Characterization of Phospholipids in Insulin Secretory Granules and Mitochondria in Pancreatic Beta Cells and Their Changes with Glucose Stimulation

The lipid composition of insulin secretory granules (ISG) has never previously been thoroughly characterized. We characterized the phospholipid composition of ISG and mitochondria in pancreatic beta cells without and with glucose stimulation. The phospholipid/protein ratios of most phospholipids containing unsaturated fatty acids were higher in ISG than in whole cells and in mitochondria. The concentrations of negatively charged phospholipids, phosphatidylserine, and phosphatidylinositol in ISG were 5-fold higher than in the whole cell. In ISG phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine, and sphingomyelin, fatty acids 12:0 and 14:0 were high, as were phosphatidylserine and phosphatidylinositol containing 18-carbon unsaturated FA. With glucose stimulation, the concentration of many ISG phosphatidylserines and phosphatidylinositols increased; unsaturated fatty acids in phosphatidylserine increased; and most phosphatidylethanolamines, phosphatidylcholines, sphingomyelins, and lysophosphatidylcholines were unchanged. Unsaturation and shorter fatty acid length in phospholipids facilitate curvature and fluidity of membranes, which favors fusion of membranes. Recent evidence suggests that negatively charged phospholipids, such as phosphatidylserine, act as coupling factors enhancing the interaction of positively charged regions in SNARE proteins in synaptic or secretory vesicle membrane lipid bilayers with positively charged regions in SNARE proteins in the plasma membrane lipid bilayer to facilitate docking of vesicles to the plasma membrane during exocytosis. The results indicate that ISG phospholipids are in a dynamic state and are consistent with the idea that changes in ISG phospholipids facilitate fusion of ISG with the plasma membrane-enhancing glucose-stimulated insulin exocytosis.     

Qualitative and quantitative changes in phospholipid spectra of the brain of rats with corazol-induced epilepsy

The enhancement of the sum of phospholipids in seizure period was observed during initially generalized corazol seizure in white rats' cerebral cortex. This sum fell in an hour after epileptic fit. It is noted that the maintenance of lysophosphatidylcholines and phosphatidylserines enhanced during the fit, and the maintenance of phosphatidylcholines and phosphatidylethanolamines fell either in cerebral cortex, or in cerebellum. At the same time the enhancement of the mixed fraction of sphingomyelin and phosphatidylinositide level and cardiolipins-1 in the cerebral cortex was observed. The maintenance of the latter in cerebellum fell during an hour after the attack. The quantity of cardiolipins-2 in cerebellum enhanced during the attack. It is suggested that the effect of corazol seizure leads to the enhancement of lisoforms at the expense of the intensification of disintegration of other fractions of lisoforms.     

Activation of arachidonoyl-phosphatidylinositol and phosphatidylcholine turnover by K+-evoked stimulation of brain synaptosomes

The turnover of arachidonoyl groups in synaptosomal phospholipids after stimulation by K⁺ was examined. Raising the K⁺ concentration in the incubation medium from 5 to 55 mM caused a rapid hydrolysis of labeled arachidonate from the synaptosomal phospholipids. Under this condition, radioactivity released from phosphatidylinositols was proportionally higher than that from phosphatidylcholines. Hydrolysis of arachidonoyl group from phospholipids was correlated to an increase in radioactivity in the free fatty acid-ion complex which appeared in the interphase after extraction with chloroform-methanol 2:1 (v/v). The K⁺-evoked phospholipid hydrolysis and the formation of fatty acid-ion complex, were Ca²⁺-dependent. Phospholipid deacylation activity was localized mainly in synaptic vesicles and synaptic plasma membranes but not in the mitochondria. The stimulated turnover of synaptosomal phospholipids appeared to be mediated by the deacylation-reacylation mechanism, because similar treatment with high K⁺ stimulated the incorporation of labeled arachidonate into phosphatidylinositols and phosphatidylcholines of synaptosomes. The possible physiological implication of membrane lipid involvement in synaptic processes is discussed.     

