Characterization and quantification of proteins in lecithins

Several methods for extraction and quantification of proteins from lecithins were compared. Extraction with hexane-2-propanol-water followed by amino acid analysis is the most suitable method for isolation and quantification of proteins from lecithins. The detection limit of the method is 15 mg protein/kg lecithin, and the quantification limit is 50 mg protein/kg. The relative repeatability limits for samples containing 0-500 and 500-5000 mg protein/kg sample were 12.6 and 7.5%, respectively. The protein recovery ranged between 101 and 123%. The protein content has been determined in different kinds of lecithins. The results were as follows: standard soy lecithins (between 232 and 1338 mg/kg), deoiled soy lecithin (342 mg/kg), phosphatydylcholine-enriched soy lecithins (not detectable and 163 mg/kg), sunflower lecithins (892 and 414 mg/kg), and egg lecithin (50 mg/kg). The sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein patterns of the standard soy and sunflower lecithins are very similar to those of soy flour. The protein profile of the egg lecithin shows several bands with a broad range of molecular masses. The molecular masses of the main proteins of soy lecithins and soy flour have been determined by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and ranged from 10.5 to 52.2 kDa. Most of the major proteins from soy and sunflower lecithins identified by MALDI-MS and electrospray tandem MS belong to the 11S globulin fraction, which is one of the main fractions of soy and sunflower seeds. In addition, the seed maturation protein P34 from the 7S globulin fraction of soy proteins has also been identified in soy lecithins. This protein has been reported as the most allergenic protein in soybean.     

Landscape composition influences roe deer habitat selection at both home range and landscape scales

Understanding how patterns of habitat selection vary in relation to landscape structure is essential to predict ecological responses of species to global change and inform management. We investigated behavioural plasticity in habitat selection of roe deer (Capreolus capreolus) in relation to variable habitat availability across a heterogeneous agricultural landscape at the home range and landscape scales. As expected, woodland was heavily selected, but we found no functional response for this habitat, i.e. no shift in habitat selection with changing habitat availability, possibly due to the presence of hedgerows which were increasingly selected as woodlands were less abundant. Hedgerows may thus function as a substitutable habitat for woodlands by providing roe deer with similar resources. We observed a functional response in the use of hedgerows, implying some degree of landscape complementation between hedgerows and open habitats, which may in part compensate for lower woodland availability. We also expected selection for woodland to be highest at the wider spatial scale, especially when this habitat was limiting. However, our results did not support this hypothesis, but rather indicated a marked influence of habitat composition, as both the availability and distribution of resources conditioned habitat selection. There was no marked between-sex difference in the pattern of habitat selection at either scale or between seasons at the landscape scale, however, within the home range, selection did differ between seasons. We conclude that landscape structure has a marked impact on roe deer habitat selection in agricultural landscapes through processes such as landscape complementation and supplementation.     

Environmental,historical, and contextual determinants of vegetation cover: a landscape perspective

We formulated and tested models of relationships among determinants of vegetation cover in two agroforested landscapes of eastern North America (Haut Saint-Laurent, Quebec, Canada) that differed by the spatial arrangement of their geomorphic features and intensity of agricultural activities. Our landscape model compared the woody plots of each landscape in terms of the relative influence of environmental attributes, land use history (1958 – 1997), and spatial context (i.e., proximity of similar or contrasting land cover). Our vegetation model evaluated the relative contribution of the same sets of variables to the distributions of herbs, trees, and shrubs. Relationships were assessed using partial Mantel tests and path analyses. Significant environmental and contextual differences were found between the vegetation plots of the two landscapes, but disturbance history was similar. Our vegetation model confirms the dominant effect of historical factors on vegetation patterns. Whereas land-use history overrides environmental and contextual control for trees, herbaceous and shrub species are more sensitive to environmental conditions. Context is determinant only for understory species in older, less-disturbed plots. Results are discussed in relevance to vegetation dynamics in a landscape perspective that integrates interactions between environmental and human influences.     

Utilisation d'anticorps spécifiques d'une fraction obtenue par chromatographie d'exclusion pour une amélioration de la caractérisation sérologique deRhizoctonia solani Kühn AG3

La purification d'un extrait mycélien soluble d'une souche deR. solani AG3 par chromatographie d'exclusion permet de recueillir une fraction antigénique. Une lapine est immunisée par voie intradermique. L'immunsérum titre 0,125 par la technique de double-diffusion en agarose. Différentes techniques sérologiques (immunodiffusion, électrotransfert suivi de révélation des fractions antigéniques, coloration immunoenzymatique du mycélium) sont appliquées à des souches deR. solani AG1, AG2, AG3, AG4, AG5, AG6, AG7, AGBI et deCeratobasidium CAG1, CAG2, CAG3, CAG4 et CAG5. Le sérum se révèle spécifique des souches AG3. A soluble mycelial extract ofR. solani AG3 was purified by size exclusion chromatography. Five peaks were present in the elution profile: the first was well defined and of large volume. It was retained to immunise a female rabbit by intradermic injection. The titre of the antiserum determined by double immuno-diffusion in agarose was 0.125. The antiserum was tested on isolates ofR. solani AG1, AG2, AG3, AG4, AG5, AG6, AG7, AGB1 and ofCeratobasidium CAG1, CAG2, CAG3, CAG4 and CAG5 (Table 1) using three different methods:

