Kidney lipids: Changes caused by dietary 9-Trans, 12-Trans-octadecadienoate

Trans, trans-linoleate at 50 and 100% of dietary fat decreased kidney size and altered its composition.Trans, trans-linoleate as the sole source of dietary fat imparied growth and caused more severe symptoms of essential fatty acid deficiency than was observed with hydrogenated coconut oil (HCO). The concentration of renal cholesterol, phospholipids (PL), triglycerides (TG) and cholesteryl esters (CE) were also decreased. Linoleic (18∶2), homo-γ-linolenic acid (20∶3n6) and arachidonic acid (20∶4n6) were significantly depressed in lipid classes, especially in PL and CE, by dietarytrans, trans-linoleate. The increase in eicosatrienoate (20∶3n9), especially in PL and CE of kidneys of rats fed HCO (essential fatty acid deficient), was slight in rats fed 100%trans, trans-linoleate, indicating that thetrans, trans acid probably inhibited acyl elongation and desaturation.     

Distribution of dietary octadecenoate isomers at the 1- and 2-positions of hepatoma and liver phospholipids

Phosphatidylcholines and phosphatidylethanolamines were isolated from hepatoma 7288CTC, normal liver, and host liver of rats fed one of the following diets: fat-free diet; fat-free diet supplemented with safflower oil, safflower oil fatty acids, or partially hydrogenated safflower oil fatty acids; and commercial chow. Thecis andtrans octadecenoate fatty acids were isolated from the 1- and 2-positions of both phosphoglycerides and analyzed quantitatively for chain positional isomers. Octadecenoates from hepatoma and liver phosphoglycerides of animals fed fat-free or natural fatsupplemented diets contained almost exclusively twocis isomers: oleic and vaccenic acids. Oleic acid predominated in the 2-position octadecenoates of both phosphoglycerides from hepatoma and liver. In contrast, vaccenic acid predominated in the 1-position of normal liver phosphatidylcholine and, to a lesser extent, phosphatidylethanolamine. Host liver and hepatoma exhibited a shift to a higher percentage of oleic acid at the 1-position. Dietarytrans fatty acids were incorporated predominately in the 1-position of both phosphoglycerides of hepatoma and liver. Except for thecis Δ10 octadecenoate isomer, all of the unnatural dietarycis isomers between Δ8 and Δ14 were incorporated into the 1-position of the phospholipids, while the unnaturalcis octadecenoates at the 2-position consisted primarily of the Δ12 isomer. Hepatoma phosphoglycerides contained higher percentages of thetrans Δ10 isomer that was nearly excluded from the 1-position of the two liver phosphoglycerides. All the othertrans octadecenoate isomers were incorporated into the 1-position of both phosphoglycerides, but the small amount oftrans fatty acids incorporated into the 2-position of liver and hepatoma phosphatidylcholine consisted of four isomers, Δ9 to Δ12, including the Δ10 isomer. Phosphatidylethanolamine exhibited a similar distribution, except for the presence of the Δ13 and Δ14 isomers at the 2-position. A combination of evidence suggests that the 1-position fatty acids in phosphatidylcholine and phosphatidylethanolamine are of similar origin. The octadecenoates at the 2-position of these two phosphoglycerides appear to be of the same origin in hepatoma but not in liver. It was also revealed that the 2-position of hepatoma phosphatidylcholine contained much higher percentages of palmitate than liver.     

