Crystal structures of Leptospira interrogans FAD-containing ferredoxin-NADP+ reductase and its complex with NADP+

Abstract Background Ferredoxin-NADP(H) reductases (FNRs) are flavoenzymes that catalyze the electron transfer between NADP(H) and the proteins ferredoxin or flavodoxin. A number of structural features distinguish plant and bacterial FNRs, one of which is the mode of the cofactor FAD binding. Leptospira interrogans is a spirochaete parasitic bacterium capable of infecting humans and mammals in general. Leptospira interrogans FNR (LepFNR) displays low sequence identity with plant (34% with Zea mays) and bacterial (31% with Escherichia coli) FNRs. However, LepFNR contains all consensus sequences that define the plastidic class FNRs. Results The crystal structures of the FAD-containing LepFNR and the complex of the enzyme with NADP+, were solved and compared to known FNRs. The comparison reveals significant structural similarities of the enzyme with the plastidic type FNRs and differences with the bacterial enzymes. Our small angle X-ray scattering experiments show that LepFNR is a monomeric enzyme. Moreover, our biochemical data demonstrate that the LepFNR has an enzymatic activity similar to those reported for the plastidic enzymes and that is significantly different from bacterial flavoenzymes, which display lower turnover rates. Conclusion LepFNR is the first plastidic type FNR found in bacteria and, despite of its low sequence similarity with plastidic FNRs still displays high catalytic turnover rates. The typical structural and biochemical characteristics of plant FNRs unveiled for LepFNR support a notion of a putative lateral gene transfer which presumably offers Leptospira interrogans evolutionary advantages. The wealth of structural information about LepFNR provides a molecular basis for advanced drugs developments against leptospirosis.

A simple criterion to predict the glass forming ability of metallic alloys

A new and simple criterion with which to quantitatively predict the glass forming ability (GFA) of metallic alloys is proposed. It was found that the critical cooling rate for glass formation (R-C) correlates well with a proper combination of two factors, the minimum topological instability (lambda(min)) and the Delta h parameter, which depends on the average work function difference (Delta phi) and the average electron density difference (Delta n(ws)(1/3)) among the constituent elements of the alloy. A correlation coefficient (R-2) of 0.76 was found between R-c and the new criterion for 68 alloys in 30 different metallic systems. The new criterion and the Uhlmann's approach were used to estimate the critical amorphous thickness (Z(C)) of alloys in the Cu-Zr system. The new criterion underestimated R-C in the Cu-Zr system, producing predicted Z(C) values larger than those observed experimentally. However, when considering a scale factor, a remarkable similarity was observed between the predicted and the experimental behavior of the GFA in the binary Cu-Zr. When using the same scale factor and performing the calculation for the ternary Zr-Cu-Al, good agreement was found between the predicted and the actual best GFA region, as well as between the expected and the observed critical amorphous thickness. (C) 2012 American Institute of Physics. [doi:10.1063/1.3676196]

New insights on reaction pathway selectivity promoted by crown ether phase-transfer catalysis: Model ab initio calculations of nucleophilic fluorination

Crown ethers have the ability of solubilizing inorganic salts in apolar solvents and to promote chemical reactions by phase-transfer catalysis. However, details on how crown ethers catalyze ionic S(N)2 reactions and control selectivity are not well understood. In this work, we have used high level theoretical calculations to shed light on the details of phase-transfer catalysis mechanism of KF reaction with alkyl halides promoted by 18-crown-6. A complete analysis of the of the model reaction between KF(18-crown-6) and ethyl bromide reveals that the calculations can accurately predict the product ratio and the overall kinetics. Our results point out the importance of the K* ion and of the crown ether ring in determining product selectivity. While the K* ion favors the S(N)2 over the E2 anti pathway, the crown ether ring favors the S(N)2 over E2 syn route. The combination effects lead to a predicted 94% for the S(N)2 pathway in excellent agreement with the experimental value of 92%. A detailed analysis of the overall mechanism of the reaction under phase-transfer conditions also reveals that the KBr product generated in the nucleophilic fluorination acts as an inhibitor of the 18-crown-6 catalyst and it is responsible for the observed slow reaction rate. (C) 2012 Elsevier B.V. All rights reserved.

