Plasma free fatty acids (non-esterified fatty acids) increase in the first hour of the onset of acute myocardial ischaemia. This results from catecholamine stimulation of adipose tissue lipolysis. It can lead to a metabolic crisis in the injured myocardium with the development of ventricular arrhythmias and increased early mortality. Preconditioning, β-adrenergic blockade and glucose–insulin–potassium are possible therapeutic approaches, but anti-lipolytic agents, such as some nicotinic acid derivatives, can reduce plasma free fatty acid concentrations within minutes and have untried potential. A clinical trial of their effectiveness is needed from the first moment when a patient with an acute coronary syndrome is seen by paramedics.

IUGR (intrauterine growth restriction) increases the incidence of perinatal complications and, although several placental transport functions have been shown to be altered in pregnancies complicated by IUGR, the mechanism behind it is not well understood. The aim of the present study was to investigate factors in maternal and cord blood plasma from normal and IUGR-complicated pregnancies associated with the body weight of newborns. At the time of Caesarean section, 24 women with IUGR pregnancies were compared with a group of 30 normal controls with AGA (appropriate gestational age) fetuses who were studied at Caesarean section, which took place 5 weeks later than IUGR pregnancies, and also to a group of 25 non-delivered gestational age-matched control pregnant women (AGA-35wk). Maternal plasma retinol, γ- and α-tocopherol, NEFAs (non-esterified fatty acids), and palmitic, palmitoleic, γ-linolenic and arachidonic acids were higher in women with IUGR pregnancies than in AGA-35wk controls, whereas stearic and α-linolenic acids were lower. Smaller differences were found when comparing these variables for IUGR and AGA women. However, umbilical vein plasma γ-tocopherol, cholesterol, triacylglycerols and NEFAs were higher in the IUGR group than in the AGA group, whereas arachidonic acid was lower. Maternal plasma retinol and NEFAs were the only variables negatively correlated with birthweight when multiple linear regressions were analysed. In conclusion, the increased levels of circulating retinol and NEFAs in maternal plasma are negatively associated with birth and placental weights, which may reflect an impaired placental transfer in IUGR pregnancies. As retinoids are involved in the control of gene transcription, it is proposed that a decrease in placental transfer of retinol could underlie the metabolic dysfunction of IUGR pregnancies.

Sympathetic nervous system activation is a hallmark of several conditions associated with an adverse prognosis, including hypertension and the metabolic syndrome. Proposed mediators of increased sympathetic drive include hyperinsulinaemia, leptin, NEFAs (non-esterified fatty acids), pro-inflammatory cytokines, baroreflex impairment and others. The role of NEFAs appears to be of particular importance given the increased levels observed in human obesity and the experimental results linking the NEFA-induced pressor response to sympathetic activation. Findings from human studies have yielded conflicting results with regards to a sympathetically mediated association between NEFAs and elevated arterial blood pressure. In the present issue of Clinical Science , Florian and Pawelczyk present some interesting results obtained from a small number of healthy normotensive lean volunteers who were exposed to NEFA infusion and cardiovascular and sympathetic monitoring using state of the art methodology that appears to be in support of such a link. However, several methodological and conceptual considerations need to be taken into account when interpreting the results from this study. Put into perspective, the case for a substantial sympathetically mediated pressor response to NEFA infusion does not appear to be a very strong one.

