoglyceride (EPG) included a very large amount of alkenylacyl compounds (61.2%). Small amounts of alkenylacyl CPG and alkylacyl EPG were also observed. The occurrence of a high amount of alkylacyl CPG may be related to the synthesis or release of platelet-activating factor (PAF) from macrophages. Fatty chains at the 1- and 2-positions in each lipid class of CPG or CPG or alkenylacyl EPG were several other. Particularly, the levels of 20∶4 (arachidonic acid) in alkylacyl CPG or alkenylacyl EPG were several times higher than those in corresponding diacyl phospholipids. Large portions of 20∶4-containing species have alkenyl or alkyl ether moieties at their 1-position in both CPG (73.6%) and EPG (85.9%). These results suggest the importance of ether-containing phospholipids in rabbit alveolar macrophages.

The effect of vitamin C on in vivo lipid peroxidation in guinea pigs as measured by pentane and ethane production.

Abstract: Measurements of pentane and ethane as indices of in vivo lipid peroxidation were made on samples of breath from vitamin C-sufficient and vitamin C-deficient guinea pigs injected with 23 μl carbon tetrachloride (CCl4)/100 g body wt. Vitamin C-deficient animals produced significantly more pentane and ethane after CCl4 treatment than did vitamin C-sufficient guinea pigs. Pretreatment of vitamin C-deficient animals with 75 mg ascorbic acid/100 g body wt significantly lowered both pentane and ethane evolution. Protection against in vivo lipid peroxidation similar to that provided by ascorbic acid was also found when vitamin C-deficient guinea pigs were pretreated with isoascorbic acid, reduced glutathione, α-tocopherol or β-carotene. When animals were pretreated with the radical scavenger mannitol, a protective effect was also observed as measured by pentane evolution.

1‐O‐alkyl‐linked phosphoglycerides of human platelets: Distribution of arachidonate and other acyl residues in the ether‐linked and diacyl species

Abstract: In this study, the 1-O-alkyl-2-acyl-sn-glycero-3-phosphocholine content of human platelets was determined. The distribution of arachidonate among the 1,2-diacyl, 1-O-alkyl-2-acyl, and 1-O-alk-l′-enyl-2-acyl classes of choline- and ethanolamine-containing phosphoglycerides was also assessed. The major platelet phospholipids were choline-containing phosphoglycerides (38%), ethanolamine-containing phosphoglycerides (25%) and sphingomyelin (18%), with smaller amounts of phosphatidylserine (11%) and phosphatidylinositol (4%). In addition to the diacyl class, the choline-linked fraction was found to contain both 1-O-alkyl-2-acyl (10%) and 1-O-alk-l′-enyl-2-acyl (9%) species. The ethanolamine-linked fraction, on the other hand, had an elevated level of the 1-O-alk-l′-enyl-2-acyl (60%) species and a small amount of the 1-O-alkyl-2-acyl component (4%). The major fatty acyl residues found in all classes of the choline and ethanolamine phospholipids were 16∶0, 18∶0, (Δ9), 18∶2(n−6) and 20∶4(n−6). The 1-O-alk-l and 1-O-alk-l′-enyl fraction of the ethanolamine-linked phospholipids also contained substantial amounts of 22∶4(n−6), 22∶5(n−3) and 22∶6(n−3) acyl chains. Arachidonate comprised 44% of the acyl residues in thesn-2 position of 1-O-alkyl-2-acyl-sn-glycero-3-phosphocholine. Corresponding values for the diacyl and 1-O-alk-l′-enyl-2-acyl species were 23% and 25%, respectively, based on all 20∶4(n−6) being linked to thesn-2 position of all classes. In the ethanolamine-linked phosphoglycerides, arachidonate constituted 60%, 20% and 68% of the acyl groups in thesn-2 position of the 1,2-diacyl, 1-O-alkyl-2-acyl and 1-O-alk-l′-enyl-2-acyl classes, respectively. The content of 1-O-alkyl-2-acyl-sn-glycero-3-phosphocholine appears sufficient to support the synthesis of platelet activating factor by a deacylation-reacylation pathway in platelets. Our findings also demonstrate that human platelets contain a significant amount of 1-O-alkyl-2-arachidonyl-sn-glycero-3-phosphocholine that could possibly serve as a precursor of both platelet activating factor and bioactive arachidonate metabolites.

