Project DyAdd: Fatty acids in adult dyslexia, ADHD, and their comorbid combination

In project DyAdd, we compared the fatty acid (FA) profiles of serum phospholipids in adults with attention deficit hyperactivity disorder (ADHD) (n=26), dyslexia (n=36), their comorbid combination (n=9), and healthy controls (n=36). FA proportions were analyzed in a 2×2 design with Bonferroni corrected post hoc comparisons. A questionnaire was used to assess dietary fat quality and use of supplements. Results showed that ADHD and dyslexia were not associated with total saturated FAs, monounsaturated FAs, or n-3 polyunsaturated FAs (PUFAs). However, those with ADHD had elevated proportions of total n-6 PUFAs (including γ-linolenic and adrenic acids) as compared to those without ADHD. Dyslexia was related to a higher proportion of monounsaturated nervonic acid and a higher ratio of n-6/n-3 PUFAs. Among females none of the associations were significant. However in males, all the original associations observed in all subjects remained and ADHD was associated with elevated nervonic acid and n-6/n-3 PUFA ratio like dyslexia. Controlling for poorly diagnosed reading difficulties, education, dietary fat quality, or use of FA supplements did not generally remove the originally observed associations.     

Fatty acids,resin acids and phenols in Pinus muricata

Acetone-soluble extractives of “blue” and “green” strain Pinus muricata D. Don were found to consist of free and “combined” fatty acids, resin acids, and phenols. The composition of the extractives from the two strains was similar though “green” strain P. muricata contained more Δ^8(9),15 isopimaric acid than “blue” strain. This difference may be used to identify these muricata strains if the age of the wood precludes a monoterpene examination.     

Fatty acids, alcohol and fatty acid ethyl esters: Toxic Ca signal generation and pancreatitis

Pancreatitis, a potentially fatal disease in which the pancreas digests itself as well as its surroundings, is a well recognized complication of hyperlipidemia. Fatty acids have toxic effects on pancreatic acinar cells and these are mediated by large sustained elevations of the cytosolic Ca²⁺ concentration. An important component of the effect of fatty acids is due to inhibition of mitochondrial function and subsequent ATP depletion, which reduces the operation of Ca²⁺-activated ATPases in both the endoplasmic reticulum and the plasma membrane. One of the main causes of pancreatitis is alcohol abuse. Whereas the effects of even high alcohol concentrations on isolated pancreatic acinar cells are variable and often small, fatty acid ethyl esters – synthesized by combination of alcohol and fatty acids – consistently evoke major Ca²⁺ release from intracellular stores, subsequently opening Ca²⁺ entry channels in the plasma membrane. The crucial trigger for pancreatic autodigestion is intracellular trypsin activation. Although there is still uncertainty about the exact molecular mechanism by which this Ca²⁺-dependent process occurs, progress has been made in identifying a subcellular compartment – namely acid post-exocytotic endocytic vacuoles – in which this activation takes place.     

Fatty acids and glycerol or lactate are required to induce gluconeogenesis from alanine in isolated rabbit renal cortical tubules

Summary In isolated rabbit renal cortical tubules, glucose synthesis from 1 mM alanine is negligible, while the amino acid is metabolized to glutamine and glutamate. The addition of 0.5 mM octanoate plus 2 mM glycerol induces incorporation of [U-¹⁴C]Alnine into glucose and decreases glutamine synthesis, whereas oleate and palmitate in the presence of glycerol are less potent than octanoate. Gluconeogenesis is also significantly accelerated when glycerol is substituted by lactate. In view of an increase in¹⁴CO₂ fixation and elevation of both cytosolic and mitochondrial NADH/NAD⁺ ratios, the activation of glucose formation from alanine upon the addition of glycerol and octanoate is likely due to (i) stimulation of pyruvate carboxylation, (ii) increased availability of NADH for glyceraldehyde-3-phosphate dehydrogenase and (iii) elevation of mitochondrial redox state causing a diminished provision of ammonium for glutamine synthesis. The induction of gluconeogenesis in the presence of alanine, glycerol and octanoate is not related to cell volume changes. The results presented in this paper show the importance of free fatty acids and glycerol for regulation of renal gluconeogenesis from alanine. The possible physiological significance of the data is discussed.     

