Availability Of Fatty Acids Research Articles

Determination of Substrate Preferences for Desaturases and Elongases for Production of Docosahexaenoic Acid from Oleic Acid in Engineered Canola

Production of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in plant seed oils has been pursued to improve availability of these omega-3 fatty acids that provide important human health benefits. Canola (Brassica napus), through the introduction of 10 enzymes, can convert oleic acid (OLA) into EPA and ultimately DHA through a pathway consisting of two elongation and five desaturation steps. Herein we present an assessment of the substrate specificity of the seven desaturases and three elongases that were introduced into canola by expressing individual proteins in yeast. In vivo feeding experiments were conducted with 14 potential fatty acid intermediates in an OLA to DHA pathway to determine the fatty acid substrate profiles for each enzyme. Membrane fractions were prepared from yeast expression strains and shown to contain active enzymes. The elongases, as expected, extended acyl-CoA substrates in the presence of malonyl-CoA. To distinguish between enzymes that desaturate CoA- and phosphatidylcholine-linked fatty acid substrates, we developed a novel in vitro method. We show that a delta-12 desaturase from Phytophthora sojae, an omega-3 desaturase from Phytophthora infestans and a delta-4 desaturase from Thraustochytrium sp., all prefer phosphatidylcholine-linked acyl substrates with comparatively low use of acyl-CoA substrates. To further validate our method, a delta-9 desaturase from Saccharomyces cerevisiae was confirmed to use acyl-CoA as substrate, but could not use phosphatidylcholine-linked substrates. The results and the assay methods presented herein will be useful in efforts to improve modeling of fatty acid metabolism and production of EPA and DHA in plants.

Repeated Prolonged Exercise Decreases Maximal Fat Oxidation in Older Men.

Fat metabolism and muscle adaptation was investigated in six older trained men (age, 61 ± 4 yr; V˙O2max, 48 ± 2 mL·kg·min) after repeated prolonged exercise). A distance of 2706 km (1681 miles) cycling was performed over 14 d, and a blood sample and a muscle biopsy were obtained at rest after an overnight fast before and 30 h after the completion of the cycling. V˙O2max and maximal fat oxidation were measured using incremental exercise tests. HR was continuously sampled during cycling to estimate exercise intensity. The daily duration of exercise was 10 h and 31 ± 37 min, and the mean intensity was 53% ± 1% of V˙O2max. Body weight remained unchanged. V˙O2max and maximal fat oxidation rate decreased by 6% ± 2% (P = 0.04) and 32% ± 8% (P < 0.01), respectively. The exercise intensity that elicits maximal fat oxidation was not significantly decreased. Plasma free fatty acid (FA) concentration decreased (P < 0.002) from 500 ± 77 μmol·L to 160 ± 38 μmol·L. Plasma glucose concentration as well as muscle glycogen, myoglobin, and triacylglycerol content remained unchanged. Muscle citrate synthase and ß-hydroxy-acyl-CoA-dehydrogenase activities were unchanged, but the protein expression of HKII, GLUT4, and adipose triacylglycerol lipase were significantly increased. Overall, the decreased maximal fat oxidation was probably due to lower exogenous plasma fatty acid availability and the muscle adaptation pattern indicates an increased glucose transport capacity and an increased muscle lipolysis capacity supporting an increased contribution of exogenous glucose and endogenous fat during exercise.

Muscle Lipid Metabolism: Role of Lipid Droplets and Perilipins.

Skeletal muscle is one of the main regulators of carbohydrate and lipid metabolism in our organism, and therefore, it is highly susceptible to changes in glucose and fatty acid (FA) availability. Skeletal muscle is an extremely complex tissue: its metabolic capacity depends on the type of fibers it is made up of and the level of stimulation it undergoes, such as acute or chronic contraction. Obesity is often associated with increased FA levels, which leads to the accumulation of toxic lipid intermediates, oxidative stress, and autophagy in skeletal fibers. This lipotoxicity is one of the most common causes of insulin resistance (IR). In this scenario, the “isolation” of certain lipids in specific cell compartments, through the action of the specific lipid droplet, perilipin (PLIN) family of proteins, is conceived as a lifeguard compensatory strategy. In this review, we summarize the cellular mechanism underlying lipid mobilization and metabolism inside skeletal muscle, focusing on the function of lipid droplets, the PLIN family of proteins, and how these entities are modified in exercise, obesity, and IR conditions.