Previous studies have shown that acute increases in plasma NEFAs (non-esterified fatty acids) raise SVR (systemic vascular resistance) and BP (blood pressure). However, these studies have failed to distinguish between CNS (central nervous system) mechanisms that raise sympathetic activity and paracrine mechanisms that increase SVR directly, independent of CNS involvement. The aim of the present study was to directly determine whether the sympathetic nervous system contributes to the pressor response to NEFAs. On 2 days separated by at least 2 weeks, 17 lean healthy volunteers (ten male/seven female; age, 22±1 years; body mass index, 23±1 kg/m 2 ; values are means±S.E.M.) received a 4-h intravenous infusion of 20% Intralipid® or placebo (in a single-blind randomized balanced order). MSNA (muscle sympathetic nerve activity), HR (heart rate), BP (oscillometric brachial measurement) and (cardiac output; acetylene rebreathing) were measured before and throughout infusion. The change in HR (+8.2±1.0 and +2.4±1.2 beats/min), systolic BP (+14.0±1.6 and +3.2±2.5 mmHg) and diastolic BP (+8.2±1.0 and −0.1±1.7 mmHg) were significantly greater after the 4-h infusion of Intralipid® compared with placebo ( P <0.001). The change in BP with Intralipid® resulted from an increase in SVR ( /mean arterial pressure; P <0.001) compared with baseline, without a change in . MSNA burst frequency increased during Intralipid® infusion compared with baseline (+4.9±1.3 bursts/min; P <0.05), and total MSNA (frequency×amplitude) was augmented 65% ( P <0.001), with no change during placebo infusion. Lipid infusion increased insulin, aldosterone and F 2 -isoprostane, but not leptin, concentrations. On the basis of the concomitant increase in BP, MSNA and SVR, we conclude that central sympathetic activation contributes to the pressor response to NEFAs. Previous studies have shown that acute increases in plasma NEFAs (non-esterified fatty acids) raise SVR (systemic vascular resistance) and BP (blood pressure). However, these studies have failed to distinguish between CNS (central nervous system) mechanisms that raise sympathetic activity and paracrine mechanisms that increase SVR directly, independent of CNS involvement. The aim of the present study was to directly determine whether the sympathetic nervous system contributes to the pressor response to NEFAs. On 2 days separated by at least 2 weeks, 17 lean healthy volunteers (ten male/seven female; age, 22±1 years; body mass index, 23±1 kg/m 2 ; values are means±S.E.M.) received a 4-h intravenous infusion of 20% Intralipid® or placebo (in a single-blind randomized balanced order). MSNA (muscle sympathetic nerve activity), HR (heart rate), BP (oscillometric brachial measurement) and (cardiac output; acetylene rebreathing) were measured before and throughout infusion. The change in HR (+8.2±1.0 and +2.4±1.2 beats/min), systolic BP (+14.0±1.6 and +3.2±2.5 mmHg) and diastolic BP (+8.2±1.0 and −0.1±1.7 mmHg) were significantly greater after the 4-h infusion of Intralipid® compared with placebo ( P <0.001). The change in BP with Intralipid® resulted from an increase in SVR ( /mean arterial pressure; P <0.001) compared with baseline, without a change in . MSNA burst frequency increased during Intralipid® infusion compared with baseline (+4.9±1.3 bursts/min; P <0.05), and total MSNA (frequency×amplitude) was augmented 65% ( P <0.001), with no change during placebo infusion. Lipid infusion increased insulin, aldosterone and F 2 -isoprostane, but not leptin, concentrations. On the basis of the concomitant increase in BP, MSNA and SVR, we conclude that central sympathetic activation contributes to the pressor response to NEFAs. Previous studies have shown that acute increases in plasma NEFAs (non-esterified fatty acids) raise SVR (systemic vascular resistance) and BP (blood pressure). However, these studies have failed to distinguish between CNS (central nervous system) mechanisms that raise sympathetic activity and paracrine mechanisms that increase SVR directly, independent of CNS involvement. The aim of the present study was to directly determine whether the sympathetic nervous system contributes to the pressor response to NEFAs. On 2 days separated by at least 2 weeks, 17 lean healthy volunteers (ten male/seven female; age, 22±1 years; body mass index, 23±1 kg/m 2 ; values are means±S.E.M.) received a 4-h intravenous infusion of 20% Intralipid® or placebo (in a single-blind randomized balanced order). MSNA (muscle sympathetic nerve activity), HR (heart rate), BP (oscillometric brachial measurement) and (cardiac output; acetylene rebreathing) were measured before and throughout infusion. The change in HR (+8.2±1.0 and +2.4±1.2 beats/min), systolic BP (+14.0±1.6 and +3.2±2.5 mmHg) and diastolic BP (+8.2±1.0 and −0.1±1.7 mmHg) were significantly greater after the 4-h infusion of Intralipid® compared with placebo ( P <0.001). The change in BP with Intralipid® resulted from an increase in SVR ( /mean arterial pressure; P <0.001) compared with baseline, without a change in . MSNA burst frequency increased during Intralipid® infusion compared with baseline (+4.9±1.3 bursts/min; P <0.05), and total MSNA (frequency×amplitude) was augmented 65% ( P <0.001), with no change during placebo infusion. Lipid infusion increased insulin, aldosterone and F 2 -isoprostane, but not leptin, concentrations. On the basis of the concomitant increase in BP, MSNA and SVR, we conclude that central sympathetic activation contributes to the pressor response to NEFAs.

