Bp-13 Pla2: Purification And Neuromuscular Activity Of A New Asp49 Toxin Isolated From Bothrops Pauloensis Snake Venom.

A new PLA2 (Bp-13) was purified from Bothrops pauloensis snake venom after a single chromatographic step of RP-HPLC on μ-Bondapak C-18. Amino acid analysis showed a high content of hydrophobic and basic amino acids and 14 half-cysteine residues. The N-terminal sequence showed a high degree of homology with basic Asp49 PLA2 myotoxins from other Bothrops venoms. Bp-13 showed allosteric enzymatic behavior and maximal activity at pH 8.1, 36°-45°C. Full Bp-13 PLA2 activity required Ca(2+); its PLA2 activity was inhibited by Mg(2+), Mn(2+), Sr(2+), and Cd(2+) in the presence and absence of 1 mM Ca(2+). In the mouse phrenic nerve-diaphragm (PND) preparation, the time for 50% paralysis was concentration-dependent (P < 0.05). Both the replacement of Ca(2+) by Sr(2+) and temperature lowering (24°C) inhibited the Bp-13 PLA2-induced twitch-tension blockade. Bp-13 PLA2 inhibited the contractile response to direct electrical stimulation in curarized mouse PND preparation corroborating its contracture effect. In biventer cervicis preparations, Bp-13 induced irreversible twitch-tension blockade and the KCl evoked contracture was partially, but significantly, inhibited (P > 0.05). The main effect of this new Asp49 PLA2 of Bothrops pauloensis venom is on muscle fiber sarcolemma, with avian preparation being less responsive than rodent preparation. The study enhances biochemical and pharmacological characterization of B. pauloensis venom.

Functional specialization of Eucalyptus fine roots: contrasting potential uptake rates for nitrogen, potassium and calcium tracers at varying soil depths

1. Little is known about the role of deep roots in the nutrition of forest trees and their ability to provide a safety-net service taking up nutrients leached from the topsoil. 2. To address this issue, we studied the potential uptake of N, K and Ca by Eucalyptus grandis trees (6 years of age - 25 m mean height), in Brazil, as a function of soil depth, texture and water content. We injected NO(3)(-)- (15)N, Rb(+) (analogue of K(+)) and Sr(2+) (analogue of Ca(2+)) tracers simultaneously in a solution through plastic tubes at 10, 50, 150 and 300 cm in depth in a sandy and a clayey Ferralsol soil. A complete randomized design was set up with three replicates of paired trees per injection depth and soil type. Recently expanded leaves were sampled at various times after tracer injection in the summer, and the experiment was repeated in the winter. Soil water contents were continuously monitored at the different depths in the two soils. 3. Determination of foliar Rb and Sr concentrations and (15)N atom % made it possible to estimate the relative uptake potential (RUP) of tracer injections from the four soil depths and the specific RUP (SRUP), defined as RUP, per unit of fine root length density in the corresponding soil layer. 4. The highest tracer uptake rates were found in the topsoil, but contrasting RUP distributions were observed for the three tracers. Whilst the RUP was higher for NO(3)(-)- (15)N than for Rb(+) and Sr(2+) in the upper 50 cm of soil, the highest SRUP values for Sr(2+) and Rb(+) were found at a depth of 300 cm in the sandy soil, as well as in the clayey soil when gravitational solutions reached that depth. 5. Our results suggest that the fine roots of E. grandis trees exhibit contrasting potential uptake rates with depth depending on the nutrient. This functional specialization of roots might contribute to the high growth rates of E. grandis trees, efficiently providing the large amounts of nutrients required throughout the development of these fast-growing plantations.

