Phospholipase PLA2G7 is complementary to GPX4 in mitigating punicic-acid-induced ferroptosis in prostate cancer cells.

Ferroptosis is a cell death pathway that can be promoted by peroxidizable polyunsaturated fatty acids in cancer cells. Here, we investigated the mechanisms underlying the toxicity of punicic acid (PunA), an isomer of conjugated linolenic acids (CLnAs) bearing three conjugated double bonds highly prone to peroxidation, on prostate cancer (PCa) cells. PunA induced ferroptosis in PCa cells and triggered massive lipidome remodeling, more strongly in PC3 androgen-negative cells than in androgen-positive cells. The greater sensitivity of androgen-negative cells to PunA was associated with lower expression of glutathione peroxidase 4 (GPX4). We then identified the phospholipase PLA2G7 as a PunA-induced ferroptosis suppressor in PCa cells. Overexpressing PLA2G7 decreased lipid peroxidation levels, suggesting that PLA2G7 hydrolyzes hydroperoxide-containing phospholipids, thus preventing ferroptosis. Importantly, overexpressing both PLA2G7 and GPX4 strongly prevented PunA-induced ferroptosis in androgen-negative PCa cells. This study shows that PLA2G7 acts complementary to GPX4 to protect PCa cells from CLnA-induced ferroptosis.

Dietary methylmercury and fatty acids affect the lipid metabolism of adipose tissue and liver in rainbow trout.

Methylmercury (MeHg) is a pervasive environmental contaminant in aquatic ecosystems that can reach elevated concentrations in fish of high trophic levels, such as salmonids. The present study aims at investigating the individual and combined impacts of dietary MeHg and fatty acids on lipid metabolism in juvenile rainbow trout (Oncorhynchus mykiss) with a focus on two key organs, adipose tissue and liver. MeHg and fatty acids are both known to act on energy homeostasis although little is known about their interplay on lipid metabolism in fish. Fish were fed diets enriched in linoleic acid (LA, 18:2 n-6), α-linolenic acid (ALA, 18:3 n-3), eicosapentaenoic acid (EPA, 20:5 n-3) or docosahexaenoic acid (DHA, 22:6 n-3) for ten weeks, with the addition of MeHg to the diets during the last six weeks (0, 2.4 or 5.5 mg MeHg/kg dry matter). LA and ALA are polyunsaturated fatty acids (PUFA) typical of plant-derived oils whereas EPA and DHA are n-3 long chain PUFA largely found in fish oil, all used in feed formulation in aquaculture. The results showed that the LA-enriched diet induced a higher whole-body lipid content compared to the three other diets. On the contrary, the addition of MeHg led to a significant reduction of the whole-body lipid content, regardless of the diet. Interestingly, the adipocytes were larger both in presence of LA, compared to EPA and DHA, or MeHg, indicating a lipogenic effect of these two compounds. No effect was, however, observed on lipid accumulation per gram of adipose tissue. The fatty acid composition of adipose tissue and liver was significantly modified by the dietary lipids, reflecting both the fatty acid composition of the diets and the high bioconversion capacity of the rainbow trout. Exposure to MeHg selectively led to a release of n-6 PUFA from the hepatic membranes of fish fed the LA-enriched diet, showing a disruption of the pathways using n-6 PUFA. This study highlights the significant impact of MeHg exposure and dietary fatty acids on lipid metabolism in fish. Further investigation is needed to elucidate the underlying mechanisms and to explore the potential involvement of other organs.

Beyond the Lab: What We Can Learn about Cancer from Wild and Domestic Animals.

