The effect of free fatty acids on the thermotropic phase transition of dimyristoyl glycerophosphocholine

The effect of free fatty acids on the phase transition characteristics and fluidity of bilayers of dimyristoyl glycerophosphocholine were studied by pyrene eximer fluorescence and differential scanning calorimetry. High melting saturated fatty acids with chain lengths of 12–18 carbon atoms raise the phase transition temperature and enhance the ability of pyrene to form clusters in the gel state while not affecting the fluidity of the membrane in the liquid crystal state. Low melting unsaturated fatty acids lower the phase transition temperature and decrease the ability of pyrene to form clusters in the gel state while not affecting the fluidity of the membrane in the liquid crystal state. The effects of the very long chain fatty acids, arachidic (C 20) and behenic (C 22) appear to be similar to those of cholesterol in that they cause a broadening of the phase transition with a lowering of the transition enthalpy but have little effect on the temperature at which the phase transition occurs.     

Effect of Macrophomina phaseolina Infection on the Physico-Chemical Components of Groundnut Seed

Various aspects of groundnut seed deterioration due to M. phaseolina infection were studied. Fungal infection caused both quantitative and qualitative damage to the seed. It resulted in characteristic discoloration of pod and kernel. There was marked reduction in the pod and kernel yield, shelling percentage and oil content. The oil extracted from healthy kernels was barium yellow, whereas, oil from diseased kernels was amber yellow in color. The diseased kernels contained lesser amounts ofstearic and behenic acids but higher quantities of palmitic, oleic, linoleic and arachidic acids. Four fatty acids, namely, lauric, myristic, palmitoleic and eicosenoic acids were absent in diseased kernels.     

Determination of saturated and unsaturated Fatty acids amount in leukocyte membranes from subjects fed with solid and fluid oils

Modifications in dietary fatty acid intake might lead to a modification in membrane phospholipid fatty acid composition. The purpose of this study was to investigate relationship between different type of oil consumption and leukocyte membrane phospholipid composition. This study was carried out in subjects utilizing butter (n = 15), margarine (n = 15), fluid oil (n = 15) and mixed types of oils (n = 15) in total 60 subjects. Leukocytes were separated from total blood by dextran sedimentation method. Membrane lipids and proteins were isolated following the cell disruption. Fatty acids of membrane phospholipids were isolated by hydrolysation with phospholipase B under ultrasonic dismembranator. Free fatty acids were identified with gas chromatography at chloroform phase. The results obtained were compared with data obtained by chromatograms of the standards. Results more prominent values of arachidic, dihomo-gamma-linolenic and palmitoleic acids were found in butter-or mixed oil-user groups; eicosadienoic, eicosamonoenoic, dihomo-gamma-linolenic and behenic acids in fluid oil heptanoic, valeric, eicosadienoic and linolenic acids in margarine groups. The fatty acid composition of mixed oil; was similar to butter, while other two oils were so different. From this study, it was concluded that the type of oil consumption might have an influence on phospholipid components of plasma membranes.     

Studies on methanol-oxidizing yeast

The yeast strain 11Bh was studied from the aspect of qualitative and qunatitative composition of lipids formed in cells during growth on methanol, synthetic ethanol and glucose. The strain was found to form some 3% free fatty acids toward the end of the growth phase. More esterified fatty acids are formed on ethanol and glucose (2.75 and 2.86%, respectively) than on methanol (1.6%). The composition of lipids and representation of the various fatty acids in the lipids is similar on all three substrates. The cell lipids contain over 40 rel.% oleic and about 16 rel.% each of palmitoleic, palmitio and linolenic acid. Odd-numbered fatty acids are present after growth on any of the three substrates, amounting to at most 4 rel.%. Of the extracellular fatty acids formed toward the end of growth of cells on methanol, propionic and acetic acid occurred in largest amounts in the medium. An erratum to this article is available at .     