Understanding the impact of divalent cation substitution on hydroxyapatite: An in vitro multiparametric study on biocompatibility

Hydroxyapatite (HA), a stable and biocompatible material for bone tissue therapy, may present a variable stoichiometry and accept a large number of cationic substitutions. Such substitutions may modify the chemical activity of HA surface, with possible impact on biocompatibility. In this work, we assessed the effects of calcium substitution with diverse divalent cations (Pb(2+), Sr(2+), Co(2+), Zn(2+), Fe(2+), Cu(2+), or Mg(2+)) on the biological behavior of HA. Physicochemical analyses revealed that apatite characteristics related to crystallinity and calcium dissolution/uptake rates are very sensitive to the nature of cationic substitution. Cytocompatibility was evaluated by mitochondrial activity, membrane integrity, cell density, proapoptotic potential, and adhesion tests. With the exception of Zn-HA, all the substituted HAs induced some level of apoptosis. The highest apoptosis levels were observed for Mg-HA and Co-HA. Cu-HA was the only material to impair simultaneously mitochondrial activity, membrane integrity, and cell density. The highest relative cell densities after exposure to the modified HAs were observed for Mg-HA and Zn-HA, while Co-HA significantly improved cell adhesion onto HA surface. These results show that changes on surface dissolution caused by cationic substitution, as well as the increase of metal species released to biological media, were the main responsible factors related to alterations on HA biocompatibility. (C) 2011 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 98A: 351-358, 2011.

Produção de poligalacturonase, pelo termofílico Bacillus sp. e algumas de suas propriedades

A produção de poligalacturonase pelo termofílico Bacillus sp. SMIA-2, cultivado em meio líquido contendo pectina cítrica como única fonte de carbono, alcançou a sua máxima atividade enzimática em 30 horas, com níveis de 42 IU.mL-1. Entre as várias fontes orgânicas e inorgânicas de nitrogênio testadas, o sulfato de amônio foi a que proporcionou maior atividade da poligalacturonase. O aumento da concentração da pectina cítrica, no meio de cultura, acima de 0,5% não proporcionou um aumento da atividade da enzima. O microrganismo foi capaz de utilizar uma variedade de fontes de carbono, mas a atividade da poligalacturonase variou com cada fonte. Pectina de maçã foi a melhor fonte de carbono para a secreção da poligalacturonase (56 IU.mL-1), enquanto frutose e maltose não foram muito efetivas. Galactose, rafinose e glicose inibiram a síntese da enzima. Estudos sobre a caracterização da poligalacturonase revelaram que a temperatura ótima dessa enzima foi 70 ºC e que ela manteve 62 e 58% de sua atividade máxima quando incubada por 2 horas a 40 e 90 ºC, respectivamente. O pH ótimo para atividade da enzima foi 7,0. A enzima manteve 90 e 75% de sua atividade máxima quando incubada a pH 8,0 e 8,5, respectivamente, por 24 horas, à temperatura ambiente. A atividade enzimática foi estimulada pelos íons Mg2+ e Zn2+. Por outro lado, foi inibida pelos íons Cs+2, Hg+2, Li+2 e Sr+2.

La Controverse (Lyon)

Périodicité : Bimensuel

Connectivité hydrologique et signature géochimique à l'échelle événementielle dans un bassin versant forestier

Dans un bassin versant, la connectivité hydrologique constitue un état hydrologique qui lie le versant à la zone riveraine. Ses impacts sur la production du débit et le transfert des éléments dissous vers le cours d’eau sont présumés substantiels. L’étude vise à 1) détecter les hydrotopes et les connexions hydrologiques à l’aide d’un réseau de puits qui permet la mesure des fluctuations de la nappe phréatique (NP); 2) identifier la variabilité spatio-temporelle et la signature géochimique des sources potentielles en eau à l’aide des éléments majeurs et traces et 3) examiner la contribution spatio-temporelle respective des sources en eau du bassin lors d’un événement de précipitation. L’étude s’effectue dans un bassin versant forestier du Bouclier canadien (l’Hermine). Nous démontrons l’existence de quatre hydrotopes représentant un gradient de convergence de l’eau, soulignant la diversité de comportement de NP. Les connexions hydrologiques se caractérisent par des coefficients de Spearman élevés des relations entre la profondeur de la NP et le débit, dans leur partie en aval, et s’enclenchent par le fill and spill. Le comportement de NP est influencé par la distance aux limites du bassin, l’horizonation du sol et la topographie souterraine. En somme, trois sources en eau se connectent à partir du versant vers la zone riveraine durant l’événement pluvial de manière chronologique: 1) les horizons B et la NP de l’ensemble du bassin (Sr); 2) les horizons LFH des zones de convergence (Ba et Zn) et 3) une dépression de sol humide sur le versant nord (Co et Mn).

