Analysis of the Organic Hydroperoxide Response of Chromobacterium violaceum Reveals That OhrR Is a Cys-Based Redox Sensor Regulated by Thioredoxin

Organic hydroperoxides are oxidants generated during bacterial-host interactions. Here, we demonstrate that the peroxidase OhrA and its negative regulator OhrR comprise a major pathway for sensing and detoxifying organic hydroperoxides in the opportunistic pathogen Chromobacterium violaceum. Initially, we found that an ohrA mutant was hypersensitive to organic hydroperoxides and that it displayed a low efficiency for decomposing these molecules. Expression of ohrA and ohrR was specifically induced by organic hydroperoxides. These genes were expressed as monocistronic transcripts and also as a bicistronic ohrR-ohrA mRNA, generating the abundantly detected ohrA mRNA and the barely detected ohrR transcript. The bicistronic transcript appears to be processed. OhrR repressed both the ohrA and ohrR genes by binding directly to inverted repeat sequences within their promoters in a redox-dependent manner. Site-directed mutagenesis of each of the four OhrR cysteine residues indicated that the conserved Cys21 is critical to organic hydroperoxide sensing, whereas Cys126 is required for disulfide bond formation. Taken together, these phenotypic, genetic and biochemical data indicate that the response of C. violaceum to organic hydroperoxides is mediated by OhrA and OhrR. Finally, we demonstrated that oxidized OhrR, inactivated by intermolecular disulfide bond formation, is specifically regenerated via thiol-disulfide exchange by thioredoxin (but not other thiol reducing agents such as glutaredoxin, glutathione and lipoamide), providing a physiological reducing system for this thiol-based redox switch.

Novel properties of melanins include promotion of DNA strand breaks, impairment of repair, and reduced ability to damage DNA after quenching of singlet oxygen

Melanins have been associated with the development of melanoma and its resistance to photodynamic therapy (PDT). Singlet molecular oxygen (102), which is produced by ultraviolet A solar radiation and the PDT system, is also involved. Here, we investigated the effects that these factors have on DNA damage and repair. Our results show that both types of melanin (eumelanin and pheomelanin) lead to DNA breakage in the absence of light irradiation and that eumelanin is more harmful than pheomelanin. Interestingly, melanins were found to bind to the minor grooves of DNA, guaranteeing close proximity to DNA and potentially causing the observed high levels of strand breaks. We also show that the interaction of melanins with DNA can impair the access of repair enzymes to lesions, contributing to the perpetuation of DNA damage. Moreover, we found that after melanins interact with 102, they exhibit a lower ability to induce DNA breakage; we propose that these effects are due to modifications of their structure. Together, our data highlight the different modes of action of the two types of melanin. Our results may have profound implications for cellular redox homeostasis, under conditions of induced melanin synthesis and irradiation with solar light. These results may also be applied to the development of protocols to sensitize melanoma cells to PDT. (c) 2012 Elsevier Inc. All rights reserved.

Exercise Training Restores Cardiac Protein Quality Control in Heart Failure

Exercise training is a well-known coadjuvant in heart failure treatment; however, the molecular mechanisms underlying its beneficial effects remain elusive. Despite the primary cause, heart failure is often preceded by two distinct phenomena: mitochondria dysfunction and cytosolic protein quality control disruption. The objective of the study was to determine the contribution of exercise training in regulating cardiac mitochondria metabolism and cytosolic protein quality control in a post-myocardial infarction-induced heart failure (MI-HF) animal model. Our data demonstrated that isolated cardiac mitochondria from MI-HF rats displayed decreased oxygen consumption, reduced maximum calcium uptake and elevated H2O2 release. These changes were accompanied by exacerbated cardiac oxidative stress and proteasomal insufficiency. Declined proteasomal activity contributes to cardiac protein quality control disruption in our MI-HF model. Using cultured neonatal cardiomyocytes, we showed that either antimycin A or H2O2 resulted in inactivation of proteasomal peptidase activity, accumulation of oxidized proteins and cell death, recapitulating our in vivo model. Of interest, eight weeks of exercise training improved cardiac function, peak oxygen uptake and exercise tolerance in MI-HF rats. Moreover, exercise training restored mitochondrial oxygen consumption, increased Ca2+-induced permeability transition and reduced H2O2 release in MI-HF rats. These changes were followed by reduced oxidative stress and better cardiac protein quality control. Taken together, our findings uncover the potential contribution of mitochondrial dysfunction and cytosolic protein quality control disruption to heart failure and highlight the positive effects of exercise training in re-establishing cardiac mitochondrial physiology and protein quality control, reinforcing the importance of this intervention as a nonpharmacological tool for heart failure therapy.

