A strategy for the rapid identification of fungal metabolites and the discovery of the antiviral activity of pyrenocine a and harzianopyridon

The isolation and identification of bioactive metabolites from complex extracts obtained from microbial growth media is a time consuming, costly, and labor-intensive task. A strategy to rapidly identify secondary metabolites isolated from extracts obtained from the culture media of marine-derived and endophytic fungal strains is described. Identification was achieved by HPLC-UV-MS and 1H NMR analyses in combination with data obtained from the Dictionary of Natural Products. Among the compounds identified, (-)-naphthoquinoneimine, citreorosein, emodin, pyrenocine A and harzianopyridone displayed moderate to potent antiviral activity. (-)-Naphthoquinoneimine was isolated as the enantiomer of its previously reported dextrorotatory congener, while 6,7-dihydroxy-2,2-dimethyl-4-chromanone is herein reported for the first time as a natural product396720731CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPBEX 4498-14-32010/50190-2; 2013/50228-8; 2011/08064-2; 2008/00331-9; 2013/23153-

A cyclopalladated complex interacts with mitochondrial membrane thiol-groups and induces the apoptotic intrinsic pathway in murine and cisplatin-resistant human tumor cells

<p>Abstract</p> <p>Background</p> <p>Systemic therapy for cancer metastatic lesions is difficult and generally renders a poor clinical response. Structural analogs of cisplatin, the most widely used synthetic metal complexes, show toxic side-effects and tumor cell resistance. Recently, palladium complexes with increased stability are being investigated to circumvent these limitations, and a biphosphinic cyclopalladated complex {Pd₂[<it>S_(-)</it>C², N-dmpa]₂(μ-dppe)Cl₂} named C7a efficiently controls the subcutaneous development of B16F10-Nex2 murine melanoma in syngeneic mice. Presently, we investigated the melanoma cell killing mechanism induced by C7a, and extended preclinical studies.</p> <p>Methods</p> <p>B16F10-Nex2 cells were treated <it>in vitro </it>with C7a in the presence/absence of DTT, and several parameters related to apoptosis induction were evaluated. Preclinical studies were performed, and mice were endovenously inoculated with B16F10-Nex2 cells, intraperitoneally treated with C7a, and lung metastatic nodules were counted. The cytotoxic effects and the respiratory metabolism were also determined in human tumor cell lines treated <it>in vitro </it>with C7a.</p> <p>Results</p> <p>Cyclopalladated complex interacts with thiol groups on the mitochondrial membrane proteins, causes dissipation of the mitochondrial membrane potential, and induces Bax translocation from the cytosol to mitochondria, colocalizing with a mitochondrial tracker. C7a also induced an increase in cytosolic calcium concentration, mainly from intracellular compartments, and a significant decrease in the ATP levels. Activation of effector caspases, chromatin condensation and DNA degradation, suggested that C7a activates the apoptotic intrinsic pathway in murine melanoma cells. In the preclinical studies, the C7a complex protected against murine metastatic melanoma and induced death in several human tumor cell lineages <it>in vitro</it>, including cisplatin-resistant ones. The mitochondria-dependent cell death was also induced by C7a in human tumor cells.</p> <p>Conclusions</p> <p>The cyclopalladated C7a complex is an effective chemotherapeutic anticancer compound against primary and metastatic murine and human tumors, including cisplatin-resistant cells, inducing apoptotic cell death via the intrinsic pathway.</p

Evaluation of the process of reabilitation of a stretch of Riparian forest in Itapemirim river watershed - ES

