Recuperação e purificação de ramnolipídeos produzidos por pseudomonas aeruginosa P029-GVIIA utilizando melaço de cana como substrato

Biosurfactants are molecules produced by microorganisms mainly bacteria as Pseudomonas and Bacillus. Among the biosurfactants, rhamnolipids play an important role due to their tensoactive as well as emulsifying properties. Besides can be produced in a well consolidated way the production costs of biosurfactants are quite expansive mainly if downstream processing is goning to be considered. Actually, attention has been given to identification of biosurfactants as well as optimization of its fermentative processes including downstream ones. This work deals with the development of strategies to recovery and purification of rhamnolipids produced by Pseudomonas aeruginosa P029-GVIIA using sugar-cane molasses as substrate. Broth free of cells was used in order to investigate the best strategies to recovery and purification produced by this system. Between the studied acids (HCl and H2SO4) for the acid precipitation step, HCl was the best one as has been showed by the experimental design 24. Extraction has been carried out using petroleum ether and quantification has been done using the thioglycolic acid method. Adsorption studies were carried out with activated carbon in a batch mode using a 24 experimental design as well as combined with an hydrophobic resin Streamline Phenyl aiming to separate the produced biosurfactant. Biosurfactant partial identification was carried out using High Performance Liquid Chromatography (HPLC). Experiments in batch mode showed that adsorption has been controlled mainly by pH and temperature. It was observed a reduction of 41.4% for the liquid phase and the solid phase it was possible to adsorb up to 15 mg of rhamnolipd/g of activated carbon. The kinetics of adsorption has been well fitted to a pseudo-first order reaction with velocity constant (k1) of 1.93 x 10-2 min-1. Experiments in packed bed ranging concentration on eluent (acetone) has been shown the highest recovery factor of 98% when pure acetone has been used. The combined effect if using activated carbon with an hydrophobic resin Streamline Phenyl has been shown successful for the rhamnolipids purification. It has been possible to purify a fraction of the crude broth with 98% of purity when the eluted of activated carbon packed bed was used with pure acetone

Mutagenicity induced by the hydroalcoholic extract of the medicinal plant Plathymenia reticulata Benth

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Análise do potencial genotóxico da superfície de titânio modificada por plasma

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Notes on Newton-Euler formulation of robotic manipulators

The equations corresponding to Newton-Euler iterative method for the determination of forces and moments acting on the rigid links of a robotic manipulator are given a new treatment using composed vectors for the representation of both kinematical and dynamical quantities. It is shown that Lagrange equations for the motion of a holonomic system are easily found from the composed vectors defined in this note. Application to a simple model of an industrial robot shows that the method developed in these notes is efficient in solving the dynamics of a robotic manipulator. An example is developed, where it is seen that with the application of appropriate control moments applied to each arm of the robot, starting from a given initial position, it is possible to reach equilibrium in a final pre-assigned position.

Notes on vibration control of a micro/macromanipulator mounted on a flexible structure

The objective of this study is to describe the design and the implementation of an experimental set-up used to study the dynamics, the experimental identification, and the active vibration control of a flexible structure mounted manipulator system. The system consists of a three-degree-of-freedom cylindrical manipulator system with a flexible link on its tip. A two-degree-of-freedom polar rigid manipulator is mounted on the flexible macromanipulator. The dynamic modelling and experimental modal analysis identification in the frequency domain are being applied to design active digital control strategies for the micro-manipulator system to damp the mechanical vibrations of the flexible structure on the tip of the macro-manipulator system.

