Structure of myotoxin II, a catalytically inactive Lys49 phospholipase A2 homologue from Atropoides nummifer venom

Lys49 snake-venom phospholipase A2 (PLA2) homologues are highly myotoxic proteins which, although lacking catalytic activity, possess the ability to disrupt biological membranes, inducing significant muscle-tissue loss and permanent disability in severely envenomed patients. Since the structural basis for their toxic activity is still only partially understood, the structure of myotoxin II, a monomeric Lys49 PLA2 homologue from Atropoides nummifer, has been determined at 2.08 Å resolution and the anion-binding site has been characterized. © 2006 International Union of Crystallography. All rights reserved.

cDNA cloning and 1.75 Å crystal structure determination of PPL2, an endochitinase and N-acetylglucosamine-binding hemagglutinin from Parkia platycephala seeds

Parkia platycephala lectin 2 was purified from Parkia platycephala (Leguminosae, Mimosoideae) seeds by affinity chromatography and RP-HPLC. Equilibrium sedimentation and MS showed that Parkia platycephala lectin 2 is a nonglycosylated monomeric protein of molecular mass 29 407 ± 15 Da, which contains six cysteine residues engaged in the formation of three intramolecular disulfide bonds. Parkia platycephala lectin 2 agglutinated rabbit erythrocytes, and this activity was specifically inhibited by N-acetylglucosamine. In addition, Parkia platycephala lectin 2 hydrolyzed β(1-4) glycosidic bonds linking 2-acetoamido-2-deoxy-β-d-glucopyranose units in chitin. The full-length amino acid sequence of Parkia platycephala lectin 2, determined by N-terminal sequencing and cDNA cloning, and its three-dimensional structure, established by X-ray crystallography at 1.75 Å resolution, showed that Parkia platycephala lectin 2 is homologous to endochitinases of the glycosyl hydrolase family 18, which share the (βα) 8 barrel topology harboring the catalytic residues Asp125, Glu127, and Tyr182. © 2006 The Authors.

Lead-cadmium fluorogermanate glasses and transparent glassceramics. Spectroscopy and structure

In Lead-cadmium fluorogermanate glasses (PbF2-CdF 2-PbGeO3) the addition of metal fluorides to the base PbGeO3 glass leads to a decrease of the glass transition temperature (Tg) and to an enhancement of the ionic conductivity properties. Based on different spectroscopic techniques (19F NMR, Ge K-edge X-ryas absorption and Raman scattering) an heterogeneous glass structure is proposed at the molecular scale, which can be described by fluoride rich regions permeating the metagermanate chains. The temperature dependence of the 19F NMR lineshapes and relaxation times exhibits the qualitative and quantitative features associated with the high fluoride mobility in these systems. Eu 3+ emission and vibronic spectra are used to follow the crystallization process leading to transparent glass ceramics.

Dispersed lipid liquid crystalline phases stabilized by a hydrophobically modified cellulose

Aqueous dispersions of monoolein (MO) with a commercial hydrophobically modified ethyl hydroxyethyl cellulose ether (HMEHEC) have been investigated with respect to the morphologies of the liquid crystalline nanoparticles. Only very low proportions of HMEHEC are accepted in the cubic and lamellar phases of the monoolein-water system. Due to the broad variation of composition and size of the commercial polymer, no other single-phase regions were found in the quasi-ternary system. Interactions of MO with different fractions of the HMEHEC sample induced the formation of lamellar and reversed hexagonal phases, identified from SAXD, polarization microscopy, and cryogenic TEM examinations. In excess water (more than 90 wt %) coarse dispersions are formed more or less spontaneously, containing particles of cubic phase from a size visible by the naked eye to small particles observed by cryoTEM. At high polymer/MO ratios, vesicles were frequently observed, often oligo-lamellar with inter-lamellar connections. After homogenization of the coarse dispersions in a microfluidizer, the large particles disappeared, apparently replaced by smaller cubic particles, often with vesicular attachments on the surfaces, and by vesicles or vesicular particles with a disordered interior. At the largest polymer contents no proper cubic particles were found directly after homogenization but mainly single-walled defected vesicles with a peculiar edgy appearance. During storage for 2 weeks, the dispersed particles changed toward more well-shaped cubic particles, even in dispersions with the highest polymer contents. In some of the samples with low polymer/MO ratio, dispersed particles of the reversed hexagonal type were found. A few of the homogenized samples were freeze-dried and rehydrated. Particles of essentially the same types, but with a less well-developed cubic character, were found after this treatment. © 2007 American Chemical Society.