Evolving Concepts of Myocardial Energy Metabolism: More Than Just Fats and Carbohydrates.

Although mitochondrial fatty acid and carbohydrate oxidation are the major source of ATP production in the heart, it is becoming increasingly clear that oxidation of other energy substrates, such as ketones and branched chain amino acids (BCAAs), can also contribute to energy production. Of importance is that alterations in cardiac ketone and branched chain amino acids metabolism may also impact the severity of heart failure through alterations in cellular signaling, despite these fuels providing a lower contribution to overall energy production. Dating back to the first studies investigating glucose uptake in the isolated rabbit heart by Locke and Rosenheim in 1907 to the first studies in humans in the late 1940s by Richard Bing and colleagues assessing cardiac substrate extraction via measurement of arterial coronary sinus differences, cardiac energy metabolism has been a topic of interest for cardiologists and scientists alike. The importance of energy metabolism in the heart has become widely appreciated, as has the dynamic nature of the heart’s ability to metabolize a wide range of energy substrates to meet its energy requirements. Fatty acids are recognized as a key source of energy for the heart, as well as carbohydrates, such as glucose and lactate (Figure).1 In 1961, Shipp et al2 were the first to demonstrate that increasing fatty acid availability to the heart results in a marked inhibition of glucose oxidation, though credit for the reciprocal relationship between fatty acids and glucose metabolism (glucose/fatty acid cycle) is attributed to the work of Randle et al.3 Confirmation of Randle’s glucose/fatty acid cycle in the rodent heart has since been demonstrated in the human heart.1 Figure. Interaction of energy substrates for oxidative energy production and branched chain amino acid (BCAA)/ketone body regulation of cellular signaling in the heart .The figure depicts how energy substrates compete …

Lake eutrophication and brownification downgrade availability and transfer of essential fatty acids for human consumption

Fish are an important source of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) for birds, mammals and humans. In aquatic food webs, these highly unsaturated fatty acids (HUFA) are essential for many physiological processes and mainly synthetized by distinct phytoplankton taxa. Consumers at different trophic levels obtain essential fatty acids from their diet because they cannot produce these sufficiently de novo. Here, we evaluated how the increase in phosphorus concentration (eutrophication) or terrestrial organic matter inputs (brownification) change EPA and DHA content in the phytoplankton. Then, we evaluated whether these changes can be seen in the EPA and DHA content of piscivorous European perch (Perca fluviatilis), which is a widely distributed species and commonly consumed by humans. Data from 713 lakes showed statistically significant differences in the abundance of EPA- and DHA-synthesizing phytoplankton as well as in the concentrations and content of these essential fatty acids among oligo-mesotrophic, eutrophic and dystrophic lakes. The EPA and DHA content of phytoplankton biomass (mgHUFAg−1) was significantly lower in the eutrophic lakes than in the oligo-mesotrophic or dystrophic lakes. We found a strong significant correlation between the DHA content in the muscle of piscivorous perch and phytoplankton DHA content (r=0.85) as well with the contribution of DHA-synthesizing phytoplankton taxa (r=0.83). Among all DHA-synthesizing phytoplankton this correlation was the strongest with the dinoflagellates (r=0.74) and chrysophytes (r=0.70). Accordingly, the EPA+DHA content of perch muscle decreased with increasing total phosphorus (r2=0.80) and dissolved organic carbon concentration (r2=0.83) in the lakes. Our results suggest that although eutrophication generally increase biomass production across different trophic levels, the high proportion of low-quality primary producers reduce EPA and DHA content in the food web up to predatory fish. Ultimately, it seems that lake eutrophication and brownification decrease the nutritional quality of fish for human consumers.

