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Fatty acid synthetase from Brevibacterium ammoniagenes: formation of monounsaturated fatty acids by a multienzyme complex.

A multienzyme fatty acid synthetase complex isolated from Brevibacterium ammoniagenes has been purified to a specific activity of 1440 nmol of malonyl-CoA incorporated per min/mg. The enzyme is homogeneous, as judged by gel electrophoresis on agarose gels, and has a molecular weight of 1.2 X 10(6). Both NADPH and NADH are required for activity. In contrast to other fatty acid synthetase complexes, the enzyme catalyzes the synthesis of both long-chain saturated and monounsaturated fatty acids from malonyl-CoA and acetyl-CoA. The formation of unsaturated fatty acids is oxygen-independent and sharply reduced by 3-decynoyl-N-acetylcysteamine, a known inhibitor of Escherchia coli beta-hydroxydecanoyl thioester dehydrase (EC 4.2.1.60).

Aerobiosis

[The Iberian pig: an animal with fat with a high content of monounsaturated fatty acids].

We have analyzed the fatty acid composition of subcutaneous fat in the Iberian pork feed with acorns using gas chromatography. We have found a 59.1% content in oleic acid (18:1) and 62% of total monounsaturated fatty acids. This figures statistically does differs from the data about fat composition in pig taken from others food composition sources. We conclude that Iberian pork feed with acorns have a very high content in monounsaturated fatty acids and can no be considered as harmful as other animals fats.

Adipose Tissue

Comparative effects of a recommended lipid-lowering diet vs a diet rich in monounsaturated fatty acids on serum lipid profiles in healthy young adults.

This crossover study investigated the effects of two fat-reduced diets, one rich in monounsaturated fatty acids (MUFAs), the other rich in polyunsaturated fatty acids (PUFAs), on serum lipid profiles in 38 healthy young adults initially on a typical western diet. After being randomly assigned to two groups, the subjects received the MUFA or PUFA diet for 3-wk and then the other diet for 3 wk. Both test diets led to significant reductions in serum cholesterol, LDL cholesterol, and HDL cholesterol (P less than 0.001). Both reduced apolipoprotein B (P less than 0.001) and apolipoprotein A-I concentrations (P less than 0.01 for the MUFA, P less than 0.001 for the PUFA diet). Apolipoprotein A-I was significantly higher on the MUFA than on the PUFA diet. The ratio of apolipoprotein A-I to B significantly increased on both diets. Thus, a low-fat, MUFA-rich diet is as effective as a low-fat, PUFA-rich diet in lowering total and LDL cholesterol, but both also lowered HDL cholesterol concentrations. The MUFA-rich diet may be more advantageous than the PUFA-rich one because it does not lower apolipoprotein A-I concentrations as much as the PUFA-rich diet.

Adult

Identification of some uncommon monounsaturated fatty acids of bacteria.

Location of the double-bond position of monounsaturated fatty acids of various bacteria was accomplished with combined gas chromatography-mass spectrometry analysis of dimethyl disulfide (DMDS) derivatives. The monoenoic fatty acids from whole cells were converted to methyl esters and then to DMDS adducts and analyzed by capillary gas chromatography-mass spectrometry. The mass spectra of DMDS adducts gave an easily recognizable molecular ion and two major diagnostic ions attributable to fragmentation between the two CH3S groups located at the original site of unsaturation. Twenty-one relatively novel monoenoic fatty acids were identified among the bacteria studied. All Flavobacterium species contained i17:1 omega 8c, Bacillus alvei contained i16:1 omega 11c and i17:1 omega 12c, and Psychrobacter immobilis contained 12:1 omega 9c. Resolution of cis and trans isomers with capillary gas chromatography and subsequent mass spectrometry permitted positive identification of 16:1 omega 7c and 16:1 omega 7t in Arcobacter (Campylobacter) cryaerophila and 16:1 omega 9c and 16:1 omega 9t in Aerococcus viridans.

Bacteria

Effects of avocado as a source of monounsaturated fatty acids on plasma lipid levels.

