The structure of a native L-amino acid oxidase, the major component of the Vipera ammodytes ammodytes venomic, reveals dynamic active site and quaternary structure stabilization by divalent ions

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

Vochysiaceae: secondary metabolites, ethnopharmacology and pharmacological potential

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

Avaliação da atividade mutagênica do extrato etanólico bruto de Paepalanthus latipes (Eriocaulaceae) e dos compostos flavonoídicos 7-metoxilados relacionados

Paepalanthus latipes (Silveira) é uma das plantas brasileiras nativas da Serra do Cipó, Minas Gerais, conhecida como sempre-vivas e muito exportada para os Estados Unidos, Europa e Japão, por seu valor ornamental. Poucos resultados são relatados quanto à atividade biológica destas plantas. Este trabalho teve como objetivo avaliar a atividade mutagênica do extrato etanólico bruto de P. latipes e de dois flavonóides 7-metoxilados dele isolados (7-metilquercetagetina e 7-metilquercetagetina-4'-O-β-D-glicopiranosídeo), através do teste de Ames, utilizando linhagens mutantes de Salmonella typhimurium TA98 e TA100. A atividade mutagênica das amostras testadas, em concentrações variando de 62,5 a 375 μg/placa, foi determinada com ativação metabólica (utilizando-se a fração microssomal S9) e na ausência dessa. Os resultados demonstraram que, tanto o extrato etanólico bruto de P. latipes quanto os flavonóides 7-metoxilados testados, não induziram atividade mutagênica nas linhagens TA98 e TA100.

Indução de Metabólitos Bioativos em Culturas de Células de Maytenus Ilicifolia

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

Carbon dioxide efflux in a rhodic hapludox as affected by tillage systems in southern Brazil

Os solos agrícolas podem atuar como dreno ou fonte de C atmosférico, dependendo do sistema de manejo adotado. Este estudo foi desenvolvido em experimento de longa duração (22 anos), durante o período de 30 dias do outono, com o objetivo de avaliar o impacto de sistemas de preparo de solo (preparo convencional-PC e plantio direto-PD) nas emissões de C-CO2 de um Latossolo Vermelho distrófico, em Cruz Alta, RS. As emissões de C-CO2 do solo foram avaliadas com câmaras dinâmica (Flux Chamber 6400-09, Licor) e estática (com captação em solução alcalina), imediatamente após a colheita da soja. A temperatura e a umidade do solo foram registradas, concomitantemente com as emissões de C-CO2, por meio de sensor de temperatura e TDR manual, respectivamente, integrantes do Licor-6400. Estimou-se que, em 30 dias, uma quantidade equivalente a menos de 30 % do C aportado pelos resíduos de soja foi emitida na forma de C-CO2. As emissões de C-CO2 no solo em PD foram similares às emissões do solo em PC, independentemente do tipo de câmara utilizada. Diferenças entre sistemas de preparo quanto à emissão de C-CO2, avaliadas com a câmara dinâmica, foram verificadas somente a curto prazo (leituras diárias), com o PD apresentando maiores emissões do que o PC no início do período experimental e menores no final. A câmara dinâmica foi mais eficiente do que a estática em captar as alterações das emissões de C-CO2 em função da variação da temperatura e a porosidade preenchida por água (PPA) no solo em PD, as quais explicaram 83 e 62 % das emissões de C-CO2, respectivamente. O fator Q10, que avalia a sensibilidade da emissão de C-CO2 à temperatura do solo, foi estimado em 3,93, indicando alta sensibilidade da atividade microbiana à temperatura do solo durante o outono. As emissões de C-CO2 registradas no solo em PD com a câmara estática foram correlacionadas às da câmara dinâmica, porém com valores subestimados em relação àquela notadamente nos maiores valores de fluxo. em condições de baixa temperatura e PPA, o preparo de solo induziu limitado incremento de emissão de C-CO2.

