Unsaturated free fatty acids regulate cAMP relay and cell-cell adhesion duringDictyostelium discoideum aggregation

Summary Cell-cell adhesion and cAMP-stimulated cAMP production (cAMP relay) regulate the aggregation that occurs early in theDictyostelium discoideum developmental cycle. Increasing the concentration of neutral lipids inD. discoideum membranes inhibits both cell-cell adhesion and cAMP relay. Fractionation experiments revealed that it was the free fatty acids, one group of the neutral lipids, that inhibited both cAMP relay and cell-cell adhesion. Work with commercially-available free fatty acids demonstrated that the addition of saturated free fatty acids, palmitic acid and stearic acid, did not alter cell-cell adhesion or cAMP relay. The addition of unsaturated free fatty acids inhibited both cAMP relay and cell-cell adhesion in a dose-dependent, saturable manner. To test the physiological significance of these observations, the concentrations of endogenous unsaturated fatty acids were modified by altering theD. discoideum diet. Decreasing the amount of fatty acids 18 carbons long with 2 double bonds enhanced cAMP relay and cell-cell adhesion. These results suggest that fatty acids may be important regulators of cAMP responsiveness and cell-cell adhesion duringD. discoideum aggregation, and therefore may play important roles in development.     

Growth temperature affects accumulation of exogenous fatty acids and fatty acid composition in <Emphasis Type="Italic">Schizosaccharomyces pombe</Emphasis>

The incorporation of exogenously supplied fatty acids, palmitic acid, palmitoleic acid, oleic acid and linoleic acid, was examined in the yeast Schizosaccharomyces pombe at two growth temperatures, 20 °C and 30 °C. Fatty acids supplied to S. pombe in the growth medium were found to be preferentially incorporated into the cells, becoming a dominant species. The relative increase in exogenous fatty acids in cells came at the expense of endogenous oleic acid as a proportion of total fatty acids. Lowering the temperature at which the yeast were grown resulted in decreased levels of incorporation of the fatty acids palmitic acid, palmitoleic acid and linoleic acid compared to cells supplemented at 30 °C. In addition, the relative amount of the endogenously produced unsaturated fatty acid oleic acid, while greatly reduced compared to unsupplemented cells, was increased in cells supplemented with fatty acids at 20 °C compared to supplemented cells at 30 °C. The differential production of oleic acid in S. pombe cells indicates that regulation of unsaturated fatty acid levels, possibly by control of the stearoyl-CoA desaturase, is an important control point in membrane composition in response to temperature and diet in this species.     

Responses to oleic,linoleic and α-linolenic acids in high-carbohydrate,high-fat diet-induced metabolic syndrome in rats

We investigated the changes in adiposity, cardiovascular and liver structure and function, and tissue fatty acid compositions in response to oleic acid-rich macadamia oil, linoleic acid-rich safflower oil and α-linolenic acid-rich flaxseed oil (C18 unsaturated fatty acids) in rats fed either a diet high in simple sugars and mainly saturated fats or a diet high in polysaccharides (cornstarch) and low in fat. The fatty acids induced lipid redistribution away from the abdomen, more pronounced with increasing unsaturation; only oleic acid increased whole-body adiposity. Oleic acid decreased plasma total cholesterol without changing triglycerides and nonesterified fatty acids, whereas linoleic and α-linolenic acids decreased plasma triglycerides and nonesterified fatty acids but not cholesterol. α-Linolenic acid improved left ventricular structure and function, diastolic stiffness and systolic blood pressure. Neither oleic nor linoleic acid changed the left ventricular remodeling induced by high-carbohydrate, high-fat diet, but both induced dilation of the left ventricle and functional deterioration in low fat-diet-fed rats. α-Linolenic acid improved glucose tolerance, while oleic and linoleic acids increased basal plasma glucose concentrations. Oleic and α-linolenic acids, but not linoleic acid, normalized systolic blood pressure. Only oleic acid reduced plasma markers of liver damage. The C18 unsaturated fatty acids reduced trans fatty acids in the heart, liver and skeletal muscle with lowered stearoyl-CoA desaturase-1 activity index; linoleic and α-linolenic acids increased accumulation of their C22 elongated products. These results demonstrate different physiological and biochemical responses to primary C18 unsaturated fatty acids in a rat model of human metabolic syndrome.     