To examine the effects of avocado on plasma lipid concentrations, a three-diet trial involving 16 healthy volunteers was carried out. A diet rich in monounsaturated fatty acids using avocado as their major source (30% of the total energy was consumed as fat: 75% of the total fat from the avocado), with restriction of saturated fats and less than 300 mg of cholesterol per day was evaluated. Subjects also were in a free-diet period with the addition of the same amount of avocado. Finally, volunteers received a low-saturated fat diet without avocado. The first and third diets were designed to simulate a usual diet and volunteers carried on their normal activities during the trial, only the three daily meals were eaten in our clinical unit. Diets lasted 2 weeks and they were assigned in a randomized order. In both rich-monounsaturated fat (RMF) and low-saturated fat (LSF) diets, there were similar reductions in the plasma total cholesterol and low-density lipoprotein cholesterol levels. The levels of high-density lipoprotein cholesterol significantly decreased (p < 0.05) after 2 weeks of the LSF and free monounsaturated-enriched (FME) diets. The plasma triacyglycerol levels lessened after RMF and FME diets, while LSF diet increased them. In total cholesterol and in low-lipoprotein cholesterol levels, there were statistically significant differences between the FME and the LSF diet periods. Avocado is an excellent source of monounsaturated fatty acid in diets designed to avoid hyperlipidemia without the undesirable effects of low-saturated fat diets on HDL-cholesterol and triacylglycerol concentrations.

Adolescent

Comparison of the effects on lipoproteins and apolipoproteins of a diet high in monounsaturated fatty acids, enriched with avocado, and a high-carbohydrate diet.

A randomized trial was conducted to compare effects of a diet high in monounsaturated fatty acids, enriched with avocado (AE), and a high-complex-carbohydrate diet (AHA-III) on blood lipid concentrations. Fifteen females between 37 and 58 y of age were allocated either to AE followed by AHA-III, or vice versa. Each dietary phase lasted 3 wk. Both diets were found to lower total cholesterol compared with baseline values. AE was more effective, with an 8.2% decrease (P less than 0.05) whereas AHA-III was associated with a 4.9% decrease (NS). Low-density-lipoprotein cholesterol and apolipoprotein B decreased significantly on AE but not on AHA-III (P less than 0.05). The high-density-lipoprotein (HDL) concentration did not change on AE but decreased 13.9% on AHA-III (P less than 0.01). It is concluded that AE is more effective than is AHA-III in decreasing total cholesterol, and, unlike AHA-III, it does not decrease HDL concentrations.

Adult

Geometrical and positional isomer content of the monounsaturated fatty acids from 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 normal rats maintained on a laboratory diet has been determined. All of the octadecenoates and most of the hexadecenoates from the lipid classes of all the tissues consisted of more than 95% cis isomers. Generally, palmitoleic was the predominant hexadeceonate, but many of the tissue phospholipids contained relatively high percentages of the delta 6 and delta 7 isomers. Oleate and vaccenate were the predominant octadecenoates in all tissues. Except for brain and adipose tissues, the oleate to vaccenate ratios differed for each lipid class, as well as between most tissues. In contrast to the monoenes of the phospholipids, the triglyceride monoenes exhibited the same approximate: percentage composition; percentage of geometrical isomers; and percentage distribution of hexadecenoate and octadecenoate positional isomers. These data add to our basic information about the percentage distribution of geometrical and positional isomers of naturally occurring unsaturated fatty acids in the major lipid classes of various normal tissues. Some new concepts were advanced as possible explanations to some of the observed positional isomer distributions.

Animals

Value of Food and Agriculture Organization data on food-balance sheets as a data source for dietary fat intake in epidemiologic studies.

The relationships between the per person supply (expressed as percent of total energy supply) of saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), and polyunsaturated fatty acids (PUFAs) obtained from 1979-1981 Food and Agriculture Organization (FAO) data, and the per person intake obtained from 52 individual dietary surveys performed in 19 countries, were examined. In particular, the ratio of PUFAs to SFAs (P:S) and the ratio of unsaturated fatty acids (MUFAs and PUFAs) to SFAs (U:S) obtained from both data sources were examined. Significant correlations (P less than 0.001) were found between the two data sources for the P:S, U:S, SFAs, and MUFAs and PUFAs (P less than 0.02) in 19 countries. It is concluded that the data on fat intake from the FAO are valid for use in epidemiologic studies. In view of the variability of the data, it is recommended to use them either expressed as percentage of energy or as ratios of the different components.