Genetically encoded sensors for ions and metabolites

Many of the genes encoding important trans porters and metabolic enzymes have been identified over the last ten years. Moreover it has been possible to study the biochemical properties of the corresponding proteins in great detail. It is expected that by 2,010 biochemical functions will have been assigned to many of the products of the approximately 30,000 Arabidopsis genes. We will get closer to understanding the biological function of the gene products by systematic analysis of mutants using knock-out and TILLING approaches. Metabolomics initiatives complement these approaches by providing insight into the changes in cellular ion and metabolite profiles in the mutants, thus giving information essential for the construction of cellular and whole plant models. However, one important dataset especially relevant to multicellular organisms is lacking: the knowledge of the spatial and temporal profiles of ions and metabolite levels at cellular and subcell ular levels. To address this issue, we have developed protein-based nanosensors for several metabolites, providing a set of tools for the determination of cytosolic and subcellular ion (e.g. iron and zinc) and metabolite levels in real time using fluorescence-based microscopy. The prototypes of these sensors were shown to function in vitro and also in vivo, i.e. in yeast and in mammalian cell cultures. One future goal is to expand the set of sensors to a wider spectrum of targets by using the natural spectrum of periplasmic binding proteins from bacteria and by computational design of proteins with altered binding pockets. Application of nanosensor technology to plant cells and tissues will help to elucidate the special and temporal distribution of ions and metabolites.     

Spatial attributes of fire regime in eastern Canada: influences of regional landscape physiography and climate

The characterization of the fire regime in the boreal forest rarely considers spatial attributes other than fire size. This study investigates the spatial attributes of fires using the physiography of the landscape as a spatial constraint at a regional scale. Using the Canadian National Fire Database, the size, shape, orientation and eccentricity were assessed for 1,136 fires between 1970 and 2010 in Quebec’s boreal forest and were summarized by ecodistrict. These spatial metrics were used to cluster 33 ecodistricts into homogeneous fire zones and then to determine which environmental variables (climate, topography, hydrography, and surficial deposits) influence the spatial attributes of fires. Analyses showed that 28 out of 33 ecodistricts belonging to a given fire zone were spatially contiguous, suggesting that factors driving the spatial attributes of fire are acting at a regional scale. Indeed, the orientation and size of fires vary significantly among the zones and are driven by the spatial orientation of the landscape and the seasonal regional climate. In some zones, prevailing winds during periods conducive to fire events parallel to the orientation of the landscape may favour the occurrence of very large fires (>100,000 ha). Conversely, an orientation of the landscape opposite to the prevailing winds may act as a natural firebreak and limit the fire size and orientation. This study highlights the need to consider the synergistic relationship between the landscape spatial patterns and the climate regime over the spatial attributes of fire at supra-regional scale. Further scale-dependant studies are needed to improve our understanding of the spatial factors controlling the spatial attributes of fire.     

Family Variation in the Morphology and Physiology of White Spruce (Picea glauca) Seedlings in Response to Elevated CO2 and Temperature

Tree improvement programs aim to develop families that are well-adapted to future growing conditions. To gain insight into the stability of the family genetic response to climate change, white spruce (Picea glauca) seedlings from 60 full-sib families were subjected to a combination of two temperature regimes and two levels of CO₂ over two growing seasons. There was positive effect of warmer temperatures and higher CO₂ on some growth variables but no significant family × treatment interactions. Instantaneous water use efficiency was the only physiological trait that was affected positively by the CO₂ treatment, showing a 51% increase that was consistent across families.     

Unusual Glycosaminoglycans from a Deep Sea Hydrothermal Bacterium Improve Fibrillar Collagen Structuring and Fibroblast Activities in Engineered Connective Tissues

Biopolymers produced by marine organisms can offer useful tools for regenerative medicine. Particularly, HE800 exopolysaccharide (HE800 EPS) secreted by a deep-sea hydrothermal bacterium displays an interesting glycosaminoglycan-like feature resembling hyaluronan. Previous studies demonstrated its effectiveness to enhance in vivo bone regeneration and to support osteoblastic cell metabolism in culture. Thus, in order to assess the usefulness of this high-molecular weight polymer in tissue engineering and tissue repair, in vitro reconstructed connective tissues containing HE800 EPS were performed. We showed that this polysaccharide promotes both collagen structuring and extracellular matrix settle by dermal fibroblasts. Furthermore, from the native HE800 EPS, a low-molecular weight sulfated derivative (HE800 DROS) displaying chemical analogy with heparan-sulfate, was designed. Thus, it was demonstrated that HE800 DROS mimics some properties of heparan-sulfate, such as promotion of fibroblast proliferation and inhibition of matrix metalloproteinase (MMP) secretion. Therefore, we suggest that the HE800EPS family can be considered as an innovative biotechnological source of glycosaminoglycan-like compounds useful to design biomaterials and drugs for tissue engineering and repair.     

Evaluation of sample pre-treatment and potential contamination of soluble carbon in soil extracts during lyophilisation

Lyophilisation of K₂SO₄ soil extracts has been proposed as a sample preparation technique before elemental analysis of carbon or nitrogen. However, previous measurements, based on wet oxidation or catalytic combustion, indicated that C measurements in lyophilised samples not always proved to be accurate. To determine whether the C analysis was affected by the lyophilisation process, an exploratory study was conducted to investigate the potential effects of the sample pre-treatment and of the lyophilisation process itself. This paper puts forward that the use of soil extracts, previously stored at −20 °C, may affect the recovery of salt in the samples and that contamination of the soluble carbon with exogenous C during lyophilisation is feasible. Therefore we recommend to use freshly prepared soil extracts for lyophilisation and always to include an internal standard among the unknown samples to account for a possible contamination.