Influence of the nature of the sterol on the behavior of palmitic acid/sterol mixtures and their derived liposomes

The phase behavior of mixtures formed with palmitic acid (PA) and one of the following sterols (dihydrocholesterol, ergosterol, 7-dehydrocholesterol, stigmasterol and stigmastanol), in a PA/sterol molar ratio of 3/7, has been characterized by IR and ²H NMR spectroscopy at different pH. Our study shows that it is possible to form liquid-ordered (lo) lamellar phases with these binary non-phospholipid mixtures. The characterization of alkyl chain dynamics of PA in these systems revealed the large ordering effect of the sterols. It was possible to extrude these systems, using standard extrusion techniques, to form large unilamellar vesicles (LUVs), except in the case of ergosterol-containing mixture. The resulting LUVs displayed a very limited passive permeability consistent with the high sterol concentration. In addition, the stability of these PA/sterol self-assembled bilayers was also found to be pH-sensitive, therefore, potentially useful as nanovectors. By examining different sterols, we could establish some correlations between the structure of these bilayers and their permeability properties. The structure of the side chain at C17 of the sterol appears to play a prime role in the mixing properties with fatty acid.     

Melittin-induced leakage from phosphatidylcholine vesicles is modulated by cholesterol: a property used for membrane targeting

Melittin, an amphiphathic peptide, affects the permeability of vesicles. This can be demonstrated using the dye release technique. Calcein, a fluorescent marker, is trapped in large unilamellar 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) vesicles and melittin-induced leakage of the dye can be monitored directly by increasing fluorescence intensity. First, we characterized the effect of increasing cholesterol content in the membrane on melittin-induced leakage and our results reveal that cholesterol inhibits the lytic activity of the peptide. Using intrinsic fluorescence of the single tryptophan of melittin and ²H-NMR of headgroup deuterated phosphatidylcholine, we demonstrated that the affinity of melittin for phosphatidylcholine vesicles is reduced in the presence of cholesterol; this is associated with the tighter lipid packing of the cholesterol-containing bilayer. This reduced binding is responsible for the reduced melittin-induced leakage from cholesterol-containing membranes. The pathway of release was determined to be an all-or-none mechanism. Finally, we investigated the possibility of achieving specific membrane targeting with melittin, when vesicles of different lipid composition are simultaneously present. Melittin incubated together with vesicles made of pure POPC and POPC containing 30(mol)% cholesterol can empty nearly all the cholesterol-free vesicles while the cholesterol-containing vesicles remain almost intact. Owing to the preferential interaction of melittin with the pure POPC vesicles, we were able to achieve controlled release of encapsulated material from a specific vesicle population. Received: 8 May 1996 / Accepted: 12 September 1996     

Cholesterol sulfate and Ca(2+) modulate the mixing properties of lipids in stratum corneum model mixtures

The influence of cholesterol sulfate (CS) and calcium on the phase behavior of lipid mixtures mimicking the stratum corneum (SC) lipids was examined using vibrational spectroscopy. Raman microspectrocopy showed that equimolar mixtures of ceramide, palmitic acid, and cholesterol underwent a phase transition in which, at low temperatures, lipids formed mainly a mosaic of microcrystalline phase-separated domains, and above 45 degrees C, a more fluid and disordered phase in which the three lipid species were more miscible. In the presence of Ca(2+), there was the formation of fatty acid-Ca(2+) complexes that led to domains stable on heating. Consequently, these lipid mixtures remained heterogeneous, and the fatty acid molecules were not extensively involved in the formation of the fluid lipid phase, which included mainly ceramide and cholesterol. However, the presence of CS displaced the association site of Ca(2+) ions and inhibited the formation of domains formed by the fatty acid molecules complexed with Ca(2+) ions. This work reveals that CS and Ca(2+) modulate the lipid mixing properties and the lipid order in SC lipid models. The balance in the equilibria involving Ca(2+), CS, and fatty acids is proposed to have an impact on the organization and the function of the epidermis.     

