Design of novel iron compounds as potential therapeutic agents against tuberculosis

In the search for new therapeutic tools against tuberculosis two novel iron complexes, [Fe(L-H)₃], with 3-aminoquinoxaline-2-carbonitrile N¹,N⁴-dioxide derivatives (L) as ligands, were synthesized, characterized by a combination of techniques, and in vitro evaluated. Results were compared with those previously reported for two analogous iron complexes of other ligands of the same family of quinoxaline derivatives. In addition, the complexes were studied by cyclic voltammetry and EPR spectroscopy. Cyclic voltammograms of the iron compounds showed several cathodic processes which were attributed to the reduction of the metal center (Fe(III)/Fe(II)) and the coordinated ligand. EPR signals were characteristic of magnetically isolated high-spin Fe(III) in a rhombic environment and arise from transitions between m_S = ± 1/2 (g_eff ~ 9) or m_S = ± 3/2 (g_eff ~ 4.3) states. Mössbauer experiments showed hyperfine parameters that are typical of high-spin Fe(III) ions in a not too distorted environment. The novel complexes showed in vitro growth inhibitory activity on Mycobacterium tuberculosis H₃₇Rv (ATCC 27294), together with very low unspecific cytotoxicity on eukaryotic cells (cultured murine cell line J774). Both complexes showed higher inhibitory effects on M. tuberculosis than the “second-line” therapeutic drugs.     

Fluorescent 3-hydroxy-4-pyridinone hexadentate iron chelators: intracellular distribution and the relevance to antimycobacterial properties

Abstract  

We report the synthesis and characterization of a fluorescent iron chelator (4), shown to be effective in inhibiting the growth of Mycobacterium avium in macrophages, together with the synthesis and characterization of two unsuccessful analogues selected to facilitate identification of the molecular properties responsible for the antimicrobial activity. Partition of the chelators in liposomes was investigated and the compounds were assessed with respect to uptake by macrophages, responsiveness to iron overload/iron deprivation and intracellular distribution by flow cytometry and confocal microscopy. The synthesis of the hexadentate chelators is based on a tetrahedral structure to which three bidentate 3-hydroxy-4-pyridinone chelating units are linked via amide bonds. The structure is synthetically versatile, allowing further addition of functional groups such as fluorophores. Here, we analyse the non-functionalized hexadentate unit (3) and the corresponding rhodamine B (4) and fluorescein (5) labelled chelators. The iron(III) stability constant was determined for 3 and the values log β = 34.4 and pFe³⁺ = 29.8 indicate an affinity for iron of the same order of magnitude as that of mycobacteria siderophores. Fluorescence properties in the presence of liposomes show that 4 strongly interacts with the lipid phase, whereas 5 does not. Such different behaviour may explain their distinct intracellular localization as revealed by confocal microscopy. The flow cytometry and confocal microscopy studies indicate that 4 is readily engulfed by macrophages and targeted to cytosol and vesicles of the endolysosomal continuum, whereas 5 is differentially distributed and only partially colocalizes with 4 after prolonged incubation. Differential distribution of the compounds is likely to account for their different efficacy against mycobacteria.     

Fusarium verticillioides (Saccardo) Nirenberg Associated with Hardlock of Cotton

The development of new immune potentiators for human vaccines is an important and expanding field of research. In the present study, the ability of the capsular polysaccharide from Neisseria meningitidis serogroup A (CPS-A), a mannose-containing carbohydrate, to enhance the antibody production against a co-administered model vaccine antigen, is examined. A protein-meningococcal serogroup C capsular polysaccharide (CPS-C) conjugate was selected as the model antigen for this study. After subcutaneous immunization of Balb/C mice, the conjugate mixed with CPS-A induced higher anti-CPS-C IgG and IgG_2a antibody levels and higher anti-meningococcal serogroup C bactericidal titers than the conjugate alone or mixed with CPS-C. The immuno-stimulatory properties exhibited by CPS-A and the fact that vaccines based on purified CPS-A has been safely used during decades to fight the serogroup A meningococcal disease, support the proposal to use CPS-A as immune potentiator for human vaccination studies.     

