Investigation of the pore blockage of a Brazilian dolomite during the sulfation reaction

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

Can sieve-element plastids in Panicum maximum (Poaceae) leaves act in the blockage of injured sieve-tube elements?

The ultrastructural features and the plastid changes caused by sample preparation were studied in sieve elements of Panicum maximum leaves. Samples of expanded leaves, taken near the ligule region, were fixed and processed by common light and transmission electron microscopy methods. In mature sieve-tube elements, the protoplast is electron-translucent and plastids are the most frequent organelles. Mitochondria and smooth endoplasmic reticulum segments are also visible and occupy a parietal position within the cell. The plastids are globular and show electron-dense proteinaceous inclusions in the stroma. The protein crystals are predominantly cuneate, but thin crystalloids and amorphous and/or filamentous proteins also occur. The presence of intact plastids plus others in different phases of plastid envelope rupture were interpreted as evidence that this rupture is a normal event in response to injury. This plastid envelope rupture is possibly activated by the release of pressure in the sieve-tube element. After plastid membrane vesiculation, the stroma and the protein crystals are dispersed within the sieve-element ground cytoplasm. The vesicles originating from the plastid envelope move to one cell pole, while protein crystalloids move to the opposite pole and agglomerate in the sieve-plate region. Our findings indicate that these protein crystalloids, which deposit in the sieve plate, may act in sieve-plate pores occlusion, preventing the release of phloem sap, similar to the role of P-protein in dicotyledons. (c) 2008 Elsevier GmbH. All rights reserved.

Androgen receptor in the Mongolian gerbil ventral prostate: Evaluation during different phases of postnatal development and following androgen blockage

The normal growth, differentiation and maintenance of the morphofunctional integrity of the prostate gland are dependent on the interaction of constant levels of androgens with their receptors. The need to study the responses to hormones under several conditions and the effect of their blockage is due to the fact that the human prostate is the site of a great number of age-related diseases, and the ones with a major medical importance are prostate cancer (Cap) and benign prostatic hyperplasia (BPH), which can both be treated with androgen suppression. Seventy-five male gerbils were divided, randomly, into 3 groups of 25 animals each, where each group corresponded to one phase of postnatal development. In each phase, it was possible to morphologically and stereologically analyze the compartments of prostatic ventral lobe, as well as to immunohistochemically analyze the degree of expression of androgen receptors (ARs) after the androgen blockage therapies. In addition, it was possible to establish the hormonal dosage of serum testosterone levels given the comparative approach of the expression of androgen receptors. There is a pattern of AR distribution in the prostatic ventral lobe throughout postnatal development, in which the younger the animal is the higher, the interaction of circulating androgens that stimulate the AR expression in both the epithelial and stromal compartments. The androgen blockage therapies decreased AR expression in the prostatic compartments, but the androgen reposition after these blockages was not sufficient to recover the glandular structure or stimulate the AR expression up to normal physiological conditions. Both the regulation and distribution of androgen receptors along the gerbil prostatic tissues are complex mechanisms that are likely to be genetically regulated by androgens prenatally or by other factors that are still unknown. This rodent species seems to be a valuable model in the attempt to improve the understanding of the morphophysiological and pathological behavior of this important gland in humans throughout aging and to stimulate new therapeutic ideas to fight prostate cancer. (C) 2008 Elsevier Ltd. All rights reserved.

A thermoporometry and small-angle x-ray scattering study of wet silica sonogels as the pore volume fraction is varied

Silica sonogels with different porosities were prepared by acid sono-hydrolysis of tetraethoxysilane. Wet sonogels were studied using small-angle x-ray scattering (SAXS) and differential scanning calorimetry (DSC). The DSC shows a broad thermal peak below the normal water melting point associated with the melting of confined ice nanocrystals, or nanoporosity. The nanopore size distribution was determined from the Gibbs-Thomson equation. As the porosity is increased, a second sharp DSC thermal peak with onset temperature at the water melting point is apparent, which was associated with the melting of ice macrocrystals, or macroporosity. The DSC result could be causing misinterpretation of the macroporosity because water may not be exactly confined in very feeble silica network regions in sonogels with high porosity. The structure of the wet gels can be described fairly well as mutually self-similar mass fractal structures with characteristic length. increasing from similar to 1.8 to similar to 5.4 nm and mass fractal dimension D diminishing discretely from similar to 2.6 to similar to 2.3 as the porosity increases in the range studied. More specifically, such a structure could be described using a two-parameter correlation function gamma(r) similar to r(D-3) exp(-r/xi), which is limited at larger scale by the cut-off distance xi but without a well-defined small scale cut-off distance, at least up to the maximum angular domain probed using SAXS in the present study.