Spatial and Temporal Variability in Growing-Season Net Ecosystem Carbon Dioxide Exchange at a Large Peatland in Ontario,Canada

We measured net ecosystem exchange of carbon dioxide (CO2) (NEE) during wet and dry summers (2000 and 2001) across a range of plant communities at Mer Bleue, a large peatland near Ottawa, southern Ontario, Canada. Wetland types included ombrotrophic bog hummocks and hollows, mineral-poor fen, and beaver pond margins. NEE was significantly different among the sites in both years, but rates of gross photosynthesis did not vary spatially even though species composition at the sites was variable. Soil respiration rates were very different across sites and dominated interannual variability in summer NEE within sites. During the dry summer of 2001, net CO2 uptake was significantly smaller, and most locations switched from a net sink to a source of CO2 under a range of levels of photosynthetically active radiation (PAR). The wetter areas--poor fen and beaver pond margin--had the largest rates of CO2 uptake and smallest rates of respiratory loss during the dry summer. Communities dominated by ericaceous shrubs (bog sites) maintained similar rates of gross photosynthesis between years; by contrast, the sedge-dominated areas (fen sites) showed signs of early senescence under drought conditions. Water table position was the strongest control on respiration in the drier summer, whereas surface peat temperature explained most of the variability in the wetter summer. Q 10 temperature-respiration quotients averaged 1.6 to 2.2. The ratio between maximum photosynthesis and respiration ranged from 3.7:1 in the poor fen to 1.2:1 at some bog sites; it declined at all sites in the drier summer owing to greater respiration rates relative to photosynthesis in evergreen shrub sites and a change in both processes in sedge sites. Our ability to predict ecosystem responses to changing climate depends on a more complete understanding of the factors that control NEE across a range of peatland plant communities.     

Developing dynamic mechanistic species distribution models: predicting bird-mediated spread of invasive plants across northeastern North America

Species distribution models are a fundamental tool in ecology, conservation biology, and biogeography and typically identify potential species distributions using static phenomenological models. We demonstrate the importance of complementing these popular models with spatially explicit, dynamic mechanistic models that link potential and realized distributions. We develop general grid-based, pattern-oriented spread models incorporating three mechanisms--plant population growth, local dispersal, and long-distance dispersal--to predict broadscale spread patterns in heterogeneous landscapes. We use the model to examine the spread of the invasive Celastrus orbiculatus (Oriental bittersweet) by Sturnus vulgaris (European starling) across northeastern North America. We find excellent quantitative agreement with historical spread records over the last century that are critically linked to the geometry of heterogeneous landscapes and each of the explanatory mechanisms considered. Spread of bittersweet before 1960 was primarily driven by high growth rates in developed and agricultural landscapes, while subsequent spread was mediated by expansion into deciduous and coniferous forests. Large, continuous patches of coniferous forests may substantially impede invasion. The success of C. orbiculatus and its potential mutualism with S. vulgaris suggest troubling predictions for the spread of other invasive, fleshy-fruited plant species across northeastern North America.     

The Use of Magnetic Resonance Spectroscopy as a Tool for the Measurement of Bi-hemispheric Transcranial Electric Stimulation Effects on Primary Motor Cortex Metabolism

Transcranial direct current stimulation (tDCS) is a neuromodulation technique that has been increasingly used over the past decade in the treatment of neurological and psychiatric disorders such as stroke and depression. Yet, the mechanisms underlying its ability to modulate brain excitability to improve clinical symptoms remains poorly understood ³³. To help improve this understanding, proton magnetic resonance spectroscopy (¹H-MRS) can be used as it allows the in vivo quantification of brain metabolites such as γ-aminobutyric acid (GABA) and glutamate in a region-specific manner ⁴¹. In fact, a recent study demonstrated that ¹H-MRS is indeed a powerful means to better understand the effects of tDCS on neurotransmitter concentration ³⁴. This article aims to describe the complete protocol for combining tDCS (NeuroConn MR compatible stimulator) with ¹H-MRS at 3 T using a MEGA-PRESS sequence. We will describe the impact of a protocol that has shown great promise for the treatment of motor dysfunctions after stroke, which consists of bilateral stimulation of primary motor cortices ^27,30,31. Methodological factors to consider and possible modifications to the protocol are also discussed.     

Persistence of Water within Perched Basins of the Peace-Athabasca Delta, Northern Canada

The Peace-Athabasca Delta (PAD) is one of the world’s largest freshwater deltas. Many of its shallow (<1.5 m) lakes and wetlands are perched above surrounding waterways. The delta has experienced a number of wetting and drying intervals. The latest drying trend ended in 1996 when high waters, generated under ice-jam and open-water conditions, recharged a number of the perched basins. The objective of this study was to determine the relative importance of hydroclimatic components on the persistence of water. A water-balance model was developed to simulate water-level responses following a flood event. Basin response was tested against a range of historical hydroclimatic conditions that have occurred in the delta during the 20th century. Ponded water duration in a 0.8 m deep perched basin was 5 years for the cool-dry period of the 1920s, slightly longer for the post-1974 flood era, and up to 9 years for the wet conditions of the 1940s and 1950s. Water drawdown occurred in almost every year and was almost exclusively due to evaporation exceeding precipitation. Net groundwater flux was minimal. Given the overlying importance of the floodwater component in the water balance of perched basins, the next step is to investigate the causes, spatial sources and frequency of flooding. This water balance model presented in this paper offers a useful tool for the management of the duration of water in perched wetlands of the PAD, which can help preserve essential habitat for wildlife.     

