Interaction of α-Lipoic Acid with the Human Na+/Multivitamin Transporter (hSMVT)

The human Na⁺/multivitamin transporter (hSMVT) has been suggested to transport α-lipoic acid (LA), a potent antioxidant and anti-inflammatory agent used in therapeutic applications, e.g. in the treatment of diabetic neuropathy and Alzheimer disease. However, the molecular basis of the cellular delivery of LA and in particular the stereospecificity of the transport process are not well understood. Here, we expressed recombinant hSMVT in Pichia pastoris and used affinity chromatography to purify the detergent-solubilized protein followed by reconstitution of hSMVT in lipid bilayers. Using a combined approach encompassing radiolabeled LA transport and equilibrium binding studies in conjunction with the stabilized R-(+)- and S-(−)-enantiomers and the R,S-(+/−) racemic mixture of LA or lipoamide, we identified the biologically active form of LA, R-LA, to be the physiological substrate of hSMVT. Interaction of R-LA with hSMVT is strictly dependent on Na⁺. Under equilibrium conditions, hSMVT can simultaneously bind ∼2 molecules of R-LA in a biphasic binding isotherm with dissociation constants (K_d) of 0.9 and 7.4 μm. Transport of R-LA in the oocyte and reconstituted system is exclusively dependent on Na⁺ and exhibits an affinity of ∼3 μm. Measuring transport with known amounts of protein in proteoliposomes containing hSMVT in outside-out orientation yielded a catalytic turnover number (k_cat) of about 1 s⁻¹, a value that is well in agreement with other Na⁺-coupled transporters. Our data suggest that hSMVT-mediated transport is highly specific for R-LA at our tested concentration range, a finding with wide ramifications for the use of LA in therapeutic applications.     

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.     

Structural–functional approach to identify post-disturbance recovery indicators in forests from northwestern Patagonia: A tool to prevent state transitions

The disruption of the natural post-disturbance recovery process, either by changes in disturbance regime or by another disturbance, can trigger transitions to alternative degraded states. In a scenario of high disturbance pressure on ecological systems, it is essential to detect recovery indicators to define the period when the system needs more protection as well as the period when the system supports certain use pressure without affecting its resilience. Recovery indicators can be identified by non-linear changes in structural and functional variables. Fire largely modulates the dynamic and stability of plant communities worldwide, and is this the case in northwestern (NW) Patagonia. The ultimate goal of this study is to propose a structural–functional approach based on a reference system (i.e. chronosequence) as a tool to detect post-disturbance recovery indicators in forests from NW Patagonia. In NW Patagonia (40–42°S), we sampled 25 Austrocedrus chilensis and Nothofagus spp. communities differing in post-fire age (0.3–180 years). In each community we recorded structural (woody species cover and height, solar radiation, air temperature, relative humidity) and functional (annual recruitment of woody and tree species) attributes. We modeled these attributes in function of post-fire age and analized the relationship between a functional attribute and a Structural Recovery Index (SRI). Communities varying in time-since-last-fire were structurally and functionally different. Moreover, response variables showed non-linear changes along the chronosequence, allowing the selection of recovery indicators. We suggest to use vegetation variables instead of environmental variables as structural recovery indicators. Horizontal and Vertical Vegetation Heterogeneity indices provided the information necessary to describe vegetation spatial reorganization after fire. Tree species annual recruitment was a good indicator of the functional recovery of forest communities. The relationship between a functional attribute and SRI allowed us to detect phases with high- and low-risk of degradation during post-fire succession. High-risk phases (<36 years old) had the highest horizontal vegetation heterogeneity and scarce tree seedling density (<7000 seedlings ha⁻¹ year⁻¹). Whereas, low-risk phases (>36 years old) had the highest vertical vegetation heterogeneity and tree species seedling density (>10,000 seedlings ha⁻¹ year⁻¹). Due to the low structural–functional levels, communities at high-risk phases would be more vulnerable to antropic pressure (e.g. livestock raising, logging) than communities at low-risk phases. The proposed approach contributes to the sustainable management of forest communities because it allows to estimate the minimum structural–functional levels from which forest communities could be harvested.     