Diet Surveys

Effects of tumor necrosis factor-alpha on peroxidation of plasma lipoprotein lipids in experimental animals and patients.

Changes in the plasma lipid composition are observed in patients and animals with malignancy and certain other diseases that are consistent with peroxidation of plasma lipoprotein lipids. These changes can be observed with water-suppressed proton (H-1) and carbon-13 (C-13) nuclear magnetic resonance spectroscopy (NMR) and gas chromatography. Gas chromatography provides evidence of a decrease in polyunsaturated fatty acids relative to monounsaturated fatty acids. This evidence is consistent with that observed by C-13 NMR spectroscopy. Mediators for these effects were sought. Cytokines, known to be released in response to malignant tumor cells and to affect lipid metabolism, were injected into normal mice and their effects on the H-1 and C-13 NMR spectra of plasma lipids were observed. Mouse recombinant tumor necrosis factor-alpha (mr-TNF-alpha) significantly decreased the H-1 methyl and methylene lipid linewidths, and the C-13 spectra indicated a decrease in the relative concentration of polyunsaturated fatty acids. The same changes were directly confirmed by gas chromatographic analysis, showing decreases in the amount of linoleic and arachidonic acids and other polyunsaturated fatty acids relative to monounsaturated fatty acids and in the ratio of polyunsaturated to monounsaturated fatty acids. Serial plasma samples from volunteers receiving an infusion of endotoxin showed similar changes in their C-13 NMR spectroscopy at times when peak TNF-alpha values were measured. In addition, in these samples the C-13 NMR spectra showed direct evidence of lipid peroxidation products. These changes were similar to those observed commonly in the plasma of cancer patients. Other cytokines (human recombinant interleukin-1 alpha [hr-IL-1 alpha], hr-IL-2, mouse recombinant interferon-gamma) did not produce these effects. We conclude that TNF-alpha is a mediator (but not necessarily the only one) of changes in plasma lipoprotein lipid composition due to peroxidation and that this is a mechanism for the changes observed in the NMR spectra of plasma from cancer patients and from normal animals injected with TNF-alpha.

Animals

Effect of dietary cis and trans fatty acids on serum lipoprotein[a] levels in humans.

Serum lipoprotein[a] (Lp[a]) is a strong risk factor for coronary heart disease. We therefore examined the effect of dietary fatty acid composition on serum Lp[a] levels in three strictly controlled experiments with healthy normocholesterolemic men and women. In Expt. I, 58 subjects consumed a control diet high in saturated fatty acids for 17 days. For the next 36 days, 6.5% of total energy intake from saturated fatty acids was replaced by monounsaturates plus polyunsaturates (monounsaturated fatty acid diet; n = 29) or by polyunsaturates alone (polyunsaturated fatty acid diet; n = 29). Both diets caused a slight, nonsignificant, increase in median Lp[a] levels, with no difference between diets. In Expt. II, 10% of energy from the cholesterol-raising saturated fatty acids (lauric, myristic, and palmitic acid) was replaced by oleic acid or by trans-monounsaturated fatty acids. Each of the 59 participants received each diet for 3 weeks in random order. The median level of Lp[a] was 26 mg/l on the saturated fatty acid diet; it increased to 32 mg/l (P less than 0.020) on the oleic acid diet and to 45 mg/l (P less than 0.001) on the trans-fatty acid diet. The difference in Lp[a] between the trans-fatty acid and the oleic acid diets was also highly significant (P less than 0.001). Expt. III involved 56 subjects; all received 8% of energy from stearic acid, from linoleic acid, or from trans-monounsaturates, for 3 weeks each. All other nutrients were equal.(ABSTRACT TRUNCATED AT 250 WORDS)

Dietary Fats

Modification of plasma and hepatic lipids of guinea pigs by feeding high oleic acid pork compared with regular pork.

