The effect of endotoxin on membrane fatty acid composition in BCG-sensitized mice

Summary The effects of endotoxin on mouse liver phospholipid fatty acid composition have been investigated. Administration of endotoxin fromSalmonella abortus equi led to a decrease in the polyunsaturated fatty acid content of livers from mice sensitized with Bacille Calmette Guérin (GCG). The content of arachidonic acid fell significantly in both the phosphatidylcholine and phosphatidylinositol fractions whereas in the phosphatidylethanolamine fraction the linoleic acid content was significantly reduced. The polyunsaturated fatty acids were replaced by increased amounts of oleic acid and palmitic acid, leading to a reduction in the polyunsaturated to saturated fatty acid ratio.     

Fatty acid profiles of major food sources of howler monkeys (Alouatta palliata) in the neotropics

Wild howler monkeys (Alouatta palliata) get most of their calories from carbohydrates (65%) and fats (18%) of native tropical plants, but little is known about their intake of individual fatty acids. The fatty acid composition of several natural food sources of howler monkeys collected in Panama was determined by gas-liquid chromatography. The predominant fatty acids were palmitic (30%), linoleic (23%), linoleic (23%), -linolenic (16%) and oleic (15%). Fatty acids with less than 16, and more than 18, carbon chains were uncommon (0–7%). Although total saturated fatty acids were high in some specific food sources (22–54% of total fatty acids and 8 energy %), most of the calories from fat in the animals' diets are derived from mono-and polyunsaturated fatty acids (9.75 energy %) All food sources had significant amounts of the -3 fatty acid, -linolenic acid (2.9 energy %). In terms of human diets, the howler monkey's fat consumption would not be considered atherogenic. Unless these animals show a particular adverse susceptibility to dietary fat, it is unlikely that their fat intake is the primary cause of the low, but significant, incidence of atherosclerosis that develops in these animals in the wild state.     

Fixation of polyunsaturated fatty acids by alpha fetoprotein and serum albumin in rats. Comparison with the accumulation of these acids in developing rat brain]

Quantitative data are presented on the fatty acid composition of rat alpha-fetoprotein (AFP) and serum albumin, (SA), and of brain extracts of suckling rats. In AFP and SA preparations, 40% and 13%, respectively, of total fatty acids present are polyenoic acids. Among them, docosahexaenoic acid is quantitatively the most important in AFP, while in SA, arachidonic acid is largely predominant. Both docosahexaenoic and arachidonic acids were the predominant polyenoic acids in brain extracts. The rate of accumulation of these acids in the brain of suckling rats and the rate of AFP secretion during the same period showed a maximum around 10--12 days after birth. These results suggest that AFP and SA play an important role in the transport and the incorporation of polyunsaturated fatty acids in the developing brain.     

Antiproliferative effect of polyunsaturated fatty acids and interleukin-2 on normal and abnormal human lymphocytes

The polyunsaturated fatty acids (PUFAs), linoleic acid (LA), alpha linolenic acid (ALA), gamma linolenic acid (GLA), arachidonic acid (AA), docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), showed inhibition of growth of both normal and abnormal (Molt-4) human lymphocytes, and inhibition was concentration-dependent. Interestingly, the production of the lymphokine Interleukin-2 (IL-2) was elevated in Molt-4 cells, but it was reduced in the normal human lymphocytes. Addition of GLA or IL-2 or a combination of both showed enhancement of SO ₂ ^– and of lipid peroxidation levels, which were significantly higher in Molt-4 cells than in the normal lymphocytes. Reduction of protein concentration was also observed in both types of cells during this treatment. The data showed that the antiproliferative effects of GLA and IL-2 may partly be exerted through the elevated production of superoxide free radicals and peroxidatin products. This is a novel finding and therefore, further exploitation of combinations of PUFAs and IL-2 may be a possible way of combating cancer cell growth.     

Effect of maternal under-nutrition on pup body weight and hypothalamic endocannabinoid levels

Dietary long-chain polyunsaturated fatty acids are known to influence brain levels of the endocannabinoid anandamide in newborn pigs and mice. Furthermore, endocannabinoids were shown to control pup suckling and body weight in mice, and food intake in adult rodents. Here we determined the effect of maternal under-nutrition during gestation, lactation, or both, on body weight, and on the levels of endocannabinoids and expression of cannabinoid CB₁ receptors and fatty acid amide hydrolase in the hypothalamus of rat pups at weaning (21 days old) or adult rats (4 months old). Maternal under-nutrition resulted in a striking decrease in body weight of weaning rats, paralleled by a decrease in the hypothalamic levels of the endocannabinoid anandamide, but not of 2-arachidonoylglycerol. No significant change in the hypothalamic expression of either cannabinoid CB₁ receptors or fatty acid amide hydrolase mRNA was detected in any of the three groups of weaned pups. The decrease in pup body weight and hypothalamic anandamide levels was not observable in 4-month-old rats from any of the three groups. These data suggest that maternal under-nutrition causes a decrease in hypothalamic anandamide levels and loss of body weight, and confirm a crucial role for endocannabinoid signalling in neonatal development. Received 4 November 2002; received after revision 29 November 2002; accepted 16 December 2002 RID="*" ID="*"Corresponding author.     