Effects of randomization of partially hydrogenated corn oil on fatty acid and cholesterol absorption,and tissue lipid levels in rats

Randomization of partially hydrogenated corn oil containing approximately 45% oftrans octadecenoic acid only slightly, but not significantly, increased the lymphatic fatty acid absorption in rats. No effect of randomization was observed on cholesterol absorption. When rats were fed these fats at the 8.8% level (with 1.2% safflower oil) for three weeks, the concentrations of serum cholesterol, and serum and liver phospholipid were significantly higher in randomized fat than in control fat, which was composed of 9% high-oleic safflower oil and 1% palm oil. Liver cholesterol tended to be higher in randomized fat. In contrast, nonrandomized fat was not hyperlipidemic compared to control fat. Although the fatty acid composition of liver phospholipids suggested a possible interference oftrans fatty acid with the metabolism of linoleic acid to arachidonic acid, there was no effect of randomization. In the two hydrogenated fat groups,trans octadecenoic acid was incorporated and distributed similarly in adipose tissue triacylglycerol. These observations indicated that randomization of partially hydrogenated fat is not beneficial to various lipid parameters in rats.     

Incorporation of dietarycis andtrans octadecenoate isomers in the lipid classes of various rat tissues

The percentage distribution of the geometrical and positional isomers in the hexadecenoates and octadecenoates isolated from triglycerides, phosphatidylcholines, and phosphatidylethanolamines of brain, heart, kidney, liver, lung, muscle, spleen, and adipose tissues from rats maintained four weeks on a semipurified diet supplemented with 15% partially hydrogenated safflower fatty acids, has been determined. Except for brain, octadecenoate percentages were increased in each of the lipid classes of all the tissues by the dietary fat. Although the diet did not contain detectable hexadecenoates, the 16∶1 fraction from the lipid classes of all the tissues was composed of 10–70% of thetrans isomers, indicating chain shortening of the dietary octadecenotes. Distribution ofcis andtrans positional isomers in triglyceride hexadecenoates was approximately the same in all tissues. Relatively high percentages of the Δ9, Δ10, and Δ11 isomers were observed, but the Δ8 was the predominatingtrans hexadecenoate isomer, indicating preferential chain shortening of thetrans δ10 octadecenoate.Trans octadecenoates were found in all tissues, but concentrations were dependent on tissue and lipid class. The distribution of thecis andtrans octadecenoate isomers was similar in all the tissue triglycerides, with the distribution of thetrans isomers resembling the diet. In contrast, the percentage distribution of thetrans octadecenoates in the phospholipid classes differed dramatically from the diet, and the distribution was dependent on both the tissue and lipid class. The Δ12, Δ13, and Δ14trans octadiet, suggesting an accumulation of these isomers. Although thecis Δ10 octadecenoate was a significant dietary component, this isomer was not incorporated significantly into any lipid class of any tissue. The metabolic fate of this isomer remains unknown.     

Autoxidation of linoleic acid and behavior of its hydroperoxides with and without tocopherols

The autoxidation of linoleic acid dispersed in an aqueous media and the effect of α-, γ- and δ-tocopherols were studied. The quantitative analysis of the hydroperoxide isomers (13-cis,trans; 13-trans,trans; 9-trans,cis; 9-trans,trans) by direct high-performance liquid chromatography exhibited a prooxidant activity of α-tocopherol at high concentration (3.8% by weight of linoleic acid). On the other hand, α-tocopherol at lower concentrations (0.38 and 0.038%) and γ- and δ-tocopherols at high concentration (3.8%) were antioxidant. Furthermore, the addition of tocopherols modified the distribution of the geometrical isomers. The formation of thetrans,trans hydroperoxide isomers was completely inhibited by the highest concentration of the three tocopherols independently of their antioxidant or prooxidant activity and only delayed by the lower concentrations of α-tocopherol. The addition of tocopherols to hydroperoxide isomers reduced the decomposition rate of these isomers in the order α-tocopherol < γ-tocopherol < δ-tocopherol for thecis,trans hydroperoxide isomer and α-tocopherol ≪ γ-tocopherol ⋍ δ-tocopherol for thetrans,trans hydroperoxide isomer. With these hydroperoxides, as during linoleic acid autoxidation, α-tocopherol was completely oxidized whatever its initial concentration, while γ-tocopherol underwent partial oxidation and δ-tocopherol was practically unchanged.     

