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Milk composition and apparent digestibilities of dietary fatty acids in lactating dairy cows abomasally infused with Cis or Trans fatty acids.

Fat supplementation of diets for dairy cows produces changes in nutrient supply and milk composition. The effect of abomasal infusion of either cis-C18:1 or trans-C18:1 fatty acid isomers on the digestibility of fatty acids and milk composition was determined in lactating dairy cows. Six multiparous midlactation Holstein cows were used and fed a control diet containing 50% forage and 50% concentrate. Treatments were (per day): no infusion, infusion of a 630-g fat mixture high in cis-C18:1 isomers, and infusion of a 623-g fat mixture high in trans-C18:1 isomers using two 3 x 3 Latin squares with 4-wk experimental periods. Fat infusion did not affect total dry matter intake and increased apparent digestibilities of total fatty acids. Apparent digestibilities of C18 fatty acids were directly related to the number of double bonds within isomers, and cis-C18:1 isomers were slightly more digestible than trans-C18:1 isomers. The lower yield of C12:0, C14:0, and C16:0 fatty acids in milk fat and higher milk citrate observed when cows were infused with trans-C18:1 suggests a depressed de novo milk fatty acid synthesis. Effects of trans infusion on milk fat were independent of ruminal fermentation, fatty acid apparent absorption, and fatty acid plasma concentrations. Lower milk protein yield in cows infused with fat may have been caused by a decrease in milk protein synthesis.

Abomasum↗

Differential effect of valproate and its Delta2- and Delta4-unsaturated metabolites, on the beta-oxidation rate of long-chain and medium-chain fatty acids.

Overall fatty acid oxidation rates were investigated in rat hepatocytes using [9,10-3H]-palmitic, [9,10-3H]-oleic, [9,10-3H]-myristic and [2,3-3H]-phenylpropionic acids. The effect of both valproate (VPA) (0-10 mM) and two of its unsaturated metabolites, Delta2(E)-VPA and Delta4-VPA (0-10 mM), on the overall 3H2O production rate was studied. The results give evidence of a general inhibitory effect of VPA on the beta-oxidation rate of all the tested substrates. Similar effects were observed with both VPA metabolites but these effects appeared to be dependent on the chain length of the substrate. When the effect on the oxidation of the medium-chain fatty acid 3-phenylpropionate (PPA) was studied, Delta2(E)-VPA at 0.5 mM caused a 94% inhibition of the overall beta-oxidation rate. However, with long-chain substrates, 0.5 mM Delta(4)-VPA was a more potent inhibitor (20-30% of control activity) than 0.5 mM Delta(2E)-VPA (60-80% of control activity). Our results suggest that VPA and/or its metabolites inhibit fatty acyl-CoA metabolism within the mitochondrion by two different mechanisms. The first mechanism involves CoASH sequestration, which affects the oxidation rate of all fatty acids with different chain length. The second mechanism is more specific in nature and involves selective inhibition of particular enzymes implicated in fatty acid beta-oxidation.

Animals↗

Long-chain polyunsaturated fatty acids and chemically induced diabetes mellitus: effect of omega-6 fatty acids.