Furan fatty acid steryl esters from the marine spongeDictyonella incisa which show inflammatory activity

Summary Three new furan fatty acids, (9Z, 19Z)-3, 6-epoxyhexacosa-3,5,19-tetraenoic acid (1a), (8Z, 11Z, 14Z, 17Z)-3, 6-epoxyeicosa-3,5,8,11,14,17-hexaenoic acid (2a), and (8Z, 11Z, 14Z, 17E)-3, 6-epoxyeicosa-3,5,8,11,14,17-hexaenoic acid (3a), and a new polyunsaturated fatty acid (5Z, 8Z, 11Z, 14Z, 17E)-eicosa-5,8,11,14,17-pentaenoic acid (6a), present in the spongeDictyonella incisa as the respective steryl esters, have been isolated as methyl esters and their structures have been determined by spectral and chemical analysis. The furan fatty acid esters have shown a high inflammatory activity, which suggests their potential role as feeding deterrents.     

Polyunsaturated fatty acids and cardiovascular disease

Replacing saturated with polyunsaturated (PUFAs) rather than monounsaturated fatty acids or carbohydrates results in cardiovascular prevention over a wide range of intakes. The mechanisms by which PUFAs reduce cardiovascular risk are manifold, and the extent and precise nature of their activities is the subject of several investigations, spanning from in vitro mechanistic studies to human intervention trials. This article reviews the most up-to-date evidence of the association between PUFA consumption and reduced cardiovascular mortality.     

Hypersensitivity to endotoxin hepatotoxicity in rats with inflammatory lesions

Summary The ratio of sinusoidal nonparenchymal cells to hepatocytes in rat liver was significantly increased following induction of inflammation, and decreased after subsequent exposure to endotoxin, particularly in the region around the terminal portal venules. Rats with inflammatory lesions were more sensitive to endotoxin hepatocytotoxicity than normal controls, as judged from the dose-dependent increase in activity of serum transaminases and from the extent of liver tissue injury. In addition, these animals, which were already in a state of depletion of hepatic glycogen, demonstrated marked hyperglycemia 24 h after endotoxin administration in small doses of less than 2 mg/kg.     

Über die Fettsäuren aus Spinatchloroplasten

Summary The fatty acids of two samples of chloroplasts fromSpinacia oleracea have been investigated quantitatively. They contain many polyenoic fatty acids (more than 70% of the total fatty acids); amongst these, a relatively high content of C¹⁶-trienoic acid is remarkable. Moreover, the presence of the ³-trans-hexadecenoic acid, recently detected in plants, is noteworthy.     

Hypersensitivity to endotoxin hepatotoxicity in rats with inflammatory lesions

The ratio of sinusoidal nonparenchymal cells to hepatocytes in rat liver was significantly increased following induction of inflammation, and decreased after subsequent exposure to endotoxin, particularly in the region around the terminal portal venules. Rats with inflammatory lesions were more sensitive to endotoxin hepatocytotoxicity than normal controls, as judged from the dose-dependent increase in activity of serum transaminases and from the extent of liver tissue injury. In addition, these animals, which were already in a state of depletion of hepatic glycogen, demonstrated marked hyperglycemia 24 h after endotoxin administration in small doses of less than 2 mg/kg.     

Absence of endotoxin-fever but not hyperthermia in Brattleboro rats

Summary I. c. v. administration of bacterial endotoxin produced a fever in the Long-Evans rat but not in the Brattleboro rat. Similar administration of arachidonic acid, prostaglandin E₂, prostacyclin, dibutyryl cAMP, norepinephrine, morphine and -endorphin caused hyperthermia in both Long-Evans and Brattleboro rats. Variable doses of exogenous arginine vasopressin (AVP) when centrally administered with endotoxin caused fever in the Brattleboro rat. It is suggested that AVP may play an important role in the production and release of endogenous pyrogen.     

