Homogalactanas sulfatadas da alga Codium isthmocladum com atividade anticoagulante

Since the first description of sulfated polysaccharides from seaweeds, the biological activities of these compounds have been evaluated under different aspects and experimental procedures. Among the broad biological activities presented by seaweed polysaccharides, anticoagulant action appears as a promising function. In this present study we have obtained sulfated polysaccharides from the green seaweed Codium isthmocladium by proteolytic digestion, followed by separation into five fractions (0.3, 0.5, 0.7, 0.9 and 1.2) by sequential acetone precipitation. The chemical analyses have demonstrated that all fractions are composed mainly by sulfated polysaccharides. The anticoagulant activity of these fractions was determined by activated partial thromboplastin time (aPTT) and prothrombin time test (PT) using citrate normal human plasma. None fraction has shown anticoagulant activity by PT test. Furthermore, all of them have shown anticoagulant activity by aPTT test. These results indicated that the molecular targets of these sulfated polysaccharides are mainly in the intrinsic via of the coagulation cascade. Agarose gel electrophoresis in 1,3-diaminopropane acetate buffer, pH 9.0, stained with 0.1% toluidine blue showed the presence of two or three bands in several fractions while the fraction 0.9 showed a single spot. By anion exchange chromatography, the acid polysaccharides from the 0.9 acetone fraction were separated into two new fractions eluted respectively with 2.0 and 3.0 M NaCl. These compounds showed a molecular weight of 6.4 and 7.4 kDa respectively. Chemical analyses and infrared spectroscopy showed that Gal 1 and Gal 2 are sulfated homogalactans and differ one from the other in degree and localization of sulfate groups. aPPT test demonstrated that fractions 2,0 and 3,0M (Gal1 and Gal 2, respectively) have anticoagulant activity. This is the first time that anticoagulant sulfated homogalatans have been isolated from green algae. To prolong the coagulation time to double the baseline value in the aPTT, the required amount of sulfated galactan 1 (6,3mg) was similar to low molecular heparin Clexane®, whereas only 0,7mg of sulfated galactan 2 was needed to obtain the same effect. Sulfated galactan 2 in high doses (250mg) induces platelet aggregation. These results suggest that these galactans from C. isthmocladum have a potential application as an anticoagulant drug

Novas propriedades do SKTI (Inibidor de tripsina de soja): inibição para elastase neutrofílica humana e efeitos no processo de injúria pulmonar aguda

Seeds from legumes including the Glycine max are known to be a rich source of protease inhibitors. The soybean Kunitz trypsin inhibitor (SKTI) has been well characterised and has been found to exhibit many biological activities. However its effects on inflammatory diseases have not been studied to date. In this study, SKTI was purified from a commercial soy fraction, enriched with this inhibitor, using anion exchange chromatography Resource Q column. The purified protein was able to inhibit human neutrophil elastase (HNE) and bovine trypsin. . Purified SKTI inhibited HNE with an IC50 value of 8 µg (0.3 nM). At this concentration SKTI showed neither cytotoxic nor haemolytic effects on human blood cell populations. SKTI showed no deleterious effects on organs, blood cells or the hepatic enzymes alanine amine transferase (ALT) and aspartate amino transferase (AST) in mice model of acute systemic toxicity. Human neutrophils incubated with SKTI released less HNE than control neutrophils when stimulated with PAF or fMLP (83.1% and 70% respectively). These results showed that SKTI affected both pathways of elastase release by PAF and fMLP stimuli, suggesting that SKTI is an antagonist of PAF/fMLP receptors. In an in vivo mouse model of acute lung injury, induced by LPS from E. coli, SKTI significantly suppressed the inflammatory effects caused by elastase in a dose dependent manner. Histological sections stained by hematoxylin/eosin confirmed this reduction in inflammation process. These results showed that SKTI could be used as a potential pharmacological agent for the therapy of many inflammatory diseases

Produção e caracterização de quitooligossacarídeos produzidos pelo fungo Metarhizium anisopliae e avaliação da citotoxicidade em células tumorais