OBJECTIVE: We previously showed that prior oral supplementation of oils rich in omega-3, eicosapentaenoic acid and docosahexaenoic acid, and omega-6, gamma-linolenic acid and arachidonic acid, can prevent the development of alloxan-induced diabetes mellitus in experimental animals. But the effect of individual fatty acids on chemically induced diabetes mellitus is not known. We report the results of our studies with omega-6 fatty acids. METHODS: Alloxan-induced in vitro cytotoxicity and apoptosis in an insulin-secreting rat insulinoma cell line, RIN, was prevented by prior exposure of these cells to linoleic acid, gamma-linolenic acid, and arachidonic acid (AA) but not to dihomo-gamma-linolenic acid. Cyclo-oxygenase and lipoxygenase inhibitors did not block this protective action of AA. Prior oral supplementation with gamma-linolenic acid and pre- and simultaneous treatments with AA prevented alloxan-induced diabetes mellitus. RESULTS: Even though pretreatment with linoleic acid and dihomo-gamma-linolenic acid and simultaneous treatment with linoleic acid, gamma-linolenic acid, and dihomo-gamma-linolenic acid did not prevent the development of diabetes mellitus, the severity of diabetes was much less. The saturated fatty acid stearic acid and the monounsaturated fatty acid oleic acid were ineffective in preventing alloxan-induced diabetes mellitus. gamma-Linolenic acid and AA not only attenuated chemically induced diabetes mellitus but also restored the antioxidant status to normal range in various tissues. Changes in the concentrations of various fatty acids of the phospholipid fraction of plasma that occurred as a result of alloxan-induced diabetes mellitus also reverted to normal in the AA-treated animals. CONCLUSIONS: These results suggest that polyunsaturated fatty acids can prevent chemically induced diabetes in experimental animals and attenuate the oxidant stress that occurs in diabetes mellitus.

Alloxan↗

Postexercise increase of free fatty acids. A qualitative indicator for free fatty acid utilisation during exercise?

AIM: The purpose of this study was to verify the hypothesis that the postexercise increase (PEI) of plasma free fatty acid (FFA) concentrations after cessation of exercise is a suitable qualitative indicator for the FFA utilisation during the foregoing exercise. METHODS: Fourteen, 17 and 23 healthy subjects participated in 3 test series performing several prolonged exercise protocols (PEP) on a bicycle ergometer. During and after cessation of the PEP heart frequency, lactate and FFA were measured. RESULTS: Fasting resulted in an increase of PEI (90%, p<0.05) and the synergistic use of upper body muscles during cycling caused a significant rise of PEI compared to "regular" cycling (39%, p<0.01). Increasing workload step by step produced continuously rising PEI (p<0.05), only from 42% to 50% of maximal workload PEI decreased by 17% (p=n.s.). CONCLUSION: The results support the hypothesis that PEI is a suitable qualitative indicator for FFA utilisation during a foregoing exercise. Furthermore the results indicate that there is a maximum of FFA utilisation in the legs at 40% of maximum workload during cycling and that the upper body muscles contribute substantially to total body FFA turnover at high inten-sities, an aspect to which possibly too little attention has been paid to when quantifying FFA turnover during cycling.

Adult↗

Physiologic concentrations of glucose regulate fatty acid synthase activity in HepG2 cells by mediating fatty acid synthase mRNA stability.

Carbohydrate feeding of animals results in striking increases in hepatic fatty acid synthesis but much of this induction is presumed to be hormone mediated. To clarify the mechanisms responsible for the specific effect of carbohydrate on fatty acid synthesis, the effects of D-glucose on the expression of human fatty acid synthase (FAS) in HepG2 cells cultured in serum-free medium were studied. Northern blots of total RNA from these cells showed a single FAS mRNA band of 9.3 kilobases that was increased 2.7-5.4-fold in the presence of D-glucose. Lactate and citrate but not L-glucose mimicked this effect. Glucose induction of FAS mRNA was time- and concentration-dependent and most of the increase in FAS message was detected within the range of physiologic glucose concentrations. Glucose induction of FAS mRNA, protein synthetic rate, and enzyme activity were similar suggesting that glucose regulates FAS expression at a pretranslational level in this system. Transcription run-off experiments showed that glucose feeding was associated with no change in the FAS transcription initiation rate despite a 5-fold increase in FAS mRNA. FAS mRNA stability as determined by actinomycin D chase was 7-fold greater in the presence of glucose. Differences between FAS mRNA levels in cells grown in the presence versus the absence of glucose were inhibited by cycloheximide but not puromycin, suggesting that glucose regulation of FAS mRNA stability is not dependent on translation. Glucose, at physiologic concentrations and in the absence of hormones, appears to regulate FAS gene expression in HepG2 cells predominantly by mediating FAS mRNA stability.