An eicosanoid-centric view of atherothrombotic risk factors

Cardiovascular disease is the foremost cause of morbidity and mortality in the Western world. Atherosclerosis followed by thrombosis (atherothrombosis) is the pathological process underlying most myocardial, cerebral, and peripheral vascular events. Atherothrombosis is a complex and heterogeneous inflammatory process that involves interactions between many cell types (including vascular smooth muscle cells, endothelial cells, macrophages, and platelets) and processes (including migration, proliferation, and activation). Despite a wealth of knowledge from many recent studies using knockout mouse and human genetic studies (GWAS and candidate approach) identifying genes and proteins directly involved in these processes, traditional cardiovascular risk factors (hyperlipidemia, hypertension, smoking, diabetes mellitus, sex, and age) remain the most useful predictor of disease. Eicosanoids (20 carbon polyunsaturated fatty acid derivatives of arachidonic acid and other essential fatty acids) are emerging as important regulators of cardiovascular disease processes. Drugs indirectly modulating these signals, including COX-1/COX-2 inhibitors, have proven to play major roles in the atherothrombotic process. However, the complexity of their roles and regulation by opposing eicosanoid signaling, have contributed to the lack of therapies directed at the eicosanoid receptors themselves. This is likely to change, as our understanding of the structure, signaling, and function of the eicosanoid receptors improves. Indeed, a major advance is emerging from the characterization of dysfunctional naturally occurring mutations of the eicosanoid receptors. In light of the proven and continuing importance of risk factors, we have elected to focus on the relationship between eicosanoids and cardiovascular risk factors.     

The effect of endotoxin on plasma α-aminoisobutyric acid

Summary The i.v. injection of bacterial endotoxin into dogs was found to cause a rapid increase in plasma levels of infused -aminoisobutyric acid. The findings suggest that nonmetabolic factors (tissue uptake, fluid shifts) influence amino acid distribution during endotoxemia.This investigation was supported in part by the Office of Naval Research, Contract N0001476CO132. R.E.K., Postdoctoral Fellow, supported by HL07090. Address correspondence to J.J.S.     

Hydra vulgaris ω10-lipoxygenase is used in vivo to synthesize new α-linolenic acid metabolites

Previous studies conducted in cytosolic extracts of the freshwater hydrozoanHydra vulgaris led to the finding of an abundant 11(R)-lipoxygenase catalyzing the peroxidation of polyunsaturated fatty acid (PUFAs) on the tenth carbon atom from the aliphatic end (10 peroxidation). Here we describe experiments aimed at identifying the actual metabolites generated in vivo by such enzymic activity. Homogenates ofH. vulgaris polyps were analyzed by HPLC. This showed the presence of three major components chromatographically identical to three metabolites obtained when incubating the homogenates with exogenous -linolenic acid (-LA). The presence, in extracts of polyps prelabelled with [¹⁴C]--linolenic acid, of radioactive metabolites displaying the same chromatographic properties, substantiated the hypothesis that the natural products isolated in vivo are derived from -LA. Gas chromatographic analyses revealed that this was the most abundant PUFA in both free and phosphoglyceride-bound fatty acid pools. [¹H]-NMR analysis of the endogenous substances, carried out in comparison with products obtained from exogenously incubated -LA, indicated that their structures were those of 9-hydroxy-, 9-hydroperoxy- and 9-keto-octadeca-10E-12Z-15Z-trienoic acids (9--HOTrE,-HPOTrE and-KOTrE).Hydra homogenates transformed 9--HPOTrE partly into 9--HOTrE and partly into 9--KOTrE. Chiral phase HPLC conducted on 9--HOTrE established that this metabolite was composed mostly of theR anantiomer. These observations, and the finding that the presence of exogenous arachidonic acid in incubated homogenates significantly reduces the production of -LA metabolites, provide strong evidence that these compounds are produced by an enzymic activity identical to the previously-describedH. vulgaris (R)-10-lipoxygenase. Further experiments suggested that -LA, acting as the native substrate for this enzyme, is mainly esterified on the 2 position ofHydra phosphoglycerides, and that the production of the -LA metabolites described here for the first time from natural sources, can be potentially enhanced in vivo by stimuli activating phospholipase A₂.     

Chemical studies on slow reacting substances/leukotrienes

The family of eicasanoids, biologically active metabolites of polyunsaturated C20 fatty acids such as arachidonic acid, has recently been enlarged by the recognition of a new biosynthetic pathway leading to the leukotrienes, including the compounds described two decades ago as 'slow reacting substances'. These biologically potent substances are involved in regulation of the immune response and also as mediators in various disease states. This account presents a brief history of this field, an overview of the biological relevance of leukotrienes, and a discussion of the investigations which led to the clarification of the molecular structures, pathway of biosynthesis and total chemical synthesis of the leukotrienes, including leukotrienes A, B, C, D and E (LTA-LTE). As a result of the synthetic work these rare substances are available for the first time in pure form and in quantities sufficient for biological and medical studies. Also reviewed are recent discoveries with regard to the development of inhibitors of leukotriene biosynthesis and anti-leukotrienes.     

