Amounts Of Phosphatidylethanolamine Research Articles

Effect of Quinoline on the Phospholipid Profile of Curvularia lunata and Its Microbial Detoxification.

Quinoline is an N-heterocyclic compound commonly found in wastewater, especially that derived from coal processing, chemical, and pharmaceutical industries. In the present study, the microscopic fungus Curvularia lunata IM 4417, which is known to degrade various xenobiotics, was used. The aim of the research was to study the elimination of quinoline and its influence on fungal phospholipids, which are considered to be excellent indicators of environmental monitoring. Quinoline biodegradation products and phospholipid contents were analyzed using gas chromatography–mass spectrometry and liquid chromatography–tandem mass spectrometry. C. lunata IM 4417 degraded quinoline, which led to the formation of conjugates of glucose with hydroxylated derivatives of the compound. Toxicity tests (Artoxkit M and Microtox assay) indicated that the elimination of lower concentrations of quinoline was efficient and led to a reduction in sample toxicity. The presence of quinoline also significantly affected the profile of fatty acids and phospholipids. The addition of quinoline to a culture of C. lunata IM 4417 caused an increase in the content of phosphatidylcholine (PC) and a decrease in the amount of phosphatidylethanolamine (PE), two major structural lipids. Additionally, decreases in the contents of phosphatidylinositol (PI) and phosphatidylserine (PS), which are responsible for tolerance to toxic substances, cell viability, and signal transduction, were noted. Thus, it can be concluded that the presence of quinoline modifies the membrane composition, and this change may be an important indicator of the presence of N-heterocyclic compounds or other toxins in the environment.

LDL uptake-dependent phosphatidylethanolamine translocation to the cell surface promotes fusion of osteoclast-like cells.

Osteoporosis is associated with vessel diseases attributed to hyperlipidemia, and bone resorption by multinucleated osteoclasts is related to lipid metabolism. In this study, we generated low-density lipoprotein receptor (LDLR)/lectin-like oxidized LDL receptor-1 (LOX-1, also known as Olr1) double knockout (dKO) mice. We found that, like LDLR single KO (sKO), LDLR/LOX-1 dKO impaired cell-cell fusion of osteoclast-like cells (OCLs). LDLR/LOX-1 dKO and LDLR sKO preosteoclasts exhibited decreased uptake of LDL. The cell surface cholesterol levels of both LDLR/LOX-1 dKO and LDLR sKO osteoclasts were lower than the levels of wild-type OCLs. Additionally, the amount of phosphatidylethanolamine (PE) on the cell surface was attenuated in LDLR/LOX-1 dKO and LDLR sKO preosteoclasts, whereas the PE distribution in wild-type OCLs was concentrated on the filopodia in contact with neighboring cells. Abrogation of the ATP binding cassette G1 (ABCG1) transporter, which transfers PE to the cell surface, caused decreased PE translocation to the cell surface and subsequent cell-cell fusion. The findings of this study indicate the involvement of a novel cascade (LDLR∼ABCG1∼PE translocation to cell surface∼cell-cell fusion) in multinucleation of OCLs.

Effect of the anti-inflammatory drug diclofenac on lipid composition of bacterial strain Raoultella sp. KDF8.

The strain Raoultella sp. KDF8 was cultivated on three sources of carbon and energy, glycerol, ethanol and diclofenac, for periods of time ranging from 24 to 72h. Using thin-layer chromatography, nine classes of phospholipids were detected and the amount of phosphatidylethanolamine (PtdEtn) decreased with increasing cultivation time. Conversely, the ratio of phospholipids having three or four acyls (acyl-phosphatidylglycerol (APtdGro), N-acyl-PtdEtn (NAPtdEtn) and cardiolipin (Ptd2Gro) increased during cultivation. GC-MS analysis showed that the percentage of fatty acids containing a cyclopropane ring increased almost tenfold whereas the amount of fatty acids bearing even-numbered chains dropped to less than one-third after 24h and 72h in cultures on glycerol and diclofenac, respectively. Shotgun analysis showed significant changes in the representation of molecular species of phospholipids. For instance, there was a 36-fold change in the ratio of 16:1/16:1/16:1-APtdGro to c17:0/c17:0/c17:0-APtdGro and a 12-fold ratio change for 16:1/16:1/16:1-NAPtdEtn to c17:0/c17:0/c17:0-NAPtdEtn; the Ptd2Gro ratio of 16:1 to c17:0 acids equalled 1750. Our results show that the bacteria overcome destabilization of the inner cytoplasmic cell membrane and a bacterial outer membrane by altering the geometric arrangement of acyl chains, i.e. switching from monounsaturated to cyclopropane fatty acids (16:1 versus c17:0).