Fatty acid and lipid composition in mycelia from submerged or surface culture of Streptomyces viridochromogenes

Abstract Streptomyces viridochromogenes was grown both as submerged and surface culture. Mycelia from these cultures were analysed for the composition of lipids and fatty acids. An increase in ornithinolipid content according to incubation time was observed. The addition of phosphate inhibited the ornithinolipid synthesis. A mutant strain with bald phenotype did not exhibit the phosphate inhibition. At the same time, the mutant strain had a higher content of 12-methyltetradecanoic acid.     

Competition between cis,trans and Cyclopropane Fatty Acid Formation and its Impact on Membrane Fluidity

The impact of cis, trans and cyclopropane fatty acids on membrane fluidity was investigated using batch‐grown Pseudomonas putida P8 and Comamonas testosteroni ATCC 17454. A major difference observed between the two investigated strains is the absence of the ability to synthesize trans‐unsaturated fatty acids in Comamonas. When grown exponentially at 30 °C, a shift to 35 °C increased the trans/cis ratios of the fatty acids of P. putida P8 from 0 to 0.81 and 0 to 1.07, in lipid extracts and cell hydrolyzates, respectively. After prolonged growth followed by nutrient deprivation for 48 h, both at 30 °C, trans fatty acids were absent, but the cyclo/cis ratios rose from 0.1 to 1.55 in lipid extracts, and from 0.1 to 1.54 in cell hydrolyzates. C. testosteroni ATCC 17454 contained no cyclo fatty acids when harvested in the exponential phase after 6 h, whereas after 72 h cultivation, the cyclo/cis ratios rose to 0.49 and 0.47, in lipid extracts and cell hydrolyzates, respectively. Trans fatty acids were never observed in this strain. Increased cyclo/cis and trans/cis ratios correlated with decreased fluidity measured by the fluorescence anisotropy of 1,6‐diphenyl‐1,3,5‐hexatriene (DPH) intercalated in the bilayers of liposomes and by Fourier Transform Infrared (FTIR) spectroscopy of lipids prepared from the cells. The specific effect of cyclopropane fatty acids on membrane fluidity was much smaller than that of trans fatty acids. FTIR‐measurements of intact cells of P. putida P8 confirmed the high potency of trans fatty acids to decrease the fluidity. In cells with induced cyclopropane fatty acid synthesis, the membranes remained more fluidized, indicating the lower importance of these fatty acids for homeoviscosis.     

Fatty acids of the scleractinian coral Galaxea fascicularis: effect of light and feeding

In order to investigate nutritional interactions in the symbiotic scleractinian coral-zooxanthella association, fatty acids of the coral Galaxea fascicularis were analysed in two groups of cultured microcolonies. The first group was fed with Artemia sp., while the second group was starved. After an initial 1-month period during which both groups were subjected to the same normal light conditions (constant irradiance of 125 E·cm^-2·s^-1 and 14:10 h light:dark), a light cap was used to cover the aquarium and keep all the microcolonies in permanent darkness for 20 days. During the light phase of the experiment it was shown that the nutritional status lead to large variations in the percentage of saturated, mono-unsaturated and polyunsaturated fatty acids. Palmitic acid (C16:0) was the most abundant fatty acid in both groups. Important differences between fed and starved microcolonies occurred during the dark phase of the experiment. In the fed group the dark phase was characterized by a significant increase in polyunsaturated fatty acids. Particularly arachidonic acid (C20:4 n-6) became the most important fatty acid followed by docosatrienoic acid (C22:3 n-3). A slight increase in these two fatty acids was also found in the starved group but the bulk of polyunsaturated fatty acids was significantly decreased. In this group, palmitic acid remained the most important fatty acid while an increased concentration of cis-vaccenic acid (C18:1 n-7) was found at the end of the experiment. The increased concentration of cis-vaccenic acid might indicate that bacteria serve as a source of energy. While the number of zooxanthellae per milligram of protein and the chlorophyll a to protein ratio strongly decreased in the starved microcolonies immediately after the beginning of the dark period, the decrease in fed microcolonies was delayed for about 10 days. Furthermore, after 20 days of dark incubation the chlorophyll a to protein ratio was the same as measured at the beginning of the dark period. This suggests that in the dark the metabolic requirements of the zooxanthellae are in part met from the animal host through a heterotrophic mode of nutrition.Abbreviations CZ cultured zooxanthellae - FAME fatty acid methylester(s) - FDM fed dark microcolonies - FLM fed light microcolonies - MUFA monounsaturated fatty acid(s) - PUFA polyunsaturated fatty acid(s) - SDM starved dark microcolonies - SFA saturated fatty acids - SLM starved-light microcolonies - SW sea water - TFA total fatty acids     