Chorismate synthase: An attractive target for drug development against orphan diseases

The increase in incidence of infectious diseases worldwide, particularly in developing countries, is worrying. Each year, 14 million people are killed by infectious diseases, mainly HIV/AIDS, respiratory infections, malaria and tuberculosis. Despite the great burden in the poor countries, drug discovery to treat tropical diseases has come to a standstill. There is no interest by the pharmaceutical industry in drug development against the major diseases of the poor countries, since the financial return cannot be guaranteed. This has created an urgent need for new therapeutics to neglected diseases. A possible approach has been the exploitation of the inhibition of unique targets, vital to the pathogen such as the shikimate pathway enzymes, which are present in bacteria, fungi and apicomplexan parasites but are absent in mammals. The chorismate synthase (CS) catalyses the seventh step in this pathway, the conversion of 5-enolpyruvylshikimate-3-phosphate to chorismate. The strict requirement for a reduced flavin mononucleotide and the anti 1,4 elimination are both unusual aspects which make CS reaction unique among flavin-dependent enzymes, representing an important target for the chemotherapeutic agents development. In this review we present the main biochemical features of CS from bacterial and fungal sources and their difference from the apicomplexan CS. The CS mechanisms proposed are discussed and compared with structural data. The CS structures of some organisms are compared and their distinct features analyzed. Some known CS inhibitors are presented and the main characteristics are discussed. The structural and kinetics data reviewed here can be useful for the design of inhibitors. © 2007 Bentham Science Publishers Ltd.

Estudo da influência dos íons K+, Mg2+, SO4 2- e CO3 2- na cristalização biomimética de fosfato de cálcio amorfo (ACP) e conversão a fosfato octacálcico (OCP)

The crystallization of hydroxyapatite (HA) in aqueous solution can be described by the mechanism ACP → OCP → HA. In this work, it was studied the influence of K+, Mg2+, SO4 2- AND CO3 2- ions in the formation of ACP and in its conversion to OCP, using biomimetic coatings on metallic substrates of commercially pure titanium (Ti c.p.). The results showed that Mg2+ and CO3 2- ions favored both the formation of ACP and its conversion to OCP. Differently, K+ and SO4 2- ions did not influence the formation of ACP and, consequently, interfered in the conversion to OCP.

Geocronologia do maciço Granítico Sararé no SW do Cráton Amazônico

Sararé Massif occurs in the southwest Mato Grosso state intruded into Mesoproterozoic units of the Jauru Block inside the Amazonian Craton. It presents an extension of approximately 80 km2 and NW-SE tectonic structures control the elongated shape. It is constituted by three major monzogranitic petrographic facies, represented by leucocratic, reddish, isotropic, equi-inequigranular to locally porphyritic rocks. The composition indicates S-type, peraluminous, with indicatives of late- to post-kinematic magmatism. Geochronological studies with 40Ar/39Ar in biotites and muscovites resulted in ages of 903 to 906 Ma was interpreted as massif rocks cooling period and U-PB 917 ± 18 Ma. ages points to the crystallization of the intrusive body. The massif is formed by melting of material of the upper crust, in an environment of continental collision and/or of post-collisional decompression at the ending of the Aguapeí-Sunsás event, in more stable environments of consolidation and tectonic stabilization of the Amazonian Craton.

Evolução geológica dos batólitos granitóides neoproterozóicos do sudeste do estado de são paulo

The Neoproterozoic granitogenesis related to the Central Mantiqueira Province comprise the calc alkaline to alkaline granitoid complexes of Sorocaba, San Francisco, São Roque, Ibiúna and Piedade. These complexes occur in a ruptil tectonic to tardi (Sn+3) event. The emplacement of the different facies in transtractives structures of the pull-apart type are characterized in the area by the main transcurrent shear zones of Taxaquara-Pirapora, Itu-Jundiuvira, Moreiras, Cangüera and Caucáia of ENE-WSW general direction. The massifs present complex internal architecture characterized by intrusions in restrict initial phase of intermediate equigranular nature. Also present a main phase of porfiroid monzo and sienogranite that fragments the previous phase, followed by lateral accretion of equi to inequigranular material, and in some cases by the accretion of late phases of circular bodies of porfiroid rapakivi granites, and a late to final phase of aplitic to pegmatitic composition. This magmatism grew with the intrusions of successive magmatic pulses, partially controlled by many reactivations of the shear zones. The REE also suggest that the magmatic phases are similar, synchronous and repetitive in four of the complexes in both domains, present in the São Francisco Complex. The crystallization starts from accretion processes, but compositionally quite different from the others. The variation in compositions and ages (TDM) for these granites reflect the derivation from different sources developed under different magmatic conditions, followed by processes of contamination that frequently occur in the crust.