Fatty acids and fat supplements in lactating dairy cow rations

In most Federal Milk Market Orders milk fat and protein yield are the major contributors to the price that producers receive for milk. The addition of supplemental fatty acid (FA) sources to diets is a common practice in dairy nutrition to increase dietary energy density and to support milk production. The ability to understand and model FA, the effects of individual FA, and different FA supplements on production parameters has direct impact on dairy industry recommendations and the usefulness of FA supplementation strategies. The emphasis of the current paper is on biological processes and quantitative changes during the metabolism of FA in the rumen and the effect this has on FA availability to the dairy cow, the digestibility of these FA, and their overall impact on performance. We will focus on recent research supplementing palmitic acid (C16:0) and stearic acid (C18:0)-enriched supplements, on feed intake, digestibility, milk production, and milk composition, and energy partitioning.

VLDL Triglyceride Kinetics in Lean, Overweight, and Obese Men and Women.

High-plasma very low-density lipoprotein (VLDL) triglyceride (TG) concentration and alterations in VLDL-TG metabolism are associated with cardiometabolic disease. This study sought to evaluate the interrelationships among factors purported to regulate VLDL-TG metabolism in a large cohort of men and women with a wide range in body adiposity and fat distribution but without diabetes. We assessed body composition and fat distribution, plasma insulin concentration, free fatty acid availability, and basal VLDL-TG and VLDL-apoB-100 (VLDL particle number) kinetics in 233 lean, overweight, and obese men and women. We found that: 1) plasma VLDL-TG concentration is determined primarily by VLDL-TG secretion rate (SR) in men and by VLDL-TG clearance rate in women; 2) there is a dissociation between VLDL-TG and VLDL-apoB-100 SRs, and VLDL-apoB-100 SR only explains ∼30% of the variance in VLDL-TG SR; 3) ∼50% of people with obesity have high plasma VLDL-TG concentration due to both an increased VLDL-TG SR and a decreased rate of VLDL-TG plasma clearance, and they have lower plasma high-density lipoprotein-cholesterol concentration and more intra-abdominal and liver fat than those with normal VLDL-TG concentration; and 4) fat-free mass, liver fat content and the rate of free fatty acid release into plasma are independent predictors (with a sex × race interaction) of VLDL-TG SR. The regulation of plasma VLDL-TG concentration is complex and influenced by multiple metabolic factors. Many people with obesity have normal plasma VLDL-TG concentrations and kinetics, whereas those with high plasma VLDL-TG concentrations have increased VLDL-TG SR and other markers of cardiometabolic disease risk.

The Gut-Brain Axis, BDNF, NMDA and CNS Disorders

Gastro-intestinal (GI) microbiota and the 'gut-brain axis' are proving to be increasingly relevant to early brain development and the emergence of psychiatric disorders. This review focuses on the influence of the GI tract on Brain-Derived Neurotrophic Factor (BDNF) and its relationship with receptors for N-methyl-D-aspartate (NMDAR), as these are believed to be involved in synaptic plasticity and cognitive function. NMDAR may be associated with the development of schizophrenia and a range of other psychopathologies including neurodegenerative disorders, depression and dementias. An analysis of the routes and mechanisms by which the GI microbiota contribute to the pathophysiology of BDNF-induced NMDAR dysfunction could yield new insights relevant to developing novel therapeutics for schizophrenia and related disorders. In the absence of GI microbes, central BDNF levels are reduced and this inhibits the maintenance of NMDAR production. A reduction of NMDAR input onto GABA inhibitory interneurons causes disinhibition of glutamatergic output which disrupts the central signal-to-noise ratio and leads to aberrant synaptic behaviour and cognitive deficits. Gut microbiota can modulate BDNF function in the CNS, via changes in neurotransmitter function by affecting modulatory mechanisms such as the kynurenine pathway, or by changes in the availability and actions of short chain fatty acids (SCFAs) in the brain. Interrupting these cycles by inducing changes in the gut microbiota using probiotics, prebiotics or antimicrobial drugs has been found promising as a preventative or therapeutic measure to counteract behavioural deficits and these may be useful to supplement the actions of drugs in the treatment of CNS disorders.