Australian fish-An excellent source of both arachidonic acid and ω-3 polyunsaturated fatty acids.

Abstract: The fatty acid methyl esters obtained by the esterification of total lipids extracted from 24 species of fin fish and 4 species of invertebrates caught in the rivers and coastal waters of southern Australia were analyzed by gas chromatography The lipids of most species contained significant levels of arachidonic acid (07–158%) as well as the more common marine polyunsaturate, eicosapentaenoic acid (07–159%) The major ω6 fatty acid present in most species was 20∶4; however, other fatty acids of this series, including 18∶2, 22∶4 and 22∶5, were present The level of total ω6 fatty acids ranged from 39 to 223% of the total lipid In general, the level of total ω3 polyunsaturates was higher than the total ω6 fatty acids with levels of ω3 fatty acids ranging from 96 to 482% Only 2 fish (barramundi and gurnard perch) had ω6/ω3 ratios greater than 10 Most of the Australian species examined contained low levels of fat (05–78% of fresh weight) Two species examined, callop (freshwater) and blue groper (marine) contained sufficient quantities of both fat (77 and 78%) and arachidonic acid (48 and 93%) to warrant consideration for commercial exploitation

(n-3) and (n-6) polyunsaturated fatty acids in the phosphoglycerides of salt-secreting epithelia from two marine fish species.

Abstract: Fatty acid analyses were carried out on phosphoglycerides isolated from microsomal fractions of the rectal gland of the dogfish,Scyliorthinus canicula, and gills of the cod,Gadus morhua. Ratios of (n-3)/(n-6) polyunsaturated fatty acids were ca. 10 for phosphatidylcholine, (PC), phosphatidylethanolamine (PE) and phosphatidylserine (PS) from cod gills, reflecting high concentrations of 20∶5 (n-3) and 22∶6(n-3). The ratio for phosphatidylinositol (PI) from cod gills was 1.3, reflecting high concentrations of 20∶4(n-6) as well as (n-3) polyunsaturates. PC, PE and PS from rectal glands all had much lower (n-3)/(n-6) ratios than in cod gills, reflecting higher concentrations of 20∶4(n-6), but the lowest ratio was again present in PI. The latter phospholipid had high concentrations of 18∶0 in both tissues. The relative constancy of the fatty acid composition of PI in the two salt-secreting tissues and its similarity to mammalian phospholipids is considered to reflect its specialized role in biomembranes.

Dietary lipid modulation of immune responsiveness.

Abstract: The influence of dietary fat concentration and saturation on blastogenesis, cytotoxicity, antibody response and fatty acid composition of murine splenic lymphocytes was studied Blastogenesis of lymphocytes from dietarily manipulated mice in response to alloantigens from control mice was significantly greater for those mice fed a diet containing minimal essential fatty acids (EFA) as the only fat source (EFA control) than those fed an EFA-deficient diet When the dietary fat concentration was increased, blastogenic responses decreased compared to the EFA control diet Lymphocyte-mediated cytotoxicity against allogeneic melanoma cells was greater for mice receiving diets with EFA only than for those deficient in EFA However, cytotoxicity responses of mice fed additional polyunsaturated fat (PUF) decreased as concentration increased, whereas responses of mice fed the saturated fat (SF) diets decreased only when the dietary fat concentration was greater than 8% As compared to diets with EFA control, direct plaqueforming cell (PFC) response was decreased for mice fed high levels of PUF and increased for mice fed high levels of SF; however, no difference in the percentage of IgM-positive cells was observed These changes in PFC response were inversely related to the levels of linoleic acid in the lymphocyte Thus, high levels of dietary fat, and particularly PUF, suppress lymphocyte functions when EFA requirements are met, whereas low levels (EFA control) intensify these responses EFA deficiency, however, suppresses some lymphocyte responses Thus, dietary lipids differentially modulate the levels of T- and B-cell responsiveness