Coût bioenergetique des modes de déplacements irréguliers en sport

The objective of this thesis was to quantify the physiological responses such as O2 uptake (VO2), heart rate (HR) and blood lactate ([LA]) to some types of activities associated with intermittent sports in athletes. Our hypothesis is that the introduction of accelerations and decelerations with or without directional changes results in a significative increase of the oxygen consumption, heart rate and blood lactate. The purpose of the first study was to measure and compare the VO2 and the HR of 6 on-court tennis drills at both high and low displacement speeds. These drills were done with and without striking the ball, over full and half-width court, in attack or in defense mode, using backhand or forehand strokes. Results show that playing an attacking style requires 6.5% more energy than playing a defensive style (p < 0.01) and the backhand stroke required 7% more VO2 at low speed than forehand stroke (p < 0.05) while the additional cost of striking the ball lies between 3.5 and 3.0 mL kg-1 min-1. Finally, while striking the ball, the energy expanded during a shuttle displacement on half-width court is 14% higher than running on full-width court. Studies #2 and #3 focused on different modes of displacement observed in irregular sports. The objective of the second study was to measure and compare VO2, HR and [LA] responses to randomly performed multiple fractioned runs with directional changes (SR) and without directional changes (FR) to those of in-line running (IR) at speeds corresponding to 60, 70 and 80% of the subject’s maximal aerobic speed (MAS). All results show that IR’s VO2 was significantly lower than SR’s and FR’s (p<0.05). SR’s VO2 was greater than FR’s only at speeds corresponding to 80%MAS. On the other hand, HR was similar in SR and FR but significantly higher than IR’s (p<0.05). [LA] varied between 4.2 ± 0.8 and 6.6 ± 0.9 mmol L-1 without significant differences between the 3 displacement modes. Finally, the third study’s objective was to measure and compare VO2 , HR and [LA] responses during directional changes at different angles and at different submaximal running speeds corresponding to 60, 70 and 80% MAS. Subjects randomly performed 4 running protocols 1) a 20-m shuttle running course (180°) (SR), 2) an 8-shaped running course with 90-degree turns every 20 m (90R), 3) a Zigzag running course (ZZR) with multiple close directional changes (~ 5 m) at different angle values of 91.8°, 90° and 38.6°, 4) an In-line run (IR) for comparison purposes. Results show that IR’s was lower (p<0.001) than for 90R’s, SR’s and ZZR’s at all intensities. VO2 obtained at 60 and 70%MAS was 48.7 and 38.1% higher during ZZR when compared to IR while and depending on the intensity, during 90R and SR was between 15.5 and 19.6% higher than during IR. Also, ZZR’s VO2 was 26.1 and 19.5% higher than 90R’s, 26.1 and 15.5% higher than SR’s at 60 and 70%MAS. SR’s and 90R’s VO2 were similar. Changing direction at a 90° angle and at 180° angle seem similar when compared to continuous in-line running. [LA] levels were similar in all modalities. Overall, the studies presented in this thesis allow the quantification of the specific energetic demands of certain types of displacement modes in comparison with conventional forward running. Also, our results confirm that the energy cost varies and increase with the introduction of accelerations and decelerations with and without directional changes.

