Role of PKC and CaV1.2 in Detrusor Overactivity in a Model of Obesity Associated with Insulin Resistance in Mice

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

Lobe identity in the Mongolian gerbil prostatic complex: A new rodent model for prostate study

Knowledge of structural and physiological differences among the prostatic lobes (PL) is the basis for development of experimental studies in traditional laboratory rodents. Although Mongolian gerbil reproductive organs have been increasingly investigated, its prostate structure is far from being properly known, and investigations of this organ focused on the ventral lobe (VL). Thus, the present study provides a thorough morphological description of prostatic complex in the male adult gerbil on the basis of topographic, histological, and ultrastructural analysis and ductal branching. Like other rodents, four pairs of PL were observed. However, in contrast to the rat and mouse, the VL is the least voluminous component and the dorsolateral lobe (DLL) is the most prominent and spatially isolated from remaining PL. The occurrence of a dorsal lobe (DL), hidden between bladder and insertion of seminal vesicles, has not been mentioned in previous reports with Mongolian gerbil. Collagenase digestion followed by microdissection revealed that, except for DL, which has a tubularacinar organization, all PL exhibit tubular organization and variable ductal branching. Distinct histological and ultrastructural features such as secretory epithelium, aspect of luminal secretion and stromal organization are reported for each PL and are confirmed by morphometric and stereological methods. Histological sections showed at least three intralobar segments in VL and DL. Ultrastructural analysis evidenced that, although luminal epithelial cells of PL share typical features of exocrine secretory cells, there are striking lobe phenotypical variations. Both merocrine and apocrine pathways are observed in variable rates in all PL, with the predominance of the former in the DLL and the latter in the CG. The morphological observations presented herein point to distinct structural identities for each PL, which probably reflects,specific functional compromise of seminal fluid secretion. These data also point to the gerbil as a good model for investigations concerning the regulation of prostate development and homeostasis, mainly with regard to the dorsal and dorsolateral PL.

A mass-transportation approach to a one dimensional fluid mechanics model with nonlocal velocity

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

BCS-BEC crossover in a trapped Fermi super-fluid using a density-functional equation

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

Path integral approach to the full Dicke model

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

Functional Characterization of an Aspergillus fumigatus Calcium Transporter (PmcA) that Is Essential for Fungal Infection

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

PHASE-TRANSITION IN A Q-DEFORMED LIPKIN MODEL

Assuming q-deformed commutation relations for the fermions, an extension of the standard Lipkin Hamiltonian is presented. The usual quasi-spin representation of the standard Lipkin model is also obtained in this q-deformed framework. A variationally obtained energy functional is used to analyse the phase transition associated with the spherical symmetry breaking. The only phase transitions in this q-deformed model are of second order. As an outcome of this analysis a critical parameter is obtained which is dependent on the deformation of the algebra and on the number of particles.

HIGHER-DERIVATIVE SCHWINGER MODEL

Using the operator formalism, we obtain the bosonic representation for the free fermion field satisfying an equation of motion with higher-order derivatives. Then, we consider the operator solution of a generalized Schwinger model with higher-derivative coupling. Since the increasing of the derivative order implies the introduction of an equivalent number of extra fermionic degrees of freedom, the mass acquired by the gauge field is bigger than the one for the standard two-dimensional QED. An analysis of the problem from the functional integration point of view corroborates the findings of canonical quantization, and corrects certain results previously announced in the literature on the basis of Fujikawa's technique.

Structural and functional characterization of neuwiedase, a nonhemorrhagic fibrin(ogen)olytic metalloprotease from Bothrops neuwiedi snake venom