The binding of micellar lipids to chitosan

Abstract: The lipid binding capacity of chitosan (partially deacetylated chitin) was determinined with respect to micellar solutions of bile salts, dodecyl sulfate, natural ox bile and an artificial mixed microemulsion. The stoichiometry was determined following the separation of the solid phase by filtration or centrifugation. The major variables in the extent of binding were the pH and ionic strength, suggesting that the interactions are mainly of ionic nature. It is noteworthy that under optimal conditions chitosan could bind, i.e., coprecipitate, with 4-5 times of its weight with all the lipid aggregates tested. These results have a bearing on the nutritional and pharmacological applications of chitosan. The analyses of the components from the precipitates with microemulsion and ox bile show a significant selectivily of binding caused by hydrophobic interactions.

Phospholipid studies of marine organisms: V1 new α-methoxy acids fromHigginsia tethyoides

Abstract: The phospholipids of the demospongeHigginsia tethyoides are shown to have at least 16 long-chain α-methoxy acids, which represent a new class of fatty acids. Among them are the saturated α-methoxy acids containing 19-24 carbon atoms. The monounsaturated compounds are 2-OMe-Δ(17)-24:1, 2-OMe-Δ(18)-25:1, 2-OMeΔ(19)-26:1 and 2-OMe-Δ(21)-28:1. The major diunsatured ones were shown to be 2-OMe-Δ(5, 19)-26:2 and 2-OMe-Δ(7, 21)-28:2. Small amounts of 2-OMe-23∶1, 2-OMe-26∶3; 2-OMe-27∶1 and 2-OMe-28∶3 were also encountered. Structures of the minor monounsaturated compounds were tentatively assigned as 2-methoxy-16-tricosenoic acid and 2-methoxy-20-heptacosenoic acids. The double bonds of the fatty acids show all-cis configuration. Circular dichroism measurements indicate an R-configuration for the α-methoxy acids. The major component of the total phospholipid acid mixture is 5,9,23-triacontatrienoic acid. Possible biosynthetic routes to these unusual phospholipid acids are discussed. The major phospholipids were phosphatidylethanolamine, phosphatidylglycerol and phosphatidylserine. The distribution of fatty acids among the phospholipids was also investigated.

Elevated levels of arachidonic acid in fish from northern Australian coastal waters.

Abstract: The fatty acid composition of 10 species of fish caught off the northwest coast of Australia (latitude 17°S) was examined. All species contained high levels of ω6 fatty acids (9.6–23.1% of total fatty acids) with arachidonic acid being the major ω6 fatty acid (5.9–14.8% of fatty acids). Docosatetraenoic and docosapentaenoic acids of the ω6 series accounted for 3–8% of the total fatty acids. The ratio of ω6 to ω3 fatty acids in these fish varied from 0.38 to 0.93, compared with an average ratio of 0.16 for fish from the northern hemisphere (latitude >30°N). The present data and figures from the literature indicate that the marine food chain in the southern hemisphere contains significant quantities of ω6 fatty acids.

