25-Hydroxycholecalciferol-binding protein: Partial purification from rat duodenal mucosa cells

The 25-hydroxycholecalciferol-binding protein has been partially purified (purification factor: 37) from rat duodenal cytosol, using chromatographic procedures on gel and ionic exchange resin. This partially purified protein bound 25-hydroxycholecalciferol with high affinity (K_D = 5.7 × 10^?9M) and low binding capacity (23 × 10^?12 mole/mg of protein. Using a rabbit antiserum obtained against such partially purified protein, we demonstrated that 25-hydroxycholecalciferol cytosolic binder and 25-hydroxycholecalciferol plasmatic binding share common antigenic sites.     

The sub-lip domain--a distinct pathway for myotome precursors that demonstrate rostral-caudal migration

We have previously reported that the myotome is formed by a first wave of pioneer cells generated from all along the dorsomedial portion of the epithelial somite and a second wave of cells issued from all four edges of the dermomyotome. Cells from the extreme rostral and caudal edges directly generate myofibers that elongate towards the opposite pole of each segment and along the pre-existing myotomal scaffold. In contrast, cells from the dorsomedial and ventrolateral lips first reach the extreme edges and then contribute to myofiber formation. The mechanism by which these epithelial cells translocate remained unknown and was the goal of the present study. We have found that epithelial cells along the dorsomedial and ventrolateral lips of the dermomyotome first delaminate into the immediate underlayer of the corresponding lips, the sub-lip domain, then migrate longitudinally along this pathway until reaching the extreme edges from which they differentiate into myofibers. Cells of the sub-lip domain are negative for Pax3 and desmin but express MyoD, Myf5 and FREK, suggesting that they are specific myogenic progenitors.     

The third wave of myotome colonization by mitotically competent progenitors: regulating the balance between differentiation and proliferation during muscle development

The myotome is formed by a first wave of pioneer cells originating from the entire dorsomedial region of epithelial somites and a second wave that derives from all four lips of the dermomyotome but generates myofibers from only the rostral and caudal edges. Because the precedent progenitors exit the cell cycle upon myotome colonization, subsequent waves must account for consecutive growth. In this study, double labeling with CM-DiI and BrdU revealed the appearance of a third wave of progenitors that enter the myotome as mitotically active cells from both rostral and caudal dermomyotome edges. These cells express the fibroblast growth factor (FGF) receptor FREK and treatment with FGF4 promotes their proliferation and redistribution towards the center of the myotome. Yet, they are negative for MyoD, Myf5 and FGF4, which are, however, expressed in myofibers. The proliferating progenitors first appear around the 30-somite stage in cervical-level myotomes and their number continuously increases, making up 85% of total muscle nuclei by embryonic day (E)4. By this stage, generation of second-wave myofibers, which also enter from the extreme lips is still under way. Formation of the latter fibers peaks at 30 somites and progressively decreases with age until E4. Thus, cells in these dermomyotome lips generate simultaneously distinct types of muscle progenitors in changing proportions as a function of age. Consistent with a heterogeneity in the cellular composition of the extreme lips, MyoD is normally expressed in only a subset of these epithelial cells. Treatment with Sonic hedgehog drives most of them to become MyoD positive and then to become myofibers, with a concurrent reduction in the proportion of proliferating muscle precursors.     

Cooperativity-based modeling of heterotypic DNA nanostructure assembly

DNA origami is a robust method for the fabrication of nanoscale 2D and 3D objects with complex features and geometries. The process of DNA origami folding has been recently studied, however quantitative understanding of it is still elusive. Here, we describe a systematic quantification of the assembly process of DNA nanostructures, focusing on the heterotypic DNA junction—in which arms are unequal—as their basic building block. Using bulk fluorescence studies we tracked this process and identified multiple levels of cooperativity from the arms in a single junction to neighboring junctions in a large DNA origami object, demonstrating that cooperativity is a central underlying mechanism in the process of DNA nanostructure assembly. We show that the assembly of junctions in which the arms are consecutively ordered is more efficient than junctions with randomly-ordered components, with the latter showing assembly through several alternative trajectories as a potential mechanism explaining the lower efficiency. This highlights consecutiveness as a new design consideration that could be implemented in DNA nanotechnology CAD tools to produce more efficient and high-yield designs. Altogether, our experimental findings allowed us to devise a quantitative, cooperativity-based heuristic model for the assembly of DNA nanostructures, which is highly consistent with experimental observations.     