The degree of dietary fatty acid unsaturation affects torpor patterns and lipid composition of a hibernator

Diets rich in unsaturated and polyunsaturated fatty acids have a positive effect on mammalian torpor, whereas diets rich in saturated fatty acids have a negative effect. To determine whether the number of double bonds in dietary fatty acids are responsible for these alterations in torpor patterns, we investigated the effect of adding to the normal diet 5% pure fatty acids of identical chain length (C18) but a different number of double bonds (0, 1, or 2) on the pattern of hibernation of the yellow-pine chipmunk, Eutamias amoenus. The response of torpor bouts to a lowering of air temperature and the mean duration of torpor bouts at an air temperature of 0.5°C (stearic acid C18:0, 4.5±0.8 days, oleic acid C18:1, 8.6±0.5 days; linoleic acid C18:2, 8.5±0.7 days) differed among animals that were maintained on the three experimental diets. The mean minimum body temperatures (C18:0, +2.3±0.3°C; C18:1, +0.3±0.2°C; C18:2,-0.2±0.2°C), which torpid individuals defended by an increase in metabolic rate, and the metabolic rate of torpid animals also differed among diet groups. Moreover, diet-induced differences were observed in the composition of total lipid fatty acids from depot fat and the phospholipid fatty acids of cardiac mitochondria. For depot fat 7 of 13 and for heart mitochondria 7 of 14 of the identified fatty acids differed significantly among the three diet groups. Significant differences among diet groups were also observed for the sum of saturated, unsaturated and polyunsaturated fatty acids. These diet-induced alterations of body fatty acids were correlated with some of the diet-induced differences in variables of torpor. The results suggest that the degree of unsaturation of dietary fatty acids influences the composition of tissues and membranes which in turn may influence torpor patterns and thus survival of hibernation.Abbreviations bm body mass - T _a air temperature - T _b body temperature - FA fatty acid - MR metabolic rate - MUFA monounsaturated fatty acids - PUFA polyunsaturated fatty acids - VO₂ rate of oxygen consumption - SFA saturated fatty acids - UFA unsaturated fatty acids - UI unsaturation index - SNK Student-Newman-Keuls test     

Manipulation of fatty acid composition of membrane phospholipid and its effects on cell growth in mouse LM cells

Fatty acid composition of the phospholipids of mouse LM cells grown in suspension culture in serum-free chemically defined medium was modified by supplementing the medium with various fatty acids bound to bovine serum albumin.Following supplementation with saturated fatty acids of longer than 15 carbons (100 μM) profound inhibition of cell growth occurred; this inhibitory effect was completely abolished when unsaturated fatty acids were added at the same concentration. Supplementing with unsaturated fatty acids such as linoleic acid, linolenic acid or arachidonic acid had no effect on the cell growth.Fatty acid composition of membrane phospholipids could be manipulated by addition of different fatty acids. The normal percentage of unsaturated fatty acids in LM cell membrane phospholipids (63%) was reduced to 35–41% following incorporation of saturated fatty acids longer than 15 carbon atoms and increased to 72–82% after addition of unsaturated fatty acids.A good correlation was found between the unsaturated fatty acid content of membrane phospholipids and cell growth. When incorporated saturated fatty acids reduced the percentage of unsaturated fatty acids in membrane phospholipids to less than 50%, severe inhibition of the cell growth was found. Simultaneous addition of an unsaturated fatty acid completely abolished this effect of saturated fatty acids.     

Effects of exogenous fatty acids on calcium uptake by brush-border membrane vesicles from rabbit small intestine

The uptake of a variety of fatty acids by isolated brush-border membranes from rabbit small intestine was studied. This uptake increased with acyl chain-length and was not diminished by washing of the lipid-treated membranes with 0.25 M CsBr. The binding of fatty acid was not accompanied by a decrease in endogenous acyl groups or of cholesterol and therefore corresponded to a net uptake accountable qualitatively and quantitatively by the fatty acid added to the membranes. The uptake of Ca2+ was stimulated by treatment of the membranes with low concentrations of unsaturated fatty acids (0.05 mM) as well as with various concentrations of caprylic acid (0.10-3.00 mM) and inhibited by treatment with higher concentrations of unsaturated fatty acids (0.20-0.60 mM). Saturated fatty acids had no marked effects on Ca2+ uptake. The stimulatory concentrations of unsaturated fatty acids did not change the Ca2+-binding characteristics of the membranes, whereas the higher concentrations decreased equilibrium binding of Ca2+ and very probably the number of high-affinity binding sites. The results of this study are assessed in terms of the effects of normal fatty acids found in the diet on the absorptive properties of the brush-border membranes.     