Age-related changes phospholipids of sterlet in liver and dorsal muscles.

Study of phospholipids changes peculiarities in the liver and dorsal muscles of sterlet (Acipenser ruthenus Linnaeus) may be important to determine the etiology and pathogenesis of fatty liver. We established that the content of total phospholipids in tissues of the liver and dorsal muscles of three-year-old sterlet was less than for two-year-old fish by 15% and 20% (P ≤ 0.01), respectively. The amount of phosphatidylcholine (P ≤ 0.05), phosphatidylethanolamine, phosphatidylserine (P ≤ 0.01), phosphatidylinositol (P ≤ 0.01) and cardiolipin in the liver of 3-year-old sterlet was lower than for the 2-year-old fish, while quantitative indices for lysophosphatidylcholine and sphingomyelin were slightly increased. Similarly, in the cells of the dorsal muscles, the amount of phospholipid components (except lysophosphatidylcholine) was decreased with age. A decrease in the amount of phosphatidylethanolamine and phosphatidylserine in the dorsal muscles of 3-year-old sterlet was significant. The major phospholipids respective distribution was stable, except for phosphatidylethanolamine and particularly sphingomyelin.

Bacillus lycopersici sp. nov., isolated from a tomato plant (Solanum lycopersicum L.).

A Gram-stain-positive, rod-shaped, endospore-forming bacterium (designated strain CC-Bw-5(T)) was isolated from chopped tomato stems. The isolate grew at 20-40 °C, pH 6.0-8.0 and tolerated 6% (w/v) NaCl. The most closely related strains in terms of 16S rRNA gene sequence similarity were Bacillus isabeliae (95.3%) and Bacillus oleronius (95.3%). The G+C content of the genomic DNA was 37.2 ± 3.6 mol%. Strain CC-Bw-5(T) was determined to possess C16 : 0, iso-C14 : 0, iso-C15 : 0, anteiso-C15 : 0 and iso-C16 : 0 as predominant fatty acids. The polar lipid profile consisted of predominant amounts of diphosphatidylglycerol, and moderate-to-trace amounts of phosphatidylethanolamine, phosphatidylglycerol, two unidentified glycolipids, an unidentified phospholipid and an unidentified phosphoglycolipid. The cell-wall peptidoglycan contained meso-diaminopimelic acid; menaquinone (MK-7) was the predominant respiratory quinone. According to distinct phylogenetic, phenotypic and chemotaxonomic features, strain CC-Bw-5(T) is proposed to represent a novel species within the genus Bacillus for which the name Bacillus lycopersici sp. nov. is proposed. The type strain is CC-Bw-5(T) ( = BCRC 80623(T) = JCM 19140(T)).

Value of Fatty acids in formation of thrombotic status in patients with ischemic heart disease

Interrelation between fatty acids and platelet aggregation has been a subject of interest for cardiologists in the terms of thrombogenic status. In order to study this problem we observed 42 patients with ischemic heart disease and 30 healthy persons. In patients we found elevation of arachidonic acid and lowering of docosahexaenic acid levels. Hypoxemic assay revealed in these patients lowering of prostacyclin and antithrombin III levels. In the pathogenesis of disturbances of platelet aggregation in patients with ischemic heart disease alteration of structure of platelets with increased quantity of phosphatidyl ethanolamine and corresponding change of thrombocyte asymmetry resulted in conformational reorganization of proteins and lipids.