Lipolysis sensation by white fat afferent nerves triggers brown fat thermogenesis

ObjectiveMetabolic challenges, such as a cold environment, stimulate sympathetic neural efferent activity to white adipose tissue (WAT) to drive lipolysis, thereby increasing the availability of free fatty acids as one source of fuel for brown adipose tissue (BAT) thermogenesis. WAT is also innervated by sensory nerve fibers that network to metabolic brain areas; moreover, activation of these afferents is reported to increase sympathetic nervous system outflow. However, the endogenous stimuli sufficient to drive WAT afferents during metabolic challenges as well as their functional relation to BAT thermogenesis remain unknown. MethodWe tested if local WAT lipolysis directly activates WAT afferent nerves, and then assessed whether this WAT sensory signal affected BAT thermogenesis in Siberian hamsters (Phodopus sungorus). Results2-deoxyglucose, a sympathetic nervous system stimulant, caused β-adrenergic receptor dependent increases in inguinal WAT (IWAT) afferent neurophysiological activity. In addition, direct IWAT injections of the β3-AR agonist CL316,243 dose-dependently increased: 1) phosphorylation of IWAT hormone sensitive lipase, an indicator of SNS-stimulated lipolysis, 2) expression of the neuronal activation marker c-Fos in dorsal root ganglion neurons receiving sensory input from IWAT, and 3) IWAT afferent neurophysiological activity, an increase blocked by antilipolytic agent 3,5-dimethylpyrazole. Finally, we demonstrated that IWAT afferent activation by lipolysis triggers interscapular BAT thermogenesis through a neural link between these two tissues. ConclusionsThese data suggest IWAT lipolysis activates local IWAT afferents triggering a neural circuit from WAT to BAT that acutely induces BAT thermogenesis.

Free fatty acid G-protein coupled receptor signaling in M1 skewed white adipose tissue macrophages.

Obesity is associated with the establishment and maintenance of a low grade, chronically inflamed state in the white adipose tissue (WAT) of the body. The WAT macrophage population is a major cellular participant in this inflammatory process that significantly contributes to the pathophysiology of the disease, with the adipose depots of obese individuals, relative to lean counterparts, having an elevated number of macrophages that are skewed towards a pro-inflammatory phenotype. Alterations in the WAT lipid micro-environment, and specifically the availability of free fatty acids, are believed to contribute towards the obesity-related quantitative and functional changes observed in these cells. This review specifically addresses the involvement of the five G-protein coupled free fatty acid receptors which bind exogenous FFAs and signal in macrophages. Particular focus is placed on the involvement of these receptors in macrophage migration and cytokine production, two important aspects that modulate inflammation.

Arachidonic and oleic acid exert distinct effects on the DNA methylome

ABSTRACTAbnormal fatty acid metabolism and availability are landmarks of metabolic diseases, which in turn are associated with aberrant DNA methylation profiles. To understand the role of fatty acids in disease epigenetics, we sought DNA methylation profiles specifically induced by arachidonic (AA) or oleic acid (OA) in cultured cells and compared those with published profiles of normal and diseased tissues. THP-1 monocytes were stimulated with AA or OA and analyzed using Infinium HumanMethylation450 BeadChip (Illumina) and Human Exon 1.0 ST array (Affymetrix). Data were corroborated in mouse embryonic fibroblasts. Comparisons with publicly available data were conducted by standard bioinformatics. AA and OA elicited a complex response marked by a general DNA hypermethylation and hypomethylation in the 1–200 μM range, respectively, with a maximal differential response at the 100 μM dose. The divergent response to AA and OA was prominent within the gene body of target genes, where it correlated positively with transcription. AA-induced DNA methylation profiles were similar to the corresponding profiles described for palmitic acid, atherosclerosis, diabetes, obesity, and autism, but relatively dissimilar from OA-induced profiles. Furthermore, human atherosclerosis grade-associated DNA methylation profiles were significantly enriched in AA-induced profiles. Biochemical evidence pointed to β-oxidation, PPAR-α, and sirtuin 1 as important mediators of AA-induced DNA methylation changes. In conclusion, AA and OA exert distinct effects on the DNA methylome. The observation that AA may contribute to shape the epigenome of important metabolic diseases, supports and expands current diet-based therapeutic and preventive efforts.