Influence of moderately intense strength training on flexibility in sedentary young women

The present study is the first to examine whether moderately intense resistance training improves flexibility in an exclusively young, sedentary women population. Twenty-four, young, sedentary women were divided into 3 groups as follows: agonist/antagonist (AA) training group, alternated strength training (AST) group, or a control group (CG). Training occurred every other day for 8 weeks for a total of 24 sessions. Training groups performed 3 sets of 10 to 12 repetitions per set except for abdominal training where 3 sets of 15 to 20 reps were performed. Strength (1 repetition maximum bench press) and flexibility were assessed before and after the training period. Flexibility was assessed on 6 articular movements: shoulder flexion and extension, horizontal shoulder adduction and abduction, and trunk flexion and extension. Both groups increased strength and flexibility significantly from baseline and significantly when compared with the CG (p ≤ 0.05). The AST group increased strength and flexibility significantly more than the AA group (p ≤ 0.05) in all but one measurement. This study shows that resistance training can improve flexibility in young sedentary women in 8 weeks.     

The influence of mesh size in environmental quality assessment of estuarine macrobenthic communities

The Water Framework Directive (WFD) strengthened the need for environmental quality assessment with rapid and accurate results. Studies of estuarine benthic macrofauna communities often use 0.5-mm mesh sieves in samples processing. However, this represents a considerable increase in sampling and identification effort compared to the use of 1-mm mesh sieves. Therefore, it is relevant to determine if mesh size matters in environmental quality assessments.The objectives of this study were as follows: (i) to test whether sieves with different mesh sizes provided different environmental status assessments in transitional systems, (ii) to compare the performance of different ecological indicators based on data from 0.5- and 1-mm mesh sieves and (iii) to compare the costs involved in using these two mesh sizes.Data were collected in the fall of 2007 and winter of 2008 at four sampling stations located in the Mondego Estuary, Portugal. The relative performance of Margalef and Shannon–Wiener indices, AMBI—AZTI Marine Biotic Index, Pielou, Eco-Exergy and Specific Eco-Exergy indices was analysed. Additionally, the multimetric Benthic Assessment Tool (BAT) was applied. The samples from the 1-mm mesh sieve were processed 2.9 times faster than the samples from the 0.5-mm mesh sieves. As expected, the density, biomass and number of species retained in the 0.5-mm mesh sieve were significantly higher in both seasons than the density, biomass and number of species retained in the 1-mm mesh sieve. All indicators were significantly different for the two mesh sizes in at least one season. The Pielou index was significantly different for the two mesh sizes in both seasons. Most indices showed that the 0.5-mm mesh sieve captured more information from the study system. The first BAT analysis provided different Ecological Quality Status (EQS) assessments for the two mesh sizes. To use the EQS obtained from the 1-mm mesh sieve as a proxy for the EQS for the 0.5-mm mesh sieve, further modifications were done in terms of reference conditions and class boundary thresholds. Regarding the Mondego Estuary, the use of a 1-mm mesh sieve appeared to be advantageous on routine environmental quality assessment, giving unbiased results with relatively less effort. Nevertheless, the methodology needs further validation and additional tests.     

H1N1pdm influenza infection in hospitalized cancer patients: clinical evolution and viral analysis

Background

The novel influenza A pandemic virus (H1N1pdm) caused considerable morbidity and mortality worldwide in 2009. The aim of the present study was to evaluate the clinical course, duration of viral shedding, H1N1pdm evolution and emergence of antiviral resistance in hospitalized cancer patients with severe H1N1pdm infections during the winter of 2009 in Brazil.

Methods

We performed a prospective single-center cohort study in a cancer center in Rio de Janeiro, Brazil. Hospitalized patients with cancer and a confirmed diagnosis of influenza A H1N1pdm were evaluated. The main outcome measures in this study were in-hospital mortality, duration of viral shedding, viral persistence and both functional and molecular analyses of H1N1pdm susceptibility to oseltamivir.

Results

A total of 44 hospitalized patients with suspected influenza-like illness were screened. A total of 24 had diagnosed H1N1pdm infections. The overall hospital mortality in our cohort was 21%. Thirteen (54%) patients required intensive care. The median age of the studied cohort was 14.5 years (3–69 years). Eighteen (75%) patients had received chemotherapy in the previous month, and 14 were neutropenic at the onset of influenza. A total of 10 patients were evaluated for their duration of viral shedding, and 5 (50%) displayed prolonged viral shedding (median 23, range = 11–63 days); however, this was not associated with the emergence of a resistant H1N1pdm virus. Viral evolution was observed in sequentially collected samples.