Base Sequence↗

Disruption of the Saccharomyces cerevisiae homologue to the murine fatty acid transport protein impairs uptake and growth on long-chain fatty acids.

The yeast Saccharomyces cerevisiae is able to utilize exogenous fatty acids for a variety of cellular processes including beta-oxidation, phospholipid biosynthesis, and protein modification. The molecular mechanisms that govern the uptake of these compounds in S. cerevisiae have not been described. We report the characterization of FAT1, a gene that encodes a putative membrane-bound long-chain fatty acid transport protein (Fat1p). Fat1p contains 623 amino acid residues that are 33% identical and 54% with similar chemical properties as compared with the fatty acid transport protein FATP described in 3T3-L1 adipocytes (Schaffer and Lodish (1994) Cell 79, 427-436), suggesting a similar function. Disruption of FAT1 results in 1) an impaired growth in YPD medium containing 25 microM cerulenin and 500 microM fatty acid (myristate (C14:0), palmitate (C16:0), or oleate (C18:1)); 2) a marked decrease in the uptake of the fluorescent long-chain fatty acid analogue boron dipyrromethene difluoride dodecanoic acid (BODIPY-3823); 3) a reduced rate of exogenous oleate incorporation into phospholipids; and 4) a 2-3-fold decrease in the rates of oleate uptake. These data support the hypothesis that Fat1p is involved in long-chain fatty acid uptake and may represent a long-chain fatty acid transport protein.

3T3 Cells↗

Molecular analysis of a bifunctional fatty acid conjugase/desaturase from tung. Implications for the evolution of plant fatty acid diversity.

The seed oil derived from the tung (Aleurites fordii Hemsl.) tree contains approximately 80% alpha-eleostearic acid (18:3delta(9cis,11trans,13trans)), an unusual conjugated fatty acid that imparts industrially important drying qualities to tung oil. Here, we describe the cloning and functional analysis of two closely related Delta(12) oleate desaturase-like enzymes that constitute consecutive steps in the biosynthetic pathway of eleostearic acid. Polymerase chain reaction screening of a tung seed cDNA library using degenerate oligonucleotide primers resulted in identification of two desaturases, FAD2 and FADX, that shared 73% amino acid identity. Both enzymes were localized to the endoplasmic reticulum of tobacco (Nicotiana tabacum cv Bright-Yellow 2) cells, and reverse transcriptase-polymerase chain reaction revealed that FADX was expressed exclusively within developing tung seeds. Expression of the cDNAs encoding these enzymes in yeast (Saccharomyces cerevisiae) revealed that FAD2 converted oleic acid (18:1delta(9cis)) into linoleic acid (18:2delta(9cis,12cis)) and that FADX converted linoleic acid into alpha-eleostearic acid. Additional characterization revealed that FADX exhibited remarkable enzymatic plasticity, capable of generating a variety of alternative conjugated and delta(12)-desaturated fatty acid products in yeast cells cultured in the presence of exogenously supplied fatty acid substrates. Unlike other desaturases reported to date, the double bond introduced by FADX during fatty acid desaturation was in the trans, rather than cis, configuration. Phylogenetic analysis revealed that tung FADX is grouped with delta(12) fatty acid desaturases and hydroxylases rather than conjugases, which is consistent with its desaturase activity. Comparison of FADX and other lipid-modifying enzymes (desaturase, hydroxylase, epoxygenase, acetylenase, and conjugase) revealed several amino acid positions near the active site that may be important determinants of enzymatic activity.

Aleurites↗

Amino acid, mineral and fatty acid content of pumpkin seeds (Cucurbita spp) and Cyperus esculentus nuts in the Republic of Niger.