Formation of lysophospholipids from unsaturated phosphatidylcholines under the influence of hypochlorous acid

The formation of lysophosphatidylcholines from unsaturated phosphatidylcholines upon treatment with hypochlorous acid was evaluated by means of MALDI-TOF mass spectrometry and 31P NMR spectroscopy. With an increasing number of double bonds in a fatty acid residue, the yield of lysophosphatidylcholines with a saturated fatty acid residue increased considerably in comparison to the total amount of higher molecular weight products like chlorohydrins and glycols. High amounts of lysophosphatidylcholines were formed from phospholipids containing arachidonic or docosahexaenoic acid residues. In phospholipids with monounsaturated fatty acid residues, the position of the double bond did not influence the yield of lyso-products. Besides the exclusive formation of chlorohydrin and glycol, hypochlorous acid caused the cleavage of the unsaturated fatty acid residue independent of its location at the first or second position of the glycerol backbone. In contrast, strong alkaline conditions, i.e. saponification led also to a hydrolysis of the saturated fatty acid residue from phosphatidylcholines. It is concluded that both MALDI-TOF mass spectrometry and 31P NMR spectroscopy are able to detect the formation of lysophosphatidylcholines. We conclude also that the formation of lysophospholipids from unsaturated phosphatidylcholines by hypochlorous acid can be relevant in vivo under acute inflammatory conditions.     

Reliability of Serum Metabolites over a Two-Year Period: A Targeted Metabolomic Approach in Fasting and Non-Fasting Samples from EPIC

Objective

Although metabolic profiles have been associated with chronic disease risk, lack of temporal stability of metabolite levels could limit their use in epidemiological investigations. The present study aims to evaluate the reliability over a two-year period of 158 metabolites and compare reliability over time in fasting and non-fasting serum samples.

Methods

Metabolites were measured with the AbsolueIDQp180 kit (Biocrates, Innsbruck, Austria) by mass spectrometry and included acylcarnitines, amino acids, biogenic amines, hexoses, phosphatidylcholines and sphingomyelins. Measurements were performed on repeat serum samples collected two years apart in 27 fasting men from Turin, Italy, and 39 non-fasting women from Utrecht, The Netherlands, all participating in the European Prospective Investigation into Cancer and Nutrition (EPIC) study. Reproducibility was assessed by estimating intraclass correlation coefficients (ICCs) in multivariable mixed models.

Results

In fasting samples, a median ICC of 0.70 was observed. ICC values were <0.50 for 48% of amino acids, 27% of acylcarnitines, 18% of lysophosphatidylcholines and 4% of phosphatidylcholines. In non-fasting samples, the median ICC was 0.54. ICC values were <0.50 for 71% of acylcarnitines, 48% of amino acids, 44% of biogenic amines, 36% of sphingomyelins, 34% of phosphatidylcholines and 33% of lysophosphatidylcholines. Overall, reproducibility was lower in non-fasting as compared to fasting samples, with a statistically significant difference for 19–36% of acylcarnitines, phosphatidylcholines and sphingomyelins.

Conclusion

A single measurement per individual may be sufficient for the study of 73% and 52% of the metabolites showing ICCs >0.50 in fasting and non-fasting samples, respectively. ICCs were higher in fasting samples that are preferable to non-fasting.     

Lipidomic characterization of exosomes isolated from human plasma using various mass spectrometry techniques

Ultrahigh-performance supercritical fluid chromatography - mass spectrometry (UHPSFC/MS), ultrahigh-performance liquid chromatography - mass spectrometry (UHPLC/MS), and matrix-assisted laser desorption/ionization (MALDI) - MS techniques were used for the lipidomic characterization of exosomes isolated from human plasma. The high-throughput methods UHPSFC/MS and UHPLC/MS using a silica-based column containing sub-2 μm particles enabled the lipid class separation and the quantitation based on exogenous class internal standards in <7 minute run time. MALDI provided the complementary information on anionic lipid classes, such as sulfatides. The nontargeted analysis of 12 healthy volunteers was performed, and absolute molar concentration of 244 lipids in exosomes and 191 lipids in plasma belonging to 10 lipid classes were quantified. The statistical evaluation of data included principal component analysis, orthogonal partial least square discriminant analysis, S-plots, p-values, T-values, fold changes, false discovery rate, box plots, and correlation plots, which resulted in the information on lipid changes in exosomes in comparison to plasma. The major changes were detected in the composition of triacylglycerols, diacylglycerols, phosphatidylcholines, and lysophosphatidylcholines, whereby sphingomyelins, phosphatidylinositols, and sulfatides showed rather similar profiles in both biological matrices.     