Cancer research has benefited immensely from the use of animal models. Several genetic tools accessible in rodent models have provided valuable insight into cellular and molecular mechanisms linked to cancer development or metastasis and various lines are available. However, at the same time, it is important to accompany these findings with those from alternative or non-model animals to offer new perspectives into the understanding of tumor development, prevention, and treatment. In this review, we first discuss animals characterized by little or no tumor development. Cancer incidence in small animals, such as the naked mole rat, blind mole rat and bats have been reported as almost negligible and tumor development may be inhibited by increased defense and repair mechanisms, altered cell cycle signaling and reduced rates of cell migration to avoid tumor microenvironments. On the other end of the size spectrum, large animals such as elephants and whales also appear to have low overall cancer rates, possibly due to gene replicates that are involved in apoptosis and therefore can inhibit uncontrolled cell cycle progression. While it is important to determine the mechanisms that lead to cancer protection in these animals, we can also take advantage of other animals that are highly susceptible to cancer, especially those which develop tumors similar to humans, such as carnivores or poultry. The use of such animals does not require the transplantation of malignant cancer cells or use of oncogenic substances as they spontaneously develop tumors of similar presentation and pathophysiology to those found in humans. For example, some tumor suppressor genes are highly conserved between humans and domestic species, and various tumors develop in similar ways or because of a common environment. These animals are therefore of great interest for broadening perspectives and techniques and for gathering information on the tumor mechanisms of certain types of cancer. Here we present a detailed review of alternative and/or non-model vertebrates, that can be used at different levels of cancer research to open new perspectives and fields of action.

The Catechins Profile of Green Tea Extracts Affects the Antioxidant Activity and Degradation of Catechins in DHA-Rich Oil.

This study investigated the effect of the catechins profile on the antioxidant activity of green tea extracts (GTEs) by comparing the antioxidant activity of an EGC-rich GTE (GTE1, catechin content: 58% EGC, 30.1% EGCG, 7.9% EC, and 3.9% ECG) and an EGCG-rich GTE (GTE2, catechin content: 60.6% EGCG, 17.7% EGC, 11.8% ECG, and 9.8% EC) in a DHA-rich oil. The effects of the individual catechins (EGC, EC, EGCG, and ECG) and reconstituted catechins mixtures (CatMix), prepared to contain the same amount of major catechins as in the GTEs, were also measured. All treatments (GTE1, CatMix1, GTE2, CatMix2, EGC250, EC250, EGCG250, and ECG250), each containing epistructured catechins at a concentration of 250 ppm, as well as the control (oil with no added antioxidant), were stored at 30 °C for 21 days with sampling intervals of 7 days. The antioxidant activity was assessed by measuring the peroxide value (PV) and p-anisidine value (p-AV) of oils. Changes in fatty acid content and catechins content were also monitored. Both GTEs enhanced the oxidative stability of the DHA-rich oil, but GTE1 demonstrated a stronger antioxidant activity than GTE2. No significant difference was observed between the PV of treatments with GTE1 and CatMix1 during storage, whereas the PV of oil with GTE2 was significantly higher than that with CatMix2 after 21 days. Among the individual catechins, EGC was the strongest antioxidant. Overall, the antioxidant activities of the extracts and catechins were observed in the decreasing order GTE1 ≈ EGC250 ≈ CatMix1 > GTE2 > EGCG250 ≈ CatMix2 > ECG250 > EC250. A significant change in fatty acid content was observed for the control and EC250 samples, and the catechins were most stable in GTE1-supplemented oil. Our results indicate that the EGC-rich GTE is a more potent antioxidant in DHA-rich oil than the EGCG-rich GTE.

Contribution of Membrane Vesicle to Reprogramming of Bacterial Membrane Fluidity in Pseudomonas aeruginosa.