Heat-induced geometrical isomerization of α-linolenic acid: Effect of temperature and heating time on the appearance of individual isomers

The formation of linolenic acid geometrical isomers (LAGIs) was studied in linseed oil that was heated under vacuum in sealed ampoules at different temperatures (190–260°C) for several durations (2–16 h). A temperature of about 190°C seems to be necessary to induce the formation of LAGIs. At higher temperatures, disappearance of linolenic acid follows a first-order kinetic. The formation of LAGIs increases with both heating time and temperature, degrees of isomerization of linolenic acid higher than 50–60% could easily be obtained by simply heating the oil under vacuum. Side reactions remain at a low level. The mean probabilities of isomerization of individual ethylenic bonds are similar to those determined in linolenic acid-containing oils marketed in European countries, 41.9, 4.7 and 53.3% for double bonds in positions 9, 12 and 15, respectively. The di-trans t,c,t (trans,cis,trans) isomer is formedvia the mono-trans c,c,t andt,c,c isomers by a two-step reaction. The proportions of thec,c,t andt,c,c isomers (relative to total LAGIs) decrease linearly with the heating time. The proportion of thec,t,c isomer is only slightly affected by this parameter; however, it increases with temperature. The proportion of thet,c,t isomer increases linearly with heating time at each tested temperature, at the expense of thec,c,t andt,c,c isomers. However, there is no simple relationship linking the disappearance of each of the mono-trans isomers and the formation of the di-trans isomer.     

Effect of essential fatty acid deficiency on lipid metabolism in isolated fat cells of epididymal fat pads of rats

Lipogenesis, lipolysis, and stimulation of glucose conversion into lipid by insulin or prostaglandin E₁ were studied in isolated fat cells of the epididymal fat pads of rats fed a fat-free diet or this diet supplemented with 10% hydrogenated coconut oil or 10% safflower seed oil. Changes in fatty acid composition, characteristic of an essential fatty acid deficiency, were well advanced in the neutral lipid but had only started in the polar lipid of the fat cells of the epididymal fat pads of animals 3 months after weaning. Cellularity of the epididymal fat pads, as indicated by protein to lipid ratio of the fat cells, was influenced greatly by hydrogenated coconut oil in the diet irrespective of an essential fatty acid deficiency. Lipogenesis was increased in the fat cells of the animals fed the hydrogenated coconut oil diet 5 weeks after weaning but was not significantly different from that of the safflower fed animals 3 months after weaning. Incorporation of glucose into lipid, oxidation to CO₂, and basal lipolysis were not significantly different in the fat cells of the essential fatty acid deficient animals from those fed safflower oil 3 months after weaning, except in animals of the fat-free group based upon cell lipid. However, conversion of glucose to free fatty acid was significantly greater in the isolated fat cells of animals fed either the hydrogenated coconut oil or the fat-free diet than in those of animals fed the safflower oil supplement. The incorporation of glucose into lipid by isolated fat cells was stimulated significantly by insulin in young animals fed a fat-free diet, but the effect on lipogenesis appeared to be reversed in the fat cells of animals receiving safflower seed oil 3 months after weaning. Prostaglandin E₁ also appeared to stimulate the incorporation of glucose into lipid in the fat cells of the older animals receiving safflower seed oil. Differences in osmolarity produced large differences in utilization of glucose and release of lipid from isolated fat cells, but no significant differences were observed between the cells from animals fed the fat-free diet and those from the controls fed safflower oil. The results demonstrated the effects of diets containing fat or no fat on enzyme activities and membrane properties of fat cells of the epididymal fat pads of essential fatty acid deficient rats.     