High phosphatidylcholine weights compared to lysophosphatidylcholine weights, in micellar intestinal contents, in the rat (author's transl)

Casein hydrolysat, lactose and lipids (100 mg of fatty acids) were introduced in the stomach of rats by a gastric tube: either pure tri-oleoylglycerol, or phospholipids, or phosphatidylcholines, or the mixture 9/1 to fatty acid weight of tri-oleoylglycerol-phospholipids or phosphatidylcholines. The rats were killed 2 h later. The intraluminal intestinal lipids of the oil and micellar phases were separated after microfiltration (Millipore filters) in preference to the filtration by gel chromatography on polyacrylamide agarose, as an hydrolysis of intraluminal phospholipid occurred after the column elution. 1. After a quantitative recovery of the intestinal lipids (no separation of the oil and micellar phases), a strong hydrolysis of the tri-oleoyglycerol was observed; in opposition, large amounts of intact phospholipids appeared. 2. After isolation of the micellar phases, no triglycerides were recovered, but fatty acids and partial glycerides from the hydrolysed tri-oleoylglycerol and dietary phosphatidylcholines and small quantities of lyso-phosphatidylcholines (hydrolysed forms) were present. 3. After ingestion of the tri-oleoylglycerol as lipid dietary source, the intestinal micellar phases contained endogenous phosphatidylcholines and a few amounts of lysophosphatidylcholines, which had mainly bile origin, since the fatty acid composition of these micellar phosphatidylcholines approached the bile phosphatidylcholine fatty acid composition. The micellar lysophosphatidylcholine masses represented one-fourth of the micellar phosphatidylcholine masses. 4. In these experiments the phosphatidylcholine lysophosphatidylcholine ratio was always high: this means that small quantities of exogenous and endogenous lysophosphatidylcholines appeared in the micellar phases.     

In vitro lipid metabolism in the rat pancreas. III. Effects of carbamylcholine and pancreozymin on the turnover of phosphatidylinositols, 1,2-diacylglycerols and phosphatidylcholines

1. The turnover of phosphatidylinositols and other glycerolipids was examined in rat pancreatic fragments incubated in the presence of carbamylcholine and pancreozymin used at a concentration inducing maximal alpha-amylase hypersecretion. 2. In stimulated tissue, [1-14C]acetate-labeled fatty acids were incorporated into phosphatidylinositols, 1,2-diacylglycerols, and phosphatidic acids in preference to phosphatidylcholines, phosphatidylethanolamines, triacylglycerols, monoacylglycerols, and free fatty acids. Variations in the percent distribution of 14C among fatty acids and in specific activity of individual fatty acids in each lipid class suggested that the secretagogues reduced selection of newly synthesized 1,2-diacylglycerols which occurred in the resting state before their incorporation into phosphatidylinositols. Secretagogues also promoted recycling of endogenous 1,2-diacylglycerols (produced from hydrolysis of unlabeled glycerolipids) for the biosynthesis of phosphatidylinositols. 3. Increased rate of incorporation of [1-14C]palmitate, [1-14C]linoleate, [1-14C]arachidonate and [1(3)(n)-3H]glycerol into phosphatidylinositols was detrimental to phosphatidylcholines. 4. The lipolytic effects of carbamylcholine and pancreozymin as illustrated by the release of 1,2-diacylglycerols and free fatty acids, were markedly inhibited in calcium-free medium enriched with 1 mM EGTA but increased turnover of phosphatidylinositols as determined from incorporation of radioactive precursors was only moderately affected.     

Individual molecular species of phosphatidylcholines and phosphatidylinositols in liver of rats fed bis(2-ethylhexyl)phthalate

Rats were given a diet containing 1% bis(2-ethylhexyl)phthalate (DEHP) for 3 weeks, and their hepatic lipids analyzed. Phosphatidylcholines increased by 20%, while other phospholipid classes and cholesterol remained unchanged and triglycerides fell. The composition of molecular species of phosphatidylcholines was changed. Thus, the hepatic content of the major species, 1-palmitoyl-2-oleoyl-, 1-palmitoyl-2-arachidonoyl- and 1-stearoyl-2-arachidonoylphosphatidylcholines, rose by about 150%, 90% and 70%, respectively. The content of the other major species, 1-palmitoyl-2-linoleoyl- and 1-stearoyl-2-linoleoylphosphatidylcholine fell by about 20% and 30%, respectively. The content of alkyl-acyl analogues of phosphatidylcholines increased by about 70%, but the composition of molecular species remained the same. The composition of molecular species of phosphatidylinositols was also unchanged. Thus, the analyses show that DEHP can induce selective changes in molecular species of certain phospholipids in the liver. This could be important for the functioning of membrane structures in the hepatocyte.     

