The transplantation of the gut microbiome of fat-1 mice protects against colonic mucus layer disruption and endoplasmic reticulum stress induced by high fat diet.

High-fat diets alter gut barrier integrity, leading to endotoxemia by impacting epithelial functions and inducing endoplasmic reticulum (ER) stress in intestinal secretory goblet cells. Indeed, ER stress, which is an important contributor to many chronic diseases such as obesity and obesity-related disorders, leads to altered synthesis and secretion of mucins that form the protective mucus barrier. In the present study, we investigated the relative contribution of omega-3 polyunsaturated fatty acid (PUFAs)-modified microbiota to alleviating alterations in intestinal mucus layer thickness and preserving gut barrier integrity. Male fat-1 transgenic mice (exhibiting endogenous omega-3 PUFAs tissue enrichment) and wild-type (WT) littermates were fed either an obesogenic high-fat diet (HFD) or a control diet. Unlike WT mice, HFD-fed fat-1 mice were protected against mucus layer alterations as well as an ER stress-mediated decrease in mucin expression. Moreover, cecal microbiota transferred from fat-1 to WT mice prevented changes in the colonic mucus layer mainly through colonic ER stress downregulation. These findings highlight a novel feature of the preventive effects of omega-3 fatty acids against intestinal permeability in obesity-related conditions.

Monitoring HSP70 exosomes in cancer patients' follow up: a clinical prospective pilot study.

Exosomes are nanovesicles released by all cells that can be found in the blood. A key point for their use as potential biomarkers in cancer is to differentiate tumour-derived exosomes from other circulating nanovesicles. Heat shock protein-70 (HSP70) has been shown to be abundantly expressed by cancer cells and to be associated with bad prognosis. We previously showed that exosomes derived from cancer cells carried HSP70 in the membrane while those from non-cancerous cells did not. In this work, we opened a prospective clinical pilot study including breast and lung cancer patients to determine whether it was possible to detect and quantify HSP70 exosomes in the blood of patients with solid cancers. We found that circulating exosomal HSP70 levels, but not soluble HSP70, reflected HSP70 content within the tumour biopsies. Circulating HSP70 exosomes increased in metastatic patients compared to non-metastatic patients or healthy volunteers. Further, we demonstrated that HSP70-exosome levels correlated with the disease status and, when compared with circulating tumour cells, were more sensitive tumour dissemination predictors. Finally, our case studies indicated that HSP70-exosome levels inversely correlated with response to the therapy and that, therefore, monitoring changes in circulating exosomal HSP70 might be useful to predict tumour response and clinical outcome.

Polyphosphoinositides are enriched in plant membrane rafts and form microdomains in the plasma membrane.

In this article, we analyzed the lipid composition of detergent-insoluble membranes (DIMs) purified from tobacco (Nicotiana tabacum) plasma membrane (PM), focusing on polyphosphoinositides, lipids known to be involved in various signal transduction events. Polyphosphoinositides were enriched in DIMs compared with whole PM, whereas all structural phospholipids were largely depleted from this fraction. Fatty acid composition analyses suggest that enrichment of polyphosphoinositides in DIMs is accompanied by their association with more saturated fatty acids. Using an immunogold-electron microscopy strategy, we were able to visualize domains of phosphatidylinositol 4,5-bisphosphate in the plane of the PM, with 60% of the epitope found in clusters of approximately 25 nm in diameter and 40% randomly distributed at the surface of the PM. Interestingly, the phosphatidylinositol 4,5-bisphosphate cluster formation was not significantly sensitive to sterol depletion induced by methyl-beta-cyclodextrin. Finally, we measured the activities of various enzymes of polyphosphoinositide metabolism in DIMs and PM and showed that these activities are present in the DIM fraction but not enriched. The putative role of plant membrane rafts as signaling membrane domains or membrane-docking platforms is discussed.

Metabolism of fatty acid in yeast: addition of reducing agents to the reaction medium influences beta-oxidation activities, gamma-decalactone production, and cell ultrastructure in Sporidiobolus ruinenii cultivated on ricinoleic acid methyl ester.

The sensitivity of Sporidiobolus ruinenii yeast to the use of reducing agents, reflected in changes in the oxidoreduction potential at pH 7 (Eh7) environment, ricinoleic acid methyl ester catabolism, gamma-decalactone synthesis, cofactor level, beta-oxidation activity, and ultrastructure of the cell, was studied. Three environmental conditions (corresponding to oxidative, neutral, and reducing conditions) were fixed with the use of air or air and reducing agents (hydrogen and dithiothreitol). Lowering Eh7 to neutral conditions (Eh7 = +30 mV and +2.5 mV) favoured the production of lactone more than the more oxidative condition (Eh7 = +350 mV). In contrast, when a reducing condition was used (Eh7 = -130 mV), the production of gamma-decalactone was very low. These results were linked to changes in the cofactor ratio during lactone production, to the beta-oxidation activity involved in decanolide synthesis, and to ultrastructural modification of the cell.