Pseudomonas aeruginosa is an opportunistic pathogen capable of resisting environmental insults by applying various strategies, including regulating membrane fluidity and producing membrane vesicles (MVs). This study examined the difference in membrane fluidity between planktonic and biofilm modes of growth in P. aeruginosa and whether the ability to alter membrane rigidity in P. aeruginosa could be transferred via MVs. To this end, planktonic and biofilm P. aeruginosa were compared with respect to the lipid composition of their membranes and their MVs and the expression of genes contributing to alteration of membrane fluidity. Additionally, viscosity maps of the bacterial membrane in planktonic and biofilm lifestyles and under the effect of incubation with bacterial MVs were obtained. Further, the growth rate and biofilm formation capability of P. aeruginosa in the presence of MVs were compared. Results showed that the membrane of the biofilm bacteria is significantly less fluid than the membrane of the planktonic bacteria and is enriched with saturated fatty acids. Moreover, the enzymes involved in altering the structure of existing lipids and favoring membrane rigidification are overexpressed in the biofilm bacteria. MVs of biofilm P. aeruginosa elicit membrane rigidification and delay the bacterial growth in the planktonic lifestyle; conversely, they enhance biofilm development in P. aeruginosa. Overall, the study describes the interplay between the planktonic and biofilm bacteria by shedding light on the role of MVs in altering membrane fluidity. IMPORTANCE Membrane rigidification is a survival strategy in Pseudomonas aeruginosa exposed to stress. Despite various studies dedicated to the mechanism behind this phenomenon, not much attention has been paid to the contribution of the bacterial membrane vesicles (MVs) in this regard. This study revealed that P. aeruginosa rigidifies its membrane in the biofilm mode of growth. Additionally, the capability of decreasing membrane fluidity is transferable to the bacterial population via the bacterial MVs, resulting in reprogramming of bacterial membrane fluidity. Given the importance of membrane rigidification for decreasing the pathogen's susceptibility to antimicrobials, elucidation of the conditions leading to such biophysicochemical modulation of the P. aeruginosa membrane should be considered for the purpose of developing therapeutic approaches against this resistant pathogen.

Influence of Proteins on the Absorption of Lipophilic Vitamins, Carotenoids and Curcumin - A Review.

While proteins have been widely used to encapsulate, protect, and regulate the release of bioactive food compounds, little is known about the influence of co-consumed proteins on the absorption of lipophilic constituents following digestion, such as vitamins (A, D, E, K), carotenoids, and curcumin. Their bioavailability is often low and very variable, depending on the food matrix and host factors. Some proteins can act as emulsifiers during digestion. Their liberated peptides have amphiphilic properties that can facilitate the absorption of microconstituents, by improving their transition from lipid droplets into mixed micelles. Contrarily, the less well digested proteins could negatively impinge on enzymatic accessibility to the lipid droplets, slowing down their processing into mixed micelles and entrapping apolar food compounds. Interactions with mixed micelles and proteins are also plausible, as shown earlier for drugs. This review focuses on the ability of proteins to act as effective emulsifiers of lipophilic vitamins, carotenoids, and curcumin during digestion. The functional properties of proteins, their chemical interactions with enzymes and food constituents during gastro-intestinal digestion, potentials and limitations for their use as emulsifiers are emphasized and data from human, animal, and in vitro trials are summarized.

Whey- and Soy Protein Isolates Added to a Carrot-Tomato Juice Alter Carotenoid Bioavailability in Healthy Adults.

Recent findings suggested that proteins can differentially affect carotenoid bioaccessibility during gastro-intestinal digestion. In this crossover, randomized human trial, we aimed to confirm that proteins, specifically whey- and soy-protein isolates (WPI/SPI) impact postprandial carotenoid bioavailability. Healthy adults (n = 12 males, n = 12 females) were recruited. After 2-week washout periods, 350 g of a tomato-carrot juice mixture was served in the absence/presence of WPI or SPI (50% of the recommended dietary allowance, RDA ≈ 60 g/d). Absorption kinetics of carotenoids and triacylglycerols (TAGs) were evaluated via the triacylglycerol-rich lipoprotein (TRL) fraction response, at timed intervals up to 10 h after test meal intake, on three occasions separated by 1 week. Maximum TRL-carotenoid concentration (Cmax) and corresponding time (Tmax) were also determined. Considering both genders and carotenoids/TAGs combined, the estimated area under the curve (AUC) for WPI increased by 45% vs. the control (p = 0.018), to 92.0 ± 1.7 nmol × h/L and by 57% vs. SPI (p = 0.006). Test meal effect was significant in males (p = 0.036), but not in females (p = 0.189). In males, significant differences were found for phytoene (p = 0.026), phytofluene (p = 0.004), α-carotene (p = 0.034), and ß-carotene (p = 0.031). Cmax for total carotenoids (nmol/L ± SD) was positively influenced by WPI (135.4 ± 38.0), while significantly lowered by SPI (89.6 ± 17.3 nmol/L) vs. the control (119.6 ± 30.9, p < 0.001). Tmax did not change. The results suggest that a well-digestible protein could enhance carotenoid bioavailability, whereas the less digestible SPI results in negative effects. This is, to our knowledge, the first study finding effects of proteins on carotenoid absorption in humans.