Validation of a mutant of the pore-forming toxin sticholysin-I for the construction of proteinase-activated immunotoxins

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

The membranotropic activity of N-terminal peptides from the pore-forming proteins sticholysin I and II is modulated by hydrophobic and electrostatic interactions as well as lipid composition

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efficiency of ethanol conversion induced by controlled modification of pore structure and acidic properties of alumina catalysts

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Preparation of silica with controlled pore sizes for enzyme immobilization

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

Structural characteristics of gels prepared from sonohydrolysis and conventional hydrolysis of TEOS: an emphasis on the mass fractal as determined from the pore size distribution

Silica gels were preparated from fixed proportion mixtures of tetraethoxysilane, water and hydrocloric acid, using either ultrasound stimulation (US) or conventional method (CO) in the hydrolysis step of the process. Wet gets were obtained with the same silica volume concentration and density. According to small-angle X-ray scattering, the structure of the wet gels can be described as mass fractal structures with mass fractal dimension D = 2.20 in a length scale xi = 7.9 nm, in the case of wet gels US, and D = 2.26 in a length scale 6.9 nm, in the case of wet gels CO. The mass fractal characteristics of the wet gels US and CO account for the different structures evolved in the drying of the gels US and CO in the obtaining of xerogels and aerogels. The pore structure of the dried gels was studied by nitrogen adsorption as a function of the temperature. Aerogels (US and CO) present high porosity with pore size distribution (PSD) curves in the mesopore region while xerogels (US and CO) present minor porosity with PSD curves mainly in the micropore region. The dried gels US (aerogels and xerogels) generally present pore volume and specific surface area greater than the dried gels CO. The mass fractal structure of the aerogels has been studied from an approach based on the PSD curves exclusively. (c) 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Model peptides mimic the structure and function of the N-terminus of the pore-forming toxin sticholysin II

To investigate the role of the N-terminal region in the lytic mechanism of the pore-forming toxin sticholysin II (St II), we studied the conformational and functional properties of peptides encompassing the first 30 residues of the protein. Peptides containing residues 1-30 (P1-30) and 11-30 (P11-30) were synthesized and their conformational properties were examined in aqueous solution as a function of peptide concentration, pH, ionic strength, and addition of the secondary structure-inducing solvent trifluoroethanol (TFE). CD spectra showed that increasing concentration, pH, and ionic strength led to aggregation of P1-30; as a consequence, the peptide acquired beta-sheet conformation. In contrast, P11-30 exhibited practically no conformational changes under the same conditions, remaining essentially structureless. Moreover, this peptide did not undergo aggregation. These differences clearly point to the modulating effect of the first 10 hydrophobic residues on the peptides aggregation and conformational properties. In TFE both the first ten hydrophobic peptides acquired alpha-helical conformation, albeit to a different extent, P11-30 displayed lower alpha-helical content. P1-30 presented a larger-fraction of residues in alpha-helical conformation in TFE than that found in St II's crystal structure for that portion of the protein. Since TFE mimics the membrane em,, such increase in helical content could also occur upon toxin binding to membranes and represent a step in the mechanism of pore formation. The peptides conformational properties correlated well with their functional behaviour. Thus, P1-30 exhibited much higher hemolytic activity than P11-30. In addition, P11-30 was able to block the toxin's hemolytic activity. The size of pores formed in red blood cells by P 1-30 was estimated by measuring the permeability PEGs of different molecular mass. The pore radius (0.95 +/- 0.01 nm) was very similar to that of the PEGs of different pore formed by the toxin. The results demonstrate that the synthetic peptide P1-30 is a good model of St 11 conformation and function and emphasize the contribution of the toxin's N-terminal region, and, in particular, the hydrophobic residues 1-10 to pore formation. (c) 2005 Wiley Periodicals, Inc.