Back-scattered electron imaging for leakage analysis of four retrofilling materials

This study aimed to evaluate, ex vivo, the nanoleakage in dentinal tubules, the linear infiltration of silver nitrate in the dentin wall/root-end filling material interface, and the presence of gaps in this interface in root-end cavities filled with 4 filling materials. Forty-eight disto-buccal root canals of maxillary molars were instrumented and filled. Retrograde cavities were prepared with ultrasonic points (apical 2 mm). The samples were divided into 2 control groups (n = 4) and 4 experimental groups (n = 10): Group I white mineral trioxide aggregate (MTA); Group II Super EBA; Group III Portland cement; and Group IV Sealer 26. After 1 week, the specimens were subjected to silver nitrate and prepared for SEM (backscattered electrons). In the apical-apical segment, an area with significantly higher leakage was observed for Super EBA, followed by Portland cement, MTA, and Sealer 26 (P = 0.0054). In the medium and cervical segments, all materials showed the same leakage behavior (P = 0.1815 and P = 0.1723, respectively). The linear infiltration at the dentin wall/root-end filling material interface was higher with Super EBA than the other groups. No differences in the percentage of gaps along the 3 mm of dentin wall/root-end filling material interface between the 4 materials were evident (P > 0.05). Nanoleakage occurred mainly in the apical segment of the samples, and Super EBA showed the highest values. The area and linear leakage were lower in the middle and coronal segments, regardless of the root-end filling material. No material perfectly sealed the root-end cavities, which allowed for the leakage occurrence. Microsc. Res. Tech. 75:796800, 2012. (C) 2011 Wiley Periodicals, Inc.

Silicon location through backscattered electron imaging and X-ray microanalysis in leaves of Cyperus ligularis L. and Rhynchospora aberrans C. B. Clarke (Cyperaceae)

(Silicon location through backscattered electron imaging and X-ray microanalysis in leaves of Cyperus ligularis L. and Rhynchospora aberrans C. B. Clarke (Cyperaceae)). The Cyperaceae show the ability to incorporate silicon by depositing colloidal silica, which is recorded by the occurrence of projections in the form of cones, in inner tangential walls of some epidermal cells or "silica cells". Leaves of C. ligularis and R. aberrans were analyzed through the technique of electron backscatter. Cyperus ligularis accumulates silica, in addition to "silica cells", in some stomata, trichomes and the cell walls that surround the cavities of the aerenchyma. The silica in the latter occurs in various forms; however, the cells located near the vascular bundles have conical projections, similar to those of the epidermis. Rhynchospora aberrans presents "silica cells" whose projections have tapered "satellites". In this species, silica also occurs in stomata and certain epidermal cells adjacent to them. It appears that the silicon deposition occurs in combination with the wall (with no apparent structural changes), and structures of secretion, or projections of the wall. These structural changes in the species, and location, are probably related to functional and environmental factors, especially the soil, in addition to relation with taxonomic groups.

Aspergillus fumigatus mitochondrial electron transport chain mediates oxidative stress homeostasis, hypoxia responses and fungal pathogenesis

We previously observed that hypoxia is an important component of host microenvironments during pulmonary fungal infections. However, mechanisms of fungal growth in these in vivo hypoxic conditions are poorly understood. Here, we report that mitochondrial respiration is active in hypoxia (1% oxygen) and critical for fungal pathogenesis. We generated Aspergillus fumigatus alternative oxidase (aoxA) and cytochrome C (cycA) null mutants and assessed their ability to tolerate hypoxia, macrophage killing and virulence. In contrast to ?aoxA, ?cycA was found to be significantly impaired in conidia germination, growth in normoxia and hypoxia, and displayed attenuated virulence. Intriguingly, loss of cycA results in increased levels of AoxA activity, which results in increased resistance to oxidative stress, macrophage killing and long-term persistence in murine lungs. Thus, our results demonstrate a previously unidentified role for fungal mitochondrial respiration in the pathogenesis of aspergillosis, and lay the foundation for future research into its role in hypoxia signalling and adaptation.