The effect of unsaturated and saturated dietary lipids on the pattern of daily torpor and the fatty acid composition of tissues and membranes of the deer mouse Peromyscus maniculatus

Summary Dieary lipids strongly influence the pattern of torpor and the body lipid composition of mammalian hibernators. The object of the present study was to investigate whether these diet-induced physiological and biochemical changes also occur in species that show shallow, daily torpor. Deer mice, Peromyscus maniculatus, were fed with rodent chow (control diet) or rodent chow with either 10% sunflower seed oil (unsaturated diet) or 10% sheep fat (saturated diet). Animals on the unsaturated diet showed a greater occurrence of torpor (80–100% vs 26–43%), longer torpor bouts (4.5 vs 2.25 h), a lower metabolic rate during torpor (0.96 vs 2.25 ml O₂·g^-1·h^-1), and a smaller loss of body mass during withdrawal of food (2.35 vs 3.90 g) than animals on the saturated diet; controls were intermediate. These diet-induced physiological changes were associated with significant alterations in the fatty acid composition of depot fat, leg muscle and brain total lipids, and heart mitochondrial phospholipids. Significant differences in the total unsaturated fatty acid (UFA) content between animals on saturated and unsaturated diet were observed in depot fat (55.7% vs 81.1%) and leg muscle (56.4% vs 72.1%). Major compositional differences between diet groups also occurred in the concentration of n6 and/or n3 fatty acids of brain and heart mitochondria. The study suggests that dietary lipids may play an important role in the seasonal adjustment of physiology in heterothermic mammals.Abbreviations EDTA ethylenediaminetetra-acetic acid - HEPES N-2 hydroxyethylpiperazine-N¹-2-ethanesulphonic acid - MUFA monounsaturated fatty acids - PUFA polyunsaturated fatty acids - RMR Testing metabolic rate - SD standard deviation - SFA saturated fatty acids - SNK Student-Newman-Keuls test - T₁ air temperature - T_b body temperature - UFA unsaturated fatty acids - rate of oxygen consumption Dedicated to the late John K. Raison     

Influence of Host Diet on the Fatty Acid Composition and Content of Rumen Protozoa in Cattle

The fatty acid profiles and contents of protozoa from the rumen fluid of cattle varied according to the type of diet consumed by their host. Changing from a high-quality hay diet to a low-quality hay diet (DA) decreased the proportions of saturated acids and increased the proportions of the unsaturated acids 18:1 cis-9, 18:2 and 18:3 in neutral lipids (NL) and phospholipids (PL). Adding sucrose, urea and sulphur (SUS) to DA increased the proportions of branched chain acids in PL while addition of safflower oil increased polyunsaturated acids in PL and 18:1 trans-11 in NL. Diet did not alter the PL fatty acid content of protozoa but oil supplement of DA resulted in a 10-fold increase in the content of free fatty acids. The defaunating effect of oil supplement was partly reversed by SUS suggesting that factors other than the fatty acid content of cells are important in determining the toxicity of oil to rumen protozoa. The results indicate that the amounts of individual long-chain fatty acids taken up by rumen ciliates are largely determined by their concentrations in rumen digesta.     

Influence of host diet on the fatty acid composition and content of rumen protozoa in cattle

The fatty acid profiles and contents of protozoa from the rumen fluid of cattle varied according to the type of diet consumed by their host. Changing from a high-quality hay diet to a low-quality hay diet (DA) decreased the proportions of saturated acids and increased the proportions of the unsaturated acids 18:1 cis-9, 18:2 and 18:3 in neutral lipids (NL) and phospholipids (PL). Adding sucrose, urea and sulphur (SUS) to DA increased the proportions of branched chain acids in PL while addition of safflower oil increased polyunsaturated acids in PL and 18:1 trans-11 in NL. Diet did not alter the PL fatty acid content of protozoa but oil supplement of DA resulted in a 10-fold increase in the content of free fatty acids. The defaunating effect of oil supplement was partly reversed by SUS suggesting that factors other than the fatty acid content of cells are important in determining the toxicity of oil to rumen protozoa. The results indicate that the amounts of individual long-chain fatty acids taken up by rumen ciliates are largely determined by their concentrations in rumen digesta.     

