Intestinal metabolism of plasma free fatty acids in streptozotocin diabetic rats

Moderate insulin deficiency was reported to be accompanied by an increased production of intestinal very low density lipoprotein (VLDL) triglyceride in the rat. Because plasma free fatty acids (FFA) are incorporated into triglyceride by intestinal mucosa of rats and humans and plasma FFA are increased in insulin-deficient diabetes mellitus, we investigated several aspects of the intestinal metabolism of plasma FFA in diabetic rats. All experiments were performed on the third day following the i.v. injection of streptozotocin (45 mg/kg body weight) or buffer alone. A (¹⁴C)palmitic acid-rat serum complex was rapidly injected intravenously and its initial uptake by small bowel mucosa, the intracellular incorporation into lipids and water soluble metabolites and the specific radioactivity of triglycerides of mucosal homogenates was determined. No significant differences could be found between diabetic and control rats at 2 and 5 min after¹⁴C-palmitate i.v., suggesting that neither the influx of plasma free fatty acids into intestinal mucosal cells nor their initial intracellular metabolic pathways are significantly altered in moderately diabetic rats. A pronounced decrease in intestinal mucosal triglyceride at 10 min after¹⁴C-palmitate i.v. might be interpreted as indirect evidence for an enhanced triglyceride efflux from intestinal mucosa into mesenteric lymph in diabetic rats.     

Origin of the high saturated fatty acid content of rat fecal lipids

The biohydrogenation of unsaturated fatty acids and the preferential absorption of unsaturated fatty acids over long chain saturated fatty acids from the gut have been investigated to find the origin of the high saturated fatty acid content of the facal lipids of rats fed soybean oil. Label from dietary (1-¹⁴C)-linoleic acid was recovered in the saturated and monounsaturated fatty acids of the fecal lipid. However, when (9,10-³H)-stearic acid and (1-¹⁴C)-linoleic acid were fed together, the isotope ratio (³H/¹⁴C) of the fecal lipid was 1.9 times that of the diet. It is concluded that both processes occur.     

Neutral lipids of chickpea flour and protein isolates

The neutral lipids composition of defatted chickpea flour and two types of protein isolates has been studied. The main compounds in neutral lipids are triacylglycerols, free fatty acids, and diacylglycerols. Other compounds present are wax esters, free fatty alcohols, and free sterols. The main fatty acids in neutral lipids are C_18:2 and C_18:1 among the unsaturated, and C_16:0 and C_18:0 among the saturated acids. Free and esterified alcohols range from C_16:0 to C_28:0, the majority being those with an even number of carbon atoms. Sterols observed are β-sito-sterol, campesterol, stigmasterol, and δ-5-avenasterol. Triacyl-glycerols are partially hydrolyzed, and the amounts of unsaturated sterols and unsaturated fatty acids are reduced as a result of the chemical treatment during production of the protein isolates.     

Studies on biosynthesis of waxes by developing jojoba seed tissue

Slices of developing jojoba cotyledons incorporated a variety of precursors in to wax, free alcohols, and polar lipids.¹⁴C-Decanoic and¹⁴C-lauric acids were elongated and desaturated, whereas¹⁴C-myristic and¹⁴C-longer chain fatty acids, although incorporated into wax, were insignificantly modified. Exogenously added¹⁴C-acetate contributed mainly to chain elongation of endogenous oleic acid, whereas¹⁴C from added glucose was uniformly distributed throughout the acyl chain of the fatty acids. These data suggest the existence of metabolically separate pools of acetate and/or sites for de novo synthesis and elongation of acyl chains.     

Skin surface lipids of the dog

The skin surface lipid of the dog has been reported to contain a high proportion of diol diesters having a lower mobility on thin layer chromatography than diesters from other species in spite of containing similar fatty acid and diol components. In the present study, dog skin surface lipid was separated by preparative thin layer chromatography into sterol esters (42%), wax diesters (32%), free sterols (9%), polar lipids (7%), and unidentified components (10%). The diesters contained 1,2-diols, each esterified with one long chain fatty acid and one isovaleric acid moiety. The diols were principally branched chain C₂₁ and C₂₂ compounds while the long chain fatty acids esterified with them were mainly C₂₀ and C₂₁ branched compounds. The fatty acids from the sterol esters were mostly saturated, branched chain C₁₉ to C₂₃, together with 7% of straight chain monoenoic acids, principally C₂₁ and C₂₂. There were only trace amounts of free sterols other than cholesterol, while the esterified sterols contained 96% cholesterol and 4% lathosterol.     