Fatty acids composition in erythrocyte membranes of athletes after one and after a series of whole body cryostimulation sessions

Whole body cryotherapy (WBC) is a treatment often used by athletes as part of biological renewal. Despite the large interest in this form therapy there is still a lack of information on the effects of WBC on the concentration of fatty acids in erythrocyte membranes. Our study aimed at comparing the fatty acids (FA) composition of erythrocyte membranes of athletes after one session and after a series of sessions of whole body cryostimulation. In our study small changes in the level of total cholesterol (decrease) were observed 24 h after a single session. After the twelfth session of whole body cryostimulation, the level of saturated fatty acids (SFA), mainly palmitic acid (C16:0) and n-3 fatty acid eicosapentaenoic (EPA, C20:5n-3) increased almost two-times fold in the red blood cell membranes. The level of n-6 polyunsaturated fatty acids (PUFA n-6), mainly gamma-linolenic acid (C18:3n-6) as well as trans fatty acids (elaidic acid) decreased in the erythrocyte membranes from men after a series of session in a cryochamber, when compared to the control sample. The n-3/n-6 FA ratio in the erythrocyte membranes was higher after twelfth session in a cryochamber in comparison to the control sample.The data obtained during our study will be important for further research regarding the biochemistry of lipids in men after sessions of whole body cryostimulation.     

Fatty acids and plasmalogens of the phospholipids of the sperm membranes and their relation with the post-thaw quality of stallion spermatozoa

Fatty acids and plasmalogens were extracted from the phospholipids of the plasma membrane of stallion spermatozoa, to determine their relation with sperm quality after freezing and thawing. Sperm quality was rated using a quality index that combined the results of the analysis of sperm motility and velocity (CASA analysis), membrane status and mitochondrial membrane potential (flow cytometry) post thaw. Receiving operating system (ROC) curves were used to evaluate the value of specific lipid components of the sperm membrane herein studied as forecast of potential freezeability. From all parameters studied the ratio of percentage of C16 plasmalogens related to total phospholipids was the one with the better diagnostic value. For potentially bad freezers, the significant area under the ROC-curve was 0.74, with 75% sensitivity and 79.9% specificity for a cut off value of 26.9. Also the percentage of plasmalogens respect to total phospholipids gave good diagnostic value for bad freezers. On the other hand, the percentage of C18 fatty aldehydes related to total phospholipids of the sperm membrane properly forecasted freezeability with an area under the ROC curve of 0.70 with 70% sensitivity and 62.5% specificity for a cut off value of 0.32.     

Fatty acids in liver microsomal lipids of rats exposed to hypoxia, tetrachloromethane, or both

Arachidonic and docosahexaenoic acid in hepatic microsomal lipids from male Sprague-Dawley rats are greatly lowered when the animals have been exposed to tetrachloromethane; at the same time, palmitic, oleic and linoleic acid are significantly increased. Hypoxia alone causes similar derangements, but to a lesser extent. These are largely corrected 18 h after exposure; the effects induced by tetrachloromethane are persistent. The increases in 16:0, 18:1 and 18:2 suggest that in both cases microsomal enzymes involved in fatty acid metabolism are inhibited, either reversibly or irreversibly. Reduction of oxygen partial pressure during tetrachloromethane exposure has little effect upon hepatotoxicity as judged by hepatic enzymes in serum; only the onset of their release into the bloodstream is earlier.     