Green Synthesis of Iron-Doped Cobalt Oxide Nanoparticles from Palm Kernel Oil via Co-Precipitation and Structural Characterization.

In this study, a bio-derived precipitating agent/ligand, palm kernel oil, has been used as an alternative route for the green synthesis of nanoparticles of Fe-doped Co3O4 via the co-precipitation reaction. The palm oil was extracted from dried palm kernel seeds by crushing, squeezing and filtration. The reaction of the palm kernel oil with potassium hydroxide, under reflux, yielded a solution containing a mixture of potassium carboxylate and excess hydroxide ions, irrespective of the length of saponification. The as-obtained solution reacts with an aqueous solution containing iron and cobalt ions to yield the desired metallo-organic precursor, iron cobalt carboxylate. Characterization of the precursors by IR and gas chromatography (GC) attests to the presence of carboxylate fatty acids in good agreement with the proportion contained in the oil, and ICP confirms that the metallic ratios are in the proportion used during the synthesis. Analysis of the products thermally decomposed between 400 °C and 600 °C by XRD, EDX, TEM and ToF-SIMS, established that cobalt iron oxide nanoparticles (Co(1-x)Fex)3O4 were obtained for x ≤ 0.2 and a nanocomposite material (Co(1-x)Fex)3O4/Fe3O4 for x ≥ 0.2, with sizes between 22 and 9 nm. ToF-SIMS and XRD provided direct evidence of the progressive substitution of cobalt by iron in the Co3O4 crystal structure for x ≤ 0.2.

Gastric lipase can significantly increase lipolysis and carotenoid bioaccessibility from plant food matrices in the harmonized INFOGEST static in vitro digestion model.

Gastrointestinal digestion of carotenoids has received much attention, as these lipophilic compounds have been related to several health benefits. Most commonly, static digestion models such as the consensus INFOGEST model are employed to study their bioaccessibility from test matrices. However, an aspect that has been much neglected is the use of gastric lipase. Its inclusion to gastro-intestinal (GI) digestion is expected to foster emulsification of lipophilic constituents prior to their incorporation into mixed micelles. In this study, we compared the effect of various lipases from R. niveus, R. oryzae, and rabbit gastric extracts (RGE), at different concentrations (0, 30, and 60 U mL-1), on carotenoid bioaccessibility from several food matrices (tomato juice, spinach, and carrot juice). We also investigated whether co-digestion of pure proteins (whey and soy protein isolates) at 0, 25, and 50% of the equivalent recommended dietary allowance, would interact with carotenoid bioaccessibility in presence or absence of RGE. Lipolysis was also studied. Considering all matrices combined, lipases significantly improved the bioaccessibility of carotenoids (p < 0.001). Compared to other lipases, RGE consistently increased carotenoid bioaccessibility in all tested matrices, by up to 182% (p < 0.001), this effect was partly maintained in the presence of co-digested proteins. Unexpectedly, all 3 lipases improved gastric lipolysis in all matrices, by an average of 10-fold (p < 0.001). In conclusion, only RGE contributed significantly to improving both lipolysis extent and carotenoid bioaccessibility in all tested matrices, while the presence of proteins mitigated the positive effect of lipases on carotenoid bioaccessibility.

Punicic Acid Triggers Ferroptotic Cell Death in Carcinoma Cells.

