Cambios en los acidos grasos de membranas de Microbacterium esteraromaticum GNP-5 con diferentes temperaturas y osmolaridades / Changes in Membrane Fatty Acids of Microbacterium esteraromaticum GNP5 with Changes of Temperature and Osmolarity

Microbacterium esteraromaticum es un microorganismo que se aísla con frecuencia de landfarming o procesos de biorremediación de hidrocarburos en la meseta de la Patagonia central (Argentina) y se halla sometido a variaciones de temperatura y a cambios de salinidad que se producen naturalmente. Su adaptabilidad a esos cambios climáticos indujo al estudio de las modificaciones que se producen en su membrana celular para resistirlos. En este trabajo se estudió el efecto conjunto de la temperatura y la concentración de cloruro de sodio sobre la composición de los ácidos grasos de membrana en la cepa de Microbacterium esteraromaticum GNP5b. M. esteraromaticum utiliza, frente al incremento de la temperatura, la estrategia es aumentar la longitud de sus ácidos grasos de cadenas ramificada impar (17 átomos de carbono) con disminución de 15:0 anteiso, así como el porcentaje de ácidos grasos 15:0 iso (de mayor punto de fusión) a partir del respectivo anteiso. El aumento de la salinidad modifica la composición de ácidos grasos siguiendo patrones diferentes según sea la temperatura de incubación. A 14 y 28 °C incrementa los 15:0 iso y 17:0 iso en detrimento de sus homólogos anteiso. A 37 °C este grupo de ácidos grasos no sigue los mismos patrones anteriores. La longitud de cadena, expresada como el índice C15/C17, es errática con el aumento de la salinidad.

Erythrocyte membrane fatty acid composition in cancer patients

Essential fatty acids (EFA) have an important role in complex metabolic reactions. The metabolism of essential polyunsaturated fatty acids (PUFA) appears to be one of the critical targets in the complex metabolic stages that lead to, or are associated with cancer. The goal of our research was to analyze the erythrocyte specific types of membrane fatty acid content, level and distribution in cancer patients as compared to non-cancer patients. Changes in fatty acid composition may affect different aspects of cell structure and function, including proliferation. Analyses of RBCs membrane fatty acids were performed for 255 patients with different types of cancer (breast, prostate, liver, pancreas, colon, and lung), 2,800 non-cancer patients and 34 healthy volunteers. Our research study demonstrated a lower level of stearic acid and an increased content of oleic acid in RBC of cancer patients in comparison with control and non-cancer patients. According to the results of this investigation, the ratio of Eicosa pentaenoic acid (EPA) and Decosa hexaenoic acid (DHA) to Alpha-linolenic acid (ALA) may be useful to estimate PUFA imbalances in cancer patients. EPA and DHA acid may be recommended as supplementation and in addition to current therapy during cancer treatment

Influencia de la concentración salina y la temperatura en la composición de ácidos grasos de membrana de Pseudomonas fluorescens GNP-OHP-3 / Influence of salinity and temperature on fatty acid composition of Pseudomonas fluorescens GNP-OHP-3 membrane

Las bacterias responden a los cambios ambientales modificando su composición, para evitar el daño que dichos cambios podrían ejercer. Una de las modificaciones más importantes es la variación de la composición de los ácidos grasos de las membranas celulares, que le permite mantener la homeoviscosidad ante situaciones de estrés. Trabajos previos han estudiado la acción de la temperatura, presión hidrostática y diferentes solventes sobre cepas de Pseudomonas putida. En este trabajo se estudió la acción conjunta de la temperatura y la salinidad sobre la composición de ácidos grasos de membranas celulares de Pseudomonas fluorescens GNP-OHP-3, una cepa bacteriana aislada de un hábitat contaminado con petróleo. Pseudomonas fluorescens GNP-OHP-3 respondió a las variaciones de temperatura modificando los ácidos grasos de sus membranas de manera similar a lo descripto en otros integrantes de su género: ante el aumento de temperatura se observó un incremento de ácidos grasos saturados y una disminución de los ácidos grasos insaturados. En el rango de concentraciones salinas ensayadas las variaciones de los ácidos grasos mayoritarios fueron en general erráticas. La respuesta de los ácidos grasos ciclo propano pudo expresarse con ecuaciones matemáticas que permitieron predecir el porcentaje de estos ácidos en relación a la concentración de cloruro de sodio.