Dried seeds and nuts are widely consumed by indigenous populations of the western Sahel, especially those who inhabit rural areas. In light of the need for quantitative information regarding the content of particular nutrients in these plant foods, we collected dried pumpkin (Cucurbita spp) seeds and nuts of Cyperus esculentus in the Republic of Niger and analyzed them for their content of essential amino acids, minerals and trace elements, and fatty acids. On a dry weight basis, pumpkin seed contained 58.8% protein and 29.8% fat. However, the lysine score of the protein was only 65% relative to the FAO/WHO protein standard. The pumpkin seed contained useful amounts of linoleic (92 microg/g dry weight) and the following elements (on a microg per g dry weight basis): potassium (5,790), magnesium (5,690), manganese (49.3), zinc (113), selenium (1.29), copper (15.4), chromium (2.84), and molybdenum (0.81), but low amounts of calcium and iron. Except for potassium (5,573 microg/g dry weight) and chromium (2.88 microg/g dry weight), the C. esculentis nuts contained much less of these same nutrients compared to pumpkin seeds. In conclusion, pumpkin seeds represent a useful source of many nutrients essential to humans. The data in this report should of practical value to public health officials in rural areas of sub-Saharan Africa.

Amino Acids↗

Trans fatty acids and coronary heart disease risk. Report of the expert panel on trans fatty acids and coronary heart disease.

This review critically evaluates the scientific data on trans fatty acids and coronary heart disease (CHD) risk. Trans fatty acids are present in a variety of foods but they contribute only 4-12% of total dietary fat intake (2-4% of total energy intake) in the United States. The physical properties of trans fatty acids are intermediate between cis and saturated fatty acids, but a trans double bond is chemically less reactive than a cis double bond. Biochemical data indicate that trans fatty acids are subject to the same metabolic control mechanisms that regulate the metabolism of saturated and cis-isomeric fatty acids. Equivocal results have been reported in observational studies of trans fatty acid intake and CHD because of numerous methodologic limitations, including the difficulties inherent in quantifying trans fatty acid intake. Studies in hamsters indicate that trans fatty acids have a neutral effect on low-density-lipoprotein (LDL)-receptor activity, LDL-cholesterol production rate, and plasma LDL-cholesterol concentration. Other animal studies show no differences in atherosclerosis incidence or severity between diets containing hydrogenated and native vegetable oils. In clinical studies partially hydrogenated oils lower total and LDL-cholesterol concentrations when substituted for animal or vegetable fats rich in saturates but raise total and LDL-cholesterol concentrations when substituted for the unhydrogenated native oil. The effects of trans fatty acids on high-density lipoprotein cholesterol and lipoprotein(a) concentrations are unclear because of limited and conflicting clinical data. Data supporting a relation between trans fatty acid intake and CHD risk are equivocal compared with extensive data from studies in animals and humans linking saturated fat intake to CHD. Additional research is needed to resolve questions about the independent effects of trans fatty acids on plasma lipoproteins and their mechanisms of action.

Animals↗

Proteins regulating the biosynthesis and inactivation of neuromodulatory fatty acid amides.

Fatty acid amides (FAAs) represent a growing family of biologically active lipids implicated in a diverse range of cellular and physiological processes. At present, two general types of fatty acid amides, the N-acylethanolamines (NAEs) and the fatty acid primary amides (FAPAs), have been identified as potential physiological neuromodulators/neurotransmitters in mammals. Representative members of these two subfamilies include the endocannabinoid NAE anandamide and the sleep-inducing FAPA oleamide. In this Chapter, molecular mechanisms proposed for the biosynthesis and inactivation of FAAs are critically evaluated, with an emphasis placed on the biochemical and cell biological properties of proteins thought to mediate these processes.

Amides↗

Effect of dietary n-3 fatty acids on the composition of long- and very-long-chain polyenoic fatty acid in rat retina.

The effect of dietary supplementation with n-3 fatty acids, primarily docosahexaenoic acid (DHA) with high purity, on the fatty acid composition, especially very-long-chain fatty acids (VLCFA) longer than DHA, with four or six double bonds, in the rod outer segment (ROS) membranes of young Sprague-Dawley rats was investigated. After several weeks of feeding, diets high in n-3 fatty acids increased the DHA level significantly, while there were decreased levels of most n-6 fatty acids, such as arachidonic acid and 22:5n-6. Six kinds of VLCFA were detected by gas chromatography-mass spectrometry (GC-MS). Feeding a high n-3 fatty acid diet significantly increased the content of some n-3 VLCFAs such as 26:4n-3 and 30:4n-3 in ROS membranes, but not all detected n-3 VLCFAs. This study demonstrates that the dietary level of n-3 fatty acids not only affects the level of DHA, but also the levels of VLCFA in ROS membranes.