Abstract 400: Determining the Contribution of Endothelial Lipase-mediated Lipolysis to Brain Phospholipid Metabolism

Polyunsaturated fatty acids (PUFA) are essential constituents of the cell membrane. Brain PUFA content is critical for synaptic function and neuroinflammation and PUFA deficiency may underlie several neurological disorders. Prior work in murine models has showed that the predominant source of central nervous system (CNS) PUFAs is 2-acyl-lysophophatidylcholine (2-lysoPC), a class of phospholipids (PLs) circulating on blood lipoproteins that is formed from the enzymatic activity of phospholipases. Endothelial lipase (EL) is a phospholipase that is a critical determinant of HDL metabolism in humans and the predominant generator of 2-lysoPC on HDL particles. While the contribution of EL to circulating HDL metabolism is well-established, its contribution to CNS fatty acid availability and brain function has not been explored. In order to investigate the physiological contribution of EL to brain FA availability in vivo, we studied PL-derived PUFA uptake in mice with EL deficiency. We adopted a method for measuring EL activity using a fluorescent synthetic HDL particle containing sn2-TopFluor-PC, a fluorescent precursor of 2-lysoPC and demonstrated its utility in vitro. Next, we tested the clearance of this synthetic HDL-derived 2-lysoPC in C57BL/6 (WT) vs EL KO mice (n=12/group). Circulating plasma 2-lysoPC clearance was significantly impaired in in EL KO mice (-32%, p=0.024) and was accompanied by a slower remodeling of the HDL particles. Measurement of labeled PL content in tissues showed a significant reduction in hepatic uptake in KO mice (22% decrease, p=0.002). No significant differences were observed in total brain PL uptake, but the distribution of PL uptake differed between WT and KOs. Ongoing studies are evaluating the physiological consequences of reduced circulating 2-lysoPC clearance, reduced hepatic uptake, and altered distribution of brain PL uptake in EL KO mice. Collectively, our data demonstrate that EL is a key physiological regulator of systemic PL catabolism.

Measuring spatial variation in secondary production and food quality using a common consumer approach in Lake Erie.

Lake Erie is a large lake straddling the border of the USA and Canada that has become increasingly eutrophic in recent years. Eutrophication is particularly focused in the shallow western basin. The western basin of Lake Erie is hydrodynamically similar to a large estuary, with riverine inputs from the Detroit and Maumee Rivers mixing together and creating gradients in chemical and physical conditions. This study was driven by two questions: (1) How does secondary production and food quality for consumers vary across this large mixing zone? and (2) Are there correlations between cyanobacterial abundance and secondary production or food quality for consumers? Measuring spatial and temporal variation in secondary production and food quality is difficult for a variety of logistical reasons, so here a common consumer approach was used. In a common consumer approach, individuals of a single species are raised under similar conditions until placed in the field across environmental gradients of interest. After some period of exposure, the response of that common consumer is measured to provide an index of spatial variation in conditions. Here, a freshwater mussel (Lampsilis siliquoidea) was deployed at 32 locations that spanned habitat types and a gradient in cyanobacterial abundance in the western basin of Lake Erie to measure spatial variation in growth (an index of secondary production) and fatty acid (FA) content (an index of food quality). We found secondary production was highest within the Maumee river mouth and lowest in the open waters of the lake. Mussel tissues in the Maumee river mouth also included more eicosapentaenoic and docosapentaenoic fatty acids (EPA and DPA, respectively), but fewer bacterial FAs, suggesting more algae at the base of the food web in the Maumee river mouth compared to open lake sites. The satellite-derived estimate of cyanobacterial abundance was not correlated to secondary production, but was positively related to EPA and DPA content in the mussels, suggesting more of these important FAs in locations with more cyanobacteria. These results suggest that growth of secondary consumers and the availability of important fatty acids in the western basin are centered on the Maumee river mouth.