Hypertriacylglycerolaemia is an important risk factor for cardiovascular disease. In men, we have shown that the effects of evening exercise on basal VLDL (very-low-density lipoprotein) metabolism are dose-dependent: a single prolonged bout of aerobic exercise [2 h at 60% of V ̇ O 2 peak (peak oxygen consumption)] reduces fasting plasma TAG [triacylglycerol (triglyceride)] concentrations, via enhanced clearance of VLDL-TAG from the circulation, whereas the same exercise performed for 1 h has no effect on VLDL-TAG metabolism and concentration. We hypothesized that women are more sensitive to the TAG-lowering effect of exercise because they reportedly use more intramuscular TAG as an energy source during exercise, and depletion of muscle TAG stores has been linked to reciprocal changes in skeletal muscle LPL (lipoprotein lipase) activity. To test our hypothesis, we measured basal VLDL-TAG and VLDL-apoB-100 (apolipoprotein B-100), and plasma NEFA [non-esterified fatty acid (‘free fatty acid’)] kinetics, by using stable isotope-labelled tracer techniques, on the morning after a single session of evening exercise of moderate duration and intensity (1 h at 60% of V ̇ O 2 peak ) in eight sedentary pre-menopausal women (age, 28±3 years; body mass index, 27±2 kg/m 2 ; body fat, 34±3%; values are means±S.E.M.). Compared with an equivalent period of evening rest, exercise had no effect on post-absorptive NEFA concentrations and the rate of appearance in plasma, VLDL-TAG and VLDL-apoB-100 concentrations, hepatic VLDL-TAG and VLDL-apoB-100 secretion and plasma clearance rates (all P >0.05). We conclude that, in women, as in men, a single session of exercise of moderate intensity and duration is not sufficient to bring about the alterations in VLDL metabolism that have been linked to post-exercise hypotriacylglycerolaemia.

The relevance of PPARγ (peroxisome-proliferator-activated receptor γ) as an important therapeutic target for the treatment of diabetes arises from its hypoglycaemic effects in diabetic patients and also from the critical role in the regulation of cardiovascular functions. From a clinical perspective, differences between current FDA (Food and Drug Administration)-approved PPARγ drugs have been observed in terms of atherosclerosis and cardiac and stroke events. The adverse effects of PPARγ-specific treatments that hamper their cardiovascular protective roles, affirm the strong need to evaluate the efficacy of the current drugs. Therefore active research is directed towards high-throughput screening and pharmacological testing of a plethora of newly identified natural or synthetic compounds. In the present review we describe the rationale behind drug design strategies targeting PPARγ, based on current knowledge regarding the effects of such drugs in experimental animal models, as well as in clinical practice. Regarding endogenous PPARγ ligands, several fatty acid derivatives bind PPARγ with different affinities, although the physiological relevance of these interactions is not always evident. Recently, NO-derived unsaturated fatty acids were found to be potent agonists of PPARs, with preferential affinity for PPARγ, compared with oxidized fatty acid derivatives. Nitroalkenes exert important bioactivities of relevance for the cardiovascular system including anti-inflammatory and antiplatelet actions, and are important mediators of vascular tone. A new generation of insulin sensitizers with PPARγ function for the treatment of diabetes may serve to limit patients from the increased cardiovascular burden of this disease.