Avaliação do caráter oxidante da violaceína

Violacein is a violet pigment isolated from many gram-negative bacteria, especially from Chromobacterium violaceum, a betaproteobacterium found in the Amazon River in Brazil. It has potential medical applications as an antibacterial, fungicide, anti-tryptanocidal, anti-ulcerogenic and anti-cancer drug, among others. Furthermore, its pro-oxidant activity has been suggested, but only in two specific tumor lineages. Thus, in the present study, the prooxidant effects of violacein were investigated in both normal and tumor cells, seeking to evaluate the cell responses. The evaluation of violacein cytotoxicity using the Trypan blue dye exclusion method indicated that CHO-K1 cells were more resistant than tumor HeLa cells. The oxidative stress induced by violacein was manifested as an increase in intracellular SOD activity in CHO-K1 and MRC-5 cells at a specific concentration range. Nevertheless, a decrease was detected specifically at 6-12 μM in HeLa and MRC-5 cells. Interestingly, the increase in SOD activity was not followed by a concomitant increase in catalase activity. Regarding to oxidative stress biomarkers, increased protein carbonylation and lipid hydroperoxides levels were detected respectively in CHO-K1 and MRC-5 cells treated with violacein at 1.5-3 μM and 3 μM, which may be an evidence that this compound causes oxidative stress specifically in these conditions. Additionally, it is believed that the decline in cell viability observed in MRC-5 cells and HeLa treated with violacein at 6-12 M is due to mechanisms not related to oxidative stress. Moreover, the results suggested that violacein might cause oxidative stress by increasing endogenous levels of O2 -, since the occurrence of an expressive change in SOD activity. In addition, in order to evaluate the antioxidant activity of violacein in the absence of a biological system, the total antioxidant and iron chelating activity were evaluated, so that antioxidant activities were detected at 30 and 60 μM of violacein. Altogether, the results indicate that although oxidative stress is triggered by incubation with violacein, it did not seem to be high enough to cause serious damage to cell biomolecules in HeLa cells and only at specific concentrations in CHOK-1 and MRC-5 cells. Comparing the results obtained in cell culture and the in vitro antioxidant activity evaluation, the results confirmed that violacein presents opposing oxidant features when in presence or absence of a biological system and the antioxidant character only occurs at high concentrations of the pigment.

Anti-clastogenic effect of beta-glucan extracted from barley towards chemically induced DNA damage in rodent cells

beta-Glucan (BG) was tested in vitro to determine its potential clastogenic and/or anti-clastogenic activity, and attempts were made to elucidate its possible mechanism of action by using combinations with an inhibitor of DNA polymerase. The study was carried out on cells deficient (CHO-k1) and cells proficient (HTC) in phases I and II enzymes, and the DNA damage was assessed by the chromosomal aberration assay. BG did not show a clastogenic effect, but was anti-clastogenic in both cell lines used, and at all concentrations tested (2.5, 5 and 10 mg/mL) in combination with damage inducing agents (methylmethane sulfonate in cell line CHO-k1, and methylmethane sulfonate or 2-aminoanthracene in cell line HTC). BG also showed a protective effect in the presence of a DNA polymerase beta inhibitor (cytosine arabinoside-3-phosphate, Ara-C), demonstrating that BG does not act through an anti-mutagenic mechanism of action involving DNA polymerase beta.

Determination of the antimutagenicity of an aqueous extract of Rhizophora mangle L. (Rhizophoraceae), using in vivo and in vitro test systems

An aqueous extract of Rhizophora mangle L. bark is used as raw material in pottery making in the State of Espirito Santo, Brazil. This extract presents large quantities of tannins, compounds possessing antioxidant properties. Tannin antioxidant activity, as a plant chemical defense mechanism in the process of stabilizing free radicals, has been an incentive to studies on anti-mutagenicity. The present work aimed to evaluate possible antimutagenic activity of a R. mangle aqueous extract, using the Allium cepa test-system and micronuclear (MN) assay with blockage of cytokinesis in Chinese hamster ovary cells (CHO-K1). The Allium cepa test-system indicated antimutagenic activity against the damage induced by the mutagenic agent methyl methanesulfonate. A reduction in both MN cell frequency and chromosome breaks occurred in both the pre and post-treatment protocols. The MN testing of CHO-K1 cells revealed anti-mutagenic activity of the R. mangle extract against methyl methanesulfonate and doxorubicin in pre, simultaneous and post-treatment protocols. These results suggest the presence of phyto-constituents in the extract presenting demutagenic and bio-antimutagenic activities. Since the chemical constitution of Rhizophora mangle species presents elevated tannin content, it is highly probable that these compounds are the antimutagenic promoters themselves.

Free actions of abelian p-groups on the n-torus

In this work we make some contributions to the theory of actions of abelian p-groups on the n-Torus T-n. Set congruent to Z(pk1)(h1) x Z(pk2)(h2) x...x Z(pkr)(hr), r >= 1, k(1) >= k(2) >=...>= k(r) >= 1, p prime. Suppose that the group H acts freely on T-n and the induced representation on pi(1)(T-n) congruent to Z(n) is faithful and has first Betti number b. We show that the numbers n, p, b, k(i) and h(i) (i = 1,..,r) satisfy some relation. In particular, when H congruent to Z(p)(h), the minimum value of n is phi(p) + b when b >= 1. Also when H congruent to Z(pk1) x Z(p) the minimum value of n is phi(p(k1)) + p - 1 + b for b >= 1. Here phi denotes the Euler function.