1,4-Diamino-2-butanone, a putrescine analogue, promotes redox imbalance in Trypanosoma cruzi and mammalian cells

The putrescine analogue 1,4-diamino-2-butanone (DAB) is highly toxic to various microorganisms, including Trypanosoma cruzi. Similar to other a-aminocarbonyl metabolites. DAB exhibits pro-oxidant properties. DAB undergoes metal-catalyzed oxidation yielding H2O2, NH4+ ion, and a highly toxic alpha-oxoaldehyde. In vitro. DAB decreases mammalian cell viability associated with changes in redox balance. Here, we aim to clarify the DAB pro-oxidant effects on trypomastigotes and on intracellular T. cruzi amastigotes. DAB (0.05-5 mM) exposure in trypomastigotes, the infective stage of T. cruzi, leads to a decline in parasite viability (IC50 c.a. 0.2 mM DAB; 4 h incubation), changes in morphology, thiol redox imbalance, and increased TcSOD activity. Medium supplementation with catalase (2.5 mu M) protects trypomastigotes against DAB toxicity, while host cell invasion by trypomastigotes is hampered by DAB. Additionally, intracellular amastigotes are susceptible to DAB toxicity. Furthermore, pre-treatment with 100-500 mu M buthionine sulfoximine (BSO) of LLC-MK2 potentiates DAB cytotoxicity, whereas 5 mM N-acetyl-cysteine (NAC) protects cells from oxidative stress. Together, these data support the hypothesis that redox imbalance contributes to DAB cytotoxicity in both T. cruzi and mammalian host cells. (C) 2012 Elsevier Inc. All rights reserved.

Diet-Sensitive Sources of Reactive Oxygen Species in Liver Mitochondria: Role of Very Long Chain Acyl-CoA Dehydrogenases

High fat diets and accompanying hepatic steatosis are highly prevalent conditions. Previous work has shown that steatosis is accompanied by enhanced generation of reactive oxygen species (ROS), which may mediate further liver damage. Here we investigated mechanisms leading to enhanced ROS generation following high fat diets (HFD). We found that mitochondria from HFD livers present no differences in maximal respiratory rates and coupling, but generate more ROS specifically when fatty acids are used as substrates. Indeed, many acyl-CoA dehydrogenase isoforms were found to be more highly expressed in HFD livers, although only the very long chain acyl-CoA dehydrogenase (VLCAD) was more functionally active. Studies conducted with permeabilized mitochondria and different chain length acyl-CoA derivatives suggest that VLCAD is also a source of ROS production in mitochondria of HFD animals. This production is stimulated by the lack of NAD+. Overall, our studies uncover VLCAD as a novel, diet-sensitive, source of mitochondrial ROS.

O ciclo da comunicação científica na semiosfera

Este ensaio apresenta a construção de um objeto de pesquisa com base na teoria da semiótica da cultura. São feitas reflexões sobre os sistemas modelizantes envolvidos no ciclo da comunicação científica em grupo de pesquisa de universidade, desde a busca da informação até a publicação dos resultados dos estudos. As linguagens naturais (idiomas) e artificiais (linguagem de busca em computadores e vocabulários controlados) são identificadas. A partir disso, o objeto se delineia como o conjunto de textos da cultura e a própria semiosfera, representada pelos diálogos dos sujeitos da cultura e o processo de comunicação envolvido. Alguns desafios se apresentam, como: a necessidade de aprofundamento na teoria da semiótica da cultura, a participação do pesquisador também como sujeito da pesquisa e o trabalho com a interdisciplinaridade para estudar um objeto com as vertentes da ciência da informação, biomedicina, semiótica e outras disciplinas a elas relacionadas.