Study on the effect of antioxidants in the autoxidation of methyl nonconjugated octadecadienoates

Many studies have been published on the effect of antioxidants on unsaturated fatty acid esters but the differences of the effects of antioxidants on geometric isomers have never been investigated. In this study, methylcis-9,cis-12-octadecadienoate and itstrans isomer methyltrans-9,trans-12-octadecadienoate were used as methyl nonconjugated dienoates, and BHA, BHT, PG, NDGA, 4,4′-dihydroxy-3,5,3′,5′-tetratert-butyl diphenyl methane, L-thyroxine sodium salt,a-tocopherol and sesamol were used, as antioxidants. The differences of the effects of antioxidants on both geometric isomers were investigated by determining the induction period using the weighing method. Also determined were the infrared and ultraviolet spectra, peroxide values, conjugated diene contents, isolatedtrans double bond contents and molecular weights for the controls and the samples containing antioxidants. Thecis,cis isomer was more easily autoxidized and had a shorter induction period than thetrans,trans form. By the end of the induction period, no isolatedtrans double bond forms in thecis,cis isomer, but a considerable amount of isolatedtrans double bond decreased in thetrans,trans isomer. In general, the effects of antioxidants, except NDGA, on thecis,cis isomer were larger than thetrans,trans form.     

Effects of dietary saturated andTrans fatty acids on tissue lipid composition and serum LCAT activity in the rat

Groups of rats were fed from weaning with diets containing 5% by wt of hydrogenated coconut oil (HCO), safflower oil, or a concentrate of ethyl elaidate and linolelaidate (TRANS) as the sole source of dietary fat. Fatty acid composition of the lipid classes from serum, liver, heart, and kidney was determined, and the serum lecithin: cholesterol acyl transferase (LCAT) activities were assayed for each animal. Serum LCAT activity was increased by both the HCO and TRANS diets in the early stages of the development of an essential fatty acid (EFA) deficiency but was suppressed in the animals of the TRANS group as they became older. The HCO and TRANS groups exhibited changes in tissue lipid fatty acid composition, as well as reduced growth, characteristic of an EFA deficiency. Conversion of oleic acid to eicosatrienoic acid was impaired in the animals fed the TRANS diet, greatly increasing the octadecenoic acid content of the tissue lipids at the expense of eicosatrienoic acid. The TRANS diet also suppressed incorporation of eicosatrienoic acid into cholesteryl esters of tissue and serum, indicating that, when fed as the sole source of unsaturated fat,trans fatty acids influenced the metabolism of unsaturated fatty acids and cholesterol.     

Content and distribution oftrans-18:1 acids in ruminant milk and meat fats. Their importance in european diets and their effect on human milk

Thetrans-18:1 acid content and distribution in fats from ewe and goat milk, beef meat and tallow were determined by a combination of capillary gas-liquid chromatography and argentation thin-layer chromatography of fatty acid isopropyl esters. Thetrans isomers account for 4.5 ± 1.1% of total fatty acids in ewe milk fat (seven samples) and 2.7±0.9% in goat milk fat (eight samples). In both species, as in cow, the main isomer is vaccenic (trans-11 18:1) acid. The distribution profile oftrans-18:1 acids is similar among the three species. The contribution of ewe and goat milk fat to the daily intake oftrans-18:1 acids was estimated for people from southern countries of the European Economic Community (EEC): France, Italy, Greece, Spain, and Portugal. It is practically negligible for most of these countries, but in Greece, ewe and goat milk fat contributeca. 45% of the daily consumption oftrans-18:1 acids from all dairy products (0.63 g/person/day for a total of 1.34 g/person/day). Thetrans-18:1 acid contents of beef meat fat (ten retail cuts, lean part) and tallow (two samples) are 2.0 ± 0.9% and 4.6%, respectively, of total fatty acids (animals slaughtered in winter). Here too, the main isomer is vaccenic acid. Othertrans isomers have a distribution pattern similar to that of milk fat. Beef meat fat contributes less than one-tenth of milk fat to thetrans-18:1 acid consumed. The daily per capita intake oftrans-18:1 acids from ruminant fats is 1.3–1.8 g for people from most countries of the EEC, Spain and Portugal being exceptions (ca. 0.8 g/person/day). In France, the respective contributions of ruminant fats and margarines to the daily consumption oftrans-18:1 acids are 1.7 and 1.1 g/person/day (60 and 40% of total, respectively). These proportions, based on consumption data, were confirmed by the analysis of fat from milk of French women (ten subjects). The mean content oftrans-18:1 acids in human milk is 2.0 ± 0.6%, with vaccenic acid being the major isomer. Based on the relative levels of thetrans-16 18:1 isomer, we could confirm that milk fat is responsible for the major part of the daily intake oftrans-18:1 acids by French people. The daily individual intake oftrans-18:1 isomers from both ruminant fats and margarines for the twelve EEC countries varies from 1.5 g in Spain to 5.8 g in Denmark, showing a well-marked gradient from the southwest to the northeast of the EEC.     