Modulation of prostaglandin synthesis in rat peritoneal macrophages with ω-3 fatty acids,

Abstract: In view of the findings that ω3 fatty acids inhibit the synthesis of prostaglandins (PG) from arachidonic acid (20∶4ω6) and that among immunologically active cells, the macrophage, is a major producer of PG, we undertook a study of the effect of dietary α-linolenic acid (18∶3ω3) on PG synthesis in the macrophage. Rats were fed purified diets containing either 10% corn oil (CO) or linseed oil (LO), providing either a low (1/32) or high (3.5/1) ratio of 18∶3ω3 to 18∶2ω6, respectively, for 6 weeks. Fatty acid analysis of macrophage phospholipids showed that there was an appreciable increase in the percentage of ω3 fatty acids and a decrease in the ω6 fatty acids in macrophages from rats fed the LO diet. The changes in fatty acid composition were associated with a significant decrease in the synthesis of prostaglandin E (PGE) by macrophages from rats fed the LO diet. Macrophages from rats fed the 2 dietary, oils did not differ in their ability to degrade PG, thus the difference in PG production appeared to be a consequence of decreased synthesis only. The dietarily induced changes in PGE synthesis were readily overcome in vitro by culturing macrophages with complexes of fat-free bovine serum albumin and either 20∶4ω6 or 20∶5ω3.

Photosensitized oxidation of methyl linoleate monohydroperoxides: hydroperoxy cyclic peroxides, dihydroperoxides, keto esters and volatile thermal decomposition products

Abstract: Previous studies of lipid secondary oxidation products have been extended to 6-membered hydroperoxy cyclic peroxides from the singlet oxygenation of a mixture of 9- and 13-hydroperoxides from autoxidized methyl linoleate. The oxidation product was fractionated by silicic acid chromatography with diethyl ether/hexane mixtures, and selected fractions were separated by polar phase high performance liquid chromatography. Products characterized by thin layer chromatography, gas liquid chromatography, ultraviolet, infrared, nuclear magnetic resonance and mass spectrometry included: 6-membered cyclic peroxides (13-hydroperoxy-9,12-epidioxy-10- and 9-hydroperoxy-10,13-epidioxy-11-octadecenoates), dihydroperoxides (8,13- and 9,14-dihydroperoxyoctadecadienoates) and keto dienes (9- and 13-oxooctadecadienoates). The 6-membered hydroperoxy cyclic peroxides are apparently formed by 1,4-addition of singlet oxygen to 9- and 13-hydroperoxides withtrans, trans-conjugated diene systems. Thermal decomposition of the 6-membered hydroperoxy cyclic peroxides at 200 C produced methyl 9-oxononanoate and hexanal as the major volatiles. Other volatiles included 2-pentylfuran, pentane, 4-oxo-2-nonenal, methyl furanoctanoate and methyl 9,12-dioxo-10-dodecenoate.

Use of antioxidants in the analysis of vatamins A and E in mammalian plasma by high performance liquid chromatography.

Abstract: A simple, sensitive, quantitative method for the simultaneous assay of retinol, α-tocopherol and γ-tocopherol in rat, guinea pig, monkey and human plasma was developed by using high performance liquid chromatography. It was found that antioxidant was required to stabilize the fat-soluble vitamins in the plasma of rats. The effect of several antioxidants on the recovery of fat-soluble vitamins was evaluated. Results showed that 0.125% butylated hydroxytoluene (BHT) in ethanol and 0.025% BHT in heptane yielded recoveries >95% in 0.1 ml plasma.

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

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

The Effect of Dietary Partially Hydrogenated Marine Oils on Desaturation of Fatty Acids in Rat Liver Microsomes

Abstract: The influence of dietary partially hydrogenated marine oils on distribution of phospholipid fatty acids in rat liver microsomes was studied with particular reference to the metabolism of linoleic acid. Five groups of weanling rats were fed diets containing 20% (w/w) peanut oil (PO), partially hydrogenated peanut oil (HPO), partially hydrogenated Norwegian capelin oil (HCO), partially hydrogenated herring oil (HHO), and rapeseed oil (RSO) for 10 weeks. The partially hydrogenated oils were supplemented with linoleic acid corresponding to 4.6 cal % in the diets. Accumulation of linoleic acid and reduced amount of total linoleic acid metabolites were observed in liver microsomal phospholipids from rats fed partially hydrogenated oils as compared to PO feeding. The most striking effects on the distribution of omega 6-polyunsaturated fatty acids was obtained after feeding HHO, a marine oil with a moderate content of trans fatty acids in comparison with HPO but rich in isomers of eicosenoic and docosenoic acids. Liver microsomal delta 6- as well as delta 5-desaturase activities as measured in vitro were reduced in rats kept on HHO as compared to PO dietary treatment. The results obtained suggest that the dietary influence of partially hydrogenated marine oils on the metabolism of linoleic acid might be better related to the intake of isomeric eicosenoic and docosenoic acids than to the total intake of trans fatty acids.