A novel binding assay identifies high affinity ligands to the rosiglitazone binding site of mitoNEET

A novel outer mitochondrial membrane protein containing [2Fe-2S] clusters, mitoNEET was first identified through its binding to the anti-diabetic drug pioglitazone. Pioglitazone belongs to a family of drugs that are peroxisome proliferator-activated receptor (PPAR) gamma agonists, collectively known as glitazones. With the lack of pharmacological tools available to fully elucidate mitoNEET's function, we developed a binding assay to probe the glitazone binding site with the aim of developing selective and high affinity compounds. We used multiple thiazolidine-2,4-dione (TZD), 2-thioxothiazolidin-4-one (TTD), and 2-iminothiazolidin-4-one (ITD) compounds to establish several trends to enhance ligand development for the purpose of elucidating mitoNEET function.     

Spatial match between Planktothrix rubescens and whitefish in a mesotrophic peri-alpine lake: Evidence of toxins accumulation

Lake Bourget (France) provides drinking water and is a place for professional and recreational fishing. Since the mid 1990s, the lake has been exhibiting blooms of the filamentous cyanobacterium Planktothrix rubescens. This species is able to produce microcystin-LR and RR, toxins that contaminate different fish tissues and, if concentrated in the liver, can induce fish mortality. However, data on fish exposure to these toxins in a natural environment are scare and comparisons of spatial distribution between P. rubescens and exploited fish are needed to determine whether these fish avoid or converge in zones affected by the cyanobacteria. From June to November 2009, diurnal data on P. rubescens and whitefish (Coregonus lavaretus) spatial distributions have been monitored by hydroacoustic and BBE probe sampling. For all water samples, intracellular microcystin concentration of P. rubescens was quantified by HPLC/PDA. Furthermore, an arbitrary sample of eight whitefish captured at the moment of highest P. rubescens concentration in the lake were analyzed to investigate both the presence of this cyanobacterium in the gastrointestinal tract by optical microscopy and the possible bioaccumulation of the microcystin in their tissues by liquid chromatography–tandem mass spectrometry. Results show that P. rubescens abundance was at a maximum between the end of July and the beginning of September. During this period, P. rubescens abundance was vertically stratified with a maximum around 14–22 m, depending on the sampling station. The horizontal distributions of P. rubescens were heterogeneous at the scale of the lake. Results indicate that the presence of P. rubescens, for the observed cyanobacterial abundance, does not exert a significant pressure on the distribution pattern of the whitefish. Whitefish were present in the same areas as P. rubescens maxima during daytime, and they do not avoid or seek out the zone of high P. rubescens abundance. Filaments of P. rubescens have been observed in intestinal tracts of whitefish and the presence of microcystin-LR has been detected in their intestine and liver. Consequently, because of a direct contact between these organisms, toxins can be incorporated into whitefish by ingestion of P. rubescens filaments, leading to potential adverse effects on the health of this species.     

Factors influencing the seed source and sink functions of a floodplain nature reserve in the Netherlands

Question: How do species traits and abiotic factors influence the extent of hydrochorous dispersal into and out of a small floodplain area along a free‐flowing river in The Netherlands? Location: The Kappersbult nature reserve (53°07′28″N, 6°37′14″E), which is a floodplain along the Dutch River Drentsche Aa. Methods: Seeds transported by the river were collected in fine mesh nets for 24 consecutive hours once or twice a week for 1 year, upstream and downstream of the studied floodplain. Data on the captured seeds were related to species traits and abiotic factors and species composition in the floodplain. Results: The floodplain functioned both as a seed source and sink. High levels of river water seemed to promote seed transport to or from the floodplain. Seeds of riverbank species occurred significantly more often in the river water than expected. Net source species had significantly higher seed production, taller stature and higher seed buoyancy, but lower site elevation than net sink species. Seed weight was significantly higher for sink species than for other species. Conclusion: Our study found that inundation, and therefore more natural river water management, is a prerequisite for seed transport to and from a floodplain. The restoration of target floodplain vegetation may be successful for common species that produce many seeds and grow in proximity to the river. Consequently, it is expected that the probability of restoring vegetation types that occur further from the river, such as wet grasslands, by hydrochorous dispersal is low.     