Determination of pore size distribution in soybean seed coats by mercury porosimetry

In this work we applied mercury porosimetry for the investigation of soybean seed coats. By using this method it was possible to determine the pore size distribution and also the pore size dispersion that is present in seed coats. The results showed that for the studied soybean genotype the seed coats had a characteristic pore diameter, but deviation of this size was not negligible. Finally, the results were confirmed by electron microscopy.

Differences and similarities in the pore size distribution of soybean seed coats

In this work we apply the mercury porosimetry technique to determine the pore size distribution in soybean seed coats of different varieties. The analyses show that the porosity of soybean seed coats is different when seeds of different genotypes are compared. This result points the possibility of using pore size distribution to varietal discrimination.

Actively transcribed GAL genes can be physically linked to the nuclear pore by the SAGA chromatin modifying complex

Recent work has demonstrated that some actively transcribed genes closely associate with nuclear pore complexes (NPC) at the nuclear periphery. The Saccharomyces cerevisiae Mlp1 and Mlp2 proteins are components of the inner nuclear basket of the nuclear pore that mediate interactions with these active genes. To investigate the physical link between the NPC and active loci, we identified proteins that interact with the carboxyl-terminal globular domain of Mlp1 by tandem affinity purification coupled with mass spectrometry. This analysis led to the identification of several components of the Spt-Ada-Gcn5-acetyltransferase ( SAGA) histone acetyltransferase complex, Gcn5, Ada2, and Spt7. We utilized co-immunoprecipitation and in vitro binding assays to confirm the interaction between the Mlp proteins and SAGA components. Chromatin immunoprecipitation experiments revealed that Mlp1 and SAGA components associate with the same region of the GAL promoters. Critically, this Mlp-promoter interaction depends on the integrity of the SAGA complex. These results identify a physical association between SAGA and the NPC, and support previous results that relied upon visualization of GAL loci at the nuclear periphery by microscopy ( Cabal, G. G. Genovesio, A., Rodriguez-Navarro, S., Zimmer, C., Gadal, O., Lesne, A., Buc, H., Feuerbach- Fournier, F., Olivo-Marin, J.-C., Hurt, E. C., and Nehrbass, U. ( 2006) Nature 441, 770-773). We propose that a physical interaction between nuclear pore components and the SAGA complex can link the actively transcribed GAL genes to the nuclear pore.

On-line preconcentration and speciation analysis of Cr(III) and Cr(VI) using baker's yeast cells immobilised on controlled pore glass

A study was undertaken to evaluate Saccharonzyces cerevisiae as a substrate for the biosorption of Cr(III) and Cr(VI) aiming to the selective determination of these species in aqueous solutions. The yeast cells were covalently immobilised on controlled pore glass (CPG), packed in a minicolumn and incorporated in an on-line flow injection system. The effect of chemical and physical variables affecting the biosorption process was tested in order to select the optimal analytical conditions for the Cr retention by S. cerevisiae. Cr(III) was retained by the immobilised cells and Cr(VI) were retained by CPG. The speciation was possible by selective and sequential elution of Cr(III) with 0.05 mol L-1 HCl and 2.0 mol L-1 HNO3 for Cr(VI). The influence of some concomitant ions up to 20 mg L-1 was also tested. Quantitative determinations of Cr were carried out by means of inductively coupled plasma optical emission spectrometry (ICP OES). Preconcentration factors of 12 were achieved for Cr(III) and 5 for Cr(VI) when 1.7 mL of sample were processed reaching detection limits of 0.45 for Cr(III) and 1.5 mu g L-1 for Cr(VI). The speciation of inorganic Cr in different kinds of natural waters was performed following the proposed method. Spiked water samples were also analysed and the recoveries were in all cases between 81 and 103%. (c) 2005 Elsevier B.V. All rights reserved.