Chitosan is a natural polymer, biodegradable, nontoxic, high molecular weight derived from marine animals, insects and microorganisms. Oligomers of glucosamine (GlcN) and N-acetylglucosamine (GlcNAc) have interesting biological activities, including antitumor effects, antimicrobial activity, antioxidant and others. The alternative proposed by this work was to study the viability of producing chitooligosaccharides using a crude enzymes extract produced by the fungus Metarhizium anisopliae. Hydrolysis of chitosan was carried out at different times, from 10 to 60 minutes to produce chitooligosaccharides with detection and quantification performed by High Performace Liquid Chromatography (HPLC). The evaluation of cytotoxicity of chitosan oligomers was carried out in tumor cells (HepG2 and HeLa) and non-tumor (3T3). The cells were treated for 72 hours with the oligomers and cell viability investigated using the method of MTT. The production of chitosan oligomers was higher for 10 minutes of hydrolysis, with pentamers concentration of 0.15 mg/mL, but the hexamers, the molecules showing greater interest in biological properties, were observed only with 30 minutes of hydrolysis with a concentration of 0.004 mg/mL. A study to evaluate the biological activities of COS including cytotoxicity in tumor and normal cells and various tests in vitro antioxidant activity of pure chitosan oligomers and the mixture of oligomers produced by the crude enzyme was performed. Moreover, the compound with the highest cytotoxicity among the oligomers was pure glucosamine, with IC50 values of 0.30; 0.49; 0.44 mg/mL for HepG2 cells, HeLa and 3T3, respectively. Superoxide anion scavenging was the mainly antioxidant activity showed by the COS and oligomers. This activity was also depending on the oligomer composition in the chitosan hydrolysates. The oligomers produced by hydrolysis for 20 minutes was analyzed for the ability to inhibit tumor cells showing inhibition of proliferation only in HeLa cells, did not show any effect in HepG2 cells and fibroblast cells (3T3)

Papel da localização da carga em determinação dos parâmetros do canal iônico

To aureus α-HL channel, we used the cysteine-scanning mutagenesis technique. Twenty-four mutants were produced from the substitution of a single aminoacid of the primary structure of the α-HL pro this yzed after the incorporation of a mutant channel in planar lipid bilayer membranes. The modified proteins were studied in the absence and presence of watersoluble specific sulphydryl-specific reagents, in order to introduce a strong positive or negative harge at positions of substitution. The introduction of a negative charge in the stem region onverted the selectivity of the channel from weak anionic to more cationic. However, the troduction of a positive charge increased its selectivity to the anion. The degree of these alterations was inversely dependent on the channel radius at the position of the introduced harge (selectivity). As to the asymmetry of the conductance-voltage, the influence of the harge was more complex. The introduction of the negative charge in the stem region (the trans art of the pore) provoked a decrease. The intensity of these alterations depended on the radius, and on the type of free charge at the pore entrance. These results suggest that the free charge at surrounds the pore wall is responsible for the cation-anion selectivity of the channel. The istribution of the charges between the entrances is crucial for determining the asymmetry of e conductance-voltage curves. We hope that these results serve as a model for studies with other nanometric channels, in biological or planar lipid bilayer membranes or in iotechnological applications

Purificação da glicerina bruta obtida a partir da transesterificação do óleo de algodão