Dietary Crude Lecithin Increases Systemic Availability of Dietary Docosahexaenoic Acid with Combined Intake in Rats.

Crude lecithin, a mixture of mainly phospholipids, potentially helps to increase the systemic availability of dietary omega-3 polyunsaturated fatty acids (n-3 PUFA), such as docosahexaenoic acid (DHA). Nevertheless, no clear data exist on the effects of prolonged combined dietary supplementation of DHA and lecithin on RBC and plasma PUFA levels. In the current experiments, levels of DHA and choline, two dietary ingredients that enhance neuronal membrane formation and function, were determined in plasma and red blood cells (RBC) from rats after dietary supplementation of DHA-containing oils with and without concomitant dietary supplementation of crude lecithin for 2–3 weeks. The aim was to provide experimental evidence for the hypothesized additive effects of dietary lecithin (not containing any DHA) on top of dietary DHA on PUFA levels in plasma and RBC. Dietary supplementation of DHA-containing oils, either as vegetable algae oil or as fish oil, increased DHA, eicosapentaenoic acid (EPA), and total n-3 PUFA, and decreased total omega-6 PUFA levels in plasma and RBC, while dietary lecithin supplementation alone did not affect these levels. However, combined dietary supplementation of DHA and lecithin increased the changes induced by DHA supplementation alone. Animals receiving a lecithin-containing diet also had a higher plasma free choline concentration as compared to controls. In conclusion, dietary DHA-containing oils and crude lecithin have synergistic effects on increasing plasma and RBC n-3 PUFA levels, including DHA and EPA. By increasing the systemic availability of dietary DHA, dietary lecithin may increase the efficacy of DHA supplementation when their intake is combined.

Deletion of GPR40 fatty acid receptor gene in mice blocks mercaptoacetate-induced feeding.

Both increased and decreased fatty acid (FA) availability contribute to control of food intake. For example, it is well documented that intestinal FA reduces feeding by triggering enterondocrine secretion of satietogenic peptides, such as cholecystokinin (CCK) and glucagon-like peptide-1 (GLP-1). In contrast, mechanisms by which decreased FA availability increase feeding are not well understood. Over the past three decades substantial research related to FA availability and increased feeding has involved use of the orexigenic compound mercaptoacetate (MA). Because MA reportedly inhibits FA oxidation, it has been assumed that reduced FA oxidation accounts for the orexigenic action of MA. Recently, however, we demonstrated that MA antagonizes G protein-coupled receptor 40 (GPR40), a membrane receptor for long and medium chain FA. We also demonstrated that, by antagonizing GPR40, MA inhibits GLP-1 secretion and attenuates vagal afferent activation by FA. Because both vagal afferent activation and GLP-1 inhibit food intake, we postulated that inhibition of GPR40 by MA might underlie the orexigenic action of MA. We tested this hypothesis using male and female GPR40 knockout (KO) and wild-type (WT) mice. Using several testing protocols, we found that MA increased feeding in WT, but not GPR40 KO mice, and that GPR40 KO mice gained more weight than WT on a high-fat diet. Metabolic monitoring after MA or saline injection in the absence of food did not reveal significant differences in respiratory quotient or energy expenditure between treatment groups or genotypes. These results support the hypothesis that MA stimulates food intake by blocking FA effects on GPR40.

Pigment epithelium-derived factor (PEDF) regulates metabolism and insulin secretion from a clonal rat pancreatic beta cell line BRIN-BD11 and mouse islets

Pigment epithelium-derived factor (PEDF) is a multifunctional glycoprotein, associated with lipid catabolism and insulin resistance. In the present study, PEDF increased chronic and acute insulin secretion in a clonal rat β-cell line BRIN-BD11, without alteration of glucose consumption. PEDF also stimulated insulin secretion from primary mouse islets. Seahorse flux analysis demonstrated that PEDF did not change mitochondrial respiration and glycolytic function. The cytosolic presence of the putative PEDF receptor - adipose triglyceride lipase (ATGL) - was identified, and ATGL associated stimulation of glycerol release was robustly enhanced by PEDF, while intracellular ATP levels increased. Addition of palmitate or ex vivo stimulation with inflammatory mediators induced β-cell dysfunction, effects not altered by the addition of PEDF. In conclusion, PEDF increased insulin secretion in BRIN-BD11 and islet cells, but had no impact on glucose metabolism. Thus elevated lipolysis and enhanced fatty acid availability may impact insulin secretion following PEDF receptor (ATGL) stimulation.