Este estudo foi realizado em uma área de floresta ciliar em processo de recuperação mediante reabilitação.A área de estudo está localizada na sub-bacia hidrográfica do rio Itapemirim, no Município de Alegre, ES, Brasil.A ocupação e uso do solo antes da revegetação eram de pastagem com Brachiaria sp. A revegetação da áreafoi feita em 1997, com espécies autóctones e alóctones arbóreas, em arranjo de distribuição aleatório, em umaárea de 1,2 ha. Para a realização dos estudos foram feitos inventários florestais nos períodos de 2004/2005e 2005/2006, sendo medidos os indivíduos de hábito arbustivo e arbóreo com circunferência à altura do peito(CAP) > 5 cm e suas alturas totais. As espécies encontradas na área foram identificadas e classificadas de acordocom seus grupos ecológicos, síndromes de dispersão e presença silvestre, sendo calculados os parâmetros florísticos,a estrutura vertical e a dinâmica estrutural desse povoamento. O objetivo do trabalho foi avaliar o desenvolvimentodo povoamento implantado para subsidiar práticas silviculturais quanto à seleção e implantação de espécies pararevegetação de áreas de floresta ciliar degradadas, em condições semelhantes. Os resultados demonstraram quefoi implantado um povoamento florestal com grande diversidade de espécies e a estratificação em classes dealtura foi à semelhança de povoamentos heterogêneos naturais. As espécies identificadas como edificadoras darevegetação da área estudada foram: Anadenanthera colubrina, Caesalpinia leyostachia, Acacia auriculiformis,Acacia mangium, Handroanthus serratifolius, Inga edulis, Joannesia princeps, Pterogyne nitens, Enterelobiumcontortisiliquum, Tabernaemontana hystrix e Anthocephalus indicus. A distribuição em classes de tamanho dacomunidade implantada ocorre em forma de "J" reverso, havendo a predominância de indivíduos pioneiros emtodas as classes de CAP. A dinâmica da estrutura horizontal apontou que, para o sucesso, continuidade e desenvolvimentoda recuperação da área, seja monitorada a regeneração natural em relação à sua presença e à eficiência dos fatoresbióticos e abióticos que nela interferem. A não observância de indivíduos arbustivos e arbóreos regenerados naturalmente,na classe de inclusão do estudo, indica a fragilidade inicial da área rumo à sustentabilidade do sistema.The study was realized in an area of riparian forest in process by means of rehabilitation. Thestudy area is located in Rio Itapemirim, in Alegre city, Brazil. The occupation and use of the soil beforethe recovery were of pasture with Brachiaria sp. The recovery of this area was in 1997, with autochthonousand allochthonous species, in casually distribution arrangement, in an area of 1,2 ha. For this studies wasrealized forest inventories in the periods of 2004/2005 and 2005/2006, being measured the arboreal individualswith circumference at breast height (CBH) and total heights. The species found in the area were identifiedand classified in this ecological groups, dispersion syndromes and wild presence, it forms calculated thefloristic parameters, the vertical structure and the structural dynamics of this plantation. The objective ofthis work went evaluate the development of the plantation implanted to subsidize practical silviculture withrelationship to the selection and plantation of species for the revegetation of degraded areas of riparianforest, in similar conditions. The results of the studies demonstrated that a forest plantation was implantedwith great diversity of species and the bedding in height classes it went to the likeness of natural heterogeneous.The species identified as builders of the plantation of the studied area were: Anadenanthera colubrina, Caesalpinia leyostachia, Acacia auriculiformis, Acacia mangium, Handroanthus serratifolius, Inga edulis, Joannesia princeps,Pterogyne nitens, Enterelobium contortisiliquum, Tabernaemontana hystrix, and Anthocephalus indicus . Theimplanted community's distribution diametric happens in reverse J shape having the pioneer individuals'predominance in all the classes of CBH. The dynamics of the horizontal structure points that for the success,continuity and development of the recovery of the area, the natural regeneration be monitored in relationto its presence and efficiency of the biotic factors and abiotic those interfere in the same. The not tree individualsobservance in natural regeneration, in the class of inclusion of the study, indicate the initial fragility of thearea heading for sustentabilidade of the system

Neutrophil Paralysis in Plasmodium vivax Malaria

Plasmodium vivax is responsible for approximately 60–80% of the malaria cases in the world, and contributes to significant social and economic instability in the developing countries of Latin America and Asia. The pathogenesis of P. vivax malaria is a consequence of host derived inflammatory mediators. Hence, a better understanding of the mechanisms involved in induction of systemic inflammation during P. vivax malaria is critical for the clinical management and prevention of severe disease. The innate immune receptors recognize Plasmodium sp. and initiate a broad spectrum of host defense mechanisms that mediate resistance to infection. However, the innate immune response is the classic “two-edged sword”, and clinical malaria is associated with high levels of circulating pro-inflammatory cytokines. Our findings show that both monocytes and neutrophils are highly activated during malaria. Monocytes produced high levels of IL-1β, IL-6 and TNF-α during acute malaria. On the other hand, neutrophils were a poor source of cytokines, but displayed an enhanced phagocytic activity and superoxide production. Unexpectedly, we noticed an impaired chemotaxis of neutrophils towards an IL-8 (CXCL8) gradient. We proposed that neutrophil paralysis is in part responsible for the enhanced susceptibility to bacterial infection observed in malaria patients

Evaluation of the impact of orally administered carbohydrates on postprandial blood glucose levels in different pre-clinical models

ABSTRACT We developed a pre-clinical model in which to evaluate the impact of orally administered carbohydrates on postprandial blood glucose levels. For this purpose, we compared the effects of different carbohydrates with well-established glycemic indexes. We orally administered (gavage) increasing amounts (0.2, 0.4, 0.6, 0.8, and 1.0 g/kg) of sucrose and lactose to rats which had been fasted for 6 h or 15 h, respectively. In part of the experiments we administered frutose (gavagem). Three different models were compared for measuring postprandial blood glucose levels: a) evaluation of interstitial glucose concentrations by using a real time continuous glucose monitoring system; b) evaluation of glucose levels in blood obtained from the rat tail; c) evaluation of serum glucose levels in blood collected after decapitation. Our results showed that blood obtained from the tails of 15-h fasted rats was the best model in which to evaluate the effect of carbohydrates on postprandial blood glucose levels

Production of vegetable oil blends and structured lipids and their effect on wound healing

Two oil blends (sunflower/canola oils 85/15 (BL1) and canola/linseed oils 70/30 (BL2)), were prepared and enzymatically interesterified to be applied to surgically-induced wounds in rats. Following surgery, the animals were submitted to the Treatment with Physiological Saline (TPS) (control group), Blends (TBL), and Structured Lipids (TSL). The control group (TPS) received physiological saline solution for 15 days. In TBL, BL1 was administered during the inflammation phase (days 0-3) and BL2 in the tissue formation and remodeling phase (days 4-15). In TSL, Structured Lipid 1 (SL1) and Structured Lipid 2 (SL2) were used instead of BL1 and BL2, respectively. The aim of this study was to compare wound closure evolution among rats treated with the blends or structured lipids versus control rats treated with physiological saline. The wound healing process was evaluated by measuring the wound areas along the treatments and the concentrations of cytokines. An increase in the areas of wounds treated with the blends and structured lipids in the inflammatory phase was observed, followed by a steeper closure curve compared to wounds treated with physiological saline. The changes observed during the inflammatory phase suggest a potential therapeutic application in cutaneous wound healing which should be further investigated.</p