Fatty acid profiles of cells from the insect cell line iprl-21 (Spodoptera frugiperda) and of the tissue culture medium after repeated use

Summary When IPL-1 medium was used for three serial incubations of cells of the IPRL-21 insect cell line (Spodoptera frugiperda, J. E. Smith) at least 23 fatty acids were identified from the media and/or from the cells. During the first incubation only negligible changes occurred in the total fatty acid content of the medium, but after the second and third incubations the total content decreased. Seven of the 23 fatty acids (palmitic, palmitoleic, stearic, oleic, linoleic, linolenic, and arachidonic acids) comprised 92% of the total fatty acid content, but the specific concentrations varied after each 7-day incubation. During the first incubation, the concentration of the monoene fatty acids increased, and the concentration of the more highly unsaturated fatty acids decreased. During the second and third serial incubations, the specific concentrations of all fatty acids decreased, with the exception of palmitoleic acid. These changes in the total fatty acid content and in the specific concentration of individual fatty acids in the cell indicated uptake of fatty acids from the medium and/or cellular lipid biosynthesis. The fatty acid content of the cells differed during the active growth phase and the stationary phase.     

Thermal dependence of apolipoprotein A-I-phospholipid recombination

Studies of the recombination of apolipoprotein A-I (apo A-I), the major protein constituent of human high-density lipoprotein, and various synthetic phospholipids, both alone and in mixtures, have been performed. Pure diacyl phospholipids containing homologous fatty acids of the C12, C13, C14, and C15 series, as well as the two positional isomers containing C14 and C16 fatty acids in positions 1 and 2, undergo reaction with the apo A-I protein only near their gel-liquid-crystalline transition temperatures; the degree of reactivity of these phospholipids toward recombination was observed to decrease as the transition temperature increased. The presence of lysolecithin in the incubation mixtures at proportions of 5 mol/mol of protein or lower was not found to have a significant effect on the rate of recombination. Binary mixtures of dimyristoylphosphatidylcholine and dipalmitoylphosphatidylcholine at various proportions react maximally with apo A-I at the onset of the phase transition, as judged by comparison with published phase diagrams; in this case also, the rate of recombination was observed to decline for mixtures with higher phase transition temperatures. These results are interpreted in terms of protein insertion at lattice defects arising from the presence of phospholipid clusters undergoing the phase transition; these clusters are derived from the cooperative and simultaneous melting of a number of molecules, the cooperativity being dependent upon the nature of the phospholipid. It is postulated that phospholipids which melt in a more highly cooperative fashion are more capable of interacting with the apolipoproteins since these phospholipids will possess larger lattice defects during the phase transition.     

Expression of the Arabidopsis ADS1 gene in Brassica juncea results in a decreased level of total saturated fatty acids

Brassica juncea plants transformed with the Arabidopsis ADS1 gene, which encodes a plant homologue of the mammalian and yeast acyl-CoA Delta9 desaturases and the cyanobateria acyl-lipid Delta9 desaturase, were found to have a statistically significant decrease in the level of saturated fatty acids in seeds. The decrease in the level of saturated fatty acids is largely attributable to decreases in palmitic acid (16:0) and stearic acid (18:0), although arachidic acid (20:0), behenic acid (22:0) and lignoceric acid (24:0) were also decreased in the transgenic seeds compared to the negative control lines. As a result, the level of oleic acid (18:1) was slightly increased in the transgenic seed lines compared to the non-transformed controls. However, a decrease in saturated fatty acid is not always accompanied by the corresponding increase in mono-unsaturated fatty acids. For example, palmitoleic acid (16:1), gondoic acid (20:1) and nervonic acid (24:1) were all found to be decreased in transgenic seeds. The levels of linoleic acid (18:2) and linolenic acid (18:3) were also notably changed in the transgenic lines compared to the controls. The present study provides preliminary experimental data suggesting that the Arabidopsis ADS1 encodes a fatty acid Delta9 desaturase and could be useful in genetic engineering for modifying the level of saturated fatty acids in oilseed crops. However, the effect of ADS1 gene expression on seed oil fatty acid composition is beyond the changes of total saturated and mono-unsaturated fatty acids, which suggests a complex mechanism is involved in the regulation of fatty acid metabolism.     