Fatty acid compositions of arthropod and cephalopod photoreceptors: interspecific,seasonal and developmental studies

Fatty acid compositions of the compound eyes of insects (soldier-bug, Hemiptera, and silk moth, Lepidoptera), crustaceans (crayfish and grapsid crab, Decapoda) and inner and outer segments of visual cells of a squid (Cephalopoda, Mollusca) were analyzed by gaschromatography for interspecific comparison. Fatty acid compositions showed great variation among species. In insect compound eyes, 16:0 and 18:0 were the main saturated fatty acids, and 18:1 was the dominant unsaturated fatty acid. Silk moth eyes contained, in addition, considerable amounts of 18:2 and 20:5. In crustacean compound eyes, the main saturated fatty acids were 16:0 and 18:0, and 14:0 (5.0%) was only detected in grapsid crabs; the main unsaturated fatty acids were 20:4, 20:5 and 22:6. Both whole eyes and rhabdom fraction of crayfish showed similar profiles of fatty acid compositions. Both inner and outer segments of squid retinae were characterized by high amounts of unsaturated fatty acids, especially 22:6. Compound eyes of grapsid crabs were used for the experiments on seasonal changes of fatty acid compositions. UFA/SFA ratios (weight in % of unsaturated fatty acids saturated fatty acids) were lowest (1.0) in July and highest (2.5) in March, and unsaturation indexes (average number of double bonds per molecule) were lowest (1.5) in July and highest (2.3) in March. Fatty acids 18:0 and 20:1 showed a significant correlation with the changes of seasonal temperature. Fatty acid analysis of the developing compound eyes of silk moths during the pupal stage revealed that eicosapentanoic acid (20:5) increased remarkably in parallel with the development of photoreceptive membranes, the rhabdoms. This suggests that eicosapentaenoic acid may play an important role in formation and function of rhabdoms.     

Spectrum of effects of dietary long-chain fatty acids on rat intestinal glucose and lipid uptake

Isocaloric modification in the ratio of dietary polyunsaturated-to-saturated fatty acids influences intestinal uptake of actively and passively transported nutrients. This study was undertaken to determine which dietary fatty acid was responsible for these alterations in absorption. Adult female rats were fed isocaloric semisynthetic diets high in palmitic and stearic acids (SFA), oleic acid (OA), linoleic acid (LA), or linolenic acid (LNA). An in vitro technique was used to measure the uptake of varying concentrations of glucose as well as a series of fatty acids and cholesterol. Jejunal uptake of 40 mM glucose was highest in rats fed SFA and lowest in those fed LA; ileal glucose uptake was similar in OA, LA, and LNA, but was lowest in SFA. Jejunal uptake of medium-chain fatty acids (8:0-12:0) was higher in OA than in other diet groups; ileal uptake of medium-chain fatty acids was unaffected by diet. Jejunal and ileal uptake of 18:2 was higher in LNA than in SFA or OA; the uptake of the other long-chain saturated or unsaturated fatty acids was unchanged by diet. The ileal but not the jejunal uptake of cholesterol was increased in LA as compared with SFA or OA, and reduced in LNA as compared with LA. These transport changes were not explained by differences in the animals' food consumption, body weight gain, intestinal mass, or mucosal surface area. We postulate that these diet-induced transport alterations may be mediated via changes in brush border membrane phospholipid fatty acyl composition. Thus, intestinal transport of nutrients may be varied by isocaloric changes in the dietary content of individual fatty acids.     