Sex differences in search and gathering skills

The hunter–gatherer theory of sex differences states that female cognition has evolutionarily adapted to gathering and male cognition to hunting. Existing studies corroborate that men excel in hunting-related skills, but there is only indirect support for women excelling in gathering tasks. This study tested if women would outperform men in laboratory-based computer tests of search and gathering skills. In Experiment 1, men found target objects faster and made fewer mistakes than women in a classic visual search study. In Experiment 2, participants gathered items (fruits or letters presented on screen), and again, men performed significantly better. In Experiment 3, participants' incidental learning of object locations in a search experiment was studied, but no statistically significant sex differences were observed. These findings found the opposite of what was expected based on the hypothesis that female cognition has adapted to gathering. Alternative interpretations of the role of object location memory, female gathering roles and the division of labor between the sexes are discussed.     

Scale specific determinants of tree diversity in an old growth temperate forest in China

The major processes generating pattern in plant community composition depend upon the spatial scale and resolution of observation, therefore understanding the role played by spatial scale on species patterns is of major concern. In this study, we investigate how well environmental (topography and soil variables) and spatial variables explain variation in species composition in a 25-ha temperate forest in northeastern China. We used new variation partitioning approaches to discover the spatial scale (using multi-scale spatial PCNM variables) at which environmental heterogeneity and other spatially structured processes influence tree community composition. We also test the effect of changing grain of the study (i.e. quadrat size) on the variation partitioning results. Our results indicate that (1) species composition in the Changbai mixed broadleaf-conifer forest was controlled mainly by spatially structured soil variation at broad scales, while at finer scales most of the explained variation was of a spatial nature, pointing to the importance of biotic processes. (2) These results held at all sampling grains. However, reducing quadrat size progressively reduced both spatially and environmentally explained variance. This probably partly reflects increasing stochasticity in species abundances, and the smaller proportion of quadrats occupied by each species, when quadrat size is reduced. The results suggest that environmental heterogeneity (i.e. niche processes) and other biotic processes such as dispersal work together, but at different spatial scales, to build up diversity patterns.     

Evaluating environmental and geometrical constraints on endemic vertebrates of the semiarid Caatinga (Brazil)

Most hypotheses explaining species diversity are related to the current and/or past environment, but explanations that rely on geometrical constraints, such as the mid-domain effect, have been recently proposed. Under this type of model, more species are expected in the centre of a domain due to the extent of occurrence of the species being constrained by the domain's geometry. Here we evaluated the proportion of the explanatory power of space, the current environment, historical vicariance events and geometric constraints on the richness of the Caatinga's terrestrial endemic vertebrates using a partial regression approach. The current environment had the highest proportion of explanatory power for the variance of the Caatinga's endemic species, in its shared effect, explaining approximately 47%. On the other hand the geometric constraints model (GCM) had the highest proportion in its single explanatory power with 15% of explanatory power. The amount of explanation by GCM changes when analyzing richness of endemics calculated from species with different range sizes. Despite the fact that GCM is the main driver of the Caatinga's endemic species, in its single effect, this does not necessarily indicate that geometric constraints and random dispersal within species ranges drive richness. The GCM model underestimates richness in the centre of the Caatinga, suggesting that other ecological and evolutionary processes may help explain the overall pattern of decreasing richness towards the borders of the biome.