Influence of sterol structure on phospholipid phase behavior as detected by parinaric acid fluorescence spectroscopy

Phospholipid-sterol interactions were investigated using parinaric acid fluorescence spectroscopy. Cholesterol and cholesterol analogues which were modified in the sterol nucleus or side chain were added at 50 mol % to multilamellar vesicles of model phospholipids selected to be representative of major components in an LM cell plasma membrane. These included sphingomyelins and saturated and monounsaturated phosphatidylcholines and phosphatidylethanolamines. Based on the changes in cis-parinaric acid steady-state fluorescence polarization observed with addition of sterol, 50 mol % cholesterol abolished the phase transition of all the model phospholipids. Dihydrocholesterol and trans-22-dehydrocholesterol behaved like cholesterol in the two systems studied. 24-Methylcholesterols interacted well with all phospholipids except phosphatidylethanolamine which contained an unsaturated fatty acid. 24-Alkyl,trans-22-dehydrocholesterols abolished the phase transition in only two systems: sphingomyelins and phosphatidylcholines possessing relatively short saturated acyl chains. Since steady-state anisotropy is a function of fluorescence lifetime, rotational diffusion rates, and limiting anisotropy, we determined these parameters for two of the phospholipid systems. The results show that steady-state anisotropy values for phospholipid-sterol interactions correlate closely with limiting anisotropy and to a lesser extent with rotational relaxation time. The behavior of the sterols in the model phospholipids are used to interpret 1) fluorescence polarization measurements made with phospholipids extracted from LM cell plasma membranes, and 2) changes in membrane lipid composition which accompany growth of LM cells on various sterols.     

The glyceryl ethers of Paramecium phospholipids and phosphonolipids

Two glyceryl ethers, 1-O-hexadecyl glycerol and 1-O-cis-octadec-11-enyl glycerol, chimyl and paramecyl alcohol, respectively, were quantified in total phospholipids and five glycerophospholipid classes from cells and cilia of the ciliated protozoon, Paramecium tetraurelia. The ether content of 2-aminoethyl phosphonoglycerolipid was 85-90 mole %. Concentrations of ethers were greatest in the ethanolamine phosphonolipids greater than phosphatidylcholines greater than phosphatidylserines greater than phosphatidylethanolamines greater than phosphatidylinositols. The glyceryl ether concentrations in total cellular phospholipids increased with culture age in P. tetraurelia and P. multimicronucleatum cells. The glyceryl ether concentrations in the phospholipids of P. tetraurelia cilia remained constant from mid log to stationary phase of culture growth. Paramecium tetraurelia phospholipid glyceryl ether concentrations were made greater by supplementation of cultures with chimyl alcohol.     

Sphingomyelins of Rat Liver: Biliary Enrichment with Molecular Species Containing 16:0 Fatty Acids as Compared to Canalicular-Enriched Plasma Membranes

We harvested canalicular-enriched plasma membranes of hepatocytes and collected fistula bile from male rats and isolated the sphingomyelins. Following sphingomyelinase hydrolysis, we identified the sphingomyelin molecular species on the basis of their benzoylated ceramide derivatives employing high performance liquid chromatography. Sphingomyelin constitutes ≤3% of total biliary phospholipids (which are mostly sn-1 16:0 long-chain phosphatidylcholines) and approximately 30% of canalicular-enriched membranes. In both cases, the principal molecular species were composed of 16:0, 18:0, 20:0, 22:0, 23:0, 24:0, 24:1 and 24:2 fatty acid classes. However, the 16:0 fatty acid species was enriched in biliary sphingomyelin to a significantly greater degree than in sphingomyelins of canalicular-enriched plasma membranes (46% vs. 25% of total). We argue a physical-chemical case for laterally separated domains of very long chain sphingomyelins on the exoplasmic leaflet of the canalicular membrane. We bolster our hypothesis by the likelihood that the least hydrophobic, e.g., 16:0 sphingomyelin molecular species, are miscible with biliary phosphatidylcholines, and are secreted into bile. Laterally separated domains of very long chain sphingomyelins on the exoplasmic leaflet of the canalicular membrane could provide a means of sequestering cholesterol molecules prior to secretion into bile. Received: 19 March 1998/Revised: 8 October 1998     