Organic matter-clay interaction along a seawater column of the Eastern Pacific upwelling system (Antofagasta bay, Chile): Implications for source rock organic matter preservation

Natural Organic Matter (OM) preservation and sedimentary burial mechanisms are still poorly understood despite their importance in oil source rock characterization and carbon cycle issues. It is now recognized that mineral sorption contributes significantly to OM preservation by conferring steric protection and preventing enzymatic access. Most of the studies dealing with OM preservation have focused on settled OM, omitting aggregation in the water column. For the first time, we have tested the complexation of natural OM on 3 types of clays in seawater in-situ conditions. Samples of Na exchanged montmorillonite (Na MMt), synthetic montmorillonite (S Na MMt) and natural kaolinite (Kaol) were immersed in an upwelling zone (Antofagasta bay, Chile). Three depths were studied: 15m (photic zone), 40m (oxygen transition zone) and 80m (anoxic zone). The clayey samples were kept immersed for different durations and analyzed after removal, via GC/MS, Py-GC/MS, flash elementary pyrolysis, DOC analyzer and XRD. The main results are: (1) for the shortest immersion times, DOC analyzes performed on clays' proximal water indicated excessive DOC concentrations compared to seawater values. This attraction effect was however temporary, as values tended to rebalance with seawater concentrations for the longest immersion times. (2) Molecular analyses performed on seawater organic extracts indicated that fatty acids, alkanes, alkyl benzenes and methoxy-benzenes were the main dissolved lipid groups. (3) These lipids were also the main sorbed compounds identified on immersed clays, with a predominance of fatty acids. This is due to the higher availability and better affinity of fatty acids with the clayey samples. (4) Because of its higher specific area and CEC, Na MMt exhibited the best sorption rates compared to Kaol and SNa MMt. X ray analyses performed on Na MMt indicated d001spacing growths, but these growths are not attributed to OM intercalation. Sorbed OM is probably located on the external surfaces leaf edges of clays.

Mechanisms Involved in the Improvement of Lipotoxicity and Impaired Lipid Metabolism by Dietary α-Linolenic Acid Rich Salvia hispanica L (Salba) Seed in the Heart of Dyslipemic Insulin-Resistant Rats.

This study explores the mechanisms underlying the altered lipid metabolism in the heart of dyslipemic insulin-resistant (IR) rats fed a sucrose-rich diet (SRD) and investigates if chia seeds (rich in α-linolenic acid 18:3, n-3 ALA) improve/reverse cardiac lipotoxicity. Wistar rats received an SRD-diet for three months. Half of the animals continued with the SRD up to month 6. The other half was fed an SRD in which the fat source, corn oil (CO), was replaced by chia seeds from month 3 to 6 (SRD+chia). A reference group consumed a control diet (CD) all the time. Triglyceride, long-chain acyl CoA (LC ACoA) and diacylglycerol (DAG) contents, pyruvate dehydrogenase complex (PDHc) and muscle-type carnitine palmitoyltransferase 1 (M-CPT1) activities and protein mass levels of M-CPT1, membrane fatty acid transporter (FAT/CD36), peroxisome proliferator activated receptor α (PPARα) and uncoupling protein 2 (UCP2) were analyzed. Results show that: (a) the hearts of SRD-fed rats display lipotoxicity suggesting impaired myocardial lipid utilization; (b) Compared with the SRD group, dietary chia normalizes blood pressure; reverses/improves heart lipotoxicity, glucose oxidation, the increased protein mass level of FAT/CD36, and the impaired insulin stimulated FAT/CD36 translocation to the plasma membrane. The enhanced M-CPT1 activity is markedly reduced without similar changes in protein mass. PPARα slightly decreases, while the UCP2 protein level remains unchanged in all groups. Normalization of dyslipidemia and IR by chia reduces plasma fatty acids (FAs) availability, suggesting that a different milieu prevents the robust translocation of FAT/CD36. This could reduce the influx of FAs, decreasing the elevated M-CPT1 activity and lipid storage and improving glucose oxidation in cardiac muscles of SRD-fed rats.