A complete separation of lipids by three-directional thin layer chromatography

Abstract: A novel three-directional thin layer chromatography (TLC) method is reported by which all the polar and neutral lipids are isolated on a single TLC plate. Following resolution of the phospholipids by two-directional TLC, lipids are visualized by ultraviolet light after spraying with 2′,7′-dichloro-fluorescein. A line is drawn across the plate, parallel to the second direction of development, separating the resolved phospholipids and the neutral lipids concentrated along the solvent front. The TLC plate is then chromatographed in the reverse direction of the second development to resolve the neutral lipids. By exposing the lipids to HCl fumes after the first development, the plasmalogen content of the lipids may also be determined. This new technique is rapid and lends itself to qualitative and quantitative analyses of total lipids.

Fatty acid metabolism and cell proliferation: IV. Effect of prostanoid biosynthesis from endogenous fatty acid release with cyclosporin-A

Abstract: Cyclosporin-A (Cyc-A) stimulates prostanoid (PGI2) synthesis in confluent smooth muscle cells from guinea pig aorta through the release of endogenous fatty acid. Cyc-A, like other stimulatory agents for prostanoids, promotes smooth muscle cell proliferation and prostanoid synthesis in these proliferating cells. Indomethacin, a cyclooxygenase inhibitor, and exogenous arachidonic acid block the Cyc-A effect on cell proliferation.

Altered Microsomal Phospholipid Composition in the Streptozotocin Diabetic Rat

Abstract: Streptozotocin diabetes in the rat alters liver microsomal membrane fatty acid composition. The present study was undertaken to determine if such changes in fatty acid composition were due to changes in the amount of individual phosphoglycerides or to disproportionate changes in fatty acid composition in any of the individual phosphoglycerides. The diabetic animals showed a small increase in total microsomal phospholipid, which is due to a selective increase in the phosphatidylethanolamine fraction. The changes in fatty acid composition in the total lipid extract (decreased palmitoleic, oleic and arachidonic acids and increased linoleic and docosahexaenoic acids) from the diabetic animals were present in both the major phosphoglycerides, phosphatidylcholine and phosphatidylethanolamine, with very little change in fatty acid composition in the phosphatidylserine and inositol fraction. Further studies are necessary to delineate the cause of the abnormal membrane phospholipid composition in the diabetic animal.

Complex formation in sonicated mixtures of β-lactoglobulin and phosphatidylcholine

Abstract: β-Lactoglobulin, the major whey protein of bovine milk, is secreted via the endomembrane system of the mammary gland. The primary structure of β-lactoglobulin shares certain characteristics with membrane proteins, although the soluble protein assumes a globular conformation. We have prepared complexes of β-lactoglobulin and phosphatidylcholines by dissolving both in a helix-forming solvent (chloroform methanol). The complex is stable when transferred to aqueous solutions and sonicated to form vesicles. Both ionic and hydrophobic interactions appear to be involved in complex formation. We have used spectroscopy (circular dichroism, fluorescence, and nuclear magnetic resonance) and electron microscopy to study these complexes. At pH 3.7, the small, single bilayer vesicles produced by sonication are protected against aggregation by the presence of the protein. As determined by circular dichroism, the proportion of α-helix in β-lactoglobulin is increased by complexation with phosphatidylcholine. Circular dichroism and fluorescence spectra show the involvement of at least 1 tryptophan residue in the conformational change. At pH 7.2, β-lactoglobulin-phosphatidylcholine vesicles form aggregates as observed by electron microscopy and31P nuclear magnetic resonance spectroscopy. These aggregated vesicles could be resuspended by raising the pH. The ability of the partially unfolded β-lactoglobulin to interact with lipids is believed to be important to its transport through the endomembrane system.