Cytotoxicity and genotoxicity of Agaricus blazei methanolic extract fractions assessed using gene and chromosomal mutation assays

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

Trânsito gastrintestinal e digestibilidade aparente de nutrientes em eqüinos alimentados com dietas contendo grãos secos ou silagem de grãos úmidos de triticale

Cinco éguas mestiças (idade e peso corporal médios de seis anos e 480 kg PV) foram distribuídas em delineamento experimental em quadrado latino para se e avaliar a utilização de dietas contendo grãos de triticale (secos ou ensilados) em substituição ao milho na alimentação de eqüinos, por meio do ensaio de avaliação do trânsito gastrintestinal e da digestibilidade. Os tratamentos consistiram de três níveis de grãos de triticale em substituição aos grãos de milho (0, 50 e 100%) e duas formas de conservação dos grãos de triticale (secos ou ensilados). As dietas foram isoprotéicas (12,5% PB), com ingestão diária de MS pelos animais de 2,0% PV (relação volumoso : concentrado de 50:50). Os coeficientes de digestibilidade aparente (CDa) da MS, MO, PB e FDN foram determinados indiretamente. Os parâmetros de trânsito gastrintestinal avaliados foram: k1 (taxa de passagem pelo intestino grosso), k2 (taxa de passagem pelo estômago), TT (tempo de trânsito), TMR (tempo médio de retenção) e TMRT (tempo médio de retenção total). Não houve diferença para os CDa da MS, MO, PB e FDN entre as dietas experimentais, observando-se valores médios de 64,31; 65,14; 74,13 e 57,25%, respectivamente. Considerando-se a cinética das fases sólida e líquida, notou-se efeito somente para k2 na fase sólida da digesta, cujo valor nas rações contendo 100% de triticale (seco ou ensilado) foi de 19,63%/h e na dieta controle (0% de triticale), de 23,72%/h. Observou-se efeito linear crescente para o TT na fase sólida da digesta, com a elevação dos níveis de substituição dos grãos de milho pelos grãos de triticale ensilados. Concluiu-se que os concentrados para eqüinos podem ser formulados com grãos de triticale secos ou ensilados em substituição total ao milho. A inclusão de grãos de triticale na alimentação desta espécie animal promoveu trânsito lento da fase sólida da digesta.

Valor nutritivo e estudo cinético do trato digestivo de dietas contendo grãos secos ou ensilados de sorgo de baixo e alto tanino para eqüinos

Quatro éguas sem raça definida (idade e peso corporal médios de seis anos e 400 kg) foram distribuídas em delineamento experimental em quadrado latino para se avaliar o valor nutritivo e o estudo cinético do trato digestivo de grãos secos ou ensilados de sorgo de baixo e alto conteúdos de tanino na alimentação de eqüinos. Os tratamentos consistiram de dietas contendo dois híbridos de grãos de sorgo (baixo e alto níveis de tanino) e dois métodos de conservação (secos e ensilados). As dietas foram isoprotéicas (12,4% PB), com ingestão diária de MS estabelecida em 1,5% PV (relação feno:concentrado de 50: 50). Os parâmetros de trânsito gastrintestinal avaliados foram: k1 (taxa de passagem pelo intestino grosso), k2 (taxa de passagem pelo estômago), TT (tempo de trânsito), TMR (tempo médio de retenção) e TMRT (tempo médio de retenção total). Os tratamentos não afetaram os coeficientes de digestibilidade aparente (CDa) da MS e do amido, cujos valores médios foram 54,04 e 98,91%, respectivamente. Verificou-se efeito benéfico da ensilagem dos grãos de sorgo de alto conteúdo de tanino sobre a digestibilidade da PB e FDN. A CDa da PB e FDN para a dieta contendo grãos secos de sorgo de alto teor de tanino foi de 49,76 e 32,20% e para as dietas com grãos de sorgo de baixo conteúdo de tanino (seco ou ensilado) e grãos ensilados de sorgo de alto teor de tanino foi de 65,63 e 43,32%, respectivamente. Obteve-se somente efeito do método de conservação dos grãos de sorgo (secos vs ensilados) sobre o TMR, em que o valor para as dietas com silagens de grãos ensilados e secos foi, respectivamente, de 40,08 e 37,9h. Concluiu-se que os grãos de sorgo secos de alto teor de tanino não devem ser usados como principal grão energético nos concentrados para eqüinos, por diminuírem a digestibilidade da proteína e fibra.

