Differential Effect of Plant Lipids on Membrane Organization: SPECIFICITIES OF PHYTOSPHINGOLIPIDS AND PHYTOSTEROLS*

The high diversity of the plant lipid mixture raises the question of their respective involvement in the definition of membrane organization. This is particularly the case for plant plasma membrane, which is enriched in specific lipids, such as free and conjugated forms of phytosterols and typical phytosphingolipids, such as glycosylinositolphosphoceramides. This question was here addressed extensively by characterizing the order level of membrane from vesicles prepared using various plant lipid mixtures and labeled with an environment-sensitive probe. Fluorescence spectroscopy experiments showed that among major phytosterols, campesterol exhibits a stronger ability than β-sitosterol and stigmasterol to order model membranes. Multispectral confocal microscopy, allowing spatial analysis of membrane organization, demonstrated accordingly the strong ability of campesterol to promote ordered domain formation and to organize their spatial distribution at the membrane surface. Conjugated sterol forms, alone and in synergy with free sterols, exhibit a striking ability to order membrane. Plant sphingolipids, particularly glycosylinositolphosphoceramides, enhanced the sterol-induced ordering effect, emphasizing the formation and increasing the size of sterol-dependent ordered domains. Altogether, our results support a differential involvement of free and conjugated phytosterols in the formation of ordered domains and suggest that the diversity of plant lipids, allowing various local combinations of lipid species, could be a major contributor to membrane organization in particular through the formation of sphingolipid-sterol interacting domains.     

Gas chromatography-mass spectrometry determination of metabolites of conjugated cis-9,trans-11,cis-15 18:3 fatty acid

Structural determination of polyunsaturated fatty acids by gas chromatography-mass spectrometry (GC-MS) requires currently the use of nitrogen containing derivatives such as picolinyl esters, 4,4-dimethyloxazoline or pyrrolidides derivatives. The derivatization is required in most cases to obtain low energy fragmentation that allows accurate location of the double bonds. In the present work, the following metabolites of rumelenic (cis-9,trans-11,cis-15 18:3) acid, from rat livers, were identified: cis-8,cis-11,trans-13,cis-17 20:4, cis-5,cis-8,cis-11,trans-13,cis-17 20:5, cis-7,cis-10,cis-13,trans-15,cis-19 22:5, and cis-4,cis-7,cis-10,cis-13,trans-15,cis-19 22:6 acids by GC-MS as their 4,4-dimethyloxazoline and methyl esters derivatives. Specific fragmentation of the methyl ester derivatives revealed some similarity with their corresponding DMOX derivatives. Indeed, intense ion fragments at m/z=M+-69, corresponding to a cleavage at the center of a bis-methylene interrupted double bond system were observed for all identified metabolites. Moreover, intense ion fragments at m/z=M+-136, corresponding to allylic cleavage of the n-12 double bonds were observed for the C20:5, C22:5, C22:6 acid metabolites. For the long chain polyunsaturated fatty acids from the rumelenic metabolism, we showed that single methyl esters derivatives might be used for both usual quantification by GC-FID and identification by GC-MS.     

Protection of mitochondria during cold storage of liver and following transplantation: comparison of the two solutions, University of Wisconsin and Eurocollins

Injury to allografts during ischaemia/reperfusion contribute to the development of graft failure following transplantation with significant morbidity and mortality to patients. The development of University of Wisconsin solution has significantly improved the quality of graft preservation and transplant outcome relative to formerly used solutions such as Eurocollins. The aim of this study was to further characterize mitochondrial structural and functional alterations occurring in rat livers following cold storage and transplantation. Mitochondrial impairment after prolonged storage in Eurocollins included decreased cyt. c+c₁, cyt. b and cyt. a+a₃ concentration and dramatic falls in the activities of the respiratory chain enzymes ubiquinol-cyt. c oxidoreductase and cytochrome oxidase. Under the same conditions the highest hydroperoxide but lowest vitamin E concentrations were also found. Although both the Eurocollins and University of Wisconsin preservation solutions have limitations in preventing oxidative injuries following cold storage and reperfusion, our data indicate that mitochondrial impairment was higher in Eurocollins- than in University of Wisconsin-stored livers. Further improvements are necessary in maintaining the stability of mitochondria in order to optimize preservations solutions used in transplantations.     