Effect of a cholesterol-biosynthesis inhibitor on the fatty acid composition of phospholipids in the serum and tissues of rats

1. Changes produced by a cholesterol-biosynthesis inhibitor, trans-1,4-bis-(2-chlorobenzylaminomethyl)cyclohexane dihydrochloride (AY-9944), in the total fatty acids in the liver and brain, and in phospholipids in the serum, liver, heart, brain and lungs from male rats, have been studied. 2. Treatment with AY-9944 produced the following changes in the fatty acid composition: (a) a marked decrease in the percentage of linoleic acid and an increase in oleic acid in the total fatty acids in the liver; (b) in the serum, an overall decrease in the percentage of linoleic acid in neutral lipids and phospholipids; (c) an increased content of linoleic acid in the beta-acyl chain of phosphatidylcholines in the liver and in sphingomyelins in the brain and lungs; (d) an increased content of palmitic acid and oleic acid in the beta-acyl chain of phosphatidylcholine in the liver, heart and lungs; (e) an increased content of phosphatidylcholines and sphingomyelins, together with an increased percentage of saturated fatty acids in these phosphatides in the lungs. 3. Changes in the phosphatides and the production of foam cells in the lungs suggest that AY-9944 may be of use in the study of the alveolar membrane.     

Dynamics of Phospholipid Content in the Vacuolar Membrane of Red Beet Taproots Exposed to Abiotic Stress

The composition of vacuolar membrane phospholipids in the taproot of red beet (Beta vulgaris L.), cv. Modana, was determined at normal conditions and under different types of stress (hypo- and hyperosmotic and oxidative stress). The experiments have shown that, among vacuolar membrane phospholipids in red beet taproot, phosphatidylcholines and phosphatidylethanolamines dominated and accounted for 70% of total phospholipids. It is interesting that the content of phosphatidic acid was high (20% of total phospholipids of the vacuolar membrane). Stress effects brought about changes in the composition of membrane phospholipids, which may be an element of phenotypic adaptation. Under hypoosmotic stress, reliable changes in the content of phosphatidic acid were observed, hyperosmotic stress was associated with changes in the level of phosphatidylcholines and phosphatidylinositols, and oxidative stress was notable for changes in the content of phosphatidylethanolamines and phosphatidylserines. The most significant changes were observed in the classes of phospholipids that may be involved in structural modification of membranes associated with transformation of their bilayer lamellar structure into hexagonal. These phospholipids comprise phosphatidic acid, phosphatidylcholines, and phosphatidylethanolamines. Revealed changes in the content of these phospholipids may alter the ratio between lamellar bilayer and nonbilayer hexagonal lipid structures in the vacuolar membrane and act as an important adaptation mechanism ensuring protection against stress.     

Regulation of trout gill AMP deaminase by lipid bilayers. Effects of phospholipid composition and temperature

Trout gill AMP deaminase is inhibited by liposomes made of synthetic phosphatidylcholines containing higher saturated fatty acids. A preincubation of 1 hr, at 4 degrees C, was necessary to obtain the maximal effect. At 4 or 25 degrees C, these phospholipids modified essentially the substrate affinity of the enzyme by increasing the Michaelis constant proportionally to the length of the fatty acid chain. At 13 degrees C, the liposomes decreased the Hill coefficient also, thus inducing a negative cooperativity. Natural phosphatidylcholine and phosphatidylserine were without significant effect on gill AMP deaminase while natural sphingomyelin exhibited a similar effect to that shown in the presence of synthetic phosphatidylcholines. These results are discussed in relation to a possible effect of sphingomyelins in vivo.     

Fatty acid profiling of phospholipids by field desorption and fast atom bombardment mass spectrometry

Natural phosphatidylcholines, phosphatidylethanolamines and sphingomyelins have been investigated by field desorption and fast atom bombardment mass spectrometry. It is demonstrated that using these soft mass spectrometric ionization techniques, accurate, fast, and sensitive fatty acid profiling of phospholipids can be performed. With respect to the analysis of intact molecular species both ionization techniques reveal similar results. Using field desorption, a specific fragment ion provides a fast access to the total distribution of fatty acids in complex lipids. Generally, a good agreement between the mass spectrometric abundance data and those produced by gas chromatographic analysis is observed.     