Desiccation-induced Changes in Antioxidant Enzymes,Fatty Acids,and Amino Acids in the Cyanobacterium <Emphasis Type="Italic">Tolypothrix scytonemoides</Emphasis>

Tolypothrix scytonemoides subjected to desiccation exhibited increased antioxidant enzyme activities when compared to fresh cells. The activities of catalase (EC 1.11.1.6) and superoxide dismutase (EC 1.15.1.1) were enhanced in desiccated cells by 42.8% and 8.1%, respectively. The isoforms of catalase and superoxide dismutase were detected by activity staining of crude samples separated on native gels. The isoforms of superoxide dismutase were identified based on their sensitivity to hydrogen peroxide and cyanide. The changes in fatty acids and amino acids in fresh and desiccated cells were also investigated and it was found that the quantity of certain fatty acids and amino acids was greater in desiccated cells. Palmitic acid, palmitoleic acid, heptadecanoic acid, linoleic acid, and myristic acid were more in desiccated cells when compared to fresh cells. Desiccated cells synthesized myristoleic acid, eicosenoic acid and behenic acid, acids which were not synthesized by the fresh cells, whereas tricosanoic acid was synthesized by the fresh cells and not by desiccated cells. The levels of lysine, serine, glycine, proline and cysteine were also comparatively greater in the desiccated cells.     

Critical temperatures for the interaction of free fatty acids with the erythrocyte membrane

Non-esterified long-chain fatty acids reduce the extent of hypotonic hemolysis at a certain low concentration range but cause hemolysis at higher concentrations. This biphasic behavior was investigated at different temperatures (0-37 degrees C) for lauric (12:0), myristic (14:0), palmitoleic (16:1), oleic (cis-18:1) and elaidic (trans-18:1) acids. The results are summarized as follows: (A) the fatty acids examined exhibit a high degree of specificity in their thermotropic behavior; (B) oleic acid protects against hypotonic hemolysis even at the highest concentrations, up to 15 degrees C, when it becomes hemolytic, but only in a limited concentration range; (C) elaidic acid does not affect the osmotic stability of erythrocytes up to 20 degrees C, when it starts protecting: above 30 degrees C, it becomes hemolytic at the highest concentrations; (D) palmitoleic acid is an excellent protecting agent at all temperatures in a certain concentration range, becoming hemolytic at higher concentrations; (E) lauric acid protects up to 30 degrees C and becomes hemolytic only above this temperature; (F) myristic acid exhibits an extremely unusual behavior at 30 and 37 degrees C by having alternating concentration ranges of protecting and hemolytic effects; (G) there is a common critical temperature for hemolysis at 30 degrees C for saturated and trans-unsaturated fatty acids; (H) the initial slope of Arrhenius plots of percent hemolysis at the concentration of maximum protection is negative for cis-unsaturated fatty acids and positive for saturated and trans-unsaturated fatty acids.     

The effect of alterations in the physical state of the membrane lipids on the ability of Acholeplasma laidlawii B to grow at various temperatures

The physical state of the membrane lipids, as determined by fatty acid composition and environmental temperature, has a marked effect on both the temperature range within which Acholeplasma laidlawii B cells can grow and on growth rates within the permissible temperature ranges. The minimum growth temperature of 8 °C is not defined by the fatty acid composition of the membrane lipids when cells are enriched in fatty acids giving rise to gel to liquid-crystalline membrane lipid phase transitions occurring below this temperature. The elevated minimum growth temperatures of cells enriched in fatty acids giving rise to lipid phase transitions occurring at higher temperatures, however, are clearly defined by the fatty acid composition of the membrane lipids. The optimum and maximum growth temperatures are also influenced indirectly by the physical state of the membrane lipids, being significantly reduced for cells supplemented with lower melting, unsaturated fatty acids. The temperature coefficient of growth at temperatures near or above the midpoint of the lipid phase transition is 16 to 18 $\text{kcal}\text{mol}$, but this value increases abruptly to 40 to 45 $\text{kcal}\text{mol}$ at temperatures below the phase transition midpoint. Both the absolute rates and temperature coefficients of cell growth are similar for cells whose membrane lipids exist entirely or predominantly in the liquid-crystalline state, but absolute growth rates decline rapidly and temperature coefficients increase at temperatures where more than half of the membrane lipids become solidified. Cell growth ceases when the conversion of the membrane lipid to the gel state approaches completion, but growth and replication can continue at temperatures where less than one tenth of the total lipid remains in the fluid state. An appreciable heterogeneity in the physical state of the membrane lipids can apparently be tolerated by this organism without a detectable loss of membrane function.     