Meat from such monogastric animals as swine can be modified to substitute monounsaturated fatty acids for saturated fatty acids. Because monounsaturated fatty acids have a beneficial effect on serum lipids as compared with saturated fatty acids, the objective of this study was to assess the effect of modified pork as compared with regular pork on serum and hepatic lipids. Guinea pigs were fed diets containing pork from control diet-fed hogs or from hogs fed a diet containing high oleic acid sunflower oil. The pork provided almost all of the fat in the diets at the level of 4 and 15 g/100 g diet, 10 or 34% energy. The high oleic pork muscle and fat contained 26 and 46% less palmitic and stearic acids (the primary saturated fatty acids), respectively, and 31 and 29% more oleic acid (the primary monounsaturated fatty acid) than the regular pork muscle and fat, respectively. Cholesterol concentration of diets ranged from 0.06 to 0.08% of the diet. Although total serum cholesterol, HDL cholesterol and triglyceride concentrations did not differ due to type of pork, results indicated that serum LDL cholesterol was lower (15%) and hepatic cholesterol was greater (15%) in the high oleic pork, 15% fat group as compared with the control pork 15% fat group. Also, serum LDL cholesterol concentration was higher in the groups fed 15% fat compared with those fed 4% fat. In this study pork modified to have more oleic acid and less saturated fatty acids had a positive effect on tissue lipids when fed to animals.

Animals

Intramitochondrial phospholipase activity and the effects of Ca2+ plus N-ethylmaleimide on mitochondrial function.

Liver mitochondria treated with N-ethylmaleimide can accumulate Ca2+ but cannot retain it. Ca2+ loss following uptake occurs in parallel with a proton uptake and collapse of the membrane potential. Respiration is not activated during Ca2+ release and cannot be stimulated by uncoupler. After Ca2+ release and accompanying phenomena are nearly complete, the mitochondria undergo a large amplitude swelling. Nupercaine inhibits the premature release of Ca2+, proton uptake, decline in membrane potential, inhibition of uncoupler-stimulated respiration, and large amplitude swelling. Ruthenium red also prevents these effects. Neither Sr2+ or Mn2+ will substitute for Ca2+ to induce these effects in N-ethylmaleimide-treated mitochondria. The effects of N-ethylmaleimide plus Ca2+ on mitochondria are not accompanied by a significant alteration in the content or composition of phospholipids but are accompanied by small increases in the mitochondrial content of free fatty acids. Free fatty acids accumulate more rapidly in response to limited Ca2+ loading in the absence of N-ethylmaleimide than they do in its presence. In the absence of N-ethylmaleimide, polyunsaturated fatty acids and saturated plus monounsaturated fatty acids accumulate at nearly equal rates. In the presence of N-ethylmaleimide, polyunsaturated fatty acids accumulate more rapidly than saturated plus monounsaturated fatty acids. Any condition or agent tested which inhibited swelling and the other effects produced by Ca2+ plus N-ethylmaleimide also prevented the more rapid accumulation of polyunsaturated, compared to saturated plus monounsaturated, fatty acids. In the light of a positional analysis of phospholipid acyl moieties, these data suggest that 1-acyllysophospholipids accumulate in swelling mitochondria but not in response to noraml Ca2+ loading or when swelling is blocked by other agents. The free fatty acid accumulation, per se, is not responsible for swelling, but levels of exogenous palmitic acid as low as 1 nmol/mg of protein dramatically alter the dependence of swelling velocity on Ca2+ concentration, producing a shift from a sigmoidal- to a hyperbolic-like relationship. This same alteration is brought about by aging the mitochondrial preparation at 0 degrees C. Either pyruvate or DL-carnitine prevents the effect of exogenous palmitate and restores the Aa2+ swelling dependence of aged N-ethylmaleimide-treated mitochondria to that of fresh N-ethylmaleimide-treated mitochondria. Intramitochondrial acylcoenzyme A or acylcarnitine, or both, therefore, to be the modulator of Ca2+ sensitivity rather than free fatty acid. The findings are discussed in terms of the role of intramitochondrial phospholipase and other phospholipid metabolizing enzymes in the mechanisms of N-ethylmaleimide plus Ca2+ effects on mitochondria.