Distribution of omega-3 and omega-6 fatty acids in the ether- and ester-linked phosphoglycerides from tissues of the rainbow trout, Salmo gairdneri

1. Data presented here demonstrate that polyunsaturated fatty acids in the phospholipids of rainbow trout tissues are compartmentalized differently than in mammalian tissues. 2. We have determined the distribution of omega-3 (n-3) and omega-6 (n-6) fatty acids in the alkyl-, alk-1-enyl-, and diacyl- subclasses of phosphatidylcholines (PC), phosphatidyl-ethanolamines (PE), phosphatidylinositols (PI), and phosphatidylserines (PS) from gill, kidney and spleen of rainbow trout. 3. Alkyl-linked PC and alk-1-enyl-linked PE were the most abundant ether-containing phospholipids, amounting to 10-15% of each class; no ether-linked PI or PS was detected. 4. C20:4 n-6 was found in high concentrations only in PI; the n-3 fatty acids were found in highest concentration in the ether-linked phospholipids as compared with the diacyl subclasses and C20:5 n-3 was especially prevalent in 1-O-alk-1'-enyl-2-acyl-sn-glycero-3-phosphoethanolamine and C22:6 n-3 was prevalent in PS.     

Phospholipase of rat intestine: mode of action]

Phospholipase activities of rat intestinal mucosa homogenate have been determined from lysophosphatidylcholines [14C] and phosphatidylcholines [-3H-14C]. In the presence of phosphatidylcholines, at pH 6.5, the homogenate has a phospholipase B activity. At pH 8.5, a phospholipase A2 activity was shown. In the presence of lysophospatidylcholines, at pH 6.5, we notice a lysophospholipase A1 activity. A kinetic study of the reactions allows us to separate the activity B into a phospholipase A2 activity and a lysophospholipase A1 activity. Thus, it appears that the total phospholipase activity of rat intestinal mucosa would results from a phospholipase A2 activity and a lysophospholipase A1 activity.     

Lipids of fish parasites and their hosts: fatty acids of phospholipids of Paratenuisentis ambiguus and its host eel (Anguilla anguilla)

The fatty acid composition of various phospholipid classes, e.g. phosphatidylcholines, phosphatidylethanolamines, phosphatidylserines and sphingomyelins, of the fish endoparasite Paratenuisentis ambiguus and of intestinal tissue of its host, eel (Anguilla anguilla), were investigated. Phospholipids from parasite and infected host intestine show considerable differences in the fatty acid compositions. High proportions of saturated long-chain and very long-chain acyl moieties were found in phosphatidylcholines, ethanolamines and serines of the parasite. Membrane phospholipids of P. ambiguus contain remarkably high proportions of polyunsaturated acyl moieties, in particular eicosapentaenoyl moieties (20:5 n-3), which undoubtedly originate from aquatic feed or host intestine. It is suggested that the parasite's membranes are stabilized by incorporation of high proportions of saturated long-chain and very long-chain fatty acids to counterbalance the destabilizing effects of the polyunsaturated fatty acids.     

The Glyceryl Ethers of Paramecium Phospholipids and Phosphonolipids1

Two glyceryl ethers, 1-O-hexadecyl glycerol and 1-O-cis-octadec-11-enyl glycerol, chimyl and paramecyl alcohol respectively, were quantified in total phospholipids and five glycerophospholipid classes from cells and cilia of the ciliated protozoon Parammecium tetraurelia. The ether content of 2-aminoethyl phosphonoglycerolipid was 85–90 mole %. Concentrations of ethers were greatest in the ethanolamine phosphonolipids > phosphatidylcholines > phosphatidylserines > phosphatidylethanolamines > phosphatidylinositols. The glyceryl ether concentrations in total cellular phospholipids increased with culture age in P. tetraurelia and P. multimteronucleatum cells. The glyceryl ether concentrations in the phospholipids of P. tetraurelia cilia remained constant from mid log to stationary phase of culture growth. Paramecium tetraurelia phospholipid glyceryl ether concentrations were made greater by supplementation of cultures with chimyl alcohol.