Effects of unsaturated fatty acids on the peroxisomal enzyme activities of Tetrahymena pyriformis

The effects of unsaturated fatty acids on the activities of peroxisomal enzymes of Tetrahymena pyriformis were investigated. When saturated fatty acids and the corresponding unsaturated fatty acids (C18) were added to the culture medium at 0.05%, the activities of peroxisomal enzymes [fatty acyl-CoA oxidase (FAO), carnitine acetyltransferase (CAT), isocitrate lyase (ICL), and malate synthase (MS)] were significantly increased. The order of effectiveness was linoleic acid greater than oleic acid greater than stearic acid. However, alpha-linolenic acid and gamma-linolenic acid at the same concentration were lethal to the cells. The inhibitory effect on growth disappeared upon addition of an antioxidant, alpha-tocopherol. Lipid peroxides derived from unsaturated fatty acids induced marked cell lysis. In the presence of a low concentration (0.005%) of linolenic acid the production of lipid peroxide was lower and no inhibitory effect on the growth was observed, while the activities of peroxisomal enzymes participating in lipid metabolism and that of catalase were significantly increased. These results indicate that the peroxisomal enzyme systems related to the beta-oxidations of fatty acids and the glyoxylate cycle are regulated by unsaturated long-chain fatty acids, including linolenic acid, at low concentrations, as well as by saturated fatty acid in the medium.     

Sensitivity to alcohol in mice with an altered brain fatty acid composition

Ethanol-induced sleep onset times, sleep times and blood alcohol levels upon awakening were measured in mice fed an essential fatty acid deficient, Purina Chow or unsaturated fat diet for nine months. These values in animals fed the essential fatty acid deficient and Purina Chow diets did not differ, but mice fed the unsaturated fat diet had longer sleep times and lower blood alcohol levels upon awakening than mice fed essential fatty acid deficient or Purina Chow diets. Crude brain mitochondrial fractions isolated from mice fed the essential fatty acid deficient diet had decreased levels of docosahexaenoic [22:6(n-3)] and increased levels of eicosatrienoic [20:3(n-9)], docosatrienoic [22:3(n-9)] and docosapentaenoic [22:5(n-6)] acids compared to mice fed the Purina Chow diet. The unsaturated fat diet decreased 22:6(n-3) and increased 22:5(n-6) compared to the Purina Chow dietary regimen. The longer sleep times and lower blood alcohol levels found in mice fed the unsaturated fat diet probably resulted from an artifact due to the obesity of the mice fed this diet and from the hinderance of obesity to the righting reflex (our measure of ethanol potency). We conclude that the alteration of several polyunsaturated fatty acid components in the brain has little or no influence on the sensitivity of the nervous system to alcohol.     

Dietary fats,selected body temperature and tissue fatty acid composition of agamid lizards (Amphibolurus nuchalis)

The composition of tissue and membrane fatty acids in ectothermic vertebrates is influenced by both temperature acclimation and diets. If such change in body lipid composition and thermal physiology were linked, a diet-induced change in body lipid composition should result in a change in thermal physiology. We therefore investigated whether the selected body temperature of the agamid lizardAmphibolurus nuchalis (body mass 20 g) is influenced by the lipid composition of dietary fatty acids and whether diet-induced changes in thermal physiology are correlated with changes in body lipid composition. The selected body temperature in two groups of lizards was indistinguishable before dietary treatments. The selected body temperature in lizards after 3 weeks on a diet rich in saturated fatty acids rose by 2.1 °C (photophase) and 3.3 °C (scotophase), whereas the body temperature of lizards on a diet rich in unsaturated fatty acids fell by 1.5 °C (photophase) and 2.0 °C (scotophase). Significant diet-induced differences were observed in the fatty acid composition of depot fat, liver and muscle. These observations suggest that dietary lipids may influence selection of body temperature in ectotherms via alterations of body lipid composition.Abbreviations bm body mass - FA fatty acid(s) - MUFA monounsaturated fatty acids - PUFA polyunsaturated fatty acids - SFA saturated fatty acids - T _a air temperature - T _b body temperature - UFA unsaturated fatty acids     

Effects of dietary fatty acids on hepatic 3-hydroxy-3-methylglutaryl CoA reductase activity in hamsters on a high-glucose diet

Hepatic 3-hydroxy-3-methylglutaryl CoA reductase activity in hamsters given a fat-free high-glucose diet for 21 days was approximately 20 times higher than that in chow-fed hamsters. The increase in enzyme activity by dietary glucose was affected by saturated or unsaturated fatty acids or cholesterol added to the high-glucose diet. Ethyllinoleate or ethyloleate, added to the diet at a concentration of 5%, suppressed the increase in the enzyme activity. In contrast, addition of ethylpalmitate to the diet further stimulated the increase in the enzyme activity. Addition of 2% cholesterol to the high-glucose diet moderately suppressed, and addition of both cholesterol and ethyllinoleate completely prevented, the increase in the enzyme activity. The enzyme activity closely correlated with the incidence of formation of cholesterol gallstones but not with the liver cholesterol level. Marked increase in the enzyme activity was observed by feeding the high-glucose diet to starved hamsters for even a short period. On the third day after feeding was resumed, the enzyme activity was increased 500-fold compared to that during starvation. This increase in the enzyme activity was also reduced by dietary unsaturated fatty acid esters and stimulated by a dietary saturated fatty acid ester.     

