Trypanosoma cruzi iron superoxide dismutases: insights from phylogenetics to chemotherapeutic target assessment.

BACKGROUND: Components of the antioxidant defense system in Trypanosoma cruzi are potential targets for new drug development. Superoxide dismutases (SODs) constitute key components of antioxidant defense systems, removing excess superoxide anions by converting them into oxygen and hydrogen peroxide. The main goal of the present study was to investigate the genes coding for iron superoxide dismutase (FeSOD) in T. cruzi strains from an evolutionary perspective. METHODS: In this study, molecular biology methods and phylogenetic studies were combined with drug assays. The FeSOD-A and FeSOD-B genes of 35 T. cruzi strains, belonging to six discrete typing units (Tcl-TcVI), from different hosts and geographical regions were amplified by PCR and sequenced using the Sanger method. Evolutionary trees were reconstructed based on Bayesian inference and maximum likelihood methods. Drugs that potentially interacted with T. cruzi FeSODs were identified and tested against the parasites. RESULTS: Our results suggest that T. cruzi FeSOD types are members of distinct families. Gene copies of FeSOD-A (n = 2), FeSOD-B (n = 4) and FeSOD-C (n = 4) were identified in the genome of the T. cruzi reference clone CL Brener. Phylogenetic inference supported the presence of two functional variants of each FeSOD type across the T. cruzi strains. Phylogenetic trees revealed a monophyletic group of FeSOD genes of T. cruzi TcIV strains in both distinct genes. Altogether, our results support the hypothesis that gene duplication followed by divergence shaped the evolution of T. cruzi FeSODs. Two drugs, mangafodipir and polaprezinc, that potentially interact with T. cruzi FeSODs were identified and tested in vitro against amastigotes and trypomastigotes: mangafodipir had a low trypanocidal effect and polaprezinc was inactive. CONCLUSIONS: Our study contributes to a better understanding of the molecular biodiversity of T. cruzi FeSODs. Herein we provide a successful approach to the study of gene/protein families as potential drug targets.

Effect of Experimental Parameters on the Extraction of Grape Seed Oil Obtained by Low Pressure and Supercritical Fluid Extraction.

Grape seeds are an important byproduct from the grape process. The objective of this work was to evaluate the effect of experimental parameters (temperature and time of pretreatment with ultrasound) to obtain grape seed oil using low pressure (Soxhlet-Sox and Bligh Dyer-BD) and high pressure (supercritical carbon dioxide-SFE) methods. The best condition for pretreatment of samples was 30 min of sonication at 30 °C before extraction by Sox or BD. Ultrasound pretreatment was efficient to increase oil extraction yield by 32.10 (Sox), 20.31 (BD) and 12.54% (SFE), depending on the extraction method used as well as, and certainly influenced the total phenolic concentration in 311 (Sox), 234 (BD), and 184 (SFE)%. Ten fatty acids were identified in the oils, the major ones being 18:2ω-6cis (linoleic 52.39%-63.12%), 16:0 (palmitic 20.22%-26.80%) and 18:0 (stearic 8.52%-13.68%). The highest epicatechin concentration was identified in the BD sample: 30-30 (150.49 ± 5.98mg/kg), which presented a concentration of ≥3 times compared to the control (56.68 ± 1.81mg/kg). Ultrasound pretreatment also contributed positively (56% and 99% increase) in the α-tocopherol content of the SFE: 30-30 and BD: 30-30 samples, respectively. The results indicate that the ultrasound pretreatment is a suitable technology to improve the quality of the oil from the grape seed.