A fibrino(geno)lytic nonhemorrhagic metalloprotease (neuwiedase) was purified from Bothrops neuwiedi snake venom by a single chromatographic step procedure on a CM-Sepharose column, Neuwiedase represented 4.5% (w/w) of the crude desiccated venom, with an approximate Mr of 20,000 and pI 5.9, As regards the amino acid composition, neuwiedase showed similarities with other metalloproteases, with high proportions of Asx, Glx, Leu, and Ser, Atomic absorption spectroscopy showed that one mole of Zn2+ and one mole of Ca2+ were present per mole olf protein. The cDNA encoding neuwiedase was isolated by RT-PCR from venom gland RNA, using oligonucleotides based on the partially determined amino-acid sequences of this metalloprotease. The fall sequence contained approximately 594 bp, which codified the 198 amino acid residues with an estimated molecular weight of 22,375. Comparison of the nucleotide and amino acid sequences of neuwiedase with those of other snake venom metalloproteases showed a high level of sequential similarity, Neuwiedase has two highly conserved characteristics sequences H(142)E(143)XXH(146)XXG(140)XXH(152) and C164I165M166. The three-dimensional structure of neuwiedase was modeled based on the crystal structure of Crotalus adamanteus Adamalysin II. This model revealed that the zinc binding site region showed a I high structural similarity with other metalloproteases,, the proteolyitc specificity, using the B beta-chain of oxidized insulin as substrate, was shown to be directed to the Ala(14)-Leu(15) and Tyr(16)-Leu(17) peptide bonds which were preferentially hydrolyzed. Neuwiedase is a A alpha,B beta fibrinogenase, Its activity upon the A alpha chain of fibrinogen was detected within 15 min of incubation. The optimal temperature and pH for the degradation of both A alpha and B beta chains were 37 degrees C and 7.4-8.0, respectively. This activity was inhibited by EDTA and 1,10-phenantroline, Neuwiedase also showed proteolytic activity upon fibrin and some components of the extracellular matrix. However, it did not show TAME esterase activity and was not able to inhibit platelet aggregation. (C) 2000 Academic Press.

The influence of the thickness on the functional properties of cassava starch edible films

The properties of edible films are influenced by several factors, including thickness. The purpose of this paper was to study the influence of thickness on the viscoelasticity properties, water vapor permeability, color and opacity of cassava starch edible films. These films were prepared by a casting technique, the film-forming solutions were 1, 2, 3 and 4% (w/v) of starch, heated to 70degreesC. Different thicknesses were obtained by putting 15 to 70 g of each solution on plexiglass plates. After drying at 30degreesC and ambient relative humidity, these samples were placed for 6 days at RH of 75%, at 22degreesC. The sample thicknesses were determined by a digital micrometer (+/-0.001 mm), as the average of nine different points. The viscoelasticity properties were determined by stress relaxation tests with a texture analyser TA.XT2i (SMS), being applied the Burgers model of four parameters. The water vapor permeability was determined with a gravimetric method, and color and opacity were determined using a Miniscan XE colorimeter, operated according to the Hunterlab method. All the tests were carried out in duplicate at 22degreesC. Practically, the four visco-elasticity properties calculated by the Burgers model had the same behavior, increasing with the thickness of all films, according to a power law model. The water vapor permeability and the color difference increased linearly with the thickness (0.013-0.144 mm) of all films prepared with solution of 1 to 4% of starch. on the other hand, the effect of the variation of the thickness over the opacity, was more important in the films with 1 and 2% of starch. It can be concluded that the control of the thickness in the elaboration of starch films by the casting technique is of extreme importance.

COMPARISON OF THE ANOMALOUS AND NONANOMALOUS GENERALIZED SCHWINGER MODELS VIA THE FUNCTIONAL FORMALISM

We calculate the Green functions of the two versions of the generalized Schwinger model, the anomalous and the nonanomalous one, in their higher order Lagrangian density form. Furthermore, it is shown through a sequence of transformations that the bosonized Lagrangian density is equivalent to the former, at least for the bosonic correlation functions. The introduction of the sources from the beginning, leading to a gauge-invariant source term, is also considered. It is verified that the two models have the same correlation functions only if the gauge-invariant sector is taken into account. Finally, there is presented a generalization of the Wess-Zumino term, and its physical consequences are studied, in particular the appearance of gauge-dependent massive excitations.

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.