Effect of diethylstilboestrol on adipose-tissue lipids

1. The effect of diethylstilboestrol on the fatty acid composition of adipose-tissue lipids of the ox (Bos taurus) was studied. 2. The capsula adiposa (perirenal) was shown to contain more total saturated fatty acids, whereas more total unsaturated fatty acids were found in the panniculus adiposus (subcutaneous). 3. Significantly more stearic acid and linolenic acid were obtained from the capsula adiposa, whereas the panniculus adiposus contained more myristoleic acid, palmitoleic acid and oleic acid. 4. Implanting diethylstilboestrol significantly increased the deposition of the saturated fatty acids, particularly stearic acid. 5. A decrease in the deposition of total unsaturated fatty acids, myristoleic acid, palmitoleic acid and linoleic acid can also be attributed to the diethylstilboestrol treatment.     

Fluorine-19 nuclear magnetic resonance studies of lipid fatty acyl chain order and dynamics in Acholeplasma laidlawii B membranes. A direct comparison of the effects of cis and trans cyclopropane ring and double-bond substituents on orientational order

The hydrocarbon chain orientational order parameters of membranes of Acholeplasma laidlawii B, enriched with large quantities of fatty acids containing either a cis or a trans cyclopropane ring or a cis or trans double bond, plus small quantities of one of an isomeric series of monofluoropalmitic acids, were determined via fluorine-19 nuclear magnetic resonance spectroscopy over a range of temperatures spanning the corresponding gel to liquid-crystalline phase transitions (determined via differential scanning calorimetry). Membrane orientational order profiles in the liquid-crystalline state were generally similar, regardless of the particular fatty acid structure present, showing a region of relatively constant order preceding a region of progressively decreasing order toward the methyl terminus of the acyl chain. In the gel state, the order profiles in the presence of either a trans cyclopropane ring or trans double-bond substituent were similar and were characterized by a pronounced head to tail gradient of order at temperatures just below the lipid phase transition, while at temperatures far below the lipid phase transition this gradient was less pronounced, all chain positions showing a more uniformly high degree of orientational ordering. In the gel state, the order profiles in the presence of either a cis cyclopropane ring or a cis double-bond substituent were also similar but were highly unusual in that order first increased and only then subsequently decreased toward the acyl chain methyl terminus. In addition, the substituents in the cis configuration, whether a cyclopropane ring or a double bond, were overall more disordered in the gel state than the corresponding substituents in the trans configuration.(ABSTRACT TRUNCATED AT 250 WORDS)     

Phosphatidate phosphatase activity plays key role in protection against fatty acid-induced toxicity in yeast

The PAH1-encoded phosphatidate (PA) phosphatase in Saccharomyces cerevisiae is a pivotal enzyme that produces diacylglycerol for the synthesis of triacylglycerol (TAG) and simultaneously controls the level of PA used for phospholipid synthesis. Quantitative lipid analysis showed that the pah1Δ mutation caused a reduction in TAG mass and an elevation in the mass of phospholipids and free fatty acids, changes that were more pronounced in the stationary phase. The levels of unsaturated fatty acids in the pah1Δ mutant were unaltered, although the ratio of palmitoleic acid to oleic acid was increased with a similar change in the fatty acid composition of phospholipids. The pah1Δ mutant exhibited classic hallmarks of apoptosis in stationary phase and a marked reduction in the quantity of cytoplasmic lipid droplets. Cells lacking PA phosphatase were sensitive to exogenous fatty acids in the order of toxicity palmitoleic acid > oleic acid > palmitic acid. In contrast, the growth of wild type cells was not inhibited by fatty acid supplementation. In addition, wild type cells supplemented with palmitoleic acid exhibited an induction in PA phosphatase activity and an increase in TAG synthesis. Deletion of the DGK1-encoded diacylglycerol kinase, which counteracts PA phosphatase in controlling PA content, suppressed the defect in lipid droplet formation in the pah1Δ mutant. However, the sensitivity of the pah1Δ mutant to palmitoleic acid was not rescued by the dgk1Δ mutation. Overall, these findings indicate a key role of PA phosphatase in TAG synthesis for protection against fatty acid-induced toxicity.     