Plant-derived conjugated linolenic acids (CLnA) have been widely studied for their preventive and therapeutic properties against diverse diseases such as cancer. In particular, punicic acid (PunA), a conjugated linolenic acid isomer (C18:3 c9t11c13) present at up to 83% in pomegranate seed oil, has been shown to exert anti-cancer effects, although the mechanism behind its cytotoxicity remains unclear. Ferroptosis, a cell death triggered by an overwhelming accumulation of lipid peroxides, has recently arisen as a potential mechanism underlying CLnA cytotoxicity. In the present study, we show that PunA is highly cytotoxic to HCT-116 colorectal and FaDu hypopharyngeal carcinoma cells grown either in monolayers or as three-dimensional spheroids. Moreover, our data indicate that PunA triggers ferroptosis in carcinoma cells. It induces significant lipid peroxidation and its effects are prevented by the addition of ferroptosis inhibitors. A combination with docosahexaenoic acid (DHA), a known polyunsaturated fatty acid with anticancer properties, synergistically increases PunA cytotoxicity. Our findings highlight the potential of using PunA as a ferroptosis-sensitizing phytochemical for the prevention and treatment of cancer.

Impact of Protein-Enriched Plant Food Items on the Bioaccessibility and Cellular Uptake of Carotenoids.

Carotenoids are lipophilic pigments which have been associated with a number of health benefits, partly related to antioxidant effects. However, due to their poor solubility during digestion, carotenoid bioavailability is low and variable. In this study, we investigated the effect of frequently consumed proteins on carotenoid bioaccessibility and cellular uptake. Whey protein isolate (WPI), soy protein isolate (SPI), sodium caseinate (SC), gelatin (GEL), turkey and cod, equivalent to 0/10/25/50% of the recommended dietary allowance (RDA, approx. 60g/d), were co-digested gastro-intestinally with carotenoid-rich food matrices (tomato and carrot juice, spinach), and digesta further studied in Caco-2 cell models. Lipid digestion, surface tension and microscopic visualization were also carried out. Co-digested proteins positively influenced the micellization of carotenes (up to 3-fold, depending on type and concentration), especially in the presence of SPI (p < 0.001). An increased cellular uptake was observed for xanthophylls/carotenes (up to 12/33%, p < 0.001), which was stronger for matrices with an initially poor carotenoid micellization (i.e., tomato juice, p < 0.001), similar to what was encountered for bioaccessibility. Turkey and cod had a weaker impact. Significant interactions between carotenoids, lipids and proteins were observed during digestion. Co-digested proteins generally improved lipid digestion in all matrices (p < 0.001), especially for carrot juice, though slight decreases were observed for GEL. Protein impact on the surface tension was limited. In conclusion, proteins generally improved both carotenoid bioaccessibility and cellular uptake, depending on the matrices and carotenoid-type (i.e., carotene vs. xanthophylls), which may be relevant under specific circumstances, such as intake of carotenoid-rich food items low in lipids.

Green Tea Extract Enhances the Oxidative Stability of DHA-Rich Oil.

Docosahexaenoic acid (DHA) is one of the most important omega-3 polyunsaturated fatty acids, with proven health-promoting properties. However, oils with a very high content in DHA (DHAO) are extremely susceptible to oxidation, which affects shelf stability and limits incorporation in food products. Green tea extracts (GTE) are potential candidates for the protection of these oils, but their use in such oils has not been previously reported. This study investigated the effect of GTE (160 ppm, 400 ppm, 1000 ppm) and α-tocopherol (80 ppm, 200 ppm, 500 ppm) on the oxidative stability of a DHAO over a 9-week storage at 30 °C. The oxidative status was monitored during storage by the measurement of peroxide value (PV) and p-anisidine value (p-AV). Changes in eicosapentaenoic acid (EPA) and DHA content, as well as in catechins and tocopherol contents, were also evaluated. The addition of GTE enhanced the oxidative stability of DHAO by reducing the formation of peroxides and secondary oxidation products, whereas α-tocopherol had no significant effect on the PV of oil during storage but led to a significantly higher p-AV. The EPA and DHA content of DHAO was stable in GTE-supplemented samples whereas a decrease was observed in the control and α-tocopherol-supplemented samples. GTE also delayed the degradation of tocopherols initially present in the oil, while catechins resulting from the addition of GTE decreased progressively during the storage period.

The Egg Yolk Content in ω-3 and Conjugated Fatty Acids Can Be Sustainably Increased upon Long-Term Feeding of Laying Hens with a Diet Containing Flaxseeds and Pomegranate Seed Oil.