Utilisation du bois de feu par les chasseurs-cueilleurs de la « Cidade de Pedra » à partir de l’Holocène moyen (Brésil Central,Mato Grosso,Rondonópolis)

Located in the south-western Mato Grosso state (Central Brazil), the vast territory of the Cidade de Pedra was continuously occupied since the mid-Holocene by several groups of hunter-gatherers. Rock paintings, lithic industries, ceramics, ornaments, combustion remains (etc.) testify their passages in the many rock art sites discovered since 1983. In this paper are presented the results of the first anthracological analyzes conducted in four of these rock shelters. The studies were realized from macro-remains carbonized from hearths, embers and concentrations. From the taxonomic identification of charcoal, the objectives are to highlight the practices of hunter-gatherers related to the collection of wood and the environment in which they evolved for nearly 5000 years. The results indicate that they practiced an opportunistic collection mainly focused on dry wood available in the vegetation around the sites. They evolved in an environment similar to that currently characterizes the region, a typical flora of the Cerrado Biome and seasonal climate.     

Global ecosystems and fire: Multi‐model assessment of fire‐induced tree‐cover and carbon storage reduction

In this study, we use simulations from seven global vegetation models to provide the first multi‐model estimate of fire impacts on global tree cover and the carbon cycle under current climate and anthropogenic land use conditions, averaged for the years 2001–2012. Fire globally reduces the tree covered area and vegetation carbon storage by 10%. Regionally, the effects are much stronger, up to 20% for certain latitudinal bands, and 17% in savanna regions. Global fire effects on total carbon storage and carbon turnover times are lower with the effect on gross primary productivity (GPP) close to 0. We find the strongest impacts of fire in savanna regions. Climatic conditions in regions with the highest burned area differ from regions with highest absolute fire impact, which are characterized by higher precipitation. Our estimates of fire‐induced vegetation change are lower than previous studies. We attribute these differences to different definitions of vegetation change and effects of anthropogenic land use, which were not considered in previous studies and decreases the impact of fire on tree cover. Accounting for fires significantly improves the spatial patterns of simulated tree cover, which demonstrates the need to represent fire in dynamic vegetation models. Based upon comparisons between models and observations, process understanding and representation in models, we assess a higher confidence in the fire impact on tree cover and vegetation carbon compared to GPP, total carbon storage and turnover times. We have higher confidence in the spatial patterns compared to the global totals of the simulated fire impact. As we used an ensemble of state‐of‐the‐art fire models, including effects of land use and the ensemble median or mean compares better to observational datasets than any individual model, we consider the here presented results to be the current best estimate of global fire effects on ecosystems.     

MD-2 controls bacterial lipopolysaccharide hyporesponsiveness in human intestinal epithelial cells

Intestinal epithelial cells (IEC) have adapted to the presence of commensal bacteria through a state of tolerance that involves a limited response to lipopolysaccharide (LPS). Low or absent expression of two LPS receptor molecules, the myeloid differentiation (MD)-2 receptor, and toll-like receptor (TLR)4 was suggested to underlie LPS tolerance in IEC. In the present study we performed transfections of TLR4 and MD-2 alone or combined in different IEC lines derived from intestinal cancer (Caco-2, HT-29, and SW837). We found that LPS responsiveness increased more than 100-fold when IEC were transfected with MD-2 alone, but not TLR4. The release of interleukin (IL)-8, but also the expression of cyclooxygenase (Cox-)2 and the related secretion of prostaglandin (PG)E2 were coordinately stimulated by LPS in IEC transfected with MD-2 alone. Supernatants collected from MD-2-transfected IEC supported LPS activation of naïve HT-29, providing additional support to the concept that MD-2 alone endows IEC with LPS responsiveness. LPS responsiveness detected at concentrations as low as 110 pg/ml, and maximal values obtained by 10 ng/ml were clearly beyond those evoked by classical stimuli as IL-1β. In polarized cells, apical LPS stimulation was markedly more efficient than basolateral. Our data contradict previous opinion that both TLR4 and MD-2 limit IEC response to LPS, and emphasize the prominent role of MD-2 in intestinal immune responses to Gram-negative bacteria.     