反刍动物体内共轭亚油酸生物合成途径研究进展

共轭亚油酸是一组共轭双键具有不同位置和几何构象的亚油酸异构体,主要存在于反刍动物的肉及乳中。本文就其中2种具有重要生理功能的共轭亚油酸(c-9,t-11CLA和t-10,c-12CLA)的生物合成途径的研究进行综述,以期了解共轭亚油酸代谢途径及其调控路径,为今后利用合成生物学指导共轭亚油酸的合成奠定基础。     

Double enrichment of chicken eggs with conjugated linoleic acid and n-3 fatty acids through dietary fat supplementation

Yolk fat fatty acid (FA) concentrations, sensory quality and firmness of eggs and laying hen performance were evaluated with respect to the combined inclusion in the diet of conjugated linoleic acid (CLA), high n-3 oil sources and high-oleic sunflower oil (HOSO). Nine diets were arranged factorially, with three levels of n-3 FA supplementation (2.9, 3.7 and 4.5 g/kg) from three different sources (two fish oils highly concentrated in eicosapentanoic (EPA) or docosahexanoic acid (DHA) and one algae oil with a very high-DHA content) in diets added with fixed amounts of CLA (2.5 g/kg) and HOSO (30 g/kg). A commercial feed with no CLA, n-3 or HOSO added, and another one containing 4.5 g/kg of high-DHA fish oil but not CLA or HOSO were also formulated. An increase in n-3 FA supplementation had little effect on proportions of CLA, monounsaturated FA, saturated FA or total polyunsaturated FA in yolk fat, but increased (P<0.005) long-chain n-3 FA and decreased (P<0.001) long-chain n-6 FA. An increment of dietary n-3 FA also impaired linearly (P<0.001) egg acceptability by consumers. An increment in the proportion of DHA with respect to total n-3 FA from 0.28 to 0.96 increased yolk concentrations of DHA (P<0.001) and total n-3 FA (P<0.01), but decreased (P<0.001) concentrations of EPA and docosapentanoic acid FA. Current data indicate that addition of HOSO to diets supplemented with moderate amounts of CLA and n-3 FA allows the production of double enriched eggs while maintaining sensory quality for consumers at acceptable levels.     

Comparison of fatty acid composition of bacterial and protozoal fractions in rumen fluid of sheep fed diet supplemented with sunflower, rapeseed and linseed oils

The objective of this study was to investigate effects of oil supplements on the composition of fatty acids (FA), especially of trans11-C18:1 (vaccenic acid, TVA) and cis9, trans11-C18:2 conjugated linoleic acid (c9,t11-CLA), in bacterial (BF) and protozoal (PF) fractions of rumen fluid of sheep that was fractionated centrifugation. Four sheep were fed a diet consisting of meadow hay (960 g dry matter (DM)/day) and of barley grain (240 g DM/day), with sunflower oil (SO), rapeseed oil (RO) or linseed oil (LO) as supplements (60 g/day) in a Latin square design. The oils were used as they are rich in linoleic acid (SO, 533 g/kg of FA), oleic acid (RO, 605 g/kg of FA) and α-linolenic acid (LO, 504 g/kg of FA). Compared to the control (i.e., without oils), oil supplements influenced the concentration of unsaturated (UFA) and saturated fatty acids (SFA). In both BF and PF, the main fatty acids were palmitic and stearic, but PF contained a higher proportion of TVA and c9,t11-CLA than BF. In PF, TVA concentrations, ranked by oil supplement, were SO > RO > LO > Control (174, 150, 118, 74 g/kg of FA, respectively) and the c9,t11-CLA concentrations were RO > SO > LO > Control (59, 51, 27 and 15 g/kg of FA, respectively). Concentrations of c9,t11-CLA in PF were two to three times higher than in BF with all the oil supplements versus the control. Oil treatments impacted the c9,t11-CLA concentration in the fractions, especially SO and RO. The protozoal fraction contained a higher proportion of TVA and c9,t11-CLA than did the bacterial fraction, and dietary addition of SO, RO and LO resulted in a higher incorporation of TVA into both bacterial and protozoal microbial fractions, which probably positively affected TVA flow from the rumen.     