Zinc deficiency and activities of lipogenic and glycolytic enzymes in liver of rats fed coconut oil or linseed oil

In previous studies, zinc-deficient rats force-fed a diet with coconut oil as the major dietary fat developed a fatty liver, whereas zinc-deficient rats force-fed a diet with linseed oil did not. The present study was conducted to elucidate the reason for this phenomenon. In a bifactorial experiment, rats were fed zinc-adequate or zinc-deficient diets containing either a mixture of coconut oil (70 g/kg) and safflower oil (10 g/kg) (“coconut oil diet”) or linseed oil (80 g/kg) (“linseed oil diet”) as a source of dietary fat, and activities of lipogenic and glycolytic enzymes in liver were determined. In order to ensure adequate food intake, all the rats were force-fed. Zinc-deficient rats on the coconut oil diet developed a fatty liver, characterized by elevated levels of triglycerides with saturated and monounsaturated fatty acids. These rats also had markedly elevated activities of the lipogenic enzymes acetyl-CoA carboxylase, fatty acid synthase (FAS), glucose-6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase (6PGDH), and citrate cleavage enzyme, whereas activities of malic enzyme and glycolytic enzymes were not different compared with zinc-adequate rats on the coconut oil diet. In contrast, rats receiving the linseed oil diet had similar triglyceride concentrations regardless of zinc status, and activities of lipogenic enzymes and glycolytic enzymes were not different between the two groups. Zinc-deficient rats fed either type of dietary fat exhibited statistically significant correlations between activities of FAS, G6PDH, 6PGDH and concentrations of saturated and monounsaturated fatty acids in liver. The concentrations of serum lipids were elevated in zinc-deficient rats fed either type of dietary fat. These results demonstrate that fatty liver in zinc-deficient rats on the coconut oil diet is caused by elevated activities of lipogenic enzymes, and not by disturbed lipid secretion from liver. Dietary linseed oil prevents both the elevation of lipogenic enzyme activity and fatty liver in zinc-deficient rats.     

Effect of dietary fat saturation of absorption and intestinal secretion of cholesterol by the hypercholesterolemic rat

The fate of an oral dose of [4-¹⁴C] cholesterol given to rats grown on diets with 20% safflower oil or 20% hydrogenated coconut oil was determined by analysis of digestive tract, feces and tissues. The pattern of isotope distribution did not support the view that rats fed a saturated fat absorb less cholesterol than those fed an unsaturated fat. Fasted animals growth on the diet with 5% of these two fats and beef fallow showed no clear difference in the amount of digitonin-peecipitable sterol in their intestines. A shorter transit time for intestinal contents was observed with the saturated fat groups. It is concluded that neither absorption of cholesterol from the gut nor secretion of β-hydroxy sterol into the gut accounts for the hypocholesterolemic effect of polyunsaturated fat. Journal Paper No. 4951 AES, Purdue University.     