The bacteria respond to environmental changes modifying their composition. One of the most important modifications is the variation on fatty acid composition of cellular membranes to maintain the homeoviscosity. The action of temperature, hydrostatic pressure and solvents on Pseudomonas putida has been thoroughly studied. In this paper, the combined action of the temperature and salinity on fatty acid composition of cellular membranes of Pseudomonas fluorescens GNP-OHP-3, a bacterial strain isolated from a petroleum contaminated habitat, was studied. The modifications in the fatty acid composition of Pseudomonas fluorescens GNP-OHP-3 membrane were similar to those described for other members of Pseudomonas: an increase in saturated fatty acids and a decrease in unsaturated fatty acids were observed with the increase of the temperature. Variations of main fatty acids were in general erratic in the range of assayed saline concentrations. The variation of cycle propane fatty acids could be expressed with mathematic equations that allowed to predict their percentage in relation to sodium chloride concentration.

Total lipids and fatty acids of strains of "Metarhizium anisopliae"

Two growth stages, conidia (C) and mycelium (M), and two media, minimal medium (MM) and complete medium (MC), were compared in 10 strains of "M. anisopliae", and two strains of "M. anisopliae" var. "majus" were similar in percentages of total lipids. Tukey test for average of lipid content in conidia (C) and mycelia (M) cultured on minimal (MM) and complete (MC) media showed significant differences between means at the 5(per cent) level of mycelia and conidia, indicating variability in total lipid production and storage during growth. Strains 5 and 7, both variety "majus", did not present sizable differences from variety "anisopliae". For fatty acids, C18:1 and C18:2, oleic and linoleic, respectively, the differences were all highly significant (p=1(per cent)) with highest means being obtained for conidia for fatty acid C18:1 and for mycelia for fatty acid C18:2.

Isolation and characterization of the basal cell membranes of the human term placenta.

A relatively simple and rapid method is described for the isolation of basal cell membranes (BCM) from the human placenta at term which showed considerable improvement in the yield, purity and membrane characteristics as compared to the earlier described methods. The method is based on thorough washings of the syncytium in balanced salt solution, selective grinding, hypotonic lysis, sonication, incubation with EDTA and then more conventional differential centrifugation and ultracentrifugation. The isolated material showed smooth surfaced vesicular structure of various sizes as revealed by both positive and negative staining and transmission electron microscopic analysis. The membrane was highly enriched in Na+/K+, Ca2+ and Mg2+ dependent ATPase activities while the cross contamination with brush border surfaces was low as revealed by the marker enzyme assays specific for the brush border membrane (BBM) such as the disaccharide hydrolases, aminopeptidase and alkaline phosphatase. The membrane showed a relatively low lipid/protein ratio and the lipid composition represented by a variety of phospholipids (phosphatidyl choline, phosphatidyl ethanolamine, sphingomyelin, phosphatidyl inositol and phosphatidyl serine), neutral lipids (cholesterol, triacyl glycerol, free fatty acids) and glycosphingolipids (ganglioside, cerebroside and sulfatide). It also contained plasmalogens. On SDS-PAGE analysis and Coomassie blue staining reaction, the isolated membrane showed 14 major bands with as many minor ones with a molecular weight ranging between 30-110 kDa.

Role of lipids in receptor mediated signal transduction.