Conjugated eicosapentaenoic acid inhibits human topoisomerase IB with a mechanism different from camptothecin

Conjugated eicosapentaenoic acid (cEPA) has been found to have antitumor effects which has been ascribed to their ability to inhibit DNA topoisomerases and DNA polymerases. We here show that cEPA inhibits the catalytic activity of human topoisomerase I, but unlike camptothecin it does not stabilize the cleavable complex, indicating a different mechanism of action. cEPA inhibits topoisomerase by impeding the catalytic cleavage of the DNA substrate as demonstrated using specific oligonucleotide substrates, and prevents the stabilization of the cleavable complex by camptothecin. Preincubation of the inhibitor with the enzyme is required to obtain complete inhibition. Molecular docking simulations indicate that the preferred cEPA binding site is proximal to the active site with the carboxylic group strongly interacting with the positively charged K443 and K587. Taken together the results indicate that cEPA inhibitor does not prevent DNA binding but inhibits DNA cleavage, binding in a region close to the topoisomerase active site.     

Effect of dietary conjugated linoleic acid isomers on water and glycerol permeability of kidney membranes

Conjugated linoleic acid (CLA) refers to a group of positional and geometrical isomers of linoleic acid in which the double bonds are conjugated. Dietary CLA has been associated with various health benefits although details of its molecular mode of action remain elusive. The effect of CLA supplemented to palm oil-based diets in Wistar rats, as a mixture of both or isolated c9,t11 and t10,c12 isomers, was examined on water and glycerol membrane permeability of kidney proximal tubule. Although water permeability was unaltered, an increase in glycerol permeability was obtained for the group supplemented with CLA mixture, even though the activation energy for glycerol permeation remained high. This effect was correlated with an increased CLA isomeric membrane incorporation for the same dietary group. These results suggest that diet supplementation with CLA mixture, in contrast to its individual isomers, may enhance membrane fluidity subsequently raising kidney glycerol reabsorption.     

Ambient Method for the Production of an Ionically Gated Carbon Nanotube Common Cathode in Tandem Organic Solar Cells

A method of fabricating organic photovoltaic (OPV) tandems that requires no vacuum processing is presented. These devices are comprised of two solution-processed polymeric cells connected in parallel by a transparent carbon nanotubes (CNT) interlayer. This structure includes improvements in fabrication techniques for tandem OPV devices. First the need for ambient-processed cathodes is considered. The CNT anode in the tandem device is tuned via ionic gating to become a common cathode. Ionic gating employs electric double layer charging to lower the work function of the CNT electrode. Secondly, the difficulty of sequentially stacking tandem layers by solution-processing is addressed. The devices are fabricated via solution and dry-lamination in ambient conditions with parallel processing steps. The method of fabricating the individual polymeric cells, the steps needed to laminate them together with a common CNT cathode, and then provide some representative results are described. These results demonstrate ionic gating of the CNT electrode to create a common cathode and addition of current and efficiency as a result of the lamination procedure.     

酶法生产共轭亚油酸的研究进展

共轭亚油酸(Conjugated linoleic acid,CLA)具有抗癌、抗动脉粥样硬化、减肥和免疫调节等生理活性。共轭亚油酸可以通过酶法异构化获得,将底物亚油酸异构形成具有生物活性物质-共轭亚油酸的异构酶称为亚油酸异构酶。因此,通过介绍亚油酸异构酶的来源、作用机制、酶学性质和基因工程菌生产等方面的研究进展,结合不断发展的基因工程技术,旨在提高亚油酸异构酶的活性、产量和异构化效率,以扩大反应底物范围,降低生产成本,从而推进共轭亚油酸的规模化、可持续性的工业生产。