Influence of fatty acids and bovine serum albumin on the growth of human hepatoma and immortalized human kidney epithelial cells

Summary The protective influence of bovine serum albumin against growth inhibition caused by fatty acids was studied in human hepatoma (HepG2) and immortalized human kidney epithelial (IHKE) cells. In general, growth inhibition by unsaturated fatty acids (0.15 mmol/liter) increased with increasing number of double bonds. For HepG2 cells crude albumin (1g/100 ml) did not greatly modify growth inhibition by arachidonic, eicosapentaenoic, and docosahexaenoic acid. With oleic, linoleic, and linolenic acids, crude and defatted albumin stimulated cell growth. In contrast, for IHKE cells both albumins counteracted growth inhibition by unsaturated fatty acids to approximately the same extent. When HepG2 cells were cultured in the presence of saturated fatty acids (0.3 mmol/liter), C2, C6, and C8 had no or little inhibitory effect. C10 and C12 inhibited cell growth appreciably, whereas C14, and especially C16, had poor inhibitory effects. Crude albumin counteracted growth inhibition by all these fatty acids. In contrast, defatted albumin had little or no effect (except against C10 and C12), and even increased the growth inhibition by C14 and C16. With unsaturated fatty acids there seemed to be an inverse relationship between cell growth and the concentration of thiobarbituric acid reactive substances (TBARS) in media. Vitamin E abolished growth inhibition (and the increase in TBARS concentration) by unsaturated fatty acids. The complex interaction between fatty acids and albumins calls for great caution when interpreting data on growth effects.     

Possible Involvement of Palmitate in Pathogenesis of Periodontitis

Type 2 diabetes (T2D) is characterized by decreased insulin sensitivity and higher concentrations of free fatty acids (FFAs) in plasma. Among FFAs, saturated fatty acids (SFAs), such as palmitate, have been suggested to promote inflammatory responses. Although many epidemiological studies have shown a link between periodontitis and T2D, little is known about the clinical significance of SFAs in periodontitis. In this study, we showed that gingival fibroblasts have cell‐surface expression of CD36, which is also known as FAT/fatty acid translocase. Moreover, CD36 expression was increased in gingival fibroblasts of high‐fat diet‐induced T2D model mice, compared with gingival fibroblasts of mice fed a normal diet. DNA microarray analysis revealed that palmitate increased mRNA expression of pro‐inflammatory cytokines and chemokines in human gingival fibroblasts (HGF). Consistent with these results, we confirmed that palmitate‐induced interleukin (IL)‐6, IL‐8, and CXCL1 secretion in HGF, using a cytokine array and ELISA. SFAs, but not an unsaturated fatty acid, oleate, induced IL‐8 production. Docosahexaenoic acid (DHA), which is one of the omega‐3 polyunsaturated fatty acids, significantly suppressed palmitate‐induced IL‐6 and IL‐8 production. Treatment of HGF with a CD36 inhibitor also inhibited palmitate‐induced pro‐inflammatory responses. Finally, we demonstrated that Porphyromonas gingivalis (P.g.) lipopolysaccharide and heat‐killed P.g. augmented palmitate‐induced chemokine secretion in HGF. These results suggest a potential link between SFAs in plasma and the pathogenesis of periodontitis. J. Cell. Physiol. 230: 2981–2989, 2015. © 2015 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc.