Long-term feeding trials examining the incorporation of conjugated linolenic acids (CLnA) into the diet of laying hens are lacking. In the present study, we compared two diets in sixty-six red Sex-Link hens (33 hens/treatment), fed for 26 weeks. The control diet was high in oleic acid, while the test diet was high in α-linolenic acid (ALA) and punicic acid (PunA). No significant differences were observed between treatments for hens' performance, egg weight and yolk weight. In contrast, dietary ALA and PunA resulted in a significant increase in n-3 PUFA, rumenic acid (RmA) and PunA contents in egg yolk, as well as in the liver, heart, muscle and adipose tissue of the hens. Other conjugated dienes resulting from the metabolism of PunA or RmA also accumulated in the egg yolk and tissues. Unlike DHA, which was exclusively distributed in phospholipids, ALA, RmA and PunA were preferably distributed in triglycerides.

Peroxidation of n-3 and n-6 polyunsaturated fatty acids in the acidic tumor environment leads to ferroptosis-mediated anticancer effects.

Tumor acidosis promotes disease progression through a stimulation of fatty acid (FA) metabolism in cancer cells. Instead of blocking the use of FAs by acidic cancer cells, we examined whether excess uptake of specific FAs could lead to antitumor effects. We found that n-3 but also remarkably n-6 polyunsaturated FA (PUFA) selectively induced ferroptosis in cancer cells under ambient acidosis. Upon exceeding buffering capacity of triglyceride storage into lipid droplets, n-3 and n-6 PUFA peroxidation led to cytotoxic effects in proportion to the number of double bonds and even more so in the presence of diacylglycerol acyltransferase inhibitors (DGATi). Finally, an n-3 long-chain PUFA-rich diet significantly delayed mouse tumor growth when compared with a monounsaturated FA-rich diet, an effect further accentuated by administration of DGATi or ferroptosis inducers. These data point out dietary PUFA as a selective adjuvant antitumor modality that may efficiently complement pharmacological approaches.

Modulations of lipid metabolism and development of the Atlantic salmon (Salmo salar) fry in response to egg-to-fry rearing conditions.

In stocking program, the use of artificial incubation conditions in hatcheries from the fertilisation of eggs to the release of unfed fry could reduce their ability to adapt to the natural environment. This study evaluates the effects of three factors on the fitness and physiology of salmon fry at their emergence, the origin of water (river vs drilling), the type of support in the incubator (support matrix vs plastic sheets) and the type of incubators (Californian vs vertical trays), and compares them to a semi-natural incubation method in river. Key biological functions including nutritional and immune status were compared among experimental conditions using biometric parameters, lipid composition and gene expression analyses. Our findings demonstrated that fry incubated in vertical trays supplied with river water had no significant difference in growth and lipid composition compared to those in semi-natural incubators. Besides, fry incubated on a substrate matrix in Californian trays exhibited phenotypic characteristics closest to those incubated in river. This support matrix improved fish growth, lipid consumption and distribution compared to fry on plastic sheets. Moreover, the large amounts of several PUFAs in these fry could allow a better membrane fluidity ensuring a better adaptation to temperature variation under cold conditions. In addition, drilling water improved the survival rate compared to river water due to lower numbers of fine particles, known to be responsible for the clogging of eggs. To conclude, using a substrate combined with drilling water in artificial incubators could increase fry fitness and its adaption to wild life.

A Three-Month Consumption of Eggs Enriched with ω-3, ω-5 and ω-7 Polyunsaturated Fatty Acids Significantly Decreases the Waist Circumference of Subjects at Risk of Developing Metabolic Syndrome: A Double-Blind Randomized Controlled Trial.