An insight on human skin penetration of diflunisal: lipogel versus hydrogel microemulsion

Diflunisal is a NSAID used in acute and long term management of pain and inflammation associated with osteoarthritis, rheumatoid arthritis and symptoms of primary dysmenorrhea. However, its oral use is associated with side effects such as peptic ulceration, dyspepsia, gastrointestinal disturbances and bleeding. The aim of this work was to develop lecithin organogels (LO) transdermal delivery system for diflunisal and to study its human skin penetration ability in comparison with an optimized microemulsion-based hydrogel. Ternary phase diagrams were constructed using butyl lactate as an organic solvent and two commercial grades of lecithin. The formation of gel phase was lecithin concentration dependent with Phosholipion 85 G being capable of forming organogels at lower lecithin concentration than Lipoid S75. The gels prepared using butyl lactate were able to tolerate higher amounts of water than could be incorporated in the lipogels prepared with other organic solvents. All the investigated gels possessed acceptable physical properties and were able to deliver diflunisal through human skin. The lipogels delivered higher total drug amount through the skin than the hydrogel. The composition of lecithin seemed to have some effect on the skin permeability enhancement ability of the lipogel. Lecithin containing higher amount of phosphatidyl ethanolamine could provide better transdermal delivery. The elaborated lecithin organogels are potential carriers that create a good opportunity for transdermal delivery of diflunisal overcoming the side effects associating its oral route.

Altered phospholipid molecular species and glycolipid composition in brain, liver and fibroblasts of Zellweger syndrome

We studied the altered molecular species of lipids in brain and liver tissues, and fibroblasts from patients with Zellweger syndrome (ZS). ZS cerebellum samples contained a higher amount of sphingomyelin with shorter chain fatty acids compared to that in normal controls. The amount of phosphatidylethanolamine (PE) was less than half of that in controls, with the absence of the PE-type of plasmalogen. Gangliosides were accumulated in the brains and fibroblasts of ZS patients. To investigate whether or not impaired beta-oxidation of very long chain fatty acids and/or plasmalogen synthesis affects glycolipids metabolism, RNAi of peroxisomal acylCo-A oxidase (ACOX1) and glyceronephosphate O-acyltransferase (GNPAT) was performed using cultured neural cells. In neuronal F3-Ngn1 cells, ACOX1 and GNPAT silencing up-regulated ceramide galactosyltransferase (UGT8) mRNA expression, and down-regulated UDP-glucose ceramide glucosyltransferase (UGCG). These results suggest that both impaired beta-oxidation of very long chain fatty acids and plasmalogen synthesis affect glycolipid metabolism in neuronal cells.

A Lipid-mediated Quality Control Process in the Golgi Apparatus in Yeast

When heme biosynthesis is disrupted, the yeast Saccharomyces cerevisiae becomes unable to synthesize its major sterol, ergosterol, and desaturate fatty acids. We took advantage of this physiological peculiarity to evaluate the consequences of ergosterol and/or unsaturated fatty acid (UFA) depletions on the biogenesis of a model polytopic plasma membrane protein, the uracil permease Fur4p. We show that under UFA shortage, which results in low amounts of diunsaturated phospholipid species, and under ergosterol depletion, Fur4p is prematurely routed from the Golgi apparatus to the vacuolar lumen in a process that requires the ubiquitin ligase Rsp5p. Interestingly, this diversion is not correlated to Fur4p exclusion from detergent-resistant membranes. In an independent set of experiments, we show that Fur4p targeting to the plasma membrane depends on phosphatidylethanolamine amounts and more specifically on the propensity of this phospholipid to form a hexagonal phase. In light of recent literature, we propose a model in which ergosterol and diunsaturated phospholipid species maintain optimal membrane curvature for Fur4p to evade the Golgi quality control process and to be properly delivered to its normal destination.

Phosphatidylethanolamine deficiency in membrane lipids inhibits keratinocyte intercellular networks formation.

