Characterization of Molecular Determinants of the Conformational Stability of Macrophage Migration Inhibitory Factor: Leucine 46 Hydrophobic Pocket

Macrophage Migration Inhibitory Factor (MIF) is a key mediator of inflammatory responses and innate immunity and has been implicated in the pathogenesis of several inflammatory and autoimmune diseases. The oligomerization of MIF, more specifically trimer formation, is essential for its keto-enol tautomerase activity and probably mediates several of its interactions and biological activities, including its binding to its receptor CD74 and activation of certain signaling pathways. Therefore, understanding the molecular factors governing the oligomerization of MIF and the role of quaternary structure in modulating its structural stability and multifunctional properties is crucial for understanding the function of MIF in health and disease. Herein, we describe highly conserved intersubunit interactions involving the hydrophobic packing of the side chain of Leu46 onto the β-strand β3 of one monomer within a hydrophobic pocket from the adjacent monomer constituted by residues Arg11, Val14, Phe18, Leu19, Val39, His40, Val41, Val42, and Pro43. To elucidate the structural significance of these intersubunit interactions and their relative contribution to MIF’s trimerization, structural stability and catalytic activity, we generated three point mutations where Leu46 was replaced by glycine (L46G), alanine (L46A) and phenylalanine (L46F), and their structural properties, stability, oligomerization state, and catalytic activity were characterized using a battery of biophysical methods and X-ray crystallography. Our findings provide new insights into the role of the Leu46 hydrophobic pocket in stabilizing the conformational state of MIF in solution. Disrupting the Leu46 hydrophobic interaction perturbs the secondary and tertiary structure of the protein but has no effect on its oligomerization state.     

Growth Analysis of Pulsed Laser Ablated Films

The spontaneous organization on a substrate of nanoparticles (NPs) nucleated in a laser-generated plasma results in self-assembling due to both direct specific interactions and to indirect ambient-mediated ones. A number of well-differentiated film morphologies are observed, from isolated NPs to percolated island structures. The distribution of the width of gaps between NPs, or islands formed upon NP coalescence in two representative percolated Au films, is lognormal, indicating that the growth process is random. A finer inspection of similar surface morphologies allows to correlate regularity in gap sizes and surface coverage to differences in film performance as substrates for surface-enhanced Raman spectroscopy.     

Cell-Type-Specific Gene Expression Profiling in Adult Mouse Brain Reveals Normal and Disease-State Signatures

1. Download high-res image (238KB)
2. Download full-size image

     

Influence of soil texture and crop management on the productivity of miscanthus (Miscanthus × giganteus Greef et Deu.) in the Mediterranean

Biomass productivity is the main favorable trait of candidate bioenergy crops. Miscanthus × giganteus is a promising species, due to its high‐yield potential and positive traits including low nutrient requirements and potential for C sequestration in soils. However, miscanthus productivity appears to be mostly related to water availability in the soil. This is important, particularly in Mediterranean regions where the risk of summer droughts is high. To date, there have been no studies on miscanthus responses under different soil conditions, while only a few have investigated the role of different crop managements, such as irrigation and nitrogen fertilization, in the Mediterranean. Therefore, the effects of contrasting soil textures (i.e. silty‐clay‐loam vs. sandy‐loam) and alternative agricultural intensification regimes (i.e. rainfed vs. irrigated and 0, 50, 100 kg ha^?1 nitrogen fertilization), on miscanthus productivity were evaluated at three different harvest times for two consecutive years. Our results confirmed the importance of water availability in determining satisfactory yields in Mediterranean environments, and how soil and site characteristics strongly affect biomass production. We found that the aboveground dry yields varied between 5 Mg ha^?1 up to 29 Mg ha^?1. Conversely, nitrogen fertilization played only a minor role on crop productivity, and high fertilization levels were relatively inefficient. Finally, a marked decrease, of up to ?40%, in the aboveground yield occurred when the harvest time was delayed from autumn to winter. Overall, our results highlighted the importance of determining crop responses on a site‐by‐site basis, and that decisions on the optimal harvest time should be driven by the biomass end use and other long‐term considerations, such as yield stability and the maintenance of soil fertility.     

Two new species of Edmockfordia García Aldrete (Psocodea, ‘Psocoptera’, Epipsocidae), from Valle del Cauca,Colombia, and description of the female E. chiquibulensis García Aldrete

Two new species of Edmockfordia García Aldrete, from Valle del Cauca, Colombia, and the female of Edmockfordia chiquibulensis García Aldrete, are described and illustrated. A key to the species of Edmockfordia is included; the genus was previously known only from Belize. The genus is re-diagnosed to include female characters. The distribution of the genus is considerably widened, from Belize to northeastern South America.     

Petrography and high-resolution geochemical records of Lower Jurassic manganese-rich deposits from Monte Mangart,Julian Alps

Deposits with unusually high Mn contents sampled at Monte Mangart in the Julian Alps include organic-rich marlstone and black shale with interbedded manganoan and siliceous limestone, which were deposited during the early Toarcian Oceanic Anoxic Event. Mn enrichment during that period has been related to global sea-level change coincident with increasing subsidence rate. The formation of Fe–Mn nodules, marking a hardground at the base of the Monte Mangart section, seems to be triggered by release of Mn from remote hydrothermal vents into a region of relatively elevated submarine topography where oxidizing conditions prevailed. However, very high Mn contents in carbonate phases above the hardground imply an additional diagenetic source of this element in the lower part of this section. The whole stratigraphic sequence (ca 30 m) displays a transition from Mn-rich (up to 8.8%) sediments, in the lower part, to Mn-poor (less than 1.8%) sediments in the middle and upper parts. The drastic decrease in Mn content's up-section is accompanied by a clear decrease in the mean size of pyrite framboids, indicating more intense anoxia/euxinia in the water column. In the presence of Mn²⁺, conditions of high alkalinity induced precipitation of Mn carbonates during early diagenetic processes. Negative δ¹³C_carb values coincident with high Mn contents indicate involvement of organic matter in the mineralization process. The striking similarity of Ce/Ce* and Mn profiles demonstrates that, consistent with redox-chemistry of Mn and Ce under anoxic conditions, Ce³⁺ and Mn²⁺ were mobilized and released into pore water where precipitation of Mn carbonates occurred.