Distributions of conjugated linoleic acid (CLA) isomers in tissue lipid classes of pigs fed a commercial CLA mixture determined by gas chromatography and silver ion-high-performance liquid chromatography

Pigs were fed a commercial conjugated linoleic acid (CLA) mixture, prepared by alkali isomerization of sunflower oil, at 2% of the basal diet, from 61.5 to 106 kg live weight, and were compared to pigs fed the same basal diet with 2% added sunflower oil. The total lipids from liver, heart, inner back fat, and omental fat of pigs fed the CLA diet were analyzed for the incorporation of CLA isomers into all the tissue lipid classes. A total of 10 lipid classes were isolated by three-directional thin-layer chromatography and analyzed by gas chromatography (GC) on long capillary columns and by silver-ion high-performance liquid chromatography (Ag⁺-HPLC); cholesterol was determined spectrophotometrically. Only trace amounts (<0.1%; by GC) of the 9,11–18∶2 cis/trans and trans, trans isomers were observed in pigs fed the control diet. Ten and twelve CLA isomers in the diet and in pig tissue lipids were sepatated by GC and Ag⁺-HPLC, respectively. The relative concentration of all the CLA isomers in the different lipid classes ranged from 1 to 6% of the total fatty acids. The four major cis/trans isomers (18.9% 11 cis, 13 trans-18∶2; 26.3% 10 trans, 12 cis-18∶2; 20.4% 9 cis, 11 trans-18∶2; and 16.1% 8 trans, 10 cis-18∶2) constituted 82% of the total CLA isomers in the dietary CLA mixture, and smaller amounts of the corresponding cis,cis (7.4%) and trans,trans (10.1%) isomers were present. The distribution of CLA isomers in inner back fat and in omental fat of the pigs was similar to that found in the diet. The liver triacylglycerols (TAG), free fatty acids (FFA), and cholesteryl esters showed a similar patterns to that found in the diet. The major liver phospholipids showed a marked increase of 9 cis,11 trans-18∶2, ranging from 36 to 54%, compared to that present in the diet. However, liver diphosphatidylglycerol (DPG) showed a high incorporation of the 11 cis,13 trans-18∶2 isomer (43%). All heart lipid classes, except TAG, showed a high content of 11 cis,13 trans-18∶2, which was in marked contrast to results in the liver. The relative proportion of 11 cis,13 trans-18∶2 ranged from 30% in the FFA to 77% in DPG. The second major isomer in all heart lipids was 9 cis,11 trans-18∶2. In both liver and heart lipids the relative proportions of both 10 trans,12 cis-18∶2 and 8 trans,10 cis-18∶2 were significantly lower compared to that found in the diet. The FFA in liver and heart showed the highest content of trans,trans isomers (31 to 36%) among all the lipid classes. The preferential accumulation of the 11 cis,13 trans-18∶2 into cardiac lipids, and in particular the major phospholipid in the inner mitochondrial membrane, DPG, in both heart and liver, appears unique and may be of concern. The levels of 11 cis,13 trans-18∶2 naturally found in foods have not been established.     

Effects of dietarytrans acids on the biosynthesis of arachidonic acid in rat liver microsomes

Effects of dietarytrans acids on the interconversion of linoleic acid was studied using the liver microsomal fraction of rats fed a semipurified diet containing fat supplements of safflower oil (SAFF), hydrogenated coconut oil (HCO) at 5 and 20% levels or a 5% level of a supplement containing 50.3% linolelaidic and 24.3% elaidic acids devoid ofcis,cis-linoleic acid (TRANS). Growth rate was suppressed to a greater extent with the animals fed the 20% than the 5% level of the HCO-supplemented diets and still further by the TRANS diet compared to the groups fed the SAFF diets. Food intake was greater in the groups fed the HCO than the SAFF-supplemented diets, demonstrating the marked effect of an essential fatty acid (EFA) deficiency on feed efficiency. In contrast to an EFA deficiency produced by the HCO supplement, which stimulated the in vitro liver microsomal biosynthesis of arachidonic acid, diets containing the TRANS supplement exacerabated the EFA deficiency and depressed 6-desaturase activity of the liver microsomal fraction. The liver microsomal fraction of the animals receiving this supplement also was more sensitive to fatty acid inhibition of the desaturation of linoleic acid than those obtained from animals fed either the SAFF or HCO diets. It is suggested that dietarytrans acids alter the physical properties of the 6-desaturase enzyme system, suppressing its activity, which increases the saturation of the tissue lipids and, in turn, the requirement for EFA or polyunsaturated fatty acids.     