The effects of partially hydrogenated marine oils on the mitochondrial function and membrane phospholipid fatty acids in rat heart

Abstract: The influence of dietary partially hydrogenated marine oils containing docosenoic acid on rat heart mitochondrial membrane phospholipid fatty acid composition was studied with particular reference to cardiolipin and oxidative phosphorylation. Five groups of male weanling rats were fed diets containing 20% (w/w) peanut oil (PO), partially hydrogenated peanut oil (HPO), partially hydrogenated Norwegian capelin oil (HCO), partially hydrogenated herring oil (HHO), and rapeseed oil (RSO) for 10 weeks. All the cardiac phospholipids investigated were influenced by the experimental diets. An increased amount of arachidonic acid observed in phosphatidylethanolamine (PE) after feeding partially hydrogenated oils suggests a changed regulation of the arachidonic acid metabolism in comparison with PO treatment. 22∶1 originating from the dietary oils was incorporated only to a small extent into phosphatidylcholine (PC) and PE. A selective incorporation of 18∶1 isomers into the 1- and 2-positions of PC and PE with respect to geometry and position of the double bond was observed. Large amounts of 18∶1trans were incorporated into the 1-position of PC and PE, irrespective of the amount of 18∶2 supplemented to the diets, replacing a considerable proportion of stearic acid in this position. After feeding HHO and RSO, the content of 22∶1 in mitochondrial cardiolipin of rat heart was found to be 3% (mainly cetoleic acid) and 10% (mainly erucic acid), respectively, indicating a high affinity forcis isomers of 22∶1, but also a considerable resistance against incorporation oftrans isomers was observed. The ability of rat cardiac mitochondria to oxidize palmitoylcarnitine and to synthesize ATP was depressed after feeding HHO and RSO. Dietarycis isomers of 22∶1 seem to have a specific ability to interfere with cardiac ATP synthesis and also to alter the fatty acid composition of cardiolipin of rat heart.

Determination of lycopene, α- and β-carotene and retinyl esters in human serum by reversed-phase high performance liquid chromatography

Abstract: A rapid specific, microdetermination of the major human blood carotenoids by high performance liquid chromatography (HPLC) separation and quantitation at 466 nm is detailed in this paper. Serum retinyl esters can also be quantified utilizing the same separation procedure but detected at 325 nm. One hundred microliters of deproteinated serum were extracted with chloroform and injected on a reverse-phase column. Separation occurred within 16 min for all compounds of interest employing a mobile solvent of MeOH/AcN/CHCl3 (47:47:6). All compounds were quantified at the wavelengths cited by integrated peak areas using retinyl acetate as a daily standard. Analysis of serum from a hypercarotenemic anorexia nervosa patient and a person suffering from hypervitaminosis A are presented as examples of the clinical application of this procedure.

Potentiating effect of 5,8,11-eicosatrienoic acid on human platelet aggregation

Abstract: 5,8,11-Eicosatrienoic acid (20∶3ω9), a fatty acid increased in the platelet phospholipids of man and animals fed saturated fats, was either added to human platelets simultaneously with the aggregating agents, or incorporated into the platelet phospholipids by preincubation. 20∶3ω9 markedly increased the response of platelets to all aggregating agents tested when added simultaneously with the agent, but solely to thrombin and ionophore, after incorporation into the platelet phospholipids. The potentiating effects of 20∶3ω9 on thrombin aggregation do not appear to be related to prostaglandin formation, but rather to the production of a monohydroxy derivative through the lipoxygenase pathway.