Electrochemical study of hydroxychloroquine and its determination in plaquenil by differential pulse voltammetry

Hydroxychloroquine (HCQ) is a halogenated aminoquinoline that presents wide biological activity, often being used as an antimalarial drug. The electrochemical reduction of HCQ was investigated by cyclic voltammetry and chronoamperometry using glassy carbon electrodes. By cyclic voltammetry, in acid medium, only the cathodic peak was observed. The electrochemical behavior of this peak is dependent on pH and the electrodic process occurs through an ErCi mechanism. The electron number (le) consumed in the reduction of HCQ was obtained by chronoamperometry. A method for the electrochemical determination of HCQ in pharmaceutical tablets was developed using differential pulse voltammetry. The detection limit reached was 11.2 mug ml(-1) of HCQ with a relative standard deviation of 0.46%. A spectrophotometric study of HCQ has been also carried out utilizing a band at 343 nm. The obtained detection limit and the relative standard deviation were 0.1 mug ml(-1) and 0.36%, respectively. The electrochemical methods are sufficiently accurate and precise to be applied for HCQ determination, in laboratorial routine, which can be used to determine the drug at low level. (C) 2003 Elsevier B.V. B.V. All rights reserved.

Combinatorial synthesis and directed evolution applied to the production of alpha-helix forming antimicrobial peptides analogues

Antimicrobial peptides (AMPs) are effector molecules of innate immune systems found in different groups of organisms, including microorganisms, plants, insects, amphibians and humans. These peptides exhibit several structural motifs but the most abundant AMPs assume an amphipathic alpha-helical structure. The alpha-helix forming antimicrobial peptides are excellent candidates for protein engineering leading to an optimization of their biological activity and target specificity. Nowadays several approaches are available and this review deals with the use of combinatorial synthesis and directed evolution in order to provide a high-throughput source of antimicrobial peptides analogues with enhanced lytic activity and specificity.

Dissociation and electrooxidation of primaquine diphosphate as an approach to the study of anti-chagas prodrugs mechanism of action

This paper describes the voltammetric behavior of primaquine as a previous support to the further understanding of the delivery and action mechanisms of its respective synthesized prodrugs. There are few papers describing the drug behavior and most of the time no correlation between oxidation process and pH is done. Our results showed that primaquine oxidation is a one-step reaction involving two electrons with the charge transfer process being strongly pH-dependent in acid medium and pH-independent in a weak basic medium, with the neutral form being easily oxidized.This leads to the conclusion that quinoline nitrogen ring neutralization is a determinant step to the formation of the oxidized primaquine form. The existence of a relationship between the primaquine dissociation equilibrium and its electrooxidation process is shown.This work points the importance of voltammetric methodology as a tool for further studies on quantitative relationship studies between chemical structure and biological activity (QSAR) for electroactive drugs. (C) 2000 Elsevier B.V. S.A. All rights reserved.

Lithium thiazolidine-4-carboxylate: Synthesis, spectroscopic characterization and preliminary in vitro cytotoxic studies in human HeLa cells

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

Isolamento e avaliação da atividade biológica dos alcalóides majoritários de Tabernaemontana angulata Mart. ex Müll. Arg., Apocynaceae

"Isolation and evaluation of the biological activity related to the major alkaloids in Tabernaemontana angulata Mart. ex Mull Arg., Apocynaceae." Introducing-new chemotherapeutic agents is a, great demand. in the control of infections diseases.' Brazil is one of the richest countries in biodiversity and the Laboratorio de Extracao at UNIP has been collecting plants from. the Amazon and Atlantic Rain Forests with the aim of screening for new antibacterial and antitumor plant extracts. Previous studies demonstrated that the ethanol fraction obtained from the crude extract of Tabernaemontana angulata stems showed antibacterial activity against Staphylococcus aureus (ATCC 6538 in the microdilution broth assay. Two alkaloids were the major compounds in the active fraction, verified by thin layer chromatography analysis. In the present study, the total alkaloids were obtained from the crude extract and were fractionated by preparative thin layer chromatography for the isolation of the main components. The isolated. compounds were identified by GC/MS and (1)H-NMR as coronaridine,e and voacangine. The alkaloid fractions obtained from the isolation procedure were tested for antibacterial activity, but no activity was detected.

Isocoumarin from Paepalanthus bromelioides

The new isocoumarin, 9,10-dihydroxy-5, 7-dimethoxy-1H-naphtho(2,3c)pyran-1-one, was isolated from the capitula of Paepalanthus bromelioides. The structure was established by means of the spectral data of the natural product and its derivatives. The natural isocoumarin isolated has strong antibiotic activity. © 1990.

