Selective mobilisation of fatty acids from human adipose tissue.

Background: Adipose tissue is a storage organ for dietary fat. During fasting, fatty acids are released into serum as free fatty acids (FFA). Experimental studies indicate that fatty acids are selectively mobilised from adipose tissue into serum. The aim of this study was to investigate whether the composition of the serum FFA fraction reflects selective mobilisation in the fasting state in humans. Methods: The fatty acid composition of fasting serum FFA and adipose tissue were analysed from 112 patients with myocardial infarction and 107 healthy control subjects using gas-liquid chromatography. The subjects' habitual diet was analysed using a food-frequency questionnaire. Results: Significant correlations were found between serum FFA and adipose tissue, particularly for the percentage content of linoleic acid (r=0.73), eicosapentaenoic acid (r=0.68), alpha-linolenic acid (r=0.67) and palmitoleic acid (r=0.60). Percentage contents of palmitic, stearic, linoleic, alpha-linolenic, eicosapentaenoic and docosahexaenoic acid were higher in serum FFA than in adipose tissue, whereas oleic and palmitoleic acid were relatively more abundant in adipose tissue. This may indicate that the former group of fatty acids is preferentially mobilised from adipose tissue into serum. High correlations for polyunsaturated fatty acids were observed between percentage contents of dietary and adipose tissue fatty acids. The correlation of fatty acids between diet and serum FFA was weak, but a tendency towards higher correlations for polyunsaturated fatty acids was observed. Conclusions: Our findings are compatible with the hypothesis that, in the fasting state, fatty acids are selectively mobilised from adipose tissue into serum FFA.

Adipose tissue fatty acids and risk of myocardial infarction--a case-control study.

OBJECTIVES: To study the association between content in adipose tissue of very long-chain n-3 fatty acids, trans fatty acids, linoleic acid and alpha-linolenic acid and risk of a first myocardial infarction. DESIGN AND SUBJECTS: A case-control design among 100 patients and 98 population controls both men and postmenopausal women, age 45-75 y. Adipose tissue fatty acids were determined by gas-liquid chromatography. Intake data were obtained through interview using a validated food frequency questionnaire. RESULTS: Dietary intake and adipose tissue content of the fatty acids studied correlated significantly. Adipose tissue contents of eicosapentaenoic acid (20:5n-3), docosapentaenoic acid (22:5n-3) and docosahexaenoic acid (22:6n-3) were significantly lower while those of trans fatty acids, linoleic and alpha-linolenic acid were significantly higher in patients than in controls. Age and sex adjusted odds ratios (OR) were significantly reduced with increasing quintiles of very long-chain n-3 fatty acids, thus the OR in the fifth compared to the first quintile was 0.23 (95% CI 0.08-0.70). After further adjustment for waist-to-hip ratio, smoking, family history of CHD and content of trans fatty acids, the OR in the highest quintile was 0.17 (95% CI 0.04-0.76) and the P for trend 0. 016. Age and sex adjusted OR was increased in the fifth compared to the first quintile of trans fatty acids (OR 2.81, 95% CI 1.16-6.84), linoleic acid (OR 2.10, 95% CI 0.87-5.07) and alpha-linolenic acid (OR 1.96, 95% CI 0.83-4.61), and P for trend was 0.002, 0.005 and 0. 020, respectively. The trends remained significant after adjustment for waist-to-hip ratio, smoking, and family history of coronary heart disease. Trans fatty acids, linoleic acid and alpha-linolenic acid in adipose tissue were strongly correlated, indicating a common source, most likely margarine. When each of these fatty acid species were adjusted for the two others the trends were no longer significant. CONCLUSION: Intake of very long-chain n-3 fatty acids as reflected in adipose tissue content is inversely associated with risk of myocardial infarction. Trans fatty acids, linoleic and alpha-linolenic acid were intercorrelated and associated with increased risk. It is suggested that the increased risk may be connected to trans fatty acids or to some other factor associated with margarine consumption. European Journal of Clinical Nutrition 54, 618-625.

Production of oxidants in alveolar macrophages and blood leukocytes.

Increased production of oxidants subsequent to phagocyte stimulation has been associated with tissue damage in lung inflammatory disorders. The overall oxidative burden of the lung may vary with inflammatory cell composition. Flow cytometry using three different dyes, dihydroethidium (DHE), dichlorofluorescein diacetate (DCFH-DA) and dihydrorhodamine 123 (DHR), all compounds that by interaction with oxidants are transformed to fluorescent products, was used to examine the production of intracellular oxidants in alveolar macrophages (AMs), including size-defined subpopulations, monocytes (Ms) and polymorphonuclear neutrophils (PMNs) during in vitro incubation in the presence or absence of phorbol myristate acetate (PMA). PMA stimulation led to slightly increased (two-fold) (p<0.05) DHE-induced fluorescence in AMs, whereas it was greatly increased in Ms and PMNs (13-fold and 113-fold, respectively). The levels of DCFH-DA- and DHR-induced fluorescence were significantly (p<0.05) increased (four-fold and 110-fold, respectively) by PMA stimulation of PMNs, but not of AMs and Ms. Significant differences (p<0.05) in the levels of DHE- and DCFH-DA-induced fluorescence in small and large AMs were also demonstrated. The results show that the potential to increase the generation of various oxidants upon stimulation was: PMNs> Ms>AMs, suggesting that the total oxidative burden of the lungs is dependent on the type of inflammatory cells present, as well as on their state of activation.