Although specific pathogenic entities contributing to diabetic risk, such as central adiposity, ectopic fat accumulation, hyperlipidaemia and inflammation, are well-characterized, the response of cellular systems to such insults are less well understood. This short review highlights the effect of increasing fat mass on ectopic fat accumulation, the role of triacylglycerols (triglycerides) in Type 2 diabetes mellitus and cardiovascular disease pathogenesis, and selected current therapeutic strategies used to ameliorate these risk factors.

NAFLD (non-alcoholic fatty liver disease) encompasses the spectrum of fatty liver disease in insulin-resistant individuals who often display T2DM (Type 2 diabetes mellitus) and obesity. The present review highlights the pathophysiological basis and clinical evidence for a possible causal linkage between NAFLD and CVD (cardiovascular disease). The role of traditional and non-traditional CVD risk factors in the pathophysiology of NAFLD is considered in the first part of the review, with the basic science shared by atherogenesis and hepatic steatogenesis discussed in depth in the second part. In conclusion, NAFLD is not an innocent bystander, but a major player in the development and progression of CVD. NAFLD and CVD also share similar molecular mechanisms and targeted treatment strategies. On the research side, studies should focus on interventions aimed at restoring energy homoeostasis in lipotoxic tissues and at improving hepatic (micro)vascular blood supply.

Etomoxir is an inhibitor of mitochondrial CPT1 (carnitine palmitoyltransferase 1) and thereby switches energy metabolism from fatty acids to glucose oxidation. Such a metabolic change may be beneficial in CHF (congestive heart failure). The ERGO (etomoxir for the recovery of glucose oxidation) study was designed in which etomoxir was tested at a dose of 80 and 40 mg compared with placebo for a period of 6 months in patients with CHF. As the principle measure of efficacy, a maximal exercise tolerance test and a submaximal 6-min corridor walk test were used. Secondary end points were echocardiographical dimensions and quality-of-life assessment scores. A total of 350 patients were planned to be screened, with the expectation that end point data would be available from approx. 260 patients. However, the study had to be stopped prematurely, because unacceptably high liver transaminase levels were detected in four patients taking etomoxir. At the termination of the study, 121 patients were randomized to placebo, 118 to 40 mg of etomoxir and 108 to 80 mg of etomoxir. At that time, 21 patients in the placebo group, 16 in the 40 mg of etomoxir group and 14 patients in the 80 mg of etomoxir group had completed the study. The mean increases in exercise time were 3.3, 10.2 and 19.4 s for the placebo, 40 mg of etomoxir and 80 mg of etomoxir groups respectively ( P value was not significant). No changes were obvious in the 6-min corridor walk test or in echocardiographical parameters from baseline. The number of patients that completed the study was too small to demonstrate significant effects on exercise time, although there was a tendency towards an increase in exercise time. Therefore, before rejecting the hypothesis that inhibition of fatty acid oxidation might be beneficial in CHF, similar studies have to be performed using different inhibitors of fatty acid oxidation targeting CPT1 and other enzymes in this metabolic pathway.

Fatty acids (FAs) have been shown to alter leucocyte function and thus to modulate inflammatory and immune responses. In this review, the effects of FAs on several aspects of lymphocyte, neutrophil and macrophage function are discussed. The mechanisms by which FAs modulate the production of lipid mediators, activity of intracellular signalling pathways, activity of lipid-raft-associated proteins, binding to TLRs (Toll-like receptors), control of gene expression, activation of transcription factors, induction of cell death and production of reactive oxygen and nitrogen species are described in this review. The rationale for the use of specific FAs to treat patients with impaired immune function is explained. Substantial improvement in the therapeutic usage of FAs or FA derivatives may be possible based on an improvement in the understanding of the precise molecular mechanisms of action with respect to the different leucocyte types and outcome with respect to the inflammatory responses.