Conclusions

Prolonged influenza A H1N1pdm shedding was observed in cancer patients. However, oseltamivir resistance was not detected. Taken together, our data suggest that severely ill cancer patients may constitute a pandemic virus reservoir with major implications for viral propagation.     

The Biotechnology Roadmap for Sugarcane Improvement

Due to the strategic importance of sugarcane to Brazil, FAPESP, the main São Paulo state research funding agency, launched in 2008 the FAPESP Bioenergy Research Program (BIOEN, http://bioenfapesp.org). BIOEN aims to generate new knowledge and human resources for the improvement of the sugarcane and ethanol industry. As part of the BIOEN program, a Workshop on Sugarcane Improvement was held on March 18th and 19th 2009 in São Paulo, Brazil. The aim of the workshop was to explore present and future challenges for sugarcane improvement and its use as a sustainable bioenergy and biomaterial feedstock. The workshop was divided in four sections that represent important challenges for sugarcane improvement: a) gene discovery and sugarcane genomics, b) transgenics and controlled transgene expression, c) sugarcane physiology (photosynthesis, sucrose metabolism, and drought) and d) breeding and statistical genetics. This report summarizes the roadmap for the improvement of sugarcane.     

Mitochondria generated nitric oxide protects against permeability transition via formation of membrane protein S-nitrosothiols

Mitochondria generated nitric oxide (NO) regulates several cell functions including energy metabolism, cell cycling, and cell death. Here we report that the NO synthase inhibitors (L-NAME, L-NNA and L-NMMA) administered either in vitro or in vivo induce Ca²⁺-dependent mitochondrial permeability transition (MPT) in rat liver mitochondria via a mechanism independent on changes in the energy state of the organelle. MPT was determined by the occurrence of cyclosporin A sensitive mitochondrial membrane potential disruption followed by mitochondrial swelling and Ca²⁺ release. In in vitro experiments, the effect of NOS inhibitors was dose-dependent (1 to 50 µM). In addition to cyclosporin A, L-NAME-induced MPT was sensitive to Mg²⁺ plus ATP, EGTA, and to a lower degree, to catalase and dithiothreitol. In contrast to L-NAME, its isomer D-NAME did not induce MPT. L-NAME-induced MPT was associated with a significant decrease in both the rate of NO generation and the content of mitochondrial S-nitrosothiol. Acute and chronic in vivo treatment with L-NAME also promoted MPT and decreased the content of mitochondrial S-nitrosothiol. SNAP (a NO donor) prevented L-NAME mediated MPT and reversed the decrease in the rate of NO generation and in the content of S-nitrosothiol. We propose that S-nitrosylation of critical membrane protein thiols by NO protects against MPT.     

Challenging Wallacean and Linnean shortfalls: knowledge gradients and conservation planning in a biodiversity hotspot

Knowledge about biodiversity remains inadequate because most species living on Earth were still not formally described (the Linnean shortfall) and because geographical distributions of most species are poorly understood and usually contain many gaps (the Wallacean shortfall). In this paper, we developed models to infer the size and placement of geographical ranges of hypothetical non‐described species, based on the range size frequency distribution of anurans recently described in the Cerrado Biome, on the level of knowledge (number of inventories) and on surrogates for habitat suitability. The rationale for these models is as follow: (1) the range size frequency distribution of these species should be similar to the range‐restricted species, which have been most recently described in the Cerrado Biome; (2) the probability of new discoveries will increase in areas with low biodiversity knowledge, mainly in suitable areas, and (3) assuming range continuity, new species should occupy adjacent cells only if the level of knowledge is low enough to allow the existence of undiscovered species. We ran a model based on the number of inventories only, and two models combining effects of number of inventories and two different estimates of habitat suitability, for a total of 100 replicates each. Finally, we performed a complementary analysis using simulated annealing to solve the set‐covering problem for each simulation (i.e. finding the smallest number of cells so that all species are represented at least once), using extents of occurrence of 160 species (131 real anuran species plus 29 new simulated species). The revised reserve system that included information about unknown or poorly sampled taxa significantly shifted northwards, when compared to a system based on currently known species. This main result can be explained by the paucity of biodiversity data in this part of the biome, associated with its relatively high habitat suitability. As a precautionary measure, weighted by the inferred distribution data, the prioritization of a system of reserves in the north part of the biome appears to be defensible.