Effect of different root caries treatments on the sealing ability of conventional glass ionomer cement restorations

In this study we compared the microleakage of conventional glass ionomer cement (GIC) restorations following the use of different methods of root caries removal. In vitro root caries were induced in 75 human root dentin samples that were divided in five groups of 15 each according to the method used for caries removal: in group 1 spherical carbide burs at low speed were used, in group 2 a hand-held excavator was used, and in groups 3 to 5 an Er,Cr:YSGG laser was used at 2.25 W, 40.18 J/cm(2) (group 3), 2.50 W, 44.64 J/cm(2) (group 4) and 2.75 W, 49.11 J/cm(2) (group 5). The air/water cooling during irradiation was set to 55%/65% respectively. All cavities were filled with GIC. Five samples from each group were evaluated by scanning electron microscopy (SEM) and the other ten samples were thermocycled and submitted to a microleakage test. The data obtained were compared by ANOVA followed by Fisher's test (pa parts per thousand currency sign0.05). Group 4 showed the lowest microleakage index (56.65 6.30; p < 0.05). There were no significant differences among the other groups. On SEM images samples of groups 1 and 2 showed a more regular interface than the irradiated samples. Demineralized dentin below the restoration was observed, that was probably affected dentin. Group 4 showed the lowest microleakage values compared to the other experimental groups, so under the conditions of the present study the method that provided the lowest microleakage was the Er,Cr:YSGG laser with a power output of 2.5 W yielding an energy density of 44.64 J/cm(2).

Motor learning, retention and transfer after virtual-reality-based training in Parkinson's disease - effect of motor and cognitive demands of games: a longitudinal, controlled clinical study

Objectives To evaluate the learning, retention and transfer of performance improvements after Nintendo Wii Fit (TM) training in patients with Parkinson's disease and healthy elderly people. Design Longitudinal, controlled clinical study. Participants Sixteen patients with early-stage Parkinson's disease and 11 healthy elderly people. Interventions Warm-up exercises and Wii Fit training that involved training motor (shifts centre of gravity and step alternation) and cognitive skills. A follow-up evaluative Wii Fit session was held 60 days after the end of training. Participants performed a functional reach test before and after training as a measure of learning transfer. Main outcome measures Learning and retention were determined based on the scores of 10 Wii Fit games over eight sessions. Transfer of learning was assessed after training using the functional reach test. Results Patients with Parkinson's disease showed no deficit in learning or retention on seven of the 10 games, despite showing poorer performance on five games compared with the healthy elderly group. Patients with Parkinson's disease showed marked learning deficits on three other games, independent of poorer initial performance. This deficit appears to be associated with cognitive demands of the games which require decision-making, response inhibition, divided attention and working memory. Finally, patients with Parkinson's disease were able to transfer motor ability trained on the games to a similar untrained task. Conclusions The ability of patients with Parkinson's disease to learn, retain and transfer performance improvements after training on the Nintendo Wii Fit depends largely on the demands, particularly cognitive demands, of the games involved, reiterating the importance of game selection for rehabilitation purposes. (C) 2012 Chartered Society of Physiotherapy. Published by Elsevier Ltd. All rights reserved.

Selection of compositions with high glass forming ability in the Ni-Nb-B alloy system

A combination of an extension of the topological instability "λ criterion" and the "average electronegativity" has been recently reported in the literature to predict compositions with high glass-forming ability (GFA). In the present work, both criteria have been applied to select the Ni61.0Nb36.0B3 alloy with a high glass-forming ability. Ingots were prepared by arc-melting and were used to produce ribbons processed by the melt-spinning technique further characterized by differential scanning calorimetry (DSC), X-ray diffraction (XRD) and scanning electron microscopy (SEM). The Ni61.0Nb36.0B3 alloy revealed a complete amorphization and supercooled liquid region ΔTx = 68 K. In addition, wedge-shaped samples were prepared using copper mold casting in order to determine the critical thickness for amorphous formation. Scanning electron microscopy (SEM) revealed that fully amorphous samples could be obtained, reaching up to ~800 µm in thickness.