     

Total lipid and fatty acid compositions of <Emphasis Type="Italic">Lertha sheppardi</Emphasis> (Neuroptera: Nemopteridae) during its main life stages

Total lipid and the fatty acid compositions of phospholipid and triacylglycerol fractions, prepared from eggs, 3^rd instars of larvae, pupae, male and female adults of Lertha sheppardi, were analyzed by gas chromatography and gas chromatography-mass spectrometry. The effect of diet (adults’ nutrition) on fatty acid composition of L. sheppardi adults was also investigated. Total lipid of L. sheppardi considerably increased in adults compared with immature stages. There was a significant decrease in total lipid level in larval stage in contrast with egg stage. Qualitative analysis revealed the presence of 14 fatty acids during all stages. The major components were C16 and C18 saturated and unsaturated components which are ubiquitous to most animal species. In addition to these components, one odd-chain (C17:0) and prostaglandin precursor fatty acids were found. The fatty acid profiles of phospholipids and triacylglycerols were substantially different. In phospholipid fraction, monounsaturated fatty acids were the major proportion of fatty acids in both sex of adults and pupae, whereas polyunsaturated fatty acids were the most dominant fatty acids in eggs and 3^rd instars. Results of triacylglycerol fraction revealed that fatty acid composition of eggs had higher level of C16:1, C18:0 and C18:3n-3 content than that of 3^rd instars and pupae, which suggests accumulation of energetic and structural reserve materials during embryonic development. At more advanced developmental stages, mainly in adult females, the amount of C16:1 increased once again, which may be related to the need for accumulation of sufficient energy and of carbon reservoir in the developing new vitellum. Percentages of C18:1 were significantly high in adult stages compared to other stages. These findings indicate that the accumulation and consumption of fatty acids fluctuate through different development stages. Diet did not effect the fatty acid composition of L. sheppardi adults.     

Substrate specificity of <Emphasis Type="Italic">Stenotrophomonas nitritireducens</Emphasis> in the hydroxylation of unsaturated fatty acid

An isolated bacterium that converted unsaturated fatty acids to hydroxy fatty acids was identified as Stenotrophomonas nitritireducens by API analysis, cellular fatty acids compositions, sequencing the full 16S ribosomal ribonucleic acid, and evaluating its nitrite reduction ability. S. nitritireducens has unique regio-specificity for C16 and C18 cis-9 unsaturated fatty acids. These fatty acids are converted to their 10-hydroxy fatty acids without detectable byproducts. Among the cis-9-unsaturated fatty acids, S. nitritireducens showed the highest specificity for linoleic acid. The cells converted 20 mM linoleic acid to 13.5 mM 10-hydroxy-12(Z)-octadecenoic acid at 30°C and pH 7.5 with a yield of 67.5% (mol/mol).     

Metabolomics of dietary Fatty Acid restriction in patients with phenylketonuria

Background

Patients with phenylketonuria (PKU) have to follow a lifelong phenylalanine restricted diet. This type of diet markedly reduces the intake of saturated and unsaturated fatty acids especially long chain polyunsaturated fatty acids (LC-PUFA). Long-chain saturated fatty acids are substrates of mitochondrial fatty acid oxidation for acetyl-CoA production. LC-PUFA are discussed to affect inflammatory and haemostaseological processes in health and disease. The influence of the long term PKU diet on fatty acid metabolism with a special focus on platelet eicosanoid metabolism has been investigated in the study presented here.

Methodology/Principal Findings

12 children with PKU under good metabolic control and 8 healthy controls were included. Activated fatty acids (acylcarnitines C6–C18) in dried blood and the cholesterol metabolism in serum were analyzed by liquid chromatographic tandem mass spectrometry (LC-MS/MS). Fatty acid composition of plasma glycerophospholipids was determined by gas chromatography. LC-PUFA metabolites were analyzed in supernatants by LC-MS/MS before and after platelet activation and aggregation using a standardized protocol. Patients with PKU had significantly lower free carnitine and lower activated fatty acids in dried blood compared to controls. Phytosterols as marker of cholesterol (re-) absorption were not influenced by the dietary fatty acid restriction. Fatty acid composition in glycerophospholipids was comparable to that of healthy controls. However, patients with PKU showed significantly increased concentrations of y-linolenic acid (C18:3n-6) a precursor of arachidonic acid. In the PKU patients significantly higher platelet counts were observed. After activation with collagen platelet aggregation and thromboxane B₂ and thromboxane B₃ release did not differ from that of healthy controls.

Conclusion/Significance

Long-term dietary fatty acid restriction influenced the intermediates of mitochondrial beta-oxidation. No functional influence on unsaturated fatty acid metabolism and platelet aggregation in patients with PKU was detected.