Alpha-linolenic acid (ALA), docosahexaenoic acid (DHA), rumenic acid (RmA), and punicic acid (PunA) are claimed to influence several physiological functions including insulin sensitivity, lipid metabolism and inflammatory processes. In this double-blind randomized controlled trial, we investigated the combined effect of ALA, DHA, RmA and PunA on subjects at risk of developing metabolic syndrome. Twenty-four women and men were randomly assigned to two groups. Each day, they consumed two eggs enriched with oleic acid (control group) or enriched with ALA, DHA, RmA, and PunA (test group) for 3 months. The waist circumference decreased significantly (-3.17 cm; p < 0.001) in the test group. There were no major changes in plasma insulin and blood glucose in the two groups. The dietary treatments had no significant effect on endothelial function as measured by peripheral arterial tonometry, although erythrocyte nitrosylated hemoglobin concentrations tended to decrease. The high consumption of eggs induced significant elevations in plasma low-density lipoprotein (LDL)- and high-density lipoprotein (HDL)-cholesterol (p < 0.001), which did not result in any change in the LDL/HDL ratio in both groups. These results indicate that consumption of eggs enriched with ALA, DHA, RmA and PunA resulted in favorable changes in abdominal obesity without affecting other factors of the metabolic syndrome.

Impaired Cytoskeletal and Membrane Biophysical Properties of Acanthocytes in Hypobetalipoproteinemia - A Case Study.

Familial hypobetalipoproteinemia is a metabolic disorder mainly caused by mutations in the apolipoprotein B gene. In its homozygous form it can lead without treatment to severe ophthalmological and neurological manifestations. In contrast, the heterozygous form is generally asymptomatic but associated with a low risk of cardiovascular disease. Acanthocytes or thorny red blood cells (RBCs) are described for both forms of the disease. However, those morphological changes are poorly characterized and their potential consequences for RBC functionality are not understood. Thus, in the present study, we asked whether, to what extent and how acanthocytes from a patient with heterozygous familial hypobetalipoproteinemia could exhibit altered RBC functionality. Acanthocytes represented 50% of the total RBC population and contained mitoTracker-positive surface patches, indicating the presence of mitochondrial fragments. While RBC osmotic fragility, calcium content and ATP homeostasis were preserved, a slight decrease of RBC deformability combined with an increase of intracellular free reactive oxygen species were observed. The spectrin cytoskeleton was altered, showing a lower density and an enrichment in patches. At the membrane level, no obvious modification of the RBC membrane fatty acids nor of the cholesterol content were detected but the ceramide species were all increased. Membrane stiffness and curvature were also increased whereas transversal asymmetry was preserved. In contrast, lateral asymmetry was highly impaired showing: (i) increased abundance and decreased functionality of sphingomyelin-enriched domains; (ii) cholesterol enrichment in spicules; and (iii) ceramide enrichment in patches. We propose that oxidative stress induces cytoskeletal alterations, leading to increased membrane stiffness and curvature and impaired lipid lateral distribution in domains and spicules. In addition, ceramide- and spectrin-enriched patches could result from a RBC maturation defect. Altogether, the data indicate that acanthocytes are associated with cytoskeletal and membrane lipid lateral asymmetry alterations, while deformability is only mildly impaired. In addition, familial hypobetalipoproteinemia might also affect RBC precursors leading to disturbed RBC maturation. This study paves the way for the potential use of membrane biophysics and lipid vital imaging as new methods for diagnosis of RBC disorders.

Methylmercury displays pro-adipogenic properties in rainbow trout preadipocytes.

Methylmercury (MeHg) is a ubiquitous contaminant largely found in aquatic environments, especially in species at high trophic level such as salmonids. The aim of this study was to evaluate the effects of MeHg on adipocyte differentiation and lipid metabolism in rainbow trout. Primary cultured preadipocytes were exposed to increasing concentrations of MeHg during six days with or without a hormonal cocktail. Main results showed a dose-dependent intracellular accumulation of neutral lipids with a preferential uptake of n-3 polyunsaturated fatty acids. Interestingly, this accumulation occurred after a fairly low uptake of MeHg by preadipocytes and was maintained after the cellular exposure to MeHg. In membrane phospholipids, arachidonic acid (20:4 n-6) was released in a dose-dependent manner. At the transcriptional level, the expression of several adipocyte-specific genes (perilipin 2 and apolipoprotein Eb) as well as lipid-related genes (fatty acid synthase and fatty acid binding protein 11a) was up-regulated in preadipocytes exposed to MeHg. These results highlight for the first time the disrupting effect of MeHg in trout adipocyte metabolism, providing new insights regarding the role of environmental pollutants in adipose tissue dysfunction and related pathologies.