Animals↗

Prostaglandin and fatty acid omega- and (omega-1)-oxidation in rabbit lung. Acetylenic fatty acid mechanism-based inactivators as specific inhibitors.

Terminal acetylenic fatty acid mechanism-based inhibitors (Ortiz de Montellano, P. R., and Reich, N. O. (1984) J. Biol. Chem. 259, 4136-4141) were used as probes in determining the substrate specificity of rabbit lung cytochrome P-450 isozymes of pregnant animals in both microsomes and reconstituted systems. Lung microsomal and reconstituted P-450 form 5-catalyzed lauric acid omega- and (omega-1)-hydroxylase activities were inhibited by a 12-carbon terminal acetylenic fatty acid, 11-dodecynoic acid (11-DDYA), and an 18-carbon terminal acetylenic fatty acid, 17-octadecynoic acid (17-ODYA). Rabbit lung microsomal lauric acid omega-hydroxylase activity was more sensitive to inhibition by 11-DDYA than was (omega-1)-hydroxylase activity. In reconstituted systems containing purified P-450 form 5, both omega- and (omega-1)-hydroxylation of lauric acid were inhibited in parallel when either 11-DDYA or 17-ODYA was used. These data suggest the presence of at least two P-450 isozymes in rabbit lung microsomes capable of lauric acid omega-hydroxylation. This is the first report indicating the multiplicity of lauric acid hydroxylases in lung microsomes. Lung microsomal prostaglandin omega-hydroxylation, mediated by the pregnancy-inducible P-450PG-omega (Williams, D. E., Hale, S. E., Okita, R. T., and Masters, B. S. S. (1984) J. Biol. Chem. 259, 14600-14608) was subject to inhibition by 17-ODYA only, whereas 11-DDYA acid was not an effective inhibitor of this hydroxylase. We have recently developed a new terminal acetylenic fatty acid, 12-hydroxy-16-heptadecynoic acid (12-HHDYA), that contains a hydroxyl group at the omega-6 position. We show that 12-HHDYA possesses a high degree of selectivity for the inactivation of rabbit lung microsomal prostaglandin omega-hydroxylase activity which cannot be obtained with the long chain acetylenic inhibitor, 17-ODYA. In addition, 12-HHDYA has no effect on lauric acid omega- or omega-1-hydroxylation or on benzphetamine N-demethylation. The development of this new terminal acetylenic fatty acid inhibitor provides us with a useful tool with which to study the physiological role of prostaglandin omega-hydroxylation in the rabbit lung during pregnancy.

Animals↗

Interrelationship between the dietary regulation of fatty acid synthesis and the fatty acyl-CoA desaturases.

In this paper we present further evidence for the close control of fatty acid synthetase and stearoyl-CoA desaturase. Furthermore, we have established that whereas dietary palmitic acid may influence the activity of this desaturase but not of fatty acid synthetase, dietary linoleic acid appears to control both these enzymes. Finally, we have studied the influence of dietary fat and carbohydrate on the activities of the delta6 and delta5 desaturases. The former is only slightly affected by these dietary components. The delta5 desaturase activity is stimulated as the dietary fat content rises but is unaffected by dietary carbohydrate. The control of these enzymes is therefore independent of the control of fatty acid synthetase and stearoyl-CoA desaturase. From the data presented, the magnitude of the controlling effect of polyunsaturated fatty acids on fatty acid synthetase and stearoyl-CoA desaturase activity is determined and its relevance to lipogenesis in man based on daily intake of carbohydrate and linoleic acid is discussed.

Animals↗

Plasma amino acid and serum unesterified fatty acid deficits and the effect of nutritional support in chemotherapy treatment.