Climate Change Effects on Vegetation Distribution and Carbon Budget in the United States

The Kyoto protocol has focused the attention of the public and policymarkers on the earth's carbon (C) budget. Previous estimates of the impacts of vegetation change have been limited to equilibrium “snapshots” that could not capture nonlinear or threshold effects along the trajectory of change. New models have been designed to complement equilibrium models and simulate vegetation succession through time while estimating variability in the C budget and responses to episodic events such as drought and fire. In addition, a plethora of future climate scenarios has been used to produce a bewildering variety of simulated ecological responses. Our objectives were to use an equilibrium model (Mapped Atmosphere–Plant–Soil system, or MAPSS) and a dynamic model (MC1) to (a) simulate changes in potential equilibrium vegetation distribution under historical conditions and across a wide gradient of future temperature changes to look for consistencies and trends among the many future scenarios, (b) simulate time-dependent changes in vegetation distribution and its associated C pools to illustrate the possible trajectories of vegetation change near the high and low ends of the temperature gradient, and (c) analyze the extent of the US area supporting a negative C balance. Both models agree that a moderate increase in temperature produces an increase in vegetation density and carbon sequestration across most of the US with small changes in vegetation types. Large increases in temperature cause losses of C with large shifts in vegetation types. In the western states, particularly southern California, precipitation and thus vegetation density increase and forests expand under all but the hottest scenarios. In the eastern US, particularly the Southeast, forests expand under the more moderate scenarios but decline under more severe climate scenarios, with catastrophic fires potentially causing rapid vegetation conversions from forest to savanna. Both models show that there is a potential for either positive or negative feedbacks to the atmosphere depending on the level of warming in the climate change scenarios. Received 12 May 2000; accepted 22 November 2000.     

Secondary settlement of cockles Cerastoderma edule as a function of current velocity and substratum: a flume study with benthic juveniles

Some newly-settled bivalve molluscs can experience a second dispersal stage in the water column and colonize areas distant from initial settlement zone ('secondary settlement'). To document mechanisms involved in such a process, experiments were conducted in a recirculating 13-m flume, using juvenile cockles Cerastoderma edule (shell length between 0.8 and 5.7 mm). After 4 h and under current surface velocities of 10, 20 and 24 cm s^?1, all juveniles left the plexiglass substratum (site of initial introduction) and 42.4, 58.6 and 76.2% of juveniles, respectively, were retrieved from a downstream sand area (which only represented 7.7% of the total flume surface). Naked-eye observations showed that smaller individuals were borne within the water column, whereas larger individuals tended to roll or slip on the substratum. Byssus threads produced by the juveniles were often seen, sometimes covered in sand grains. These observations were confirmed by finding larger juveniles in the upstream part of the sand area. When sand was replaced by mud, a reduced proportion of cockles was retrieved in the experimental substratum (18.4, 20.0 and 16.4%, respectively). Observations showed that juveniles rarely succeeded in anchoring themselves in mud. When initially introduced on a favourable substratum (medium sand), more than 87% of juveniles were retrieved from that sand array at all flow velocities. This study shows that secondary settlement occurs for juvenile cockles up to 5.7 mm in shell length and depends not only on flow velocity but on substratum type.     

The interactions of substituted pyrido[1,2-e]purines with oligonucleotides depend on the amphiphilic properties of their side chain.

Three pyrido[1,2-e]purines of increasing hydrophilicity have been synthesized to evaluate as anticancer agents. These drugs interact quite differently with a synthetic oligodeoxynucleotide d(CGATCG)2. [1] is very hydrophobic due to a phenyl residue in its side chain. It only shows limited interactions with the minihelix without any evidence of intercalation. [2] and [3], on the other hand, have one ([2]) or two ([3]) hydroxyl groups in their acyl chain and present rather amphiphilic properties. The result is a similar intercalation of these derivatives between C and G base pairs as revealed by intermolecular nOe, 1H and 31P chemical shift variations. Models for the intercalation of [2] are proposed using energy minimizations and molecular dynamics (MD) calculations subject to restraints from nOe connectivities. Simulations and experiments indicate weak stability and thus fast exchange of [2] in its intercalation site.