Incorporation ofcis- andtrans-octadecenoic acids into the membranes of rat liver mitochondria

The incorporation of dietary isomeric fatty acids into the membranes of liver mitochondria was investigated. Three groups of rats were fed diets containing 3% sunflower seed oil plus 15%, 20%, or 25% partially hydrogenated arachis oil. A fourth group was fed 25% partially hydrogenated arachis oil, but no sunflower seed oil. All diets were given for 3, 6, or 10 weeks. After 10 weeks, the content oftrans fatty acids in the lipids of the mitochondrial membranes was 15–19% of the total fatty acids. The composition of thetrans- and thecis-octadecenoic acids in the lipids of the mitochondrial membranes was similar for all groups supplemented with sunflower seed oil (SO), irrespective of time and dietary level of partially hydrogenated arachis oil (HAO). Thecis 18∶1 (n−8), which was a major isomer of the partially hydrogenated arachis oil, was almost excluded from the mitochondrial fatty acids. Likewise, the content oftrans 18∶1 (n−8) was considerably lower in the mitochondrial lipids than in the diet. On the contrary, the content oftrans 18∶1 (n−6) was higher in the mitochondrial lipids than in the diet. In the group fed without sunflower seed oil, isomers of linoleic acid and arachidonic acid were observed in the lipids of mitochondrial membranes. Presented in part at the ISF Congress, Marseille, September 1976.     

Interrelationship between dietarytrans fatty acids and the 6- and 9-desaturases in the rat

Studies are reported on the effects of dietarytrans fatty acids on the 6- and 9-acyl desaturase activities in the liver microsomes of rats fed essential fatty acid (EFA)-deficient and non-FFA-deficient diets. In experiment I, weanling male rats were fed a semisynthetic diet with either 10% safflower oil (SAF) or 10% hydrogenated coconut oil (HCO). At the age of one year, half of the dietary fat was replaced by a supplement containing elaidate, linolelaidate andcis,trans-trans,cis-18∶2 (TRANS) for 12 weeks. In experiment II, male rats which were kept from weaning on a 10% SAF diet for one year received one of the following fat supplements for a 12-week period: 10% HCO, 9% HCO+1% TRANS, or 5% HCO+5% TRANS. Feeding TRANS depressed the 6-desaturase activity in the liver microsomes, especially in the EFA-deficient rats (HCO+TRANS group of experiment I). Unlike the 6-deaturase activity, the 9-desaturase activity was not inhibited by the dietarytrans fatty acids and was significantly stimulated in the non-EFA-deficient rats (SAF+TRANS group of experiment I and HCO+TRANS groups of experiment II). This was evidenced by incubation reactions and by comparisons of fatty acid consumptions and microsomal fatty acid levels, showing extra biosynthesis of 16∶1 and 18∶1 when TRANS was fed. The biosynthesis of essential (n−6) fatty acids was depressed by the TRANS supplement in EFA-deficient as well as in non-EFA-deficient animals.     

Determination of CLA <Emphasis Type="Italic">trans</Emphasis>,<Emphasis Type="Italic">trans</Emphasis> Positional Isomerism in CLA-Rich Soy Oil by GC–MS and Silver Ion HPLC

