Hidroperóxidos de lipídios como fontes de oxigênio molecular singlete (O2 [1 DELTA g]), detecção e danos em biomoléculas / Lipid hidroperoxides as singlet molecular oxygen precursors (O2 [1 DELTA g]), detection and damage to biomolecules

O estudo do processo da peroxidação de lipídios tem aumentado nos últimos anos, principalmente devido à implicação dos hidroperóxidos de lipídios (LOOH) em diversos processos patológicos. A decomposição destes LOOH é capaz de gerar subprodutos capazes de promover danos em biomoléculas, incluindo proteínas e DNA. No presente trabalho, utilizando hidroperóxidos de ácido linoléico isotopicamente marcado com átomo de oxigênio-18 (LA18O18OH), fomos capazes de demonstrar que estas moléculas gerararam oxigênio singlete marcado [18(1O2)] em células em cultura. A detecção de tal espécie foi possível através da utilização de uma nova metodologia utilizando um derivado antracenico. Para este propósito foi utilizado o derivado de antraceno 3,3'-(9,10-antracenodiil) bisacrilato (DADB), cujo produto especifico da reação com o 1O2 (o endoperóxido do DADB DADBO2) do pode ser facilmente detectado por HPLC-MS/MS. De forma a expandir a compreensão dos efeitos tóxicos desses LOOH, investigamos o efeito destes compostos gerados intracelularmente. Para tal, foi utilizado o Rosa bengala (RB), um fotosensibilizador que tem afinidade por espaços apolares como membranas e lisossomos. A fotosenssibilização deste composto foi capaz de induzir a morte celular, e esta morte estaria relacionada a uma maior formação de 1O2 e a um maior acumulo de peróxidos. Nestes estudos foi possível demonstrar que carotenóides e sistemas antioxidantes dependentes de glutationa foram capazes de proteger contra os efeitos tóxicos da fotosensibilização na presença de RB. Adicionalmente foram avaliados os efeitos da hemoglobina (Hb) e do hidroperóxido do ácido linoléico (LAOOH) em uma série de parâmetros toxicológicos, como citotoxicidade, estado redox, a peroxidação lipídica e dano ao DNA. Nós demonstramos que a pré-incubação das células com Hb e sua posterior exposição à LAOOH (Hb + LAOOH) levou a um aumento na morte celular, a oxidação do DCFH, formação de malonaldeído e fragmentação do DNA e que esses...

The study of the process of lipid peroxidation has increased in recent years, mainly due to the involvement of lipid hydroperoxide (LOOH) in a series of pathological processes. The decomposition of LOOH is able to generate products that can promote damage to biomolecules, including proteins and DNA. In the present work, using linoleic acid hydroperoxide isotopically labeled with 18O2 (LA18O18OH), we demonstrate that these molecules were able to generate labeled singlet oxygen [18(1O2)] in cultured cells. The detection of such species was possible using a new methodology using an anthracene derivative .For this purpose we used the anthracene derivative of 3,3'-(9,10-antracendiil) bisacrilate (DADB), whose specific reaction product with 1O2 (DADB endoperoxide DADBO2) can be easily detected by HPLC-MS/MS. In order to expand the understanding of the toxic effects of LOOH, we investigated the effect of these compounds generated intracellularly. For this porpoise, we used Rose Bengal (RB), a photosensitizer that has affinity for apolar spaces such as membranes and lysosomes. The photosensitization of this compound was able to induce cell death, and this death was related to increased formation of 1O2 and a higher accumulation of peroxides. In these studies we have shown that carotenoids and glutathione-dependent antioxidant systems were capable of protecting against the toxic effects of photosensitization in the presence of RB. Additionally, we evaluated the effects of hemoglobin (Hb) and linoleic acid hydroperoxide (LAOOH) in a series of toxicological endpoints such as cytotoxicity, redox status, lipid peroxidation and DNA damage. We demonstrated that preincubation of cells with Hb and its subsequent exposure to LAOOH (Hb + LAOOH) led to an increase in cell death, DCFH oxidation, formation of malonaldehyde and DNA fragmentation, and that these effects were related to the peroxide and the heme group. It was demonstrated that cells incubated with LAOOH and Hb showed...

Determination of lead, naphthalene, phenanthrene, anthracene and pyrene in street dust

Street dust is a potential source of lead exposure to humans, however scarce information about the pollution levels with lead and polyromatic hydrocarbons exists in Venezuela, limiting the appropriate evaluation of the levels of risk of the people. This work was aimed in the determination of the concentrations of lead, naphtalene, anthracene, phenanthrene and pyrene in the street dust of the most transited avenues and streets of Maracay city. Thirty street dust samples were collected at the streets and avenues, troll and bus main station. Lead was determined by atomic absorption spectrometry after acid digestion [Pb-total], also the fractions of lead soluble in 1 M MgCI2 and 0.5 M ammonium acetate [pH = 7] were quantified. The polyaromatic hydrocarbons concentrations were determined by capillary gas chromatography equipped with a flame ionization detector. The Pb-total ranged between 734 and 11.439 micro g/g with the higher values at the most transited streets and avenues. About 60% of samples exhibited concentrations between 1.000 and 2.500 micro g/g, similar to the values reported in the literature for soils of urban areas. The fraction of lead soluble in magnesium accounted for less than 3% of Pb-Total, while the 0.5 M ammonium acetate solution represented more than the 75% of the total loads of the pollutant. The most contaminated samples were those taken at the toll with concentrations of 695.5 and 252.1 micro g/g phenanthrene and anthracene were the most abundant, while at the bus station all compounds were detected

Presenca de compostos antracenicos em folhas de Cassia fastuosa Willd / Presence of anthracenic compounds in leaves of Cassia fastuosa Willd

Foram submetidas a tecnicas de extracao para compostos antracenicos de Cassia fastuosa Willd. Constatada a presenca de aloemodina, reina, emodina, senoside A e de senoside B, foram procedidos de doseamentos da reina e do senoside B. Alem disso, foi realizado ensaio biologico preliminar, com o extrato aquoso a 10%, para a verificacao da atividade laxante.