Ethanolamine (Etn) is required for the growth of epithelial cells in culture. Without Etn, the amount of phosphatidylethanolamine (PE) in membrane lipids is reduced, and cell proliferation stops. When the membrane lipids are deficient of PE, some extracellular signaling processes become impaired. In this study, we examined the effect of Etn deprivation on the formation of intercellular networks in immortalized human oral keratinocytes. Keratinocytes proliferate with undifferentiated morphologies in a low-calcium medium, whereas they undergo differentiation to form intercellular networks in a high-calcium medium. The cells were first cultured with or without Etn supplement in a low-calcium (0.07 mM) medium, and then the calcium concentration was raised to 1.8 mM. The localization and organization of the following proteins were examined: (1) desmogleins and plakoglobin in desmosomes, (2) E-cadherin and beta-catenin in adherens junctions and (3) actin and keratin filaments in cytoskeletons. As expected, in the Etn-supplemented cells, the elevated level of calcium induced the junctional localization of the proteins associated with desmosomes and adherens junctions and also induced the formation of keratin and actin networks. On the contrary, in the Etn-deprived cells, the elevated level of calcium induced none of the above processes. The results suggest that having a sufficient amount of PE or proper phospholipid composition in the membranes is crucial for differentiation in epithelial cells.

Effect of dietary fish oil (active EPA-30) on liver phospholipids in young and aged rats

We explored the uses of fish oil (active EPA-30) as a source of eicosapentaenate (EPA; 20:5 n-3), to young and old rats. We treated three subgroups of rats each comprising 20 young or old rats, respectively. The first group was kept on the basal ration (lab-pellet) as control diet, the second group was fed semi-purified diets contained 5% pig-fat (n-3 fatty acids deficient diet). The third group was fed a modified diet in which 50% of pig-fat was replaced by active EPA-30. Livers of young rats fed pig-fat had a drastic decrease in the amount of phosphatidylethanolamine (PE) and omega-3 polyunsaturated fatty acids (EPA, 20:5 n-3 and docosahexaenoic, DHA, 22:6 n-3) and compensatory increase of phosphatidylcholine, saturated fatty acids and n-6 polyunsaturated fatty acids in the liver phospholipids. In contrast, the liver of young rats fed active EPA-30 had large amounts of PE and concomitant enrichment in polyunsaturated fatty acids. The liver of old rats, fed on active EPA-30 supplemented diet had lower amounts of PE and there were no significant changes in the phospholipid fatty acid composition.

Ethanol-induced alterations in lipid composition of Saccharomyces cerevisiae in the presence of exogenous fatty acid

Ethanol-induced alterations in the lipid composition of Saccharomyces cerevisiae grown in the presence of exogenous fatty acids were studied. The addition of both palmitic acid and ethanol (4–8%) to the basal medium resulted in a striking increase in the palmitic acid content and decreases in the content of myristoleic, palmitoleic, and oleic acids in the phospholipid fatty acid composition, compared with their contents in the absence of ethanol. On the other hand, the addition of linoleic acid to the medium containing ethanol (4–8 %) resulted in an increase in oleic acid and corresponding decreases in myristoleic, palmitoleic acid, and palmitic acid, while the linoleic acid levels were maintained in excess of 14%. Generally, growth in the presence of ethanol resulted in a reduction in the amount of phosphatidylcholine (PC) and more obviously, in the amount of phosphatidylethanolamine (PE), leading to an increase in the PC PE ratio. A good cell survival rate in replacement cultures containing 18% ethanol was attained when cells were preincubated in the presence of ethanol (4–8%). Under the same conditions, palmitic acid-enriched cells had additional ethanol-endurability and a higher cell-membrane integrity than linoleic acid-enriched cells.

Colony Variation of Helicobacter pylori: Pathogenic Potential Is Correlated to Cell Wall Lipid Composition

Background: Differences in expression of disease after infection with Helicobacter pylori have so far been connected with host factors and bacterial interstrain variation. In this study, spontaneous and ecology-mediated intrastrain variation was examined. Methods: Four clinical isolates of H. pylori were shown to give rise to two colony forms. Bacterial morphology was examined by electron microscopy. Bacterial fractions were examined for proteins using ion exchange chromatography and SDS-PAGE; for lipids using thin-layer chromatography, lipid anion-exchange chromatography, column chromatography on silica gel, 31P-NMR, gas chromatography and mass spectrometry. Bacterial in vitro invasiveness and adhesiveness were examined in two different systems, and urease and VacA toxin were assayed by Western blot analysis. Results: H. pylori was shown to give rise to two colony forms: at normal pH the population was dominated by L colonies. One strain was chosen for further studies. Bacteria from L colonies retained VacA toxin and urease, did not invade or adhere to epithelial cells, and contained normal quantities of phosphatidylethanolamine. In a small frequency, spontaneous S colonies were formed. Bacteria from these colonies released VacA and urease, adhered to and invaded epithelial cells and contained increased amounts of lysophosphatidyl ethanolamine and phosphatidyl serine. After addition of HC1 to the culture medium (pH 6), almost only S colonies were formed. The results demonstrate that environmental factors, such as HC1, can change the bacterial cell wall, and thereby enhance expression of virulence factors of H. pylori in vitro. A similar in vivo variation would have implications for our understanding of the interaction between HC1 secretion in the gastric mucosa and H. pylori in the development of peptic ulcer disease.