A ~20% CLA-rich soy oil with low saturated fat and no cholesterol was produced by photoisomerizing soy oil linoleic acid. The oil is predominately trans,trans CLA, with the oil containing 17% trans,trans CLA. Recent studies have shown that trans,trans CLA-rich soy oil significantly reduces heart disease and diabetes risk factors in obese rats. However, the positional isomerism of these geometrical isomers has not been identified. The objectives of the study were to determine trans,trans CLA positional isomerism of CLA fatty acids in CLA-rich soy oil and determine the resolution of trans,trans CLA positional isomers by silver ion chromatography. GC–MS studies of 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD) derivatives of CLA-rich oil showed that 9,11 CLA and 10,12 CLA were the major positional isomers. These were hypothesized to be the trans,trans CLA isomers, which was confirmed by silver ion chromatography and subsequent GC–FID fatty acid and ATR-FTIR geometrical isomer analysis of the collected fractions. The identification of 9,11 trans,trans CLA and 10,12 trans,trans CLA as the major CLA isomers in CLA-rich oil then allowed the deduction of the synthetic mechanism of the photoisomerism of soy oil linoleic acid to trans,trans CLA.     

Differential effects of dietary linoleic and α-linolenic acid on lipid metabolism in rat tissues

Comparative effects of feeding dietary linoleic (safflower oil) and α-linolenic (linseed oil) acids on the cholesterol content and fatty acid composition of plasma, liver, heart and epididymal fat pads of rats were examined. Animals fed hydrogenated beef tallow were used as isocaloric controls. Plasma cholesterol concentration was lower and the cholesterol level in liver increased in animals fed the safflower oil diet. Feeding the linseed oil diet was more effective in lowering plasma cholesterol content and did not result in cholesterol accumulation in the liver. The cholesterol concentration in heart and the epididymal fat pad was not affected by the type of dietary fatty acid fed. Arachidonic acid content of plasma lipids was significantly elevated in animals fed the safflower oil diet and remained unchanged by feeding the linseed oil diet, when compared with the isocaloric control animals fed hydrogenated beef tallow. Arachidonic acid content of liver and heart lipids was lower in animals fed diets containing safflower oil or linseed oil. Replacement of 50% of the safflower oil in the diet with linseed oil increased α-linolenic, docosapentaenoic and docosahexaenoic acids in plasma, liver, heart and epididymal fat pad lipids. These results suggest that dietary 18∶2ω6 shifts cholesterol from plasma to liver pools followed by redistribution of 20∶4ω6 from tissue to plasma pools. This redistribution pattern was not apparent when 18∶3ω3 was included in the diet.     

Effect of dietary fats on some membrane-bound enzyme activities,membrane lipid composition and fatty acid profiles of rat heart sarcolemma

The effect of various dietary fats on membrane lipid composition, fatty acid profiles and membrane-bound enzyme activities of rat cardiac sarcolemma was assessed. Four groups of male weanling Charles Foster Young rats were fed diets containing 20% of groundnut, coconut, safflower or mustard oil for 16 weeks. Cardiac sarcolemma was prepared from each group and the activities of Na⁺,K⁺-ATPase, 5′-nucleotidase, Ca²⁺-ATPase and acetylcholinesterase were examined. ATPase activities were similar in all groups except the one fed coconut oil, which had the highest activities. Acetylcholinesterase activity was also similar in all the groups, however, it was significantly higher in the group fed mustard oil. No significant changes were observed among the groups in 5′-nucleotidase activity, in the cholesterol-to-phospholipid molar ratio and in sialic acid content. The coconut, safflower and mustard oil diets significantly increased cholesterol and phospholipid contents and the lipid-to-protein ratio of cardiac sarcolemma as compared to feeding the groundnut oil diet. The fatty acid composition of membrane lipids was quite different among the various groups, reflecting the type of dietary fat given. The total unsaturated-to-saturated fatty acid ratio was not different among the various groups; however, the levels of some major fatty acids such as palmitic (16∶0), oleic (18∶1) and linoleic (18∶2) acids were significantly different. Cardiac sarcolemma of the group fed safflower oil had the highest polyunsaturated fatty acid content. The results suggest that dietary fats induce changes not only in the fatty acid composition of the component lipids but also in the activities of sarcolemmal enzymes involved in the regulation of cardiac function.