Interplay between dietary lipids and cadmium exposure in rainbow trout liver: Influence on fatty acid metabolism, metal accumulation and stress response.

The present study aimed at investigating interactive effects between dietary lipids and both short- and long-term exposures to a low, environmentally realistic, cadmium (Cd) concentration. Juvenile rainbow trout were fed four isolipidic diets (31.7 g/kg) enriched in either linoleic acid (LA, 18:2n-6), alpha-linolenic acid (ALA, 18:3n-3), eicosapentaenoic acid (EPA, 20:5n-3) or docosahexaenoic acid (DHA, 22:6n-3). From the 4th week of this 10-week experiment, the lipid level of the diet was increased (120.0 g/kg) and half of the fish fed each diet were aqueously exposed to Cd (0.3 µg/L) while the other half were not exposed to Cd (control). Fish were sampled and their liver was harvested for fatty acid profile, hepatic Cd and calcium concentrations, total glutathione level and gene expression assessment, either (i) after 4 weeks of feeding and 24 h of Cd contamination (day 29) (short-term Cd exposure) or (ii) after 10 weeks of feeding and 6 weeks of Cd contamination (day 70) (long-term Cd exposure). We found that both dietary lipids and Cd exposure influenced fatty acid homeostasis and metabolism. The hepatic fatty acid profile mostly reflected that of the diet (e.g. n-3/n-6 ratio) with some differences, including selective retention of specific long chain polyunsaturated fatty acids (LC-PUFAs) like DHA and active biotransformation of dietary LA and ALA into LC-PUFAs. Cd effects on hepatic fatty acid profiles were influenced by the duration of the exposure and the nutritional status of the fish. The effects of diet and Cd exposure on the fatty acid profiles were only sparsely explained by variation of the expression pattern of genes involved in fatty acid metabolism. The biological responses to Cd were also influenced by dietary lipids. Fish fed the ALA-enriched diet seemed to be the least affected by the Cd exposure, as they showed a higher detoxifying ability against Cd with an early upregulation of protective metallothionein a (MTa) and apoptosis regulator BCL2-Like1 (BCLx) genes, an increased long-term phospholipid synthesis and turnover and fatty acid bioconversion efficiency, as well as a lower long-term accumulation of Cd in their liver. In contrast, fish fed the EPA-enriched diet seemed to be the most sensitive to a long-term Cd exposure, with an impaired growth performance and a decreased antioxidant capacity (lower glutathione level). Our results highlight that low, environmentally realistic aqueous concentrations of Cd can affect biological response in fish and that these effects are influenced by the dietary fatty acid composition.

In vitro Lipolysis and Leptin Production of Elephant Seal Blubber Using Precision-Cut Adipose Tissue Slices.

Adipose tissue plays key roles in energy homeostasis. Understanding its metabolism and regulation is essential to predict the impact of environmental changes on wildlife health, especially in fasting-adapted species. However, in vivo experimental work in wild vertebrates can be challenging. We have developed a novel in vitro approach of precision-cut adipose tissue slices from northern elephant seal (Mirounga angustirostris) as a complementary approach to whole animal models. Blubber biopsies were collected from 14 pups during early and late post-weaning fast (Año Nuevo, CA, United States), precision-cut into 1 mm thick slices and maintained in culture at 37°C for at least 63 h. The slices exhibited an efficient response to ß-adrenergic stimulation, even after 2 days of culture, revealing good in vitro tissue function. The response to lipolytic stimulus did not vary between regions of outer and inner blubber, but was higher at early than at late fast for inner blubber slices. At early fast, lipolysis significantly reduced leptin production. At this stage, inner blubber slices were also more efficient at producing leptin than outer blubber slices, especially in the non-lipolytic condition. This model will aid the study of adipose tissue metabolism and its response to environmental stressors in marine mammals.