Comparison of Helicobacter mustelae and Helicobacter pylori adhesion to eukaryotic cells in vitro

Background & Aims : Bacterial adhesion to mucosal surfaces is an important pathogenic mechanism for Helicobacter-induced gastritis. The aims of this study were to compare binding of selected Helicobacter mustelae and Helicobacter pylori strains to lipids extracted from HEp-2, Chinese hamster ovary, human embryonic lung cells, and ferret gastrointestinal tissues as well as to intact tissue culture cells and to analyze the fatty acids of the receptor. Methods : Thin-layer chromatography overlay binding and a receptor-based immunoassay detected adhesion of bacteria to commercial lipids and to individual species within the lipid extracts. H. mustelae binding to tissue culture cells was performed by whole cell bacterial adhesion assay. Results : H. mustelae and H. pylori both bound to phosphatidylethanolamine and lysophosphatidylethanolamine. Adhesion of H. mustelae to intact eukaryotic cells correlated with the amount of phosphatidylethanolamine. Binding of helicobacters was greater to lipids derived from ferret antrum compared with colon ( P < 0.05). Biochemical analysis suggested that heterogeneity in fatty acid composition of phosphatidylethanolamine could influence the degree of Helicobacter binding. Conclusions : Adhesion of Helicobacter strains correlates with the quantity of phosphatidylethanolamine present in the epithelial cell and with the differences in the fatty acid profile of the lipid.

The pss and psd genes are required for motility and chemotaxis in Escherichia coli

Mutants of Escherichia coli defective in phosphatidylserine synthase (encoded by pss) and phosphatidylserine decarboxylase (encoded by psd) make cell membranes deficient in phosphatidylethanolamine. In this report we show that wild-type pss and psd genes are required for motility and chemotaxis. Null mutants or strains with temperature-sensitive pss or psd mutations grown at high temperature (35 degrees C) were nonmotile. They lacked flagella and showed reduced rates of transcription of the flhD master operon (encoding FlhD and FlhC), the fliA operon (encoding sigma F), and the fliC operon (encoding flagellin). At low temperature (25 degrees C), the temperature-sensitive mutant cells showed motility and chemotaxis but at reduced levels. The extent of the motility and chemotaxis defects in the mutants was correlated with the amount of phosphatidylethanolamine in the membranes, suggesting a link between membrane phospholipid composition and expression of the flagellum chemotaxis regulon.

A reevaluation of the effect of auxin on phospholipids in pea stem segments

Auxin has been reported to substantially alter concentration and fatty acid composition of phospholipids in elongating pea ( Pisum sativum L. cv. Feltham First) stem segments. To further investigate this phenomenon, phospholipid composition of cv. Alaska pea segments treated with the auxin 2,4-dichlorophenoxyacetic acid (2,4-D) for 30 or 60 min was analyzed. The amount of phosphatidylethanolamine (PE) was lower in 2,4-D-treated segments at 30 min, but by 60 min, the amount was comparable to controls. Quantitative differences in phosphatidylcholine (PC) and phosphatidylinositol (PI) were not significant. Auxin did not affect phospholipid fatty acid composition. Auxin did not inhibited the incorporation of [ 14C]acetate or [ 3H]choline into the major phospholipids. These results suggest that effects of auxin on phospholipid metabolism are transient, and that shifts in fatty acid composition do not universally occur in response to auxin treatment.

Large scale production and characterization of lyophilized pyridoxalated hemoglobin-polyoxyethylene (PHP).

