Combined Strategies to Improve the Expression of Recombinant Sterol C24-Methyltransferase from <Emphasis Type="Italic">Leishmania braziliensis</Emphasis> in <Emphasis Type="Italic">E. coli</Emphasis>

Among the neglected tropical diseases, leishmaniasis stands out for its worldwide distribution and diversity of symptoms. Cutaneous leishmaniasis (CL), for instance, is endemic in 18 countries, but the available drugs to fight it have high toxicity and low patient adherence. In order to overcome this, dilemma drugs that target enzymes which are absent in the human host, such as Leishmania braziliensis sterol C24-methyltransferase (SMT-C24, EC 2.1.1.41), are needed. However, medicinal chemistry efforts toward this goal have been hampered by the low yield of soluble recombinant SMT-C24 afforded by currently available expression systems. Herein, we show that a combination of molecular biology and chromatographic strategies may increase the yield of LbSMT-C24 in up to fivefold. These results lay the ground for future investigation of this enzyme as a drug target.     

Mitochondria Bound to Lipid Droplets Have Unique Bioenergetics,Composition, and Dynamics that Support Lipid Droplet Expansion

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Phylogeography of the tree Hymenaea stigonocarpa (Fabaceae: Caesalpinioideae) and the influence of quaternary climate changes in the Brazilian cerrado

BACKGROUND AND AIMS: Hymenaea stigonocarpa (Fabaceae: Caesalpinioideae) is an endemic tree from the Brazilian cerrado (savanna vegetation), a biome classified as a hotspot for conservation priority. This study investigates the phylogeographic structure of H. stigonocarpa, in order to understand the processes that have led to its current spatial genetic pattern. METHODS: The polymorphism level and spatial distribution of variants of the plastid non-coding region between the genes psbC and trnS were investigated in 175 individuals from 17 populations, covering the greater part of the total distribution of the species. Molecular diversity indices were calculated and intra-specific relationships were inferred by the construction of haplotype networks using the median-joining method. Genetic differentiation among populations and main geographical groups was evaluated using spatial analysis of molecular variance (SAMOVA). KEY RESULTS: Twenty-three different haplotypes were identified. The level of differentiation among the populations analysed was relatively high (F(ST) = 0.692). Phylogeographic analyses showed a clear association between the haplotype network and geographic distribution of populations, revealing three main geographical groups: western, central and eastern. SAMOVA corroborated this finding, indicating that most of the variation can be attributed to differences among these three groups (58.8 %), with little difference among populations within groups (F(SC) = 0.252). CONCLUSIONS: The subdivision of the geographic distribution of H. stigonocarpa populations into three genetically differentiated groups can be associated with Quaternary climatic changes. The data suggest that during glacial times H. stigonocarpa populations became extinct in most parts of the southern present-day cerrado area. Milder climatic conditions in the north and eastern portions of the cerrado resulted in maintenance of populations in these regions. Thus it is inferred that the most southern part of the present-day cerrado was re-colonized by different lineages from northern parts of this biome, after postglacial climate amelioration.     

Taxonomical and functional microbial community selection in soybean rhizosphere

This study addressed the selection of the rhizospheric microbial community from the bulk soil reservoir under agricultural management of soybean in Amazon forest soils. We used a shotgun metagenomics approach to investigate the taxonomic and functional diversities of microbial communities in the bulk soil and in the rhizosphere of soybean plants and tested the validity of neutral and niche theories to explain the rhizosphere community assembly processes. Our results showed a clear selection at both taxonomic and functional levels operating in the assembly of the soybean rhizosphere community. The taxonomic analysis revealed that the rhizosphere community is a subset of the bulk soil community. Species abundance in rhizosphere fits the log-normal distribution model, which is an indicator of the occurrence of niche-based processes. In addition, the data indicate that the rhizosphere community is selected based on functional cores related to the metabolisms of nitrogen, iron, phosphorus and potassium, which are related to benefits to the plant, such as growth promotion and nutrition. The network analysis including bacterial groups and functions was less complex in rhizosphere, suggesting the specialization of some specific metabolic pathways. We conclude that the assembly of the microbial community in the rhizosphere is based on niche-based processes as a result of the selection power of the plant and other environmental factors.     

