Expression, purification and spectroscopic analysis of an HdrC: An iron-sulfur cluster-containing protein from Acidithiobacillus ferrooxidans

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Composição Corporal e Exigências Nutricionais em Cálcio e Fósforo para Ganho e Mantença de Cordeiros Santa Inês dos 15 kg aos 25 kg de Peso Vivo

The research was carried out to determine body composition and calcium and phosphorus requirements of Santa Ines lambs. Eighteen entire male lambs with average initial live weight of 15 kg were used. The animals were allotted to three groups: six animals were slaughtered at the beginning of the experiment, to access the amount of calcium and phosphorus present in the body, as reference animals for the comparative slaughter technique. Six animals were ad libitum fed and six were restrict fed (maintenance level plus 20%). The animals ad libitum and restrict fed started the experimental period by pairs and they were both slaughtered when the first reached 25 kg body weight. The body composition was estimated through the prediction equations obtained by regression of the logarithm of the amount of calcium and phosphorus in the empty body on the logarithm of the empty body weight. Net requirements for calcium and phosphorus for maintenance and the absorption coefficient were obtained through the correlation between the amount of each mineral consumed and retained in the animal body. The net requirements maintenance for live weight gain were obtained by means of derivation of the prediction of body composition equations. The net requirements maintainance of calcium and phosphorus for animals from 15 to 25 kg body weight were: 305 mg Ca/day and 325 mg P/day and net requirements for kg of the body weight gain for animals with 15 and 25 kg LW were 11.41 and 10.33 g Ca and 5.72 and 4.94 g P, respectively. The absorption coefficients were estimated to be .44 and 0.55 for Ca and P, respectively.

Lamé differential equations and electrostatics

The problem of existence and uniqueness of polynomial solutions of the Lamé differential equation A(x)y″ + 2B(x)y′ + C(x)y = 0, where A(x),B(x) and C(x) are polynomials of degree p + 1,p and p - 1, is under discussion. We concentrate on the case when A(x) has only real zeros aj and, in contrast to a classical result of Heine and Stieltjes which concerns the case of positive coefficients rj in the partial fraction decomposition B(x)/A(x) = ∑j p=0 rj/(x - aj), we allow the presence of both positive and negative coefficients rj. The corresponding electrostatic interpretation of the zeros of the solution y(x) as points of equilibrium in an electrostatic field generated by charges rj at aj is given. As an application we prove that the zeros of the Gegenbauer-Laurent polynomials are the points of unique equilibrium in a field generated by two positive and two negative charges. © 2000 American Mathematical Society.

Optical limiting behavior of tungstate fluorophosphate glasses

Nonlinear absorption measurements were performed on fluorophosphate glasses with high concentration of tungsten oxide. Large two-photon absorption coefficients, α2, were determined at 660 nm using nanosecond laser pulses. It was observed that α2 increases for increasing tungsten oxide concentrations and therefore the optical limiting performance of this new glass composition can be controlled.

Variance heterogeneity for milk yield in Brazilian and Colombian Holstein herds

Heterogeneity of variances for milk yield (MY) was determined for Brazilian and Colombian herds. The herds were grouped as high or low variability within each country, using as criterion the phenotypic standard deviation (PSD) of MY in the contemporary groups of cows, from the first to the sixth calving. Brazilian and Colombian herds with PSD greater than 1,168 kg or 1,012 kg, respectively, were classified as high variability while the herd groups with values lower than those were classified as low variability. The genetic parameters for MY within each herd group were estimated using bivariate analysis in an animal model and the restricted maximum likelihood method with a derivative free algorithm, using 72,280 first lactations of cows, daughters of 1,880 sires. Heterogeneous variances were found, and Brazilian herds with high PSD had the greatest additive and residual genetic variances and heritability coefficients for MY. MY genetic correlation coefficients between herds of high and low variability within each country were 0.96 and 0.93 while between countries they varied from 0.72 to 0.81, suggesting that there was a reclassification of animals in the two countries and also heterogeneity of variances. This phenomenon leads to the questioning of the strategy of imported semen usage and the need to do genetic evaluations to identify sires with greater genetic potential for (sub) tropical environmental conditions.

Time-like and space-like pion form factors within front-form dynamics

An approach for a unified description of the pion electromagnetic form factor in the space- and time-like regions, within a constituent quark model on the light front is briefly illustrated. Three main ingredients enter our approach: i) the on-shell quark-hadron vertex functions in the valence sector, ii) the dressed photon vertex where a photon decays in a quark-antiquark pair, and iii) the emission and absorption amplitudes of a pion by a quark. © 2005 American Institute of Physics.