Hydrolysis of triglycerides in the isolated perfused rat lung

The purpose of this study was to investigate the hydrolysis of saturated and unsaturated triglycerides by lung lipoprotein lipase and to measure the incorporation of triglyceride fatty acids into lung tissue lipids. Lipolytic activity was studied in the isolated ventilated rat lung, perfused for 100 min in a recycling system with Krebs Ringer bicarbonate containing bovine serum albumin, 5.6 mM glucose, and 1.5 or 10 mM triglyceride. Saturated triglycerides were hydrolyzed at significantly (p<0.05) lower rates than unsaturated triglycerides; tricaprylin, trimyristin and tripalmitin were hydrolyzed at 8.1+1.8, 5.4+1.5 and 9.5+1.8 μmol free fatty acids/g dry wt/100 min, respectively, whereas triolein and trilinolein were hydrolyzed at 20.2+1.8 and 20.6+0.3 μmol free fatty acids/g dry wt/100 min, respectively. The polyunsaturated triglycerides, trilinolein and triarachidonin were hydrolyzed at even higher rates (44.3+3.0 and 50.9+5.4 μmol free fatty acids/g dry wt/100 min, respectively). Intralipid infused at a concentration of 10 mM triglyceride was hydrolyzed at a significantly higher rate than at 1.5 mM triglyceride (58+6.3 μmol free fatty acids/g dry wt/100 min vs 16.6+1.7 μmol free fatty acids/g dry wt/100 min, respectively). Labeled unsaturated triglycerides were broken down at significantly higher rates than labeled saturated triglycerides. Incorporation of triglyceride-fatty acid into lung lipid was greater into neutral lipids than into phospholipids. The data suggest that (a) the factors that appear to affect lung lipoprotein lipase activity are composition and concentration of circulating triglyceride, (b) uptake of fatty acids into the tissue was proportional to the rate of hydrolysis of the emulsion, and (c) triglyceride-fatty acids could therefore be used by the lung for metabolic needs. The data presented in part at the Annual Meetings of the American Physiological Society, Atlanta, GA, April 1981, and the American Thoracic Society, Detroit, MI, May 1981, and published in abstract form-Fed. Proc. 40, 621 (1981), andAm. Rev. Respir. Dis. 123, 219 (1981).     

Wax esters in the cystacanths ofPolymorphus minutus (Acanthocephala)

The lipids of the cystacanth of the acanthocephalanPolymorphus minutus have been analyzed. Wax esters constituted nearly 90% of the total cystacanth lipids. The wax ester fraction contained approximately 10% steroid ester; the rest was long chain alcohols C₁₂ to C₂₀, largely saturated, esterified with fatty acids C₁₂ to C₂₂, mostly unsaturated, with C₁₈ predominating. Corresponding quantities of wax esters were not found in the adult parasite. Cholesterol was identified as the only steroid present in the cystacanth.     

Cuticular lipid constituents of cabbage seedpod weevils and host plant oviposition sites as potential pheromones

The cuticular lipids of cabbage seedpod weevils (Ceutorrhynchus assimilis Payk.) and those at their oviposition site, i.e., the seed pods of the host plant (Brassica napus L.), were analyzed by chromatographic techniques in conjunction with mass spectrometry (MS). Long chain hydrocarbons were most abundant in both lipids; however, the seed pods ofB. napus containedn-alkanes only, whereasC. assimilis showed a predominance of dimethylalkanes over internally branched methylalkanes as well as iso- andn-alkanes. The amounts of ketones, secondary alcohols and aldehydes, the usual plant components that are unique in insects, occurred in cuticular lipids of both organisms in approximately the same ratio. The composition ofn-alkanes,n-ketones, secondaryn-alcohols, iso- and anteisoaldehydes and esterified primary anteiso-alcohols of wax esters was similar betweenC. assimilis andB. napus. In both sources, qualitative but not quantitative similarities were observed in the composition ofn-aldehydes and esterified primaryn- and iso-alcohols, respectively. The esterified fatty acids of wax esters fromB. napus were composed of roughly equal proportions of saturated branched and unbranched components. The esterified fatty acids of wax esters and steryl esters fromC. assimilis consisted of major proportions of saturated as well as unsaturatedn-compounds, whereas iso- and anteiso structures were present in minor proportions only. Free fatty acids and traces of ethyl esters of fatty acids found inC. assimilis were mainly composed of unsaturatedn-compounds.     