Dipodal Ferrocene Derivatives for Nanostructured Redox-Functionalised Surfaces

Self-assembled monolayers (SAMs) on solid surfaces are of great current interest in science and nanotechnology. This thesis describes the preparation of several symmetrically 1,1’-substituted ferrocene derivatives that contain anchoring groups suitable for chemisorption on gold and may give rise to SAMs with electrochemically switchable properties. The binding groups are isocyano (-NC), isothiocyanato (-NCS), phosphanyl (-PPh2), thioether (-SR) and thienyl. In the context of SAM fabrication, isothiocyanates and phosphanes are adsorbate systems which, surprisingly, have remained essentially unexplored. SAMs on gold have been fabricated with the adsorbates from solution and investigated primarily by X-ray photoelectron spectroscopy and near-edge X-ray absorption fine structure spectroscopy. The results of these analytical investigations are presented and discussed in matters of the film quality and possible binding modes. The quality of self-assembled monolayers fabricated from 1,1’-diisocyanoferrocene and 1,1’-diisothiocyanatoferrocene turned out to be superior to that of films based on the other adsorbate species investigated. Films of those absorbates as well as of dppf afforded well-defined SAMs of good quality. All other films of this study based on sulfur containing anchoring groups exhibit chemical inhomogeneity and low orientational order of the film constituents and therefore failed to give rise to well-defined SAMs. Surface coordination chemistry is naturally related to molecular coordination chemistry. Since all SAMs described in this thesis were prepared on gold (111) surfaces, the ferrocene-based ligands of this study have been investigated in their ability for complexation towards gold(I). The sulfur-based ferrocene ligands [fc(SR)2] failed to give stable gold(I) complexes. In contrast, 1,1’-diisocyanoferrocene (1) proved to be an excellent ligand for the complexation of gold(I). Several complexes were prepared and characterised utilising a series of gold(I) acetylides. These complexes show interesting structural motifs in the solid state, since intramolecular aurophilic interactions lead to a parallel orientation of the isocyano moieties, combined with an antiparallel alignment of neighbouring units. The reaction of 1 with the gold(I) acetylide [Au(C≡C–Fc)]n turned out to be very unusual, since the two chemically equivalent isocyano groups undergo a different reaction. One group shows an ordinary coordination and the other one undergoes an extraordinary 1,1-insertion into the Au-C bond. As a sideline of the research of this thesis several ferrocene derivatives have been tested for their suitability for potential surface reactions. Copper(I) mediated 1,3-dipolar cycloadditions of azidoferrocene derivatives with terminal alkynes appeared very promising in this context, but failed to a certain extent in terms of ‘click’ chemistry, since the formation of the triazoles depended on the strict exclusion of oxygen and moisture and yields were only moderate. Staudinger reactions between dppf and azidoferrocene derivatives were also tested. The nucleophilic additions of secondary amines to 1,1’-diisothiocyanatoferrocene led to the respective thiourea derivatives in quantitative yields.

Seleção para aumento de produtividade e qualidade de frutos em abobrinha 'Piramoita' comparando dois métodos de melhoramento

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Synthesis and sintering of PZT ceramics

Lead zirconate powder, with Zr/Ti ratio of 50/50 was prepared by polymeric precursor method and doped with 3, 5 and 7 mol% of Sr+2 Or Ba+2, as well as by 0.2 to 5 mol% of Nb+5. The powder was calcined at 750 degrees C by 4 hours and milled during 1.5 h in isopropilic alcohol. Powders were characterized by surface area measurements (BET method), by infrared spectroscopy and by X-ray diffraction to characterize the crystal structure. Isostatically pressed samples were sintered in a dilatometer furnace by using a constant heating rate of 10 degrees C/min from ambient to 1200 degrees C. Synthetic air and air with water vapor were used as atmospheres. Both Sr+2 and Ba+2 substitute Pb+2 and favor the formation of rhombohedral phase. Otherwise, Nb+5 substitute preferentially Zr+4 favoring tetragonal phase. The concentration of dopants and the atmosphere influence the densification and the microstructure of the PZT, which alters the dielectric and piezoelectric properties of the ceramics.

Influence of the modifier on the short and long range disorder of stannate perovskites

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)