Structure and properties of Ti4+-Ureasil organic-inorganic hybrids

Hybrid planar waveguides were prepared from Ti4+-acetylacetone (acac)-Ureasil sols deposited on glass substrates. Structural features have been investigated by spectroscopic measurements (Fourier Transform Infrared Spectroscopy (FTIR), X-ray Photoelectron Spectroscopy (XPS) and Raman scattering) and Small Angle X-ray Scattering (SAXS). Addition of Ti 4+-acac to the ureasil (Ti:Si molar ratio 1:1) leads to the formation of bonds between the Ti complex and the siloxane groups, whereas further addition of Ti4+ (Ti:Si molar ratio 5:1) leads to the additional formation of titanium-rich nanoclusters. The optical parameters of the waveguides such as refractive index, thickness, propagating modes and attenuation coefficient were measured at 632.8, 543.5 and 1550 nm by the prism coupling technique. The refractive index can be tuned by the Ti4+ relative content. The few microns thick planar waveguides support well confined propagating modes with low attenuation loss for all compositions. ©2006 Sociedade Brasileira de Química.

Analysis of the kinetics of phase and amplitude gratings recorded in azopolymer films

In this work we use a stabilized holographic technique to study both refractive index and absorption gratings recorded in thin films made of Disperse Red 1 (DR1) embedded in an organic polymer matrix (PMMA) deposited on glass substrate. Gratings are recorded by linearly polarized illumination with the interference pattern of two crossing beams. One of the beams is phase modulated and the interference signals between the transmitted and diffracted waves are detected by a tuned lock-in amplifier. The technique allows measuring separately changes of the refractive index and the absorption coefficient during the course of the photoreaction process. The time evolution of the diffraction efficiencies during recording has shown bi-exponential kinetics for both gratings. © 2008 American Institute of Physics.

Determination of critical velocity and anaerobic capacity of running rats

The aim of this study was to validate a non-invasive protocol to determine aerobic and anaerobic capacity of treadmill running rats. Thirteen male Wistar rats (90 days old) were submitted to 4 exercise tests, consisting of running at 25, 30, 35 and 40 m min-1, continuously until exhaustion. For the critical velocity (CV) and anaerobic running capacity (ARC) estimations, the hyperbolic curve (velocity versus time to exhaustion (tlim)) was linearized to V= CV+ARC/tlim, where the CV and ARC were linear and slope coefficients, respectively. In order to verify if the CV was the maximal aerobic intensity, the rats were submitted to the maximal lactate steady state test (MLSS) composed of three 25-minute tests of continuous running trials at 15, 20 and 25 m min-1, with blood collection every 5 minutes. The CV was obtained at 22.8±0.7 m min-1 and the ARC, at 26.80±2.77 m. The MLSS was observed at 20m min-1, with blood lactate 3.84 ± 0.31 mmol L-1. There was a progressive increase in lactate concentration at 25 m min-1. The CV and MLSS were different, but presented a high and significant correlation (r=0.81). These results indicate that the non-invasive protocol can be used for physical evaluation of aerobic running rats, but the ARC should still be further investigated.

Synthesis and evaluation of diethylethylamine-chitosan for gene delivery: Composition effects on the in vitro transfection efficiency

Chitosan has been indicated as a safe and promising polycation vector for gene delivery. However its low transfection efficiency has been a challenging obstacle for its application. To address this limitation, we synthesized chitosan derivatives which had increasing amounts of diethylethylamine groups (DEAE) attached to the chitosan main chain. The plasmid DNA VR1412 (pDNA), encoding the ß-galactosidase (ß-gal) reporter gene was used to prepare nanoparticles with the chitosan derivatives, and the transfection studies were performed with HeLa cells. By means of dynamic light scattering and zeta potential measurements, it was shown that diethylethylamine-chitosan derivatives (DEAEx-CH) were able to condense DNA into small particles having a surface charge depending on the polymer/DNA ratio (N/P ratio). Nanoparticles prepared with derivatives containing 15 and 25% of DEAE groups (DEAE15-CH and DEAE25-CH) exhibited transfection efficiencies ten times higher than that observed with deacetylated chitosan (CH). For derivatives with higher degrees of substitution (DS), transfection efficiency decreased. The most effective carriers showed low cytotoxicity and good transfection activities at low charge ratios (N/P). Vectors with low DS were easily degraded in the presence of lysozyme at physiological conditions in vitro and the nontoxicity displayed by these vectors opens up new opportunities in the design of DEAE-chitosan-based nanoparticles for gene delivery. © 2013 IOP Publishing Ltd.