Investigation of the interaction between melittin and dipalmitoylphosphatidylglycerol bilayers by vibrational spectroscopy.

Melittin is shown to affect the structure of the charged phospholipid dipalmitoylphosphatidylglycerol (DPPG). In the gel phase, the presence of melittin leads to (i) an increased lipid interchain vibrational coupling, (ii) a shift of the rectangular to hexagonal lipid packing transition toward low temperatures, (iii) a very small conformational disordering effect, (iv) a decrease of the polarity or hydrogen bonding capability of the lipid ester group surrounding, (v) an important decrease of the water content in the complexes where the remaining water has a more disordered structure than bulk water, and (vi) an interlamellar repeat distance of 79 A. All these observations are rationalized by the following model: adjacent bilayers of DPPG are bridged by tetramers of melittin through electrostatic interactions inducing surface charge neutralization and partial dehydration of the complexes. Melittin also affects the thermotropic behavior of DPPG. When a small amount of the toxin is present, its affinity for charged lipids is such that a phase separation occurs, the domains being stable enough to have their own gel to liquid-crystalline phase transition. In the fluid state, a deeper penetration into the lipid matrix is proposed based on the downshift of the phase transition and the low vibrational interchain coupling. This study brings out general features of cationic species/anionic lipid complexes. The charge neutralization leads to stronger interchain coupling, and electrostatic bridging of adjacent bilayers seems to be common. The hydrophobicity of the peptide is a key factor in the modulation of the gel to liquid-crystalline phase transition and in its insertion in the fluid lipid matrix.     

Influence of lipid chain unsaturation on melittin-induced micellization.

It is well known that melittin, an amphipathic helical peptide, causes the micellization of phosphatidylcholine vesicles. In the present work, we conclude that the extent of micellization is dependent on the level of unsaturation of the lipid acyl chains. We report the results obtained on two systems: dipalmitoylphosphatidylcholine (DPPC), containing 10(mol)% saturated or unsaturated fatty acid (palmitic, oleic, or linoleic), and DPPC, containing 10(mol)% positively charged diacyloxy-3-(trimethylammonio)propane bearing palmitic or oleic acyl chains. For both systems, the presence of unsaturation in the lipid acyl chains inhibits melittin-induced micellization. Conversely, the addition of saturated palmitic acid to the DPPC matrix enhances the micellization. This modulation is proposed to be associated with the cohesion of the hydrophobic core. When the lipid chain packing of the gel-phase bilayer is already perturbed by the presence of unsaturation, it seems easier for the membrane to accommodate melittin at the interface, and the distribution of the peptide in the bilayer could be the origin of the inhibition of the micellization. The cohesion of the apolar core is shown to play an unquestionable role in melittin-induced micellization; however, this contribution does not appear to be as important as the electrostatic interactions between melittin and positively or negatively charged lipids.     

Phytoremediation of groundwater contaminated with pesticides using short-rotation willow crops: A case study of an apple orchard

The occurrence of pesticides in groundwater represents an important health issue, notably for population whose drinking water supply source is located in agricultural areas. However, few solutions have been considered with regard to this issue. We tested the efficacy of a vegetal filtering system made of shrub willows planted at a high density (16,000 plants ha^?1) to filter or degrade pesticides found in the groundwater flowing out of an apple orchard. Ethylene urea (EU), ethylene thiourea (ETU), tetrahydrophthalimide (THPI), atrazine, and desethylatrazine were monitored in the soil solution in willow and control plots over one growing season. ETU and atrazine concentrations were lower in the willow plots relative to the control plots, whereas desethylatrazine concentration was higher in the willow plots. No significant difference was detected for EU and THPI. Furthermore, pesticide concentrations displayed complex temporal patterns. These results suggest that willow filter systems can filter or degrade pesticides, notably ETU and atrazine, and could be used for phytoremediation purposes. Yet, this potential remains to be quantified with further studies using experimental settings allowing more estimation in time and space.