The deficits in plasma amino acids and serum unesterified fatty acids of cancer patients undergoing chemotherapy and/or radiation therapy were studied to delineate the special requirements of the patients and efficacy of our nutritional therapy. Seven general surgery patients and 13 patients treated by the Head-Neck Service had baseline levels measured as part of their nutritional evaluation prior to surgical treatment of their cancers. Fifteen chemotherapy outpatients maintained on their regular diets had fasting levels analyzed. Twenty-six patients who were admitted for their therapy had their intake of the regular hospital diet supplemented with a low-residue enteral diet formula (Vivonex High Nitrogen Diet); parenteral nutrition was used only if their oral intake was totally inadequate. Baseline and sequential measurements were made of plasma amino acid and serum unesterified fatty acid levels by gas liquid chromatographic techniques. Before operation the patients had normal levels of amino acids except for a significant deficiency of threonine and glycine observed in patients with head-neck tumors. Outpatients with and without hepatic metastases had significantly depressed levels of the essential amino acids valine, leucine, threonine, and methionine and the nonessential amino acids serine, glycine, and proline. The baseline levels of the patients admitted for treatment had similar deficiencies except for more evidence of lysine deficiency. Patients supported with total parenteral nutrition had rapid elevation of the amino acid levels. The patients whose intake was supplemented with the oral diets had improvement in their amino acid levels, but the deficiency in the leucine and threonine fractions persisted up to 4 weeks of therapy. Although the lysine levels were normal when first analyzed, significant differences developed in the patients without hepatic metastases after the start of chemotherapy with return to normal only after chemotherapy was discontinued. Fatty acid levels were not significantly different between the cancer groups except for preoperative elevated oleic acid levels noted in the general surgery tumor group; there were no deficiencies in the essential fatty acids. These studies indicate a need for enteral formulas with adequate branched-chain amino acids and enrichment with threonine and lysine for supplementing the nutrition of the cancer patient who is undergoing chemotherapy.

Amino Acids↗

[Effect of dietary n-6/n-3 polyunsaturated fatty acid ratio on spleen lymphocyte's function and fatty acid composition in mice].

OBJECTIVE: To study the effect of dietary n-6/n-3 PUFA ratio on spleen lymphocyte's function and fatty acid composition in mice. METHODS: Male BALb/c mice were divided into 5 groups: the dietary S : m : p was 1 : 1.5 : 1 n-6/ n-3 PUFA ratio was 1, 7.5, 15, 30 respectively, and in the control group, dietary S : M : P was 1 : 1.5 : 3.7, which was followed the AIN-93G formulation. All groups consumed purified diet based on the AIN-93G formulation. The different n-6/ n-3 PUFA ratios in diets were varied by mixing of oils. After 12 wks breeding, all mice were sacrificed, and the function and fatty acid composition of Lymphocyte, IL-2 and PGE2 concentration were measured. RESULTS: When dietary n-6/n-3 PUFA ratio approximated 1, the lymphocyte proliferation, proportions of CD4+ and CD8+ T cell, IL-2 and PGE2 concentration decreased significantly. The concentration of C18 : 2, C20 : 4, n-6 PUFA in lymphocyte decreased significantly; Meanwhile, the concentration of C22 : 6, C16 : 1, C18 : 1 and total MUFA in lymphocyte significantly higher than other groups. The concentration of C2 : 6 in lymphocytes were significantly negative correlated with the proliferation of spleen lymphocytes. The concentration of C20 : 5 in lymphocyte was significantly negative correlated with the proportions of CD4+ T cell and IL-2 level. The concentration of C16 : 1 in lymphocyte was significantly negative correlated with the proportions of CD4+ and CD8+ T cell. CONCLUSION: The fatty acid composition of diet affected that of lymphocyte in mice. Compared with the T lymphocyte function in diet with n-6/n-3 ratio approximated 30, the T lymphocyte function in mice was suppressed when dietary n-6/n-3 ratio approximated 1.

Animals↗