不同类型脂肪酸对SD大鼠血清脂肪酸及胰岛素抵抗的影响

目的分析中链饱和脂肪酸（MC-SFA,MCF组）、长链饱和脂肪酸（LC-SFA,LCF组）、n-6多不饱和脂肪酸（n-6 PUFA,SUF组）和n-3多不饱和脂肪酸（n-3 PUFA,TUF组）四种脂肪酸对大鼠血清脂肪酸及胰岛素抵抗的影响。方法雄性SD大鼠40只随机分为5组,对照组给予普通日粮,高脂组给予脂肪热量比相同的高脂日粮。喂养10周,每18 d测定空腹血糖（GLU）、血清脂肪酸、血清胰岛素水平,根据胰岛素敏感性指数（ISI）=ln1/（FPG×FINS）评定大鼠的胰岛素敏感性。结果10周后,LCF组和SUF组大鼠体重显著高于对照组和其它高脂组;LCF组血清胰岛素显著高于对照组（P﹤0.05）;LCF组、TUF组ISI显著低于对照组（P﹤0.05）;各组间血糖无明显差异（P〉0.05）。SUF组、TUF组血清LC-SFA浓度显著低于LCF组（P﹤0.05）;TUF组血清（n-3 PUFA）显著高于对照组和其它高脂组（P﹤0.05）。结论不同类型脂肪酸的高脂饲料对SD大鼠的血清脂肪酸组成和含量有显著的影响,SD大鼠脂肪沉积及胰岛素抵抗程度随血清脂肪酸代谢作用的不同而变化。     

Non-methylene-interrupted fatty acids from marine invertebrates: Occurrence,characterization and biological properties

Marine organisms, in particular invertebrates, have proved to be a major source of unique fatty acid (FA) structures originating from unusual biosynthetic pathways. Among them, non-methylene-interrupted (NMI) FA occur in various molluscs in the wide ranges of concentrations (up to 20%), such as the most often encountered 20:2 Δ5,11, 20:2 Δ5,13, 22:2 Δ7,13 or 22:2 Δ7,15. Such NMI FA have also been reported from algae, echinoderms, sponges, tropical rays, and many other invertebrates. The most intriguing marine invertebrates seem to be sponges that commonly contain very long-chain Δ5,9 FA. A third double bond can occur in the NMI FA as reported in some marine organisms, such as 20:3 Δ7,13,16 or 30:3 Δ5,9,23. Lipids of invertebrates from deep-sea hydrothermal and cold-seep vents gave rise to an intense research activity including reports on unprecedented NMI polyunsaturated FA. The bivalve molluscs are able to synthesize de novo the NMI FA but their precise biological interest is presently not well-known, although structural and functional roles in biological membranes have been suggested, in particular a higher resistance to oxidative processes and microbial lipases. Biosynthetic pathways of Δ5,9 FA in sponges were demonstrated up to C₂₆ FA structures and include particular elongation and desaturation steps. Recently, intense research effort has been conducted to investigate the biomedical potential of these unusual FA. Thus, Δ5,9 FA displayed interesting antiplasmodial activity. The most promising FA topoisomerase I inhibitors to date seem to be the long-chain Δ5,9 FA. This inhibitory activity is probably partially responsible for the toxicity displayed by some of the Δ5,9 FA towards cancer cell lines.     

Fatty acid derivatives and their use as CFPP additives in biodiesel

One of the main drawbacks of the use of biodiesel is its bad behavior at low temperatures. In this work, we show that it is possible to take profit of the presence of free fatty acids in the starting materials used for biodiesel production (i.e. reused oils) by synthesizing additives able to improve cold flow properties. The synthesis of fatty acid derivatives have been successfully carried out by esterification of stearic, oleic and linoleic acids with bulky linear and cyclic alcohols and by epoxidation of methyl oleate and subsequent ring-opening reaction with the same alcohols. The study of crystallization patterns of pure derivatives by DSC and optical microscopy revealed the improvement of cold properties of biodiesel. Blends of biodiesel with up to 5% of some of these compounds allowed a decrease of CFPP (Cold Filter Pour Point). Both observations reveal the utility of these compounds as cheap and renewable additives.     

Detection of microbial-derived fatty acids in carious dentin by gas chromatography and gas chromatography-mass spectrometry

The aim of the present study was to analyze the fatty acid content of carious and sound human dentin. Gas chromatography and gas chromatography-mass spectrometry revealed the presence of fatty acids of C₁₀-C₁₈ size in the carious dentin, whereas fatty acids of C₁₆ size were present in minute amounts in three samples of the corresponding sound dentine controls. No fatty acids were detected in the other sound dentin control samples. The source of fatty acids was considered to be microorganisms invading the dentin during the progression of the caries lesion. The presence of bacterial fatty acids in carious dentin may serve as a marker for the pathological process and thus contribute to the understanding of the mechanisms involved.     