Lyophilized PHP as an oxygen carrier is prepared from outdated red cell and dicarboxymethylated polyoxyethylene. In order to apply PHP for a clinical use, a large scale production of high quality PHP has been studied. We have set up a 20 L scale production flow of PHP88. The product was tested to confirm the quality and lot-to-lot consistency. The blood group specific materials were weakly positive in stroma-free hemoglobin (SFH), however, were found negative in the PHP of this scale. The amount of phosphatidylethanolamine (PE) in purified SFH and PHP88 reconstituted solution was 0.19 +/- 0.04 and 0.03 +/- 0.01 ppm, respectively. Contamination of viruses such as HBV and Non A non B hepatitis virus could not be observed in the final product. Elimination and inactivation of HIV was validated through a spike test. The characterizations of the final products in 20 L scale were done through MW, P50, Hill coefficient, viscosity, and molecular weight distribution by SDS-PAGE and batch to batch consistency was also confirmed. The results show that production process is appropriate to eliminate the blood group materials, PE and virus, and produce PHP of high quality. Lyophilized PHP88 can be produced by addition of maltose and can be stored over 1 year.

A Rapid Method For Phospholipid Class Separation by HPLC using an Evaporative Light-Scattering Detector

Abstract A high-performance liquid chromatographic method for the separation and quantification of major natural phospholipid classes using an evaporative light-scattering detector is described. Separation and detection of cholesterol, palmitic acid, phosphatidylethanolamine, phosphatidylserine, phosphatidylcholine and sphingomyelin was achieved in less than 15 minutes using a Spherisorb 10 cm long silica column. A simplified ternary mobile phase gradient of hexane, isopropanol and water was used to separate these six standards reproducibly and easily. A crude sample of egg yolk was also analyzed using this method and found to contain mostly cholesterol with small amounts of phosphatidylethanolamine, phosphatidylcholine and sphingomyelin. The evaporative detector gave a stable flat baseline with detection limits for most compounds in the low nanogram range.

小児と成人中耳貯留液の生化学的特徴 第１報 脂質成分

Middle ear effusions (MEEs) from adult were usually serous in nature, but those from child were mucous. MEEs contain the substances secreted by the epithelial cells of the middle ear and Eustachian tube to regulate surface tensions as well as those produced as the result of inflammation, but the biochemical bases of serous and mucous MEEs have not been clearly established. To clarify the biochemical differences giving rise to the characteristic physicochemical properties of MEE from children and from adults, all lipid components of MEEs from 7 children and 7 adults by thin-layer and gas liquid chromatographies were analyzed, and also they were compared with those from sera. Although no significant difference in lipid composition was observed between adult and pediatric sera, the amounts of phosphatidyl ethanolamine (PE) and phosphatidyl serine (PS) in pediatric MEE were higher than those in adult MEE. The relative concentration of PE in the total phospholipids of pediatric MEE was 26.5%, significantly higher than that in adult MEE (9.2%), and pediatric (6.1%) and adult (3.1%) sera. A similar high concentration of PS was also observed in pediatric MEE. Since phospholipids were the major components of surfactants secreted from the epithelial cells of the middle ear, significantly high concentrations of the charged phospholipids, PE and PS, might be responsible in part for the characteristics of pediatric MEE. To compare physical properties in different phospholipid compositions between pediatric and adult MEEs, polarization value was measured with liposomes in each phospholipid compositions. The polarization value in pediatric phospholipid composition liposomes was higher than in adult. It seemed that the microviscosity in pediatric MEE was lower than in adult.

Fluorimetric Assay of Phospholipase A Acting on Biomembrane Phospholipids.

A sensitive phospholipase A assay suitable for organelle activities is described. Activation of the enzyme produces an increase in membrane lysophospholipids. The amino groups of extracted lipids were derivatized with 4-fluoro-7-nitrobenzo-2-oxa-1,3-diazole and the amounts of phosphatidylethanolamine and its lysoform were quantitated using HPLC equipped with a SiO2 column and a fluorescence detector. The procedure was tested with porcine enzyme acting on liposomes and mitochondria, and with endogenous mitochondrial enzyme.