Seasonal Changes of Fatty Acid Compositions in Overwintering Larvae of Rice Stem Borer,Chilo suppressalis (Lepidoptera: Pyralidae)

Cold acclimation and overwintering state can affect fatty acid compositions of insects. To determine compositional change of fatty acids during nondiapause and diapause stages, an experiment was conducted to investigate fatty acid constituents from whole body of C. suppressalis larvae. Five most abundant fatty acids were found to be palmitoleic (35–58%), palmitic (18–44%), oleic (14–23%), stearic (0.5–2.5%) and linoleic acid (0.4–2%). However, linolenic, erucic, lauric and myristic acid were found at lower level. Saturated fatty acids significantly decreased and conversely unsaturated fatty acids increased from August (pre-diapause) to October (initiation of diapause). The increase in seasonal cold hardiness during cold acclimation, exposed at −15°C for 24 h, was related to degree of fatty acid unsaturation. The elevation of palmitoleic acid content at low temperature resulted in an increase in the overall degree of unsaturation in the whole body. These results indicated the importance of unsaturated fatty acids composition to prepare larvae entering diapause phase.     

Studies on the Conjugated Lipids

The chemical nature of fatty acids was studied on sphingomyelin which was prepared from horse spinal cord. Nervonic, palmitic, stearic and behenic acids were isolated from the hydrolysate of the sphingomyelin preparation. The presence of behenic acid in sphingomyelin has in this paper been reported for the first time.     

Fatty acid fingerprints of Streptococcus mutans NCTC 10832 grown at various temperatures.

Fatty acid fingerprints were determined gas chromatographically for Strepcococcus mutans NCTC 1082 which had been grown in batch brain heart infusion at a series of nine temperatures ranging from 29.0 to 40.0 degrees C. The major acids at all temperatures were n-palmitic and octadecenoic acids. Other acids detected at all temperatures included n-myristic, palmitoleic, n-stearic, and eicosenoic acids. An increase in temperature resulted in a decrease in the proportion of unsaturated to saturated fatty acids, indicating the importance of accurate temperature control in such gas-liquid chromatographic, chemotaxonomic studies.     

Incorporation of specific exogenous fatty acids into membrane lipids modulates protonophore resistance in Bacillus subtilis.

Attempts to manipulate the level of C16:1 fatty acids in membrane phospholipids were made by using Bacillus subtilis and its protonophore-resistant mutants to test the hypothesis that C16:1 fatty acid levels relate to the bioenergetic properties of the mutant strains. Growth of the three mutants in the presence of palmitoleic acid restored the level of C16:1 fatty acids in the membrane lipids to somewhat above those found in the wild type. The palmitoleic acid was preferentially incorporated into diphosphatidylglycerol (cardiolipin) and phosphatidylethanolamine and was associated with increased levels of these phospholipids. These membrane preparations showed no increase in the levels of free fatty acids. The increase in C16:1 fatty acids achieved by growth in the presence of palmitoleic acid was accompanied by secondary changes in membrane lipids as well as a pronounced diminution in the protonophore resistance of growth and ATP synthesis. Other membrane-associated properties that had been observed in these mutants, e.g., elevated ATPase levels, were not altered coordinately with protonophore resistance and C16:1 fatty acid levels. Growth of the wild type in the presence of palmitic acid caused a modest elevation of the C16:0 of the membrane lipids and a modest increase in the protonophore resistance of growth and ATP synthesis. Growth of the wild type at elevated temperatures, in the absence of fatty acid supplementation, also enhanced its resistance to protonophores. The results support the hypothesis that specific changes in membrane lipid composition underlie the bioenergetic changes associated with protonophore resistance.     

