Redox tuning of the catalytic activity of soluble fumarate reductases from Shewanella

AbstractMany enzymes involved in bioenergetic processes contain chains of redox centers that link the protein surface, where interaction with electron donors or acceptors occurs, to a secluded catalytic site. In numerous cases these redox centers can transfer only single electrons even when they are associated to catalytic sites that perform two-electron chemistry. These chains provide no obvious contribution to enhance chemiosmotic energy conservation, and often have more redox centers than those necessary to hold sufficient electrons to sustain one catalytic turnover of the enzyme. To investigate the role of such a redox chain we analyzed the transient kinetics of fumarate reduction by two flavocytochromes c3 of Shewanella species while these enzymes were being reduced by sodium dithionite. These soluble monomeric proteins contain a chain of four hemes that interact with a flavin adenine dinucleotide (FAD) catalytic center that performs the obligatory two electron–two proton reduction of fumarate to succinate. Our results enabled us to parse the kinetic contribution of each heme towards electron uptake and conduction to the catalytic center, and to determine that the rate of fumarate reduction is modulated by the redox stage of the enzyme, which is defined by the number of reduced centers. In both enzymes the catalytically most competent redox stages are those least prevalent in a quasi-stationary condition of turnover. Furthermore, the electron distribution among the redox centers during turnover suggested how these enzymes can play a role in the switch between respiration of solid and soluble terminal electron acceptors in the anaerobic bioenergetic metabolism of Shewanella

Fungal Meningitis in an Immunocompetent Patient

Cryptococcal meningitis is a rare entity among immunocompetent hosts but, when it occurs, it is associated with significant morbidity and mortality. Clinical presen- tation as well as the course of the disease is usually subtle and indolent with headache and altered mental status. The authors present the case of a 59-year-old man, who sought medical help with a 2-week history of headaches accompanied by nausea and visual and hearing disturbances. On admission the patient was afebrile, presented visual and hearing deficits and had a normal magnetic resonance image of the brain. A lumbar puncture was performed and microscopic examination of the cerebrospinal fluid revealed yeasts that were identified as Cryptococcus spp. and later, by means of molecular biology techniques, as Cryptococcus neoformans, var. grubii. The patient was treated with liposomal amphotericin B plus fluconazole for 28 weeks. At follow-up after 1 year the patient was asymptomatic and received fluconazole 400 mg/day as prophylactic therapy. The outcome of Cryptococcus infections in immunocompetent hosts is reported to be poor as a result of a delayed diagnosis and suboptimal initial antifungal therapy. The influence of the normal immune response is unclear

Análise descritiva das idades dos nadadores dos jogos olímpicos de Pequim 2008

O estudo teve como objectivo efectuar uma análise descritiva das idades e posterior comparação entre sexos, de todos os participantes nas provas de Natação Pura Desportiva nos Jogos Olímpicos de Pequim 2008

Halogenated polycyclic aromatic hydrocarbons associated with drinking water disinfection

Introduction: Disinfection by-products (DBPs) have been identified in chlorinated water. This fact justifies the growing concern about the potential health effects of emerging unregulated DBPs, some of which appear to be more genotoxic than the regulated DBPs[1]. Polycyclic aromatic hydrocarbons (PAHs) are among the most persistent contaminants detected in environmental samples such as river sediments and tap water. A few studies have already proven that water disinfection can lead to the formation of halogenated derivatives of PAHs, such as chlorinated and brominated PAHs[2] . The available toxicological studies have shown that these compounds possess, in general, greater mutagenicity than the corresponding parent PAHs. Our research group has also shown that exposure of HepG2 cells to a dose-range of 6-Cl-benzo[a]pyrene (6-ClBaP) and BaP resulted in cytotoxicity above 50 µM and that, at the equimolar doses of 100 and 125 µM, 6-ClBaP was able to induce a significantly higher level of DNA damage than BaP[3] . The present study had two main objectives: 1) identification of the major chlorinated and brominated derivatives of benzo[a]anthracene (BaA) and pyrene (Pyr) formed as disinfection by-products and 2) evaluation of their potential hazard to humans, through the characterization of their potential cytotoxic and genotoxic effects in a human cell line.The authors wish to thank Instituto Nacional de Saúde Dr. Ricardo Jorge, Lisboa for financial support by the grant BRJ-DSA/2012- Doenças Oncológica

Drinking water contaminants: toxicity of halogenated polycyclic aromatic hydrocarbons

Food may be contaminated with polycyclic aromatic hydrocarbons (PAHs) in the process of smoking or heating. These contaminants or their derivatives can also be present in drinking water when raw water contacts with discharges of untreated industrial/waste water effluents, forest fires or by solubilisation of organic material from contaminated soils. A few studies have shown that water disinfection can lead to halogenated derivatives of PAHs (HPAHs) as chlorinated and brominated derivatives, and there are evidences that these compounds may have greater mutagenicity than the parent PAHs. In this study the cytotoxic and genotoxic effects of chlorinated/brominated derivatives of pyrene (Pyr) and benzo[a]anthracene (BaA), 1-ClPyr, 1-BrPyr and 7-ClBaA, which can be formed as water disinfection by-products, were studied in HepG2 cells to assess their potential hazard to human health. The formation of 1-ClPyr, 1-BrPyr and 7-ClBaA under aqueous disinfection conditions in waters contaminated with Pyr and BaA, was confirmed with an optimized gas chromatography method. Cells exposed (24h) to several concentrations of BaA and 7-ClBaA (1 to 200μM) displayed a dose-related and significant increase of cytotoxicity (neutral red assay) with IC50 values of 3.37 and 12.63µM respectively. For Pyr, 1-ClPyr and 1-BrPyr (10 to 200μM), a lower but significant dose-related cytotoxicity was observed. At non-cytotoxic concentrations (10 and 15µM), 7-ClBaA was able to induce a significantly higher level of oxidative DNA damage in HepG2 cells than its parent compound, as assessed by the FPG-modified comet assay. Under these conditions neither Pyr nor its derivatives were genotoxic. In conclusion, the disinfection process may give rise to genotoxic HPAHs with potential impact on human health and it should be performed in raw waters with minimal content of total organic carbon. In real conditions, humans may be exposed to a mixture of these organic compounds and thus their combined toxic effects should be further evaluated