Dietary oxysterols induce in vivo toxicity of coronary endothelial and smooth muscle cells.

Dietary cholesterol oxidation products (COPs) were reported to exhibit in vitro toxicity toward vascular cells. The aim of this study was to determine whether dietary COPs induce in vivo toxicity toward coronary arteries and to evaluate their effect on the coronary reactivity. Golden Syrian hamsters were fed either a normolipidic diet or a hyperlipidic diet with or without a mixture of COPs (1.4 mg/kg/day). At the end of the feeding periods, cardiac mitochondria and cytosol were prepared to determine the subcellular distribution of cytochrome c. Oxidative phosphorylation was evaluated with glutamate, pyruvate or palmitoylcarnitine as a substrate. The main coronary artery was examined all along its length by transmission electron microscopy (TEM). Plasma sterol concentrations were determined. Furthermore, at the end of the 3-month feeding period, the hearts were perfused at constant pressure by the Langendorff method. The endothelium-dependent reactivity to acetylcholine was evaluated. The myocardial sterol concentration was also estimated. After a 15-day diet with dietary COPs, a release of cytochrome c into the cytosolic fraction of the whole heart occurred, which indicated apoptosis of one or several types of cardiac cells probably induced by excess circulating cholestanetriol. The morphological data obtained by TEM after three months of diet suggested that mainly vascular cells (endothelial and smooth muscle cells) were damaged by dietary COPs, whereas cardiomyocytes appeared healthy. Furthermore, the mitochondrial oxidation of palmitoylcarnitine was reduced and that of pyruvate was increased, suggesting some maintenance of energy metabolism. This strengthens the hypothesis of apoptosis. Several changes in coronary reactivity suggesting an increased NO production were observed. In conclusion, dietary COPs triggered in vivo apoptosis of coronary cells through the release of cytochrome c in the cytosol. This toxicity was counterbalanced by an increased endothelium-dependent dilation.

Effects of dietary oxysterols on coronary arteries in hyperlipidaemic hamsters.

The aim of this study was to evaluate the effect of dietary oxysterols on coronary atherosclerosis and vasospasm. Golden Syrian hamsters were fed three diets with different lipid contents for 3 months: (1) a normolipidaemic diet containing 25 g corn oil-fish oil (4:1, w/w)/kg (group Low L); (2) a hyperlipidaemic diet composed of the normolipidaemic diet supplemented with 150 g lard+30 g cholesterol/kg (group High L); (3) a third diet, similar to the hyperlipidaemic diet, in which 4 g cholesterol/kg was replaced by a mixture of oxysterols (group High L+OS). The oxysterol mixture contained (g/kg): 5,6alpha-epoxycholesterol 211, 5,6beta-epoxycholesterol 179, 7alpha-hydroxycholesterol 67, 7beta-hydroxycholesterol (7betaOH) 185, 7-ketocholesterol (7 K) 235; and trace amounts of 7-hydroperoxycholesterols (approximately 30 g/kg). Atherosclerosis was evaluated by measuring myocardial Ca, oxysterols and acyl-CoA cholesterol acyl transferase (ACAT) activity; furthermore, coronary reactivity to sodium nitroprusside was measured and the morphology of coronary arteries was visualized by transmission electron microscopy. Coronary spasm was determined by evaluating reactivity to serotonin. Feeding the high-lipid diet (group High L) increased the plasma level of 7betaOH, 7 K and cholestanetriol. The presence of oxysterols in the diet (group High L+OS) further increased the concentrations of 7betaOH and 7 K in the plasma. However, as evidenced by myocardial Ca, ACAT activity and coronary reactivity to sodium nitroprusside, severe atherosclerosis did not develop during the 3-month diet. 7 K was increased in myocardial lipids of groups High L and High L+OS. Electron microscopy did not show the development of atherosclerosis in group High L, whereas vascular wall thickening, endothelial damage and smooth muscle cell proliferation and migration occurred when oxysterols were present in the food. Serotonin induced exacerbated coronary vasoconstriction in group High L that was completely reversed by dietary oxysterols. In conclusion, dietary oxysterols exhibit anti-spasmodic properties, but they cannot be used as agents against excess dietary lipid-induced coronary spasm because of their atherogenic properties.