n-3PUFA对高脂SHR 大鼠血压及血浆游离脂肪酸谱的影响

目的:观察高饱和脂肪酸及n-3多不饱和脂肪酸饮食后对自发性高血压大鼠血压、静息心率、体重、血脂、血糖及游离脂肪酸谱的影响。方法:选择8周龄雄性自发性高血压大鼠(SHR)30只和同龄对照大鼠(WKY)30只,随机分为6组:SHR、WKY普通饲料组各10只,SHR、WKY高脂组各10只,SHR、WKY高脂加鱼油饮食组各10只,持续喂养至16周龄。干预期间每两周测定血压和体重,干预前后测定静息心率、血脂、血糖及血浆游离脂肪酸谱。结果:(1)血压和静息心率的变化:SHR大鼠高脂饮食组较普食组血压水平显著性增高,而高脂加鱼油饮食组较高脂饮食组血压水平显著性减低;WKY大鼠高脂饮食组较普食组血压水平显著性增高,而高脂加鱼油饮食组较高脂饮食组血压水平显著性减低;SHR大鼠高脂饮食组较普食组静息心率显著性增高(P=0.007),而高脂加鱼油饮食组较高脂饮食组静息心率有下降趋势,但差异无显著性(P=0.125),WKY大鼠静息心率各组间无明显差异。(2)血浆游离脂肪酸谱:与WKY大鼠比较,SHR大鼠中亚麻酸(Linolenic acid,ALA)、花生四烯酸(Linoleic Acid,AA)与n-6多不饱和脂肪酸(n-6 polyunsaturated fatty acids,n-6PUFA含量增高,高脂饮食增加了饱和脂肪酸(Saturated fatty acid,SFA),有显著差异(P0.05),高脂鱼油组二十二碳六烯酸(Docosahexaenoic acid,DHA)及二十碳五烯酸(Docosapentaenoic acid,EPA)增加导致n-3多不饱和脂肪酸(n-3 polyunsaturated fatty acids,n-3PUFA)含量增加(P0.05),SHR大鼠高脂鱼油组亚油酸(Linoleic Acid,LA)、AA含量减低(P0.05)。结论:膳食补充n-3PUFA可能通过影响交感神经活性和血浆脂肪酸谱的组成而改善高饱和脂肪酸所致SHR大鼠的血压升高。     

Fatty acids’ profiles as indicators of stress induced by of a common herbicide on two marine bivalves species: Cerastoderma edule (Linnaeus, 1758) and Scrobicularia plana (da Costa, 1778)

In Europe, mainly the Mediterranean region, intensive use of fertilizers and pesticides has been recorded over the past 30 years, exceeding, in some cases, the limits of contamination authorized by the European Union. The intensive use of pollutants in fields near ecological coastal wetlands has led to implementation of pesticide monitoring programs to recover aquatic systems such as the Mondego estuary (Figueira da Foz, Portugal). According to information from the agricultural cooperatives of the Mondego valley, Primextra^® Gold TZ is the most-used herbicide in corn crop fields. Biomarkers, such fatty acids (FAs), proved to be new and potentially powerful tools to detect, illustrate, and evaluate exposure to and the effects of contamination hazards. They play important roles in establishing neural levels in organisms’ biochemical and physiological responses and are considered good bio-indicators of stress and potential indicators of ecosystem health. Bivalves are currently used in ecotoxicological bioassays because of their ecological importance, wide geographic distribution, ease of handling in the laboratory and in the field, and their ability to filter and ingest large volumes of water and sediment particles. Thus, the main goal of this work was to determine the toxic and biochemical (namely fatty acid profiles) responses of two size classes (small and big) of the two marine bivalve species Cerastoderma edule and Scrobicularia plana to the herbicide Primextra^® Gold. Furthermore, we aimed to compare the fatty acid contents, and thus the nutritive values, of both species and size classes collected in the field with those under laboratory conditions. Results show S. plana is more sensitive to the herbicide than C. edule. In general, among the larger-sized specimens in the field, S. plana is more nutritive than C. edule, but among the smaller-sized specimens, the opposite tendency is seen, where C. edule presents a greater abundance of FAs.     