Accumulation of neutral lipids by human skin fibroblasts: Differential effects of saturated and unsaturated fatty acids

The accumulation of neutral lipids by human skin fibroblasts grown in medium supplemented with fatty acids has been investigated. GM-10 cells incorporated exogenous fatty acids into both phospholipids and neutral lipids. More [¹⁴C] oleate, linoleate, or linolenate was incorporated into triacylglycerol than was [¹⁴C] palmitate or stearate. Supplementation of medium containing delipidized serum with unsaturated fatty acids resulted in far more stimulation of [¹⁴C] glycerol incorporation into triacylglycerol than did supplementation with saturated fatty acids. Palmitate- and stearate-fed cells incorporated sizable amounts of [¹⁴C] fatty acids and [¹⁴C] glycerol into diacylglycerol as well as triacylglycerol, especially at higher fatty acid concentrations. Increased oleate supplementation from 10–300 μM resulted in increased triacylglycerol synthesis and accumulation of discrete cytoplasmic lipid droplets; palmitate concentrations above 70 μm were toxic. Micrographs of the palmitate-fed cells showed electron translucent slits, suggesting solid depositions of saturated fat, rather than the discrete osmiophilic droplets found in oleate-fed cells. Although GM-10 cells can synthesize fully saturated triacylglycerols, these data suggest that in cells fed saturated fatty acids, solid depositions of neutral lipids may sequester diacylglycerols and thus limit triacylglycerol synthesis.     

Preferential oxidation of linolenic acid compared to linoleic acid in the liver of catfish (Heteropneustes fossilis andClarias batrachus)

The fate of [1-¹⁴C] linoleic acid and [1-¹⁴C] linolenic acid in the liver slices and also in the liver tissues of live carnivorous catfish,Heteropneustes fossilis andClarias batrachus, was studied. Incorporation of the fatty acids into different lipid classes in the live fish differed greatly from the tissue slices, indicating certain physiological control operative in vivo. The extent of desaturation and chain elongation of linoleic and linolenic acids into long-chain polyunsaturated fatty acids was low. Linolenic acid was oxidized (thus labeling the saturated fatty acid with liberated¹⁴C-acetyl-CoA) in preference to linoleic acid, and this oxidation also seemed to be under physiological control since both of the fatty acids were poorly oxidized in the tissue slices and in the killed fish. These fish can therefore recognize the difference in the acyl chain structures of linoleate and linolenate. The higher oxidation of liolenic acid and poor capacity for its conversion to longer chain, highly unsaturated derivatives indicates a higher demand for the dietary supply of these essential fatty acids in these two species.     

Sterol synthesis from biliary squalene in the jejunal mucosa of the rat in vivo

Because bile contains substantial amounts of cholesterol precursors, e.g., squalene and differnet methyl sterols, the fate of biliary squalene was studied by incubating isolated jejunal loops of the rat in vivo with bile containing³H-squalene and¹⁴C-cholesterol. After 90 min, no radioactivity was found in plasma lipids. Closer analysis of gut epithelium revealed that both labeled compounds were preferentially taken up by the villous cells. Biliary³H-squalene was absorbed almost completely and was further cyclized to free and esterified methyl sterols and cholesterol. Whereas squalene not cyclized to sterols stayed in the mucosa, the newly synthesized sterols were transferred to lumen. The lipid patterns of gut lumen and mucosal cells were quite different, suggesting that the transfer of the newly synthesized lipid to intestinal lumen was not due to the desquamation of epithelial cells alone. The results suggest that biliary cholesterol precursors can contribute to the cholesterol production of the jejunal villous cells bypassing the rate-limiting step of the cholesterol synthesis pathway, and to the “nonexchanging” fecal neutral sterols of the rat.     

Free fatty acids in the protective coats ofSpongilla wagneri gemmules

A gas chromatographic and mass spectrometric analysis of the free fatty acids found in the resistant coat ofSpongilla wagneri gemmules was performed. The coats were found to contain 8.79% extractable lipids with 18.53% free fatty acids, ranging in chain length from C₁₀ to C₂₄. The unsaturated acids were in relatively low concentrations with the preponderant saturated being palmitic and behenic. Distinct differences in distribution were observed when compared to the total gemmule fatty acids.     