Aldosterone alters the phospholipid composition of rat colonocytes

Previous studies have shown that aldosterone treatment of amphibian epithelial cells results not only in stimulation of Na(+) absorption but also in changes in phospholipid composition which are necessary for the mineralocorticoid action of aldosterone. The present study was designed to investigate the effect of aldosterone on phospholipids of mammalian epithelia. Phospholipid and fatty acid composition was examined in colonic epithelium (mineralocorticoid target tissue) and thymus (non-mineralocorticoid but glucocorticoid target tissue) of rats which had received aldosterone or vehicle by a miniosmotic pump for 7 days. Aldosterone increased the mass of colonic phospholipids relative to cellular proteins with concomitant changes in the percentage distribution of fatty acids, whereas the relative distribution of membrane phospholipds was not changed. Phosphatidylcholine increased the content of polyunsaturated and decreased that of monounsaturated fatty acids, which predominantly reflected the accretion of arachidonic and a decrease in oleic and palmitoleic acids. Within the phosphatidylethanolamine subclass, pretreatment of rats with aldosterone decreased the content of monounsaturated fatty acids (predominantly oleic and palmitoleic acid) and of n-3 fatty acids, and increased the content of saturated fatty acids (palmitic acid). The saturated-to-nonsaturated fatty acid ratio also significantly increased after aldosterone treatment. No changes in thymic phospholipids were seen. The results are consistent with the contention that aldosterone specifically modulates phospholipid concentration and metabolism in mineralocorticoid target tissue. The changes in phospholipid content and its fatty acid composition during the fully developed effect of aldosterone may reflect a physiologically important phenomenon with long-term consequences for membrane structure and function.     

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.     

Effects of fatty acids on activity of cGMP-stimulated cyclic nucleotide phosphodiesterase from calf liver

Effects of fatty acids, prostaglandins, and phospholipids on the activity of purified cGMP-stimulated cyclic nucleotide phosphodiesterase from calf liver were investigated. Prostaglandins A2, E1, E2, F1 alpha, and F2 alpha, thromboxane B2, and most phospholipids were without effect; lysophosphatidylcholine was a potent inhibitor. Several saturated fatty acids (carbon chain length 14-24), at concentrations up to 1 mM, had little or no effect on hydrolysis of 0.5 microM [3H]cGMP or 0.5 microM [3H]cAMP with or without 1 microM cGMP. In general, unsaturated fatty acids were inhibitory, except for myristoleic and palmitoleic acids which increased hydrolysis of 0.5 microM [3H]cAMP. The extent of inhibition by cis-isomers correlated with the number of double bonds. Increasing concentrations of palmitoleic acid from 10 to 100 microM increased hydrolysis of [3H]cAMP with maximal activation (60%) at 100 microM; higher concentrations were inhibitory. Palmitoleic acid inhibited cGMP hydrolysis and cGMP-stimulated cAMP hydrolysis with IC50 values of 110 and 75 microM, respectively. Inhibitory effects of palmitoleic acid were completely or partially prevented by equimolar alpha-tocopherol. Palmitelaidic acid, the trans isomer, had only slightly inhibitory effects. The effects of palmitoleic acid (100 microM) were dependent on substrate concentration. Activation was maximal with 1 microM [3H]cAMP and was reduced with increasing substrate; with greater than 10 microM cAMP, palmitoleic had no effect. Inhibition of cGMP hydrolysis was maximal at 2.5 microM cGMP and was reduced with increasing cGMP; at greater than 100 microM cGMP palmitoleic acid increased hydrolysis slightly. Palmitoleic acid did not affect apparent Km or Vmax for cAMP hydrolysis, but increased the apparent Km (from 17 to 60 microM) and Vmax for cGMP hydrolysis with little or no effect on the Hill coefficient for either substrate. These results suggest that certain hydrophobic domains play an important role in modifying the catalytic specificity of the cGMP-stimulated phosphodiesterase for cAMP and cGMP.