A new strategy has been successfully applied to reconstitute the brain specific serotonin 5-HT1A receptor-G protein-adenylate cyclase complex. A mild method of tissue preparation gave a stable, membrane-bound form of the receptor (SBP) which retained its natural lipid content. Treatment of SBP with serotonin (1 microM) and 3-[(3-cholamidopropyl) dimethyl ammonio]-1-propanesulphonate (CHAPS) (2%) solubilized the ligand-receptor-G protein-ligand complex along with the associated phospholipids and cholesterol. Dialysis of this extract (SBDS) against buffer containing 25% ethylene glycol produced a stable, reconstituted and active preparation (SBDSE) of vesicles which upon centrifugal separation followed by gentle resuspension retained 95-100% [3H] 8-OH-DPAT binding activity as well as 60% [3H] GppNHp binding and adenylate cyclase activities of SBDSE. The reconstituted receptor preparation compared well with the membrane-bound form in displaying a similar value for KD (2.1 nM) and a single affinity state for [3H] 8-OH-DPAT binding (Bmax = 118 fmol/mg). However, in sharp contrast to the membrane-bound receptor which was negatively coupled to adenylate cyclase, agonist treatment of the solubilized and reconstituted receptor resulted in an increase in adenylate cyclase. This change in receptor-adenylate cyclase coupling following reshuffling of membrane lipids during solubilization and reconstitution suggested that membrane lipids could have a profound effect on receptor-effector coupling. To study the effect of membrane lipid composition on receptor-mediated signal transduction in a stabler and more natural system, neural cells derived from different parts of the brain (hippocampus, HN2; CNS, NCB-20; dorsal root ganglion, F-11) and a non-neural cell line (CHO), all with differing membrane lipid compositions, were selected. Since no known cell line contains the serotonin 5-HT1A receptor (5-HT1A-R), stable transfection of the selected cell lines with a DNA construct encoding the human 5-HT1A-R was carried out and this resulted in a late increase of [3H] 8-OH-DPAT binding in the stationary phase only in the cell lines of neural origin. In the non-neural cell line (CHO), which also displayed marked difference in membrane lipids, the receptor was positively coupled to the phospholipase C-IP3-[Ca2+]i cascade. Even though GPLC was present in the NCB-20 and F-11 cells as evidenced by a bradykinin receptor-mediated increase in inositol phosphates in these cells 8-OH-DPAT treatment resulted in no change in phospholipase C in any of the cell lines of neural origin.(ABSTRACT TRUNCATED AT 400 WORDS)

Studies on nonesterified fatty acids in rat intestinal cell membranes.

Lipid composition of total membrane fractions prepared from scraped rat intestinal mucosa and isolated epithelial cells were compared. Membranes prepared from mucosa had four times higher nonesterified fatty acids (NEFA) as compared to the epithelial cell membranes. Cholesterol and phospholipid contents were similar in both the membrane preparations but triglyceride content was high and di- and monoglyceride were low in epithelial cell membranes as compared to the mucosal membranes. Inclusion of p-bromophenacyl bromide, a phospholipase inhibitor, in the intestinal lumen wash solution and homogenizing buffer did not reduce the NEFA content of the scraped mucosal membranes whereas inclusion of diethyl p-nitrophenyl phosphate, a lipase inhibitor reduced it by 40%. These results suggest that NEFA are normal constituent of intestinal cell membranes.

Lipid composition of intestinal brush border membrane in alloxan induced acute experimental diabetes.

Several alterations were observed in the rat intestinal brush border membrane (BBM) lipid composition after 7 days old alloxane-induced diabetes as compared to the control animals. There was no change in the total protein contents but a significant increase in the total lipid contents was observed. Glycolipids constituting the major lipid components showed a two-fold increase. No significant difference was observed in the total phospholipid contents. A significant decline in the free cholesterol (CH) level, free fatty acids, triglycerides and sialic acid contents was observed in membranes from diabetic rats. Esterified CH, monoglycerides+diglycerides, phosphatidyl serine+phosphatidyl inositol and phosphatidyl choline levels remained unaffected. A significant increase in sphingomyelin with a parallel decrease in phosphatidyl ethanolamine was observed in BBM preparations from diabetic rats. The observed changes in intestinal BBM might be responsible for altered functions of the diabetic intestines.

Effect of high-fat diet on mice intestinal brush border membrane composition.

Effect of feeding high-fat (26% fat) diet to mice for 21 days on intestinal brush border membrane composition was evaluated by comparing with controls fed 10% fat diet. 125I-labelled lectin binding and chemical analysis of fucose, sialic acid, hexoses and hexosamines revealed essentially similar results in control and test groups. Membrane phospholipids, expressed on dry membrane basis, were significantly reduced while total cholesterol was enhanced in experimental group compared to controls. Triglyceride content was not altered under these conditions. [14C]-acetate incorporation studies showed that decrease in phospholipid content was due to reduced synthesis of phospholipid constituents, in particular, phosphatidylcholine and phosphatidylethanolamine, as a result of feeding high fat diet. The results suggest that high amount of fat in the diet of adult mice does not alter sugar content in brush border membrane but affects membrane lipid composition.