Cytotoxicity, genotoxicity and antimutagenicity of hexane extracts of Agaricus blazei determined in vitro by the comet assay and CHO/HGPRT gene mutation assay

Agaricus blazei Murrill ss. Heinem, known as the sun mushroom or himematsutake, is a basidiomycete native to Brazil, which is popular for its medicinal properties. The aim of this study was to test hexane extracts (one fraction and its four sub-fractions) of A. blazei for bioactivity in cultured mammalian cells (CHO-K1). The comet assay, the colony forming assay (CFA) and CHO/HGPRT gene mutation assay were used respectively to determine genotoxicity, cytotoxicity and antimutagenicity of these extracts at different concentrations. The cells were incubated in culture medium and treated for 3 h according to the standard protocol for each assay. The DNA damage-inducing agent ethylmethane sulfonate (EMS) was utilized as the positive control and also in combination with extracts to test for a protective effect. Statistical analysis of the data was performed using analysis of variance (ANOVA) and Tukey's test. A relationship between cytotoxicity and genotoxicity could be established and two extracts EH6B and EH6D showed a protective tendency, while the others did not, with the primary extract EH6 causing the most substantial damage to genetic material. These findings warrant more in-depth studies of the active principles of this mushroom. (c) 2005 Elsevier Ltd. All rights reserved.

Structures of the noncovalent complexes of human and bovine prothrombin fragment 2 with human PPACK-thrombin

Both human and bovine prothrombin fragment 2 (the second kringle) have been cocrystallized separately with human PPACK (D-Phe-Pro-Arg)-thrombin, and the structures of these noncovalent complexes have been determined and refined (R = 0.155 and 0.157, respectively) at 3.3-Å resolution using X-ray crystallographic methods. The kringles interact with thrombin at a site that has previously been proposed to be the heparin binding region. The latter is a highly electropositive surface near the C-terminal helix of thrombin abundant in arginine and lysine residues. These form salt bridges with acidic side chains of kringle 2. Somewhat unexpectedly, the negative groups of the kringle correspond to an enlarged anionic center of the lysine binding site of lysine binding kringles such as plasminogens K1 and K4 and TPA K2. The anionic motif is DGDEE in prothrombin kringle 2. The corresponding cationic center of the lysine binding site region has an unfavorable Arg70Asp substitution, but Lys35 is conserved. However, the folding of fragment 2 is different from that of prothrombin kringle 1 and other kringles: the second outer loop possesses a distorted two-turn helix, and the hairpin β-turn of the second inner loop pivots at Val64 and Asp70 by 60°. Lys35 is located on a turn of the helix, which causes it to project into solvent space in the fragment 2-thrombin complex, thereby devastating any vestige of the cationic center of the lysine binding site. Since fragment 2 has not been reported to bind lysine, it most likely has a different inherent folding conformation for the second outer loop, as has also been observed to be the case with TPA K2 and the urokinase kringle. The movement of the Val64-Asp70 β-turn is most likely a conformational change accompanying complexation, which reveals a new heretofore unsuspected flexibility in kringles. The fragment 2-thrombin complex is only the second cassette module-catalytic domain structure to be determined for a multidomain blood protein and only the third domain-domain interaction to be described among such proteins, the others being factor Xa without a Gla domain and Ca2+ prothrombin fragment 1 with a Gla domain and a kringle. © 1993 American Chemical Society.

Special subsets and optimum codes based on lattices and cosets

This paper introduces the concept of special subsets when applied to generator matrices based on lattices and cosets as presented by Calder-bank and Sloane. By using the special subsets we propose a non exhaustive code search for optimum codes. Although non exhaustive, the search always results in optimum codes for given (k1, V, Λ/Λ′). Tables with binary and ternary optimum codes to partitions of lattices with 8, 9 e 16 cosets, were obtained.