Formação de biofilme cristalino de Proteus mirabilis em cateter urológico

The indwelling urethral catheter has an important role for patients with urinary retention, bladder obstruction, neurological damage and other diseases. Urine contains minerals which precipitate in alkaline pH, crystallize and block the urological catheter. The crystallization of the ionic components of urine occurs in the presence of urease, an enzyme produced by Proteus mirabilis. This bacterium adheres to inanimate surfaces and forms biofilms. The aim of this study was to investigate the formation of crystalline biofilm on the luminal surface of siliconized latex catheters by means of scanning electron microscope, after channeling artificial urine infected with Proteus mirabilis. The experiment was performed in vitro using a dynamic flow system. The artificial urine compounds were salts of calcium, magnesium, phosphates, urea and egg albumin, and it was infected with Proteus mirabilis ATCC 25933. The urine flow was stopped after crystallization of the ionic components. Crystallization was observed after alkalinization of urine. Scanning electron microscopy showed the presence of crystals and morphologies typical of bacilli embedded in an amorphous mass on the internal lumen of the catheter. The present study showed that catheter encrustation may limit the use of long-term indwelling catheter. © SBEB - Sociedade Brasileira de Engenharia Biomédica.

Glass-ceramic material from the SiO 2-Al 2O 3-CaO system using sugar-cane bagasse ash (SCBA)

Brazil is the world's largest producer of alcohol and sugar from sugarcane. Currently, sugarcane bagasse is burned in boilers to produce steam and electrical energy, producing a huge volume of ash. The major component of the ash is SiO 2, and among the minor components there are some mineralizing agents or fluxing. Published works have shown the potential of transforming silicate-based residues into glass-ceramic products of great utility. This work reports the research results of SCBA use to produce glass-ceramics with wollastonite, rankinite and gehlenite as the major phases. These silicates have important applications as building industry materials, principally wollastonite, due to their special properties: high resistance to weathering, zero water absorption, and hardness among others. The glasses (frits) were prepared mixing ash, calcium carbonate and sodium or potassium carbonates as flux agents, in different concentrations. X-ray fluorescence was used to determine the chemical composition of the glasses and their crystallization was assessed by using thermal analysis (DTA/DSC/TGA) and X-ray diffraction. The crystallization kinetics was evaluated using the Kissinger method, giving activation energies ranging from 200 to 600 kJ/mol. © 2011 Ceramic Society of Japan.

Caracterização geotermobarométrica dos metabasitos de Cajamar (SP), Grupo São Roque, Cinturão Ribeira

Metamafic orthoderivate rocks, characterized by MORB signatures and paleoproterozoic ages occur in Cajamar region central east part of São Paulo State, Brazil. They are associated to philites, metasandstones and metalimestones of São Roque Group volcanosedimentary sequence, Southeastern portion of the Ribeira Belt. For the metamafic rocks the assemblage of metamorphic peak is represented by actinolitealbite- epidote-chlorite typical of regional metamorphism, greenschist facies. For a good characterization of P and T parameters of metamorphism microprobe analyses of main minerals were performed. Metamorphism did not destroy completely the primary mineralogy of original basalt and microgabbro, and augite crystals are registered in many samples, preserved together with metamorphic assemblage, and possibly indicate that the igneous temperatures of crystallization are between 900 and 1.100°C. Textures of mineral equilibrium for the peak paragenesis are not always present in the studied samples. It may be partially caused by the superposition of retrometamorphic events that is a consequence of regional shearing process. Where the equilibrium is reached, chiefly in the occurrences in the south of the area temperature and pressure determinations indicate values close to 487 ± 42°C and 8,2 ± 2.0 kbar, respectively. Other values for P and T lower than that of the peak were also obtained in central and north portion of the area and they are: 450 ± 68°C for T e 4.7 kbar for P and 315 ± 26°C for T and 3.0 kbar for P and may indicate that the mineral equilibrium was not reached in these samples or that they represent the retrometamorphic event. As a concluding remark considering the metamorphic P-T path now obtained one can say that the exhumation process in this area occurred by isothermal decompression.

Parâmetros físicos pré-eruptivos do magmatismo ácido da província magmática do Paraná: Resultados preliminares