Location of chloroplast membrane resonances in high resolution 13C-NMR spectrum of spinach leaves.

Spinach leaves were studied by high resolution 13C-NMR spectroscopy. The spectra obtained, though highly complex, showed some distinct resonances assignable to carbons of lipids of the chloroplast membranes, thereby offering a novel way for studying the chloroplast membranes in situ.

Characterization of glycosyl phosphatidylinositol lipids of parasitic protozoans: Leishmania mexicana mexicana promastigotes, Trypanosoma cruzi Peru epimastigotes and Tritrichomonas foetus.

Glycosyl phosphatidylinositol lipids of cultured L.mex, mexicana LV732 promastigotes, T. cruzi Peru epimastigotes and Tritrichomonas foetus have been isolated and characterized using metabolic labelling and chromatographic and mass spectrometric (MS) techniques. TLC of the unsaponifiable lipid fractions of L. mex. mexicana and T. cruzi obtained from DEAE Sephadex A-25 followed by Iatrobead column chromatography showed three inositol phosphate-containing lipid components. [3H]myo-inositol, [3H]palmitic acid or H3 32PO4 lipid precursors were incorporated into these three lipid components. Fraction 2 (LM2 and TCP-2) comprises inositol phosphate ceramides. The other two fractions appear to contain mono-O-alkyl and di-O-alkyl glycerol inositol phosphates. Lyso-1-O-alkyl phosphatidylinositols could be cleaved by treatment of PI-specific phosphalipase C. The di-O-alkyl-phospho inositols of these parasites being the first dialkylglycerol lipids reported from eukaryotic membranes raises the possibility of chemotherapy for leishmaniasis and trypanosomiasis based upon functional impairment of alkyl ether lipids. Tritrichomonas foetus contains two major glycophosphosphingolipids, designated TF1 and TF2, which are metabolically labelled with [3H]myo-inositol and H3 32PO4. Both lipids contained ceramides. The major ceramide contains the 18:0 and 18:1 bases and 16:0 N-acyl group. The major glycolipid fraction (TF1) contains fucose linked to inositol diphosphate; one of the phosphates being linked to the ceramide moiety, and the other to ethanolamine. TF1 appears to be a novel class of glycophosphosphingolipid, which may be a part of a membrane anchor.

Changes in lipid profiles of brain of albino rats following envenomation.

Effect of high doses of cobra venom (150 micrograms/120 +/- 20 g body weight) and viper venom (300 micrograms/120 +/- 20 g body weight) on total lipid, triglyceride, phospholipid, cholesterol, high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) of brain of albino rats was studied. Total lipid (TL) triglyceride (TG) and phospholipid (PL) are decreased in both viper and cobra venom treated groups while cholesterol (C), and LDL-C are increased in both the groups in relation to controlled ones. HDL-C content was almost unaltered. Decrease in triglyceride and phospholipid may be due to effect of lipases and phospholipases whereas increased cholesterol and LDL-C may be attributed to lysis of cell membrane.

Effect of iron deficiency on phosphoinositide levels in rat brain.

Adult rats raised on 90% iron deficient diet for 6 weeks showed a decrease in the levels of brain phosphatidylinositol (60%), phosphatidylinositol 4-phosphate (25%) and phosphatidylinositol 4,5-bisphosphate (30%), the precursors for the second messengers, diacylglycerol and inositol phosphates. Rehabilitation to normal diet for 24 days was found to restore the levels to control values. The data suggests that iron deficiency could lead to alterations in the functions related to phosphoinositide-linked receptor system(s).

Modification of electrocyte membrane lipid composition induced by denervation

1. We determined the effect of denervation on the lipid composition of the main electric organ and electrocyte postsynaptic membrane vesicles of Electrophorus electricus. 2. Lipid extracts of whole electric organ contain mainly cholesterol, triglycerides, phosphatidylcholine and phosphatidlethanolamine. After 30 days of denervation, the lipid composition of whole electric organ did not change appreciably. 3. Lipid extracts of the membrane vesicles were similar, except that they contained mainly free fatty acids rather than triglycerides. After 30 days of denervation, cholesterol concentration was increased and phsopholipids were decreased in the membrane fraction, with higher relative concentrations of phosphatidylethanolamine and cerebrosides. 4. These results suggest that electrocyte membrane fluidity and permeability will change after 30 days of denervation