Structural characteristics of P123-modified supercritical drying and hydrophobic ambient pressure drying aerogels

Supercritical drying (SCD) and hydrophobic ambient pressure drying (APD) aerogels were prepared from hydrolysis of tetraethoxysilane in solutions of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (P123) in the range of composition below the threshold for the ordered mesoporous silica precipitation. APD was carried out after silylation of wet gels with trimethylchlorosilane (TMCS) or hexamethyldisilazane (HMDZ). The samples were analyzed by small-angle X-ray scattering and nitrogen adsorption. Wet gels are formed by mass-fractal domains, with fractal dimension close to 2, and larger pores superposing the pores belonging to the fractal structure in case of high P123 concentrations. Aerogels exhibit smaller-sized mass-fractal domains with larger mass-fractal dimension accounting for some porosity elimination on drying. The pore volume of the aerogels increases significantly with the P123 amount and it is even larger in the APD aerogels than in the SCD aerogels. © 2013 Elsevier B.V. All rights reserved.

Structural and functional studies with mytoxin II from Bothrops moojeni reveal remarkable similarities and differences compared to other catalytically inactive phospholipases A2-like

Lys49-phospholipases A2 (Lys49-PLA2s) are proteins found in bothropic snake venoms (Viperidae family) and belong to a class of proteins which presents a phospholipase A2 scaffold but are catalytically inactive. These proteins (also known as PLA2s-like toxins) exert a pronounced local myotoxic effect and are not neutralized by antivenom, being their study relevant in terms of medical and scientific interest. Despite of the several studies reported in the literature for this class of proteins only a partial consensus has been achieved concerning their functional-structural relationships. In this work, we present a comprehensive structural and functional study with the MjTX-II, a dimeric Lys49-PLA2 from Bothrops moojeni venom which includes: (i) high-resolution crystal structure; (ii) dynamic light scattering and bioinformatics studies in order to confirm its biological assembly; (iii) myographic and electrophysiological studies and, (iv) comparative studies with other Lys49-PLA2s. These comparative analyses let us to get important insights into the role of Lys122 amino acid, previously indicated as responsible for Lys49-PLA2s catalytic inactivity and added important elements to establish the correct biological assembly for this class of proteins. Furthermore, we show two unique sequential features of MjTX-II (an amino acid insertion and a mutation) in comparison to all bothropic Lys49-PLA2s that lead to a distinct way of ligand binding at the toxin's hydrophobic channel and also, allowed the presence of an additional ligand molecule in this region. These facts suggest a possible particular mode of binding for long-chain ligands that interacts with MjTX-II hydrophobic channel, a feature that may directly affect the design of structure-based ligands for Lys49-PLA2s. © 2013 Elsevier Ltd.

Estimativas da radiação global incidente em superfícies inclinadas com base na razáo de insolação

Estimating equations of global radiation based on the sunshine duration were proposed for horizontal surface and with inclination of 12.85, 22.85 and 32.85° facing the North in Botucatu, SP, Brazil, in monthly, seasonal and annual groupings of data. Simple linear correlations were applied (for definition of the linear and angular coefficients of Angstrom-Prescott model), in a database measured in all three inclinations in different periods (22.85°: 04/1998 to 07/2001; 12.85°: 08/2011 to 02/2003; and 32.85°: 03/2003 to 12/2007) concomitant with horizontal measures and sunshine duration. The statistical performance of the model was analysed by the means absolute error (MBE), the square root of the mean square error (RMSE) and the index adjustment (d). The minimum global radiation transmissivity varied from 14.35% in August (12.85°) to 27.86% in December (32.85°) and the maximum transmissivity ranged between 62.10% and 78.90%, for June (32.85°) and December (12.85°). Increasing the angle of inclination surface increased the scattering and decreased the index of adjustment and performance. The worst results were found for application of the seasonal and annual models in the months of autumn and winter for 32.85° (RMSE below 42.93% and adjustment superior to 0.4693).

Estimativas das radiações direta e difusa em superfícies inclinadas com base na razáo de insolação

Statistical equations were obtained and evaluated with annual, seasonal and monthly data groupings for estimates of direct and diffuse components of solar radiation based on the sunshine duration (ratio of sunshine and photoperiod) incident on horizontal and inclined surfaces to 12.85, 22.85 and 32.85° with facing to North, in Botucatu, SP. The ratios between the two components and radiation at the top of the atmosphere were used, in a database whose inclinations were measured in three different periods (22.85°: 04/1998 to 07/2001; 12.85°: 08/2011 to 02/2003; and 32.85°: 03/2003 to 12/2007) and concomitant with horizontal measures and sunshine duration. The correlations showed a linear and second degree polynomial behavior for the direct and diffuse radiation, with higher coefficients of determination in periods of low variation in the coverage of the sky (cloudiness). The highest values of the direct and diffuse radiation were found in winter and summer, respectively for all surfaces evaluated. The increase in the inclination angle decreased the performance of equations in all groups of data with increase in scattering and decrease in index of the adjustment, however, the monthly equations allowed better performance for the two components.

Estudo das propriedades físicas de blendas de PVDF/Látex visando aplicação como biomaterial

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)