The crude glycerine is a raw material that can be used in a wide variety of products. Even with all the impurities inherent in the process of being obtained, the crude glycerin is already in a marketable product. However, the market is much more favorable to the commercialization of purified glycerine. The glycerin is a byproduct gotten from the process of transesterification of waste oils and fats in the production of biodiesel. More recently, the deployment of the new Federal Law of Brazil, related to the implementation of energy resources, forces, from 2008, the increase of 2% biodiesel in diesel common with prospects for 5% (B5). Therefore, it is indispensable that new routes of purification as well as new markets are developed. The objective of this work was to purify, through ion exchange, the crude glycerin, obtained from the reaction of transesterification of cottonseed oil. The cottonseed oil was characterized as the fatty acid composition and physical-chemical properties. The process of ion exchange was conducted in batch. In this process were used strong cation, low anion resins and a mixed resin used to de-ionize water. The purified glycerin was characterized as the content of metals. Tests were performed with activated charcoal adsorption, and for this, it was made tests of time contact with coal as well as quantity of coal used. The time of activation, the amount of the activation solution, the contact time of the glycerol solution in resins, the amount and type of resin applied were evaluated. Considering the analysis made with activated charcoal, when the glycerin solution was treated using the resins individually it was observed that in the conditions for treatment with 10 g of resin, 5 hours of contact with each resin and 50 mL of glycerin solution, its conductivity decreased to a cationic resin, increased to the anionic resin and had a variable value with respect to resin mixed. In the treatment in series, there was a constant decrease in the conductivity of the solution of glycerin. Considering two types of treatment, in series and individually, the content of glycerol in glycerin pre-purified solution with the different resins varied from 12,46 to 29.51% (diluted solution). In analysis performed without the use of activated charcoal, the behavior of the conductivity of the solution of glycerin were similar to results for treatment with activated charcoal, both in series as individually. The solution of glycerin pre-purified had a glycerol content varying from 8.3 to 25.7% (diluted solution). In relation to pH, it had a behavior in accordance with the expected: acid for the glycerin solution treated with cationic resin, basic when the glycerin solution was treated with the anionic resin and neutral when treated with the mixed resin, independent of the kind of procedure used (with or without coal, resins individually or in series). In relation to the color of the glycerin pre-purified solution, the resin that showed the best result was the anionic (colorless), however this does not mean that the solution is more in pure glycerol. The chromatographic analysis of the solutions obtained after the passage through the resins indicated that the treatment was effective by the presence of only one component (glycerol), not considering the solvent of the analysis

Síntese de zeólitas tipo ton utilizando um sal de imidazólio como agente direcionador de estrutura

This work reports the synthesis of zeolites with different compositions (pure silica, Si/Ti and Si/Al), via hydroxide and fluoride medium using the cation 1-butyl-3- methylimidazolium as structure directing agent. Initially, the cation was synthesized in chloride form and used for the synthesis in hydroxide medium. An anion-exchange (Cl- for OH-) was required for the synthesis in fluoride medium. Different reactants were used for the formation of gels synthesis, resulting in the crystallization of MFI and TON phases, the latter predominant in many compositions. The cation and synthesized zeolites obtained were characterized by different techniques such as NMR, TG/DTG, XRD, SEM, N2 adsorption and desorption, DRS and EPMA. Besides characterizing the cation and zeolites, the mother liquor of hydroxide synthesis was characterized and it was possible to observe a modification of the cation in the synthesis conditions employed. The materials synthesized in this work can be applied in catalytic reactions and adsorption

Síntese, caracterização, propriedades e aplicação dos sistemas Mg- Al, Zn-Al e Mg-Fe

The Layered Double Hydroxides has become extremely promising materials due to its range of applications, easily obtained in the laboratory and reusability after calcination, so the knowledge regarding their properties is of utmost importance. In this study were synthesized layered double hydroxides of two systems, Mg-Al and Zn-Al, and such materials were analyzed with X-ray diffraction and, from these data, we determined the volume density, planar atomic density, size crystallite, lattice parameters, interplanar spacing and interlayer space available. Such materials were also subjected to thermogravimetric analysis reasons for heating 5, 10, 20 and 25 ° C / min to determine kinetic parameters for the formation of metaphases HTD and HTB based on theoretical models Ozawa, Flynn-Wall Starink and Model Free Kinetics. In addition, the layered double hydroxides synthesized in this working ratios were calcined heating 2.5 ° C / min and 20 ° C / min, and tested for adsorption of nitrate anion in aqueous solution batch system at time intervals 5 min, 15 min, 30 min, 1h, 2h and 4h. Such calcined materials were also subjected to exposure to the atmosphere and at intervals of 1 week, 2 weeks and 1 month were analyzed by infrared spectroscopy to study the kinetics of regeneration determining structural called "memory effect"