LVH [LV (left ventricular) hypertrophy] is an independent risk factor for CHD (coronary heart disease). During LVH, the preferred cardiac energy substrate switches from FAs (fatty acids) to glucose. LPL (lipoprotein lipase) is the key enzyme in triacylglycerol (triglyceride) hydrolysis and supplies FAs to the heart. To investigate whether substrate utilization influences cardiac growth and CHD risk, we examined the association between the functional LPL S447X (rs328) variant and hypertension-induced LV growth and CHD risk. LPL-X447 has been shown to be more hydrolytically efficient and would therefore release more free FAs than LPL-S477. In a cohort of 190 hypertensive subjects, LPL X447 was associated with a greater LV mass index [85.2 (1.7) in S/S compared with 91.1 (3.4) in S/X+X/X; P =0.01], but no such association was seen in normotensive controls ( n =60). X447 allele frequency was higher in hypertensives with than those without LVH {0.14 [95% CI (confidence interval), 0.08–0.19] compared with 0.07 (95% CI, 0.05–0.10) respectively; odds ratio, 2.52 (95% CI, 1.17–5.40), P =0.02}. The association of LPL S447X with CHD risk was then examined in a prospective study of healthy middle-aged U.K. men ( n =2716). In normotensive individuals, compared with S447 homozygotes, X447 carriers were protected from CHD risk [HR (hazard ratio), 0.48 (95% CI, 0.23–1.00); P =0.05], whereas, in the hypertensives, X447 carriers had increased risk [HR, 1.54 (95% CI, 1.13–2.09) for S/S ( P =0.006) and 2.30 (95% CI, 1.53–3.45) for X447+ ( P <0.0001)] and had a significant interaction with hypertension in CHD risk determination ( P =0.007). In conclusion, hypertensive LPL X447 carriers have increased risk of LVH and CHD, suggesting that altered FA delivery constitutes a mechanism through which LVH and CHD are associated in hypertensive subjects.

The aim of the present study was to investigate whether fasting for 24 and 48 h induces apoptosis of rat mesenteric lymph node lymphocytes similar to that observed previously in diabetic patients and alloxan-induced diabetic rats. Several features of lymphocyte death were evaluated by flow cytometry. Plasma levels of glucose, NEFAs (non-esterified fatty acids) and ketone bodies (acetoacetate and β-hydroxybutyrate) were determined in rats fasted for 24 and 48 h. Lymphocytes obtained from fasted rats had an increase in DNA fragmentation and phosphatidylserine externalization after 48 h of culture, although there was no loss of membrane integrity in lymphocytes even after 48 h of culture. Cytochrome c release from the mitochondrial intermembrane space into the cytosol was increased significantly in lymphocytes from fasted rats cultured for 24 h, whereas the levels of bcl-2 and bax proteins were not affected. Activities of caspases 3, 6, 8 and 9 were increased significantly in lymphocytes from rats fasted for 24 h, whereas only an increase in caspase 3 and 9 activities were observed in rats fasted for 48 h. In conclusion, fasting for 24 and 48 h caused a significant increase in the proportion of lymphocytes undergoing apoptosis. The occurrence of apoptosis was observed by DNA fragmentation, phosphatidylserine externalization, cytochrome c release from the mitochondria and activation of the caspase cascade. These findings support the hypothesis that conditions that raise plasma fatty acids levels (e.g. diabetes and starvation) may impair immune function by causing lymphocyte death.

In the present study, the cytotoxicity of palmitic, stearic, oleic, linoleic, arachidonic, docosahexaenoic and eicosapentaenoic acids on a macrophage cell line (J774) was investigated. The induction of toxicity was investigated by changes in cell size, granularity, membrane integrity, DNA fragmentation and phosphatidylserine externalization by using flow cytometry. Fluorescence microscopy was used to determine the type of cell death (Acridine Orange/ethidium bromide assay). The possible mechanisms involved were examined by measuring mitochondrial depolarization, lipid accumulation and PPARγ (peroxisome-proliferator-activated receptor γ) activation. The results demonstrate that fatty acids induce apoptosis and necrosis of J774 cells. At high concentrations, fatty acids cause macrophage death mainly by necrosis. The cytotoxicity of the fatty acids was not strictly related to the number of double bonds in the molecules: palmitic acid>docosahexaenoic acid>stearic acid=eicosapentaenoic acid=arachidonic acid>oleic acid>linoleic acid. The induction of cell death did not involve PPARγ activation. The mechanisms of fatty acids to induce cell death involved changes in mitochondrial transmembrane potential and intracellular neutral lipid accumulation. Fatty acids poorly incorporated into triacylglycerol had the highest toxicity.