Palliative splenic irradiation for symptomatic splenomegaly in non-Hodgkin lymphoma

INTRODUCTION AND AIMS: Splenic marginal zone lymphoma, an uncommon subtype of non-Hodgkin lymphoma (NHL), is usually present with symptomatic splenomegaly. Although splenectomy has long been considered the first-line therapy in symptomatic or cytopenic patients, it can lead to significant morbidity and mortality. Splenic irradiation is an option for patients who have a poor response to systemic therapy and/or are not surgical candidates. In this paper, we present a case report of a patient who received splenic radiotherapy for symptomatic splenomegaly. METHODS: An 85-year-old Caucasian man with a 4 year history of low-grade NHL presented with progressive pancytopenia, significant weight loss and symptomatic splenomegaly (abdominal discomfort, sense of fullness and limitation of mobility due to spleen size). The patient refused splenectomy and, in December 2017, was referred to palliative splenic radiotherapy. He was initially treated with five fractions of one Grey (Gy) in order to evaluate clinical and haematology response. After that, 1.5 Gy daily, 5 days a week for 3 weeks. 3D conformal radiotherapy, multiple fields and mixed energy (6 and 15 Mv) were used. RESULTS: Radiotherapy allowed significant splenic reduction to almost half the size, resolving abdominal discomfort and improving quality of life. There was no decline of haemoglobin, leukocytes and platelet counts; in fact, there was a marginal increase. CONCLUSION: Palliative splenic irradiation was well tolerated confirming that it is a safe treatment option for palliation of symptomatic splenomegaly. Thereby, splenic irradiation should be strongly considered in the management of symptomatic splenomegaly, for selected patients who are refractory to or unsuitable for other options or when the patient refuses other treatments.info:eu-repo/semantics/publishedVersio

Crystallization and preliminary crystallographic studies of FoxE from Rhodobacter ferrooxidans SW2, an FeII oxidoreductase involved in photoferrotrophy

FoxE is a protein encoded by the foxEYZ operon of Rhodobacter ferrooxidans SW2 that is involved in Fe^II-based anoxygenic photosynthesis (`photoferrotrophy'). It is thought to reside in the periplasm, where it stimulates light-dependent Fe^II oxidation. It contains 259 residues, including two haem c-binding motifs. As no three-dimensional model is available and there is no structure with a similar sequence, crystals of FoxE were produced. They diffracted to 2.44 Å resolution using synchrotron radiation at the Fe edge. The phase problem was solved by SAD using SHELXC/D/E and the experimental maps confirmed the presence of two haems per molecule

Modificações no perfil bioenergético e biomecânico de nadadores entre dois períodos "pré-taper"

O estudo teve como objectivo analisar as alterações ao nível do perfil bioenergético e biomecânico de nadadores de elite entre dois períodos “pré-taper”. Foram analisados 7 nadadores portugueses masculinos de elevado nível competitivo. Os parâmetros bioenergéticos e biomecânicos foram obtidos em dois momentos “pré-taper” (T1 e T2) antecedentes a competições importantes (Novembro e Março da época desportiva de 2009-2010). Foram obtidos: (i) velocidade do equilíbrio máximo de lactato estimada às 4 mmol (V4), como indicador bioenergético; (ii) distância de ciclo à V4 (DC@V4), frequência gestual à V4 (FG@V4), índice de nado à V4 (IN@V4) e a eficiência propulsiva à V4 (hp@V4) como indicadores biomecânicos. A comparação entre os dois momentos de avaliação foi efectuada com recurso à estatística não paramétrica Teste de Wilcoxon. Foram verificados aumentos com significado estatístico na V4 e na FG@V4 de T1 para T2. A DC@V4e o IN@V4 apresentaram-se estáveis sem variações significativas. Do ponto de vista da análise individual 4 dos 7 nadadores aumentaram o seu IN@V4. A hp@V4 revelou uma diminuição, contudo sem diferenças estatisticamente significativas. Em síntese, existe uma tendência para o aumento da V4 ao longo da época desportiva. As alterações na V4 parecem ser decorrentes de modificações nos pressupostos biomecânicos, mais precisamente no aumento da FG@V4

Análise longitudinal das modificações no perfil biomecânico de nadadores de elite e o seu impacto na performance ao longo da época desportiva

Diversos estudos têm sido realizados no sentido de determinar os factores que mais e melhor predizem a performance em natação pura desportiva. A área da Biomecânica tem vindo a ser apontada como determinante para alcançar elevados níveis de rendimento neste desporto. Contudo, a maioria dos estudos neste âmbito são desenhos transversais, não tendo em vista as variações no perfil dos nadadores tomando em consideração o factor temporal