The Effects of Starvation and Refeeding on Oxidative Stress Parameters (MDA, SOD, GPx), Lipid Profile, Thyroid Hormones and Thyroid Histopathology in Male Wistar Rats

Objective: This study aimed to investigate the effect of starvation and re-feeding on oxidative stress markers, lipid profile, thyroid hormones, and changes in thyroid tissue and in male Wistar rats. Method: Fifty-six male Wistar rats were divided into 6 groups. After 3 months of feeding animals with specific diet, the first group was used without fasting (day zero). Other rats were exposed to fasting for 14 days. The second group was considered as a group after fasting (day 14). In groups 3 to 6, replenishment was done and then these groups were euthanized on days 16 to 22, and blood and tissue samples were taken. Results: Significant increase was observed in MDA, triglycerides, and VLDL cholesterol. The concentrations of SOD, GPx and T3 were decreased significantly. By re-feeding, SOD, GPx, triglycerides, total cholesterol, and VLDL cholesterol increased. T3 concentration was significantly increased in all groups after re-feeding. In thyroid tissue, the diameter of follicles and the amount of colloid decreased and the number of parafollicular cells increased during starvation. By days 2 and 4 post-refeeding there were follicles with different sizes and the numbers of parafollicular cells were increased. Conclusion: Starvation may act on the depletion of antioxidants, predisposing subjects to oxidative injury highlights with enhanced level of MDA. Oxidative stress, may play a critical role in the pathophysiology of starvation- refeeding syndrome. Starvation results in a decrease in the serum T3 concentration due to decreased peripheral generation of T3 from T4, because carbohydrates are major regulators of T4 deiodination. The increased free fatty acid availability to the circulation and eventually to the liver contribute to increased VLDL synthesis and elevated plasma triglyceride levels after starvation. The negative effects of oxidative stress in considerable time after re-feeding should be considered in future studies.

Altered myocardial metabolic adaptation to increased fatty acid availability in cardiomyocyte-specific CLOCK mutant mice

A mismatch between fatty acid availability and utilization leads to cellular/organ dysfunction during cardiometabolic disease states (e.g., obesity, diabetes mellitus). This can precipitate cardiac dysfunction. The heart adapts to increased fatty acid availability at transcriptional, translational, post-translational and metabolic levels, thereby attenuating cardiomyopathy development. We have previously reported that the cardiomyocyte circadian clock regulates transcriptional responsiveness of the heart to acute increases in fatty acid availability (e.g., short-term fasting). The purpose of the present study was to investigate whether the cardiomyocyte circadian clock plays a role in adaptation of the heart to chronic elevations in fatty acid availability. Fatty acid availability was increased in cardiomyocyte-specific CLOCK mutant (CCM) and wild-type (WT) littermate mice for 9weeks in time-of-day-independent (streptozotocin (STZ) induced diabetes) and dependent (high fat diet meal feeding) manners. Indices of myocardial metabolic adaptation (e.g., substrate reliance perturbations) to STZ-induced diabetes and high fat meal feeding were found to be dependent on genotype. Various transcriptional and post-translational mechanisms were investigated, revealing that Cte1 mRNA induction in the heart during STZ-induced diabetes is attenuated in CCM hearts. At the functional level, time-of-day-dependent high fat meal feeding tended to influence cardiac function to a greater extent in WT versus CCM mice. Collectively, these data suggest that CLOCK (a circadian clock component) is important for metabolic adaption of the heart to prolonged elevations in fatty acid availability. This article is part of a Special Issue entitled: Heart Lipid Metabolism edited by G.D. Lopaschuk.