Paracoccidioides brasiliensis antigen batches from the same isolate show immunological and biochemical differences

We investigated the occurrence of antigenic and biochemical variability among Paracoccidioides brasiliensis antigen batches prepared according to the same protocol. Initially (experiment 1), we analyzed two antigen lots of two human isolates (Bt1 and Bt2), cultured in two media (PYG: bactopeptone, yeast extract, glucose; MMM: McVeigh and Morton medium) in SDS-PAGE and in two immunological tests (immunodiffusion-ID and footpad swelling test-FPT). Afterwards (experiment 2), we compared the antigenic profile of three antigen hatches from three human isolates (Bt1, Bt2 and Bt3) by two-dimensional immunoelectrophoresis (2 D-IEP) against a reference system for P. brasiliensis antigens. In experiment 1, there were important intra- and inter-strain antigenic differences between batches of the fungal isolates cultured on both media. The block titration of the antigen batches for the immunological tests revealed correlation between protein concentration and biological activity in ID and no correlation in FPT. In experiment 2, the reference system for P brasiliensis showed 26 antigen peaks. There were important differences between batches prepared from the same isolate and between batches from different isolates. Our data suggested the occurrence of instability in the synthesis of antigenic components by a same P. brasiliensis isolate, under controlled incubation conditions.

Fator de crescimento derivado de plaquetas humanas obtido por uma metodologia rápida e de baixo custo

Human platelet-derived growth factor (PDGF) was purified from lysates of clinically outdated human platelets by ionic exchange chromatography in CM-Sepharose. The eluated fraction was submitted to the Immunoblot/Slot Blot assay using anti-PDGF-AA and anti-PDGF-BB polyclonal antibodies and was evaluated as to its biological activity through the test of [H 3]-thymidine incorporation in NIH/3T3 cell line fibroblasts in culture. The Immunoblot/Slot Blot assay using anti-PDGF-AA and anti-PDGF-BB antibodies proved the presence of the PDGF in chromatographic cationic fraction. The comparison of biological activities between fiblobrast stimulation assay using recombinant PDGF-AB and partially purified PDGF was demonstrated in 165.796 and 157.567 cpm, respectively. This result, proved the potent mitogenic effect of partially purified PDGF and consequently their evidence about the wound healing activity.

Estratégias biotecnológicas para a liberação controlada de antígenos e adjuvantes em vacinas através de lipossomas

Liposomes (LP) are colloidal systems with ability to compartmentalize therapeutic molecules in order to improve biological activity, decreases the potential toxicity, and to obtain prolonged effect. In this work it was discussed the role of the various liposomes types to encapsulate drug molecules able to provoke some immunological response (drugs, antigens and DNA). The effect of the liposomes and the parameters about the formation of the structures are also analyzed. Detailed literature review shows that, depending on the molecules polarity and the superficial charge of the liposome structures, the system may be efficiently used to optimize the therapeutic effects by means of the release control or through a drug delivery mechanism.

Atividade microbiana em um Latossolo degradado tratado com lodo de esgoto

The degraded soil shows, in general, poor biological activity, considering its physical characteristics, low fertility and organic matter, mainly due to removal or degradation of its superficial layer. The sewage sludge, due to its high content of easily decomposed organic matter can be an alternate source of organic residues and combined to its high content of the principal nutrients for the plants can be an important factor to promote biological activities in degraded soil. In order to study the actions of the sewage sludge in the recovery of a degraded Latosol, the carbon in the microbial biomass (C mic), the carbon released CO2 (C-CO2) and the relation between microbial and organic carbon (Cmic/Corg) were used as indicators of the effects. To do so, two doses (30 and 60 Mg ha-1) of sewer slime applied in topdressing and incorporated together with a mineral fertilizer treatment, using the eucalyptus as a test crop. A completely randomized design with 4 treatments and 4 repetitions was used. The sewage sludge promoted increase of liberated C-CO2 and the C mic, which constitute the adequate quality indicators for monitoring the soil recovery.