Structural and functional characterization of myotoxin I, a Lys49 phospholipase A(2) homologue from Bothrops moojeni (Caissaca) snake venom

Myotoxin-I (MjTX-I) was purified to homogeneity from the venom of Bothrops moojeni by ion-exchange chromatography on CM-Sepharose. Its molecular weight, estimated by SDS-PAGE, was 13,400 (reduced) or 26,000 (unreduced). The extinction coefficient (E-1.0 cm(1.0 mg/ml)) of MjTX-I was 1.145 at lambda = 278 nm, pH 7.0, and its isoelectric point was 8.2 at ionic strength mu = 0.1. When lyophilized and stored at 4 degrees C, dimeric, trimeric, and pentameric forms of the protein were identified by SDS-PAGE. This heterogeneous sample could be separated into three fractions by gel filtration on Sephadex 6-50. The fractions were analyzed by isoelectric focusing, immunoelectrophoresis, and amino acid composition, which indicated that heterogeneity was the result of different levels of self-association. Protein sequencing indicated that MjTX-I is a Lys49 myotoxin and consists of 121 amino acids (M-r = 13,669), containing a high proportion of basic and hydrophobic residues. It shares a high degree of sequence identity with other Lys49 PLA(2)-like myotoxins, but shows a significantly lower identity with catalytically active Asp49 PLA(2)s. The three-dimensional structure of MjTX-I was modeled based on the crystal structures of three highly homologous Lys49 PLA(2)-like myotoxins. This model showed that the amino acid substitutions are conservative, and mainly the beta-wing region, and the C-terminal extended random coil. MjTX-I displays local myotoxic and edema-inducing activities in mice, and is lethal by intraperitoneal injection, with an LD50 value of 8.5 +/- 0.8 mg/kg, In addition, it is cytotoxic to myoblasts/ myotubes in culture, and disrupts negatively charged liposomes. In comparison with the freshly prepared dimeric sample, the more aggregated forms showed significantly reduced myotoxic activity. However, the edema-inducing activity of MjTX-I was independent of molecular association. Phospholipase A(2) activity on egg yolk, as well as anticoagulant activity, were undetectable both in the native and in the more associated forms. His, Tyr, and Trp residues of the toxin were chemically modified by specific reagents. Although the myotoxic and lethal activities of the modified toxins were reduced by these treatments, neither its edema-inducing or Liposome-disrupting activities were significantly altered. Rabbit antibodies to native MjTX-I cross-reacted with the chemically modified forms, and both the native and modified MjTX-I preparations were recognized by antibodies against the C-terminal region 115-129 of myotoxin II from B. asper, a highly Lys49 PLA(2)-homologue with high sequencial similarity. (C) 2000 Academic Press.

Molecular model of shikimate kinase from Mycobacterium tuberculosis

Tuberculosis (TB) resurged in the late 1980s and now kills approximately 3 million people a year. The reemergence of tuberculosis as a public health threat has created a need to develop new anti-mycobacterial agents. The shikimate pathway is an attractive target for herbicides and anti-microbial agents development because it is essential in algae, higher plants, bacteria, and fungi, but absent from mammals. Homologs to enzymes in the shikimate pathway have been identified in the genome sequence of Mycobacterium tuberculosis. Among them, the shikimate kinase I encoding gene (aroK) was proposed to be present by sequence homology. Accordingly, to pave the way for structural and functional efforts towards anti-mycobacterial agents development, here we describe the molecular modeling of M. tuberculosis shikimate kinase that should provide a structural framework on which the design of specific inhibitors may be based. © 2002 Elsevier Science (USA). All rights reserved.

Thermoeconomic model considering the environment impacts on thermoelectric power plants: Natural gas versus diesel

In this paper a comparative analysis of the environmental impact caused by the use of natural gas and diesel in thermoelectric power plants utilizing combined cycle is performed. The objective is to apply a thermoeconomical analysis in order to compare the two proposed fuels. In this analysis, a new methodology that incorporates the economical engineering concept to the ecological efficiency once Cardu and Baica [1, 2], which evaluates, in general terms, the environmental impacts caused by CO2, SO2, NOx and Particulate Matter (PM), adopting as reference the air quality standards in vigour is employed. The thermoeconomic model herein proposed utilizes functional diagrams that allow the minimization the Exergetic Manufacturing Cost, which represents the cost of production of electricity incorporating the environmental impact effects to study the performance of the thermoelectric power plant [3,4], It follows that it is possible to determine the environmental impact caused by thermoelectric power plants and, under the ecological standpoint, the use of natural gas as a fuel is the best option compared to the use of the diesel, presenting ecological efficiency values of 0.944 and 0.914 respectively. From the Exergoeconomic point of view of, it was found out that the EMC (Exergetic Manufacturing Cost) is better when natural gas is used as fuel compared to the diesel fuel. Copyright © 2006 by ASME.