Fatty acids potentiate interleukin-1beta toxicity in the beta-cell line INS-1E

Evidence for "lipotoxicity," i.e., negative effects of fatty acids (FA) on pancreatic beta-cells is incomplete. Here, we tested whether non-toxic concentrations of FA potentiate toxic effects of interleukin-1beta (IL-1beta). Culture of INS-1E clonal beta-cells for 2-6 days with 70 microM docosahexaenoic acid (DHA), eicosapentaenoic acid, arachidonic acid, 0.1mM linoleic acid, or 0.1-0.2mM oleic acid exerted no or minor negative effects on cell viability (MTT assay). Viability was reduced by 0.5 ng/ml IL-1beta. All tested FA significantly aggravated this effect after 6 days of culture. IL-1beta also exerted negative effects on cellular insulin content and DHA and oleic acid aggravated these effects. L-NAME, an inhibitor of constitutive nitric oxide (NO) synthase, reduced the negative effects of IL-1beta per se but did not abolish the potentiating effects of FA. CONCLUSION: FA potentiate toxic effects of IL-1beta on beta-cells by mechanisms that include NO-independent ones.     

Cholesterol Esters in Rat Brain Infected with Acute Measles Encephalitis: Concentration and Fatty Acid Composition

The present study deals with the concentration and fatty acid composition of cholesterol esters in rat brains infected experimentally with measles virus to induce acute encephalitis. The left side of the cerebrum, as well as other portions of the brain, when inoculated percutaneously contained a large amount of cholesterol esters. The major fatty acids from the esters in the brain were C16:0, C16:1, C18:0, and C18:1; those from the serum were C18:1, C18:2, and C20:4. This result indicates that cholesterol esters may not come from serum but can be synthesized in situ, even in the brain with acute viral infection.     

Fatty acid-activated K+ channels in autonomic neurons: activation by an endogenous source of free fatty acids

Application of arachidonic acid evoked robust activation of large-conductance K+ channels in cell-attached and excised inside-out patches from acutely isolated chick ciliary ganglion neurons. A similar effect was produced by 5,8,11,14-eicosatetraynoic acid, a nonmetabolizable analogue of arachidonic acid. The unitary conductance of fatty acid-activated channels was 35-40 pS at +20 mV with physiological gradients of K+ and 165 pS at +20 mV with an extracellular K+ concentration of 37.5 mM and an intracellular K+ concentration of 150 mM. Gating of these channels in cell-attached patches was potentiated by membrane stretch. Channel gating evoked by both lipids was concentration-dependent, with detectable activation apparent at 4 microM in the majority of patches and maximal activation occurring between 32 and 64 microM. Gating was relatively voltage-independent. Large-conductance K+ channels were also activated in inside-out patches by the monounsaturated fatty acid 11-cis-eicosenoic acid but not by the fully saturated fatty acid arachidic acid. Application of 100 microM H2O2, an agent that activates cytosolic phospholipase A2, also caused activation of large-conductance K+ channels in intact neurons. The stimulatory effects of H2O2 were blocked by pretreatment with 20 microM 4-bromophenacyl bromide, an irreversible inhibitor of phospholipase A2. Therefore, mobilization of endogenous fatty acids can cause activation of large-conductance K+ channels in autonomic neurons.     

Recovery of Altered Fatty Acid Composition Induced by a Diet Devoid of n-3 Fatty Acids in Myelin, Synaptosomes, Mitochondria, and Microsomes of Developing Rat Brain

Rats were fed a semisynthetic diet containing either sunflower oil or soya oil. Half the litter fed with sunflower oil diet was changed to a soya oil diet when the pups were 15 days old (during active myelination). Fatty acid analysis was then performed on subcellular fractions of the animals fed (a) soya oil, (b) sunflower oil, and (c) soya oil replacing sunflower oil from the 15th day, to determine the speed of the recovery. All material from animals fed sunflower oil showed an important reduction in docosahexaenoic acid (22:6 n-3), compensated by an increase in docosapentaenoic acid (22:5 n-6), whereas arachidonic acid (20:4 n-6) was not affected. In all fractions examined, when sunflower oil was replaced by soya oil in 15-day-old pups the recovery started from the very first day but lasted more than 2 months (this recovery was determined by the increase of 22:6 n-3 up to the normal value and decrease of the 22:5 n-6). In addition a delay was found for myelin recovery, starting only from the 25th day.