A specific modulatory effect of PUFAs (polyunsaturated fatty acids) on gene expression of some cytokines involved in bone remodelling has been reported previously. In particular, although a direct action of AA (arachidonic acid) on bone cytokine gene expression has been shown in human osteoblastic cells, OA (oleic acid) and EPA (eicosapentaenoic acid) were ineffective. Since the NO (nitric oxide) system has also been shown to have an important modulatory activity on osteoblasts, osteoclasts and bone metabolism, in the present study we have investigated the effects of PUFAs on iNOS (inducible NO synthase) gene expression in a human osteoblast-like cell line. AA induced a significant increase in iNOS mRNA expression, whereas EPA and OA had no stimulatory effects but instead caused a significant inhibition of AA-induced iNOS gene expression. Blocking of the COX (cyclo-oxygenase) pathway did not inhibit AA-induced iNOS expression. AA action was inhibited instead by the addition of calphostin C and genistein, inhibitors of PKC (protein kinase C) and tyrosine kinases respectively. Experiments performed with specific anti-cytokine antibodies showed a significant decrease in iNOS expression in AA-treated osteoblastic cells, suggesting that both cytokine-dependent and -independent mechanisms account for the effects of AA on iNOS gene expression. In conclusion, our investigation clearly shows specific effects of PUFAs on iNOS expression in human osteoblast-like cells with a cytokine-dependent and -independent mechanism. These results might have clinical relevance and are of interest for understanding the reported beneficial effects of dietary PUFA manipulation on the prevention and/or treatment of primary and secondary bone disease.

Insulin-resistant states such as obesity can result in an increase in the function and mass of pancreatic β-cells, so that insulin secretion is up-regulated and Type II diabetes does not develop. However, expansion of β-cell mass is not indefinite and may well decrease with time. Changes in circulating concentrations of nutritional factors, such as fatty acids and/or glucose, may lead to a reduction in β-cell mass in vivo . Few previous studies have attempted to explore the interplay between glucose, amino acids and fatty acids with respect to β-cell mass and functional integrity. In the present study, we demonstrate that culture of clonal BRIN-BD11 cells for 24 h with the polyunsaturated fatty acid arachidonic acid (AA) increased β-cell proliferation and enhanced alanine-stimulated insulin secretion. These effects of AA were associated with significant decreases in the cellular consumption of D -glucose and L -alanine as well as decreased rates of production of nitric oxide and ammonia. Conversely 24 h exposure to the saturated fatty acid palmitic acid (PA) was found to decrease β-cell viability (by increasing apoptosis), increase the intracellular concentration of triacylglycerol (triglyceride), while inhibiting alanine-stimulated insulin secretion. These effects of PA were associated with significant increases in D -glucose and L -glutamine consumption as well as nitric oxide and ammonia production. However, L -alanine consumption was decreased in the presence of PA. The effects of AA, but not PA, were additionally dependent on glucose concentration. These studies indicate that AA may have a critical role in maintaining the appropriate mass and function of islet β-cells by influencing rates of cell proliferation and insulin secretion. This regulatory effect may be compromised by high circulating levels of glucose and/or PA, both of which are elevated in Type II diabetes and may impact upon dysfunctional and apoptotic intracellular events in the β-cell.

We report here that monounsaturated fatty acids and polyunsaturated fatty acids (PUFAs) provoke the accumulation of neutral lipids and apoptosis in retinoic acid-treated HL-60 cells in a concentration- and time-dependent manner. The PUFAs (arachidonic acid, docosahexanoic acid and eicosapentaenoic acid) provoked higher levels of HL-60 apoptosis compared with the monounsaturated oleic acid or the saturated palmitic acid. Cell size and granularity were also altered by fatty acid treatment. The PUFA-induced apoptosis was correlated with increased activity of caspase 3 and caspase 9. Lipid peroxidation was also increased in the presence of PUFAs, but was not responsible for activating cell apoptosis. Lipid derived metabolites may be responsible for activation of caspases and induction of cell apoptosis.