A set of 12 samples of acid rock types Palmas (ATP) and Chapecó (ATC) was used to determine the chemical composition of plagioclase and pyroxene by electron microprobe, with the purpose to get information about the pressure and temperature of crystallization of these rocks. The results show that the pyroxene of ATP rocks (3,2 ± 1,2 kbar, max = 5,1 kbar and 1028 ± 38°C) were formed under pressure conditions higher than those ATC (1,8 ± 0,9 kbar, max = 3,4 kbar and 995 ± 26oC). However, the pressures obtained from plagioclase showed higher pressures for ATC (3.2 ± 1 kbar, max = 6,4 kbar and 1033 ± 12°C) than ATP (1,9 ± 1 kbar, max = 4,8 kbar and 1043 ± 5°C), suggesting that the crystallization of rocktype ATP began with the formation of pyroxene and plagioclase almost simultaneously at a depth of around 17 km while the ATC, began with the crystallization of plagioclase at a depth of about 21 km (assuming a gradient of 3,3 kbar/km). The geothermometry of plagioclase allow us to calculate the concentration of water from about 1 ± 0,3% H2O for both acid rock types. Additional calculations allow us to get the depth of water exsolution of magmatic liquid at 30m below the surface. Although the data are still preliminary and insufficient to model the extrusion of these rocks, they point out to an effusion mechanism of a partially fluidized magma by volatile, which would spread to large areas with small friction with the surface that would increased with the increase of viscosity caused by the loss of volatile and decreasing of temperature, developing coherent structures as lava flows.

Structural insights into selectivity and cofactor binding in snake venom l-amino acid oxidases

l-Amino acid oxidases (LAAOs) are flavoenzymes that catalytically deaminate l-amino acids to corresponding α-keto acids with the concomitant production of ammonia (NH 3) and hydrogen peroxide (H 2O 2). Particularly, snake venom LAAOs have been attracted much attention due to their diverse clinical and biological effects, interfering on human coagulation factors and being cytotoxic against some pathogenic bacteria and Leishmania ssp. In this work, a new LAAO from Bothrops jararacussu venom (BjsuLAAO) was purified, functionally characterized and its structure determined by X-ray crystallography at 3.1å resolution. BjsuLAAO showed high catalytic specificity for aromatic and aliphatic large side-chain amino acids. Comparative structural analysis with prokaryotic LAAOs, which exhibit low specificity, indicates the importance of the active-site volume in modulating enzyme selectivity. Surprisingly, the flavin adenine dinucleotide (FAD) cofactor was found in a different orientation canonically described for both prokaryotic and eukaryotic LAAOs. In this new conformational state, the adenosyl group is flipped towards the 62-71 loop, being stabilized by several hydrogen-bond interactions, which is equally stable to the classical binding mode. © 2012 Elsevier Inc.

Kinetic parameters for thermal decomposition of supramolecular polymers derived from diclofenac-meglumine supramolecular adducts

Meglumine is an aminocarbohydrate able to form supramolecular adducts with organic acids. The recognition is based on hydrogen bonds and the structures resulting from the complexation have high solubility in water. This property has been exploited by the pharmaceutical industry in the improvement of existing drugs, and the successful example of this approach involves the poorly soluble non-steroidal anti-inflammatory drugs (NSAIDs). Investigation of the thermal behavior of adduct obtained from meglumine and the NSAID diclofenac revealed that a polymer-like material is formed from the self-assembly of diclofenac-meglumine adducts in the melt. This polymer showed a high molecular weight around 2.0×105kDa. The kinetic parameters for the thermal decomposition step of the polymer were determined by the Capela-Ribeiro non-linear isoconversional method. From data for the TG curves in nitrogen atmosphere and heating rates of 5, 10, 15 and 20°Cmin-1, the Eα and Bα terms could be determined, and consequently the pre-exponential factor, Aα, as well as the kinetic model, g(α). © 2012 Elsevier B.V.

Fractal dimension and Shannon's entropy analyses of the architectural complexity caused by the inflammatory reactions induced by highly crystalline polyvinyl alcohol) microspheres implanted in subcutaneous tissues of the Wistar rats

The results of the histopathological analyses after the implantation of highly crystalline PVA microspheres in subcutaneous tissues of Wistar rats are here in reported. Three different groups of PVA microparticles were systematically studied: highly crystalline, amorphous, and commercial ones. In addition to these experiments, complementary analyses of architectural complexity were performed using fractal dimension (FD), and Shannon's entropy (SE) concepts. The highly crystalline microspheres induced inflammatory reactions similar to the ones observed for the commercial ones, while the inflammatory reactions caused by the amorphous ones were less intense. Statistical analyses of the subcutaneous tissues of Wistar rats implanted with the highly crystalline microspheres resulted in FD and SE values significantly higher than the statistical parameters observed for the amorphous ones. The FD and SE parameters obtained for the subcutaneous tissues of Wistar rats implanted with crystalline and commercial microparticles were statistically similar. Briefly, the results indicated that the new highly crystalline microspheres had biocompatible behavior comparable to the commercial ones. In addition, statistical tools such as FD and SE analyses when combined with histopathological analyses can be useful tools to investigate the architectural complexity tissues caused by complex inflammatory reactions. © 2012 WILEY PERIODICALS, INC.