Animals

Olive oil, corn oil, and n-3 fatty acids differently affect lipids, lipoproteins, platelets, and superoxide formation in type II hypercholesterolemia.

To evaluate which dietary fat may provide the best response in terms of plasma lipids and lipoproteins and also of platelet aggregability and superoxide formation by white blood cells, 12 type II patients were randomly allocated to three different diets, which provided polyunsaturated fatty acids (corn oil), monounsaturated fatty acids (olive oil), and a supplementation of ethyl esters of n-3 fatty acids to a prudent diet. Olive oil and, more significantly, n-3 ethyl esters lowered total cholesterol best (-2.2% and -5.8%, respectively); the latter diet, as expected, also significantly lowered triglyceridemia (-21.4%). The corn-oil diet exerted a small, statistically significant reduction of high-density-lipoprotein cholesterol (HDL) (-4.3%), and it also lowered plasma total apo B concentrations (-3.8%). n-3 ethyl esters significantly raised both total (+3.1%) and particularly HDL2 cholesterol (+24%). Platelet reactivity was insignificantly reduced by the three regimens, but all three significantly reduced thrombin-stimulated formation of thromboxane B2. Finally, only the n-3 fatty acid supplementation significantly reduced O2- generation by adherent monocytes. Dietary unsaturated fatty acids are generally effective on the plasma lipid and lipoproteins in type II patients, but significant differences may be found between the three tested regimens.

Apolipoproteins

Long-term effects of dietary monounsaturated and polyunsaturated fatty acids on plasma lipids in dogs.

To compare the effect of monounsaturated vs. polyunsaturated fatty acids on plasma lipids, three months old dogs were studied for a period of six months. The dogs were fed a basal diet supplemented with either 14% olive oil or sunflower oil. Blood samples were taken fortnightly during this period. We have found changes in the various unsaturated fatty acids in response to the diet. Oleic and 20:3 n-9 acids were higher in the olive oil group while linoleic acid was increased in the dogs fed the sunflower oil diet. Arachidonic acid and PUFA n-3 > 18C index were nearly similar with both diets. The cholesterol levels were similar to those found in adult humans and no significant differences were brought about diets at any time. Thus a diet rich in monounsaturated fatty acids is as efficient as a polyunsaturated rich in relation to total cholesterol levels but more beneficial because of the antiatherogenic effect of HDL-cholesterol which is increased with this type of dietary fat.

Animals

The fatty acid composition of subcutaneous fat in German adults.

The fatty acid (FA) composition of subcutaneous fat (SCF) was analyzed in 47 German adults. No influences of sex or age on the FA status of the probands could be detected. SCF consisted mainly of monoenoic fatty acids. Linoleic acid was the major polyunsaturated fatty acid. Linoleic acid metabolites were found in small quantities only. In comparison to the results of studies in the USA and the Netherlands the FA composition of SCF in German probands was characterized by lower levels of linoleic acid and higher contents of palmitic acid. The trans-FA content was similar to that in Dutch women, but lower than in American male probands. However, in evaluating the differences in the FA composition of SCF in different countries, it has to be noted that there are wide ranges in the proportions of the major FA in all populations studied.

Adipose Tissue

Studies on the inhibition of the desaturases by cyclopropenoid fatty acids.

Unwashed rat liver microsomes were used to study the inhibition of the delta6 and delta9 desaturases by cyclopropenoid fatty acids with the ring structure about the 9,10 or 6,7 carbon atoms. The 9,10 cyclopropenoid acid (sterculic acid) is shown to be an effective inhibitor of only delta9 desaturase and then only in the presence of MgCl2 and coenzyme A (presumably due to the formation of sterculoyl-CoA). Two 6,7 cyclopropenoid acids of different chain lengths showed no marked inhibition of either the delta6 or delta9 desaturase. By the use of [3H]-sterculic acid, it has been shown that under conditions of high inhibition of the delta9 desaturase the inhibitor is not covalently attached to the enzyme at any point. This disproves older ideas on the mechanism of inhibition that assumed reaction between the cyclopropenoid ring and sulphydryl groups on the enzymes.

Animals