Mechanisms of lipid peroxidation: Iron catalyzed decomposition of fatty acid hydroperoxides as the basis of hydrocarbon evolution in vivo

Summary Ethane and pentane are evolved during peroxidation of tissue lipids in vivo and are believed to be derived from 3 and 6 fatty acids respectively. We present evidence supporting a scission mechanism as the means of forming these hydrocarbons during hydroperoxide decomposition. A fatty acid 6 hydroperoxide (methyl 13-hydroperoxy-6,9,11-octatrienoate), when incubated with a ferrous ion, yielded pentane as the major hydrocarbon (98%). Reaction with ferrous ion results in an alkoxy radical capable of undergoing scission to an aldehydic fatty acid and a hydrocarbon.We wish to thank Dr Ned Porter for both his generous donation of a supply of hydroperoxide and for his suggestions regarding possible reaction mechanisms. This research was supported by U.S.P.H.S. N.I.H. Grants ES00798, HL16264 and T32 GM07105.     

The effect of hunger on free fatty acid and corticosterone plasma levels in rats.

The time function of free fatty acids and of corticosterone in plasma in fed and fasted rats was investigated. Also a study of the influence of hypoxia on the concentration of plasma free fatty acids was carried out. Whereas fasting does not seem to stimulate lipolysis markedly before 12 h, 2 h of hypoxia elevate the free fatty acid level to its maximum. In the fasted condition, all values are significantly higher than corticosterone in the fed animal.     

In vitro esterification of plant sterols by the esterifying enzyme of the small intestine of rat

Summary The in vitro esterification of plant sterols,-sitosterol, campesterol and stigmasterol, by the esterifying enzyme of the small intestine of rat was studied in the presence of saturated and unsaturated fatty acids. Campesterol esterification was highest, followed by sitosterol and stigmasterol irrespective of the type of fatty acid. Both campesterol and sitosterol esterification was greater with unsaturated fatty acids than with saturated fatty acids.Acknowledgments. Financed by the Emery Goff Research Award from the American Heart Association, Louisiana, Inc. and the USPHS research grant HL-08974 from the National Institutes of Health, Bethesda, Md.     

Fatty acid modulation of antiestrogen action and antiestrogen-binding protein in cultured lymphoid cells

Nonsteroidal antiestrogens reversibly and specifically inhibited the proliferation of two estrogen receptor-negative lymphoid cell lines (EL4 and Raji) in a dose-dependent manner. [3H]Thymidine incorporation of concanavalin A-stimulated primary splenocytes was also inhibited by 10(-6) M clomiphene (1-[4-(2-diethylaminoethoxy)phenyl]-1,2-diphenyl-2-chloroethylene). The antiproliferative effect could be prevented by the simultaneous presence in the growth medium of 10(-5) M linoleic acid or 10(-5) M arachidonic acid but not by 10(-6) M estradiol. Both lymphoid cell lines had high affinity antiestrogen-binding sites whose affinity could be altered by conditions of growth. Growth of EL4 cells in RPMI 1640 medium supplemented with charcoal-pretreated 5% fetal calf serum (charcoal-stripped medium) resulted in significantly higher affinity (Kd 0.54 nM +/- 0.11 nM; n = 6) than growth in medium supplemented with untreated serum (complete medium) (Kd = 1.68 nM +/- 0.48 nM; n = 6) (p less than 0.001). This change in affinity was partly due to removal of fatty acids from the growth medium by charcoal pretreatment, since addition of 10(-5) M linoleic acid or 10(-5) M gamma-linolenic to charcoal-stripped medium decreased the affinity of the antiestrogen-binding protein. In contrast, growth in 10(-5) M stearic acid or 10(-5) M oleic acid did not significantly alter the affinity of the antiestrogen-binding protein, whereas 10(-5) M palmitic acid significantly increased its affinity. The same fatty acids were also tested for their intrinsic effects on EL4 cell proliferation. Oleic, linoleic and gamma-linolenic acids were growth stimulatory while stearic and palmitic acids were not. Thus linoleic and gamma-linolenic acids whose presence in the growth medium was associated with decreased affinity of [3H]tamoxifen (1-[4-(2-dimethylaminoethoxy)phenyl]-1,2-diphenylbut-1(Z)-ene) binding to the intracellular antiestrogen-binding protein were also growth stimulatory. Unsaturated fatty acids have previously been shown to inhibit binding of [3H]tamoxifen to the antiestrogen-binding protein in a cell-free system. The present observations demonstrate that unsaturated fatty acids also modify the affinity of the antiestrogen-binding protein in intact cells.