Endophytic bacteria in long-term in vitro cultivated “axenic” pineapple microplants revealed by PCR–DGGE

In vitro propagated plants are believed to be free of microbes. However, after 5 years of in vitro culture of pineapple plants, without evidence of microbial contamination, the use of culture-independent molecular approach [classifying heterogeneous nucleic acids amplified via universal and specific 16S rRNA gene by polymerase chain reaction (PCR)], and further analysis by denaturing gradient gel electrophoresis (DGGE) revealed endophytic bacteria in roots, young and mature leaves of such plants. The amplification of 16S rRNA gene (Bacteria domain) with the exclusion of the plant chloroplast DNA interference, confirmed the presence of bacterial DNA, from endophytic microorganisms within microplant tissues. PCR–DGGE analysis revealed clear differences on bacterial communities depending on plant organ. Group-specific DGGE analyses also indicated differences in the structures of Actinobacteria, Alphaproteobacteria and Betaproteobacteria communities in each part of plants. The results suggest the occurrence of a succession of bacterial communities colonizing actively the microplants organs. This study is the first report that brings together evidences that pineapple microplants, previously considered axenic, harbor an endophytic bacterial community encompassing members of Actinobacteria, Alphaproteobacteria and Betaproteobacteria group which is responsive to differences in organs due to plant development.     

Hydroxytyrosol administration enhances atherosclerotic lesion development in apo E deficient mice

Hydroxytyrosol is a phenol found in olive oil. To verify the effect of hydroxytyrosol on the development of atherosclerosis, two groups of apo E deficient male mice on a standard chow diet were used: the control group receiving only water, and the second group an aqueous solution of hydroxytyrosol in order to provide a dose of 10 mg/kg/day to each mouse. This treatment was maintained for 10 weeks. At the moment of sacrifice, blood was drawn and heart removed. Plasma lipids, apolipoproteins and monocyte Mac-1 expression were assayed as well as aortic atherosclerotic areas in both groups. Data showed no significant changes in HDL cholesterol, paraoxonase, apolipoprotein B or triglyceride levels. However, hydroxytyrosol administration decreased apolipoprotein A-I and increased total cholesterol, atherosclerotic lesion areas and circulating monocytes expressing Mac-1. The latter was highly correlated with lesion areas (r = 0.65, P < 0.01). These results indicate that administration of hydroxytyrosol in low cholesterol diets increases atherosclerotic lesion associated with the degree of monocyte activation and remodelling of plasma lipoproteins. Our data supports the concept that phenolic-enriched products, out of the original matrix, could be not only non useful but also harmful. Our results suggest that the formulation of possible functional foods should approximate as much as possible the natural environment in which active molecules are found.     

“Filling a gap: knowledge in health related science for middle school students in formal and informal contexts

ABSTRACT

Background: Recent studies shows lack of knowledge concerning health literacy relatively microorganism/disease and the correct use of antibiotics. These facts lead to the spreading of resistant strains bacteria.

Aims: The overarching goals of this study were a) to promote health literacy predominantly in respect to microorganism/disease, diseases transmission, antibiotic target and adequate antibiotic use; b) to show the importance of the transmission of health information in different environments; c) to motivate students for science research.

Methods: Students’ knowledge was evaluated with a survey instrument. Responses were analyzed before and after the implementation of the activity. The activity consists of an interactive lecture, slide show presentation and some practical activities. The sample includes 25 Portuguese students, 13–17 years old who attended the activity in Palácio de Cristal (Porto) and 66 students from D. Maria II School, V.N. Famalicão.

Results: In this study, significant increase in knowledge about all the topics evaluated were observed both in formal and informal environments between the pre and post-test; however the best results were detected in informal contexts.

Conclusion: The findings showed the importance of the implementation of scientific activities to middle-school students. This experience was valuable and helped to improve student knowledge on heath topics.