Modulation of adhesion molecule profiles on alveolar macrophages and blood leukocytes.

BACKGROUND: Cell adhesion molecules are believed to be essential for blood cell recruitment to the lung and for the movement of alveolar macrophages (AM) within the lung. OBJECTIVE: To investigate the expression pattern of L-selectin and beta(2) integrins on blood leukocytes and AM, including AM of various maturity. METHODS: Flow cytometry was used to study the expression of L-selectin (CD62L) and of the beta(2) integrins CD11a, CD11b, and CD11c on AM (including density-defined subpopulations), monocytes (Mo), polymorphonuclear neutrophils (PMN) and lymphocytes (Ly) sampled from healthy individuals, during incubation with and without lipopolysaccharide (LPS). RESULTS: A significantly different modulation pattern of beta(2) integrins and L-selectin was demonstrated on Mo and AM, cells of the same differentiation lineage. In contrast to AM, Mo had a marked ability to respond to LPS stimulation by increased expression of CD11a, CD11b and CD11c and decreased expression of L- selectin. These molecules were expressed to a similar degree on AM, whereas the basal levels of CD11b and L-selectin were considerably higher on Mo than on AM. A significantly different expression of CD11a as well as differences in the regulation of L-selectin during incubation were also demonstrated between density-defined subpopulations of AM. CD11a could not be upregulated on PMN, otherwise the modulation patterns of CD11b, CD11c and L-selectin were similar to that on Mo. The expression of CD11a on Ly was 3- to 6-fold higher than on Mo, PMN and AM. The level of CD11b decreased significantly upon incubation (uninfluenced by LPS stimulation), and CD11c was hardly expressed on Ly. The level of L-selectin on Ly was higher than on Mo, AM and PMN and was not decreased during incubation. CONCLUSION: Developmental origin, degree of cell differentiation (maturity) as well as different environmental conditions all heavily influence the expression and modulation pattern of beta(2) integrins and L-selectin on leukocytes and Mo-derived AM.

Heterogeneity of procoagulant activity and cytokine release in subpopulations of alveolar macrophages and monocytes.

We have studied the expression of tissue factor (TF) and fibrinopeptide A (FPA) generation as well as the release capacity of TNF-alpha, IL-1beta, and IL-6 in density-defined subpopulations of alveolar macrophages (AM) and monocytes (Mo). TF was equally expressed on all AM subpopulations and Mo, while the FPA-forming capacity was at the same level in low density AM as in Mo and was significantly (P < 0.05) higher in low density AM than in high density AM. The lipopolysaccharide (LPS)-induced release of TNF-alpha was higher (P < 0.05) in high density AM than in low density AM and in Mo. IL-1beta release was undetectable in unstimulated AM and in LPS-stimulated low density AM, while the LPS-induced IL-1beta release in high density AM was low compared to the levels demonstrated in Mo. LPS-stimulated IL-6 release was not distinctively different in the AM subpopulations and Mo. The presented study showed that FPA generation and LPS-stimulated TNF-alpha release were dependent on the density (i.e., maturity) of AM. This implies that a skewed distribution of AM subpopulations induced by disease processes may profoundly influence the inflammatory reactions, including extravascular activation of coagulation.

CD14 expression and binding of lipopolysaccharide to alveolar macrophages and monocytes.

We have studied the expression of the lipopolysaccharide (LPS) receptor CD14 on monocytes (Mo) and alveolar macrophages (AM), including density- and size-defined subpopulations. Bronchoalveolar lavage (BAL) was performed on eleven healthy non-smokers and blood sampled from 5 of them, and the levels of cell CD14 expression was investigated using flow cytometry. The influence of LPS stimulation on the CD14 expression of AM was studied at various intervals during prolonged incubation. Further, the relationship between CD14 expression and LPS binding to Mo and subpopulations of AM was studied by measuring fluorescein isothiocyanate (FITC)-LPS binding (flow cytometry) and binding of radioiodinated LPS (125I-LPS). The CD14 expression was 13-fold higher (P < 0.02) on Mo than on unfractionated and high density AM. The CD14 level on the latter was higher than on low density AM, and also higher (P < 0.05) on small AM compared to large (flow cytometrically defined) AM. LPS stimulation had a downregulating effect on AM CD14 level, but after several hours of continuing decreased expression, an increased (P < 0.05) CD14 expression was demonstrated, indicating de novo synthesis. The binding of LPS to subpopulations of AM and isolated Mo was not significantly different, but the binding of FITC-LPS to Mo in whole blood was higher than to AM (P < 0.02). The presented results indicate that AM of different size and maturity have different and variable (activation dependent) CD14 levels. The LPS binding capacity was, however, not proportional to the CD14 expression, indicating that LPS binding mechanisms unrelated to CD14 levels were also operable.