Incorporation of [1-14C] acetate into fatty acids of the crustaceansDaphnia magna andCyclops strenus in relation to temperature

Daphnia magna andCyclops strenus were maintained in aquaria containing sodium [1-¹⁴C] acetate and the effect of temperature on labeling of their lipids was investigated. Incorporation of radioactivity in total lipids was slowed by a factor of 4 in cold-exposed (5C) specimens compared to those incubated at 25 C. There was no significant difference in the distribution of label in the lipid classes of animals incubated at the two extreme temperatures. Decrease of the temperature from 25 to 5 C brought about a considerable reduction in the formation of palmitic and stearic acids and an increase in labeling of monounsaturated (18∶1) fatty acids inD. magna. Docosapolyenoic acids were absent from lipids of this crustacean.C. strenus directed a higher proportion of radioactivity into both oleic and docosahexaenoic acids upon cold exposure. In response to decrease of the temperature,D. magna formed a less unsaturated fatty acid population, as judged from dpm ratios of total saturated to total unsaturated fatty acids, thanC. strenus. Inability to form and accumulate docosapolyenoic fatty acids byD. magna might be related to their poor survival at reduced temperatures.     

Dietary fat effect on incorporation and release of lipids and cholesterol by rat intestinal slices

The effect of saturated and unsaturated fats on in vitro formation and release of lipids and cholesterol from¹⁴C acetate by rat intestinal tissue was investigated. The rats were fed a basal diet enriched with either 25% corn oil or lard and then sacrificed after a 10- or 25-day feeding period. It was observed that a similar¹⁴C lipid content but a greater¹⁴C cholesterol content was found in the intestinal tissue of rats fed corn oil than in rats fed lard for 10 days. After a longer period of feeding of 25 days, the intestinal tissue¹⁴C cholesterol level was decreased in the corn oil fed rats without any significant effect on other lipids. These data suggest that corn oil in some way influences cholesterol biosynthesis depending upon its degree of unsaturation and the period of time for which it is fed. The decrease at the later time might involve some mechanism which aids in getting rid of accumulated tissue cholesterol. Less¹⁴C lipid and¹⁴C cholesterol were released by the intestinal tissue of rats fed the unsaturated fat as compared with those fed the saturated fat, suggesting a possible role in vivo in reducing blood lipids and blood cholesterol levels. Robert A. Welch Foundation.     

Onset and persistence of changes in intestinal transport following dietary fat manipulation

In this study we determined the time-course for the onset and the loss of the effect of short-term feeding rats isocaloric semisynthetic diets containing a high content of saturated (HS) or polyunsaturated (HP) fatty acids on the jejunal and ileal uptake of medium- and long chain fatty acids, cholesterol and glucose. Animals were fed HP or HS for 3, 7 or 14 days; then the diet was switched to standard Purina^® rat chow for a further 3, 7 or 14 days. The uptake of medium chain fatty acids was unchanged. The differences between HP and HS in glucose uptake occurred within 3 days, but persisted for 14 days, whereas there were qualitative as well as quantitative changes in the pattern of lipid uptake: differences in uptake of stearic, oleic, linoleic and linolenic acids and cholesterol occurred after 7 days of feeding HP or HS. Jejunal uptake of linoleic acid was greater in HP than HS on day 7, but HS was greater than HP on day 14. The effect of diet on lipid uptake was similar in the jejunum and ileum. The altered uptake of stearic and oleic acids persisted after the rats were switched back to chow, whereas the uptake of the other nutrients became similar. Thus, (i) changes in dietary content of saturated and polyunsaturated fatty acids have early effects on intestinal transport function; (ii) some of these changes persist even when animals are returned to feeding on chow; and (iii) glucose transport is rapidly altered by dietary changes, whereas lipid uptake changes only after 7 days. We conclude that the transport function of the intestine is responsive to changes in dietary fatty acids.     

Measurement oftrans and other isomeric unsaturated fatty acids in butterand margarine

A procedure is described for gas liquid chromatographic determination of cis andtrans isomers of unsaturated fatty acids after fractionation of the saturated, monenoic, dienoic, and polyenoic fatty acid methyl esters by argentation thin layer chromatography. To test its reliability, the procedure was used for quantitative measurement of transisomers of unsaturated fatty acids in a known mixture of simple triglycerides containing saturated fatty acids from 4:0 to 24:0 andcis andtrans isomers of 14:1. 16:1, 18:1, and 18:2. Results of the analyses of five margarine and five butter samples are presented, together with results of infrared spectrophotometric analyses fortrans fatty acid concentrations, ultraviolet spectrophotometric analyses for conjugated fatty acid concentrations, and enzymatic analyses forcis-cis-methylene interrupted fatty acid concentrations. The combined argentation thin layer and gas Chromatographic procedure is suitable for determination of the principal fatty acids in complex food lipids such as milk fat.     

Effects of Dielectric Barrier Ambient Air Plasma on Two Brassicaceae Seeds: Arabidopsis thaliana and Camelina sativa

We investigated low-temperature plasma effects on two Brassicaceae seeds (A. thaliana and C. sativa) using dielectric barrier discharge in air. Comparisons of plasma treatments on seeds showed distinct responses on germination rate and speed. Optimal treatment time giving optimal germination is 15 min for A. thaliana with 85% increase compared to control after 48 h of germination and 1 min for C. sativa with 75% increase compared to control after 32 h of germination. Such germination increases are associated with morphological changes shown by SEM of seed surface. For better understanding at the biochemical level, seed surfaces were analyzed using gas chromatography-mass spectrometry which underlined changes of lipidic composition. For both treated seeds, there is a decrease of saturated (palmitic and stearic) fatty acids while treated C. sativa showed a decrease of unsaturated (oleic and linoleic) acids and treated A. thaliana an increase of unsaturated ones. Such lipid changes, specifically a decrease of hydrophobic saturated fatty acids, are coherent with the other analyses (SEM, water uptake and contact angle). Moreover, an increase in A. thaliana of unsaturated acids (very reactive) probably neutralizes plasma RONS effects thus needing longer plasma exposure time (15 min) to reach optimal germination. For C. sativa, 1 min is enough because unsaturated linoleic acid becomes lower in treated C. sativa (1.2 × 10⁷) compared to treated A. thaliana (3.7 × 10⁷).     

A simple photometric method for microdetermination of fatty acids in lipids

A single step photometric micromethod for determination of fatty acids in lipids in benzene solution, using rhodamine 6G reagent, has been developed. The method eliminates the disadvantage of formation of a biphasic system and is applicable in the presence of glycerides, sterols, epoxy compounds, hydrocarbons and long chain hydroxy compounds such as long chain fatty alcohols. The long chain fatty acids from C₁₂ to C₂₂, both saturated and unsaturated, can be determined with reasonable accuracy in the concentration range of 0.08–0.25 μmole/ml. The method is simple, rapid, and requires relatively inexpensive chemicals.     

Fatty acid biosynthesis and incorporation into lipid classes in seeds and seed tissues of flax

Fatty acid metabolism in developing flaxseeds was studied by incubating whole seeds or isolated seed tissues in buffered solutions of 1-¹⁴C-acetate, 2-¹⁴C-malonate and¹⁴CO₂. Lipid classes were separated by thin layer chromatography, and fatty acid labeling in phospholipids, diglycerides and triglycerides was determined by combined thin layer and gas liquid chromatographic techniques. Incorporation of¹⁴C from acetate into embryo lipids was very rapid with phospholipids and 1,2-diglycerides becoming highly labeled in treatment times as short as 5 min. Triglycerides were labeled more slowly. Phospholipid radioactivity was largely associated with the phosphatidyl choline fraction. Oleic acid had the highest specific activity of all major fatty acids in short treatment periods. This was followed in decreasing order of activity by palmitic, linoleic, stearic and linolenic acids. As the treatment period was lengthened to 90 min or longer, linoleic and linolenic activities were markedly increased. Use of malonate or CO₂ rather than acetate as the substrate increased the labeling of the saturated acids. Incorporation of¹⁴C from acetate into lipids of endosperm tissues and whole flax seeds was slower than incorporation into embryo lipids. Stearate had the highest specific activity of the fatty acids in endosperm and whole seeds. Presented in part at the AOCS Meeting in New York, October 1968.     

Fatty acid composition of seed oils of some members of the meliaceae and combretaceae families

Seed oils of some members of the Meliceae (six) and Combretaceae (three) were analyzed for their fatty acid composition. In oils of members of both families palmitic acid was the most abundant saturated acid. Trace amounts of short chain (C12–C14) and long chain (C20–C22) saturated acids were detected in some members of the two families. Oleic acid was the most abundant unsaturated acids in the oils of four members of the Meliaceae. However, in the oils ofCedrella odorata andLovoa trichilloides, dienoic acid (C18:2) was the major unsaturated acid. Strikingly high levels of trienoic (C18:3) and monoenoic (C16:1) acids were detected in the seed oils ofC. odorata andEnthandrophragma angolense, respectively. Oleic acid also was the most abundant unsaturated acid in the Combretaceae. The nutritional value and industrial potentials of these oils are given.