Redox regulation of the proteasome via S-glutathionylation

The proteasome is a multimeric and multicatalytic intracellular protease responsible for the degradation of proteins involved in cell cycle control, various signaling processes, antigen presentation, and control of protein synthesis. The central catalytic complex of the proteasome is called the 20S core particle. The majority of these are flanked on one or both sides by regulatory units. Most common among these units is the 19S regulatory unit. When coupled to the 19S unit, the complex is termed the asymmetric or symmetric 26S proteasome depending on whether one or both sides are coupled to the 19S unit, respectively. The 26S proteasome recognizes poly-ubiquitinylated substrates targeted for proteolysis. Targeted proteins interact with the 19S unit where they are deubiquitinylated, unfolded, and translocated to the 20S catalytic chamber for degradation. The 26S proteasome is responsible for the degradation of major proteins involved in the regulation of the cellular cycle, antigen presentation and control of protein synthesis. Alternatively, the proteasome is also active when dissociated from regulatory units. This free pool of 20S proteasome is described in yeast to mammalian cells. The free 20S proteasome degrades proteins by a process independent of poly-ubiquitinylation and ATP consumption. Oxidatively modified proteins and other substrates are degraded in this manner. The 20S proteasome comprises two central heptamers (β-rings) where the catalytic sites are located and two external heptamers (α-rings) that are responsible for proteasomal gating. Because the 20S proteasome lacks regulatory units, it is unclear what mechanisms regulate the gating of α-rings between open and closed forms. In the present review, we discuss 20S proteasomal gating modulation through a redox mechanism, namely, S-glutathionylation of cysteine residues located in the α-rings, and the consequence of this post-translational modification on 20S proteasomal function.

Evaluación clínica del dolor torácico y su seguimiento : experiencia en el Hospital General de Lanzarote : tesis doctoral

Programa de Doctorado: Biomedicina Aplicada a la Práctica Clínica

Nefropatía diabética : intervención desde atención primaria : tesis doctoral

Programa de Doctorado en Biomedicina Aplicada a la Práctica Clínica

Validación de las reglas del tobillo de Ottawa: repercusión en los costes y calidad asistencial

Programa de doctorado: Biomedicina aplicada a la práctica clínica.

Incidencia de la sordera genética en Fuerteventura en la población de 3 a 6 años : tratamiento y seguimiento

Programa de doctorado: Biomedicina aplicada a la práctica clínica

Combinación de 4 días de alto volumen de ejercicio y dieta muy baja en calorías como nuevo modelo en la lucha contra la obesidad

Programa de Doctorado en Investigación en Biomedicina. Grupo de Investigación en Rendimiento Humano, Actividad Física y Salud.

La globalizzazione della bioetica: Analisi della bioetica latinoamericana

La tesi esplora la riflessione bioetica latino-americana e la sua evoluzione, valutandone i legami, intenzionali e non, con le posizioni bioetiche più consolidate e tendenzialmente dominanti del panorama internazionale. La trattazione indaga lo sviluppo della bioetica nel sub-continente latinoamericano, ponendo in evidenza lo sviluppo delle teorie morali che definiscono l'orizzonte interpretativo del dibattito contemporaneo, come pure le peculiarità della produzione sudamericana in materia di bioetica e la casistica tipica di questi territori. Per una comprensione delle caratteristiche di un pensiero bioetico, tipico di un territorio in via di sviluppo, la tesi presenta la trattazione di due precisi case studies: A- Il progetto peruviano: Propuesta de incorporación del enfoque intercultural y de representantes indígenas en los comités de ética en países multiculturales; B- La biomedicina e gli xenotrapianti in Messico. L'analisi dei casi studio è funzionale alla messa in luce delle caratteristiche costitutive del pensiero latinoamericano e della casistica tipica del territorio. La bioetica Latinoamericana, infatti, sviluppatasi circa con vent'anni di ritardo rispetto ai luoghi di nascita di questa disciplina (Gran Bretagna e USA), rivendica l'esigenza di un dibattito e di una prassi bioetica autoctone, capaci quindi di riassumere in sé stesse concetti e principi declinabili all'interno delle esperienze e delle concrete esigenze che prendono forma nella realtà culturale e socio-economica. Per questo motivo si discosta dalle cosiddette teorie bioetiche classiche rivendicando una bioetica di contestazione che si vincola costitutivamente al linguaggio dei diritti umani. La tesi propone inoltre l'utilizzo dei dati emersi dai casi studio per sviluppare una riflessione sulle modalità e su alcuni esiti della globalizzazione della bioetica. L’analisi si avvale di strumenti e categorie proprie della sociologia, dell’economia e della politica al fine di mettere in evidenza le diverse componenti costitutive e le criticità della bioetica globalizzata.

Integración de la Medicina Tradicional y del Sistema de Salud Formal en las Estrategias Familiares Andinas

La medicina tradicional no solamente sigue vigente en los Andes bolivianos, sino constituye la opción de primera elección para las familias rurales. Un estudio realizado en dos comunidades de la Subcentral Waka Playa, Municipio Tapacarí, muestra que los campesinos quechua-hablantes de los valles interandinos integran el sistema tradicional con el sistema de salud formal en el marco de sus estrategias familiares, reflejando un diálogo ínter-ontológico entre diferentes concepciones de salud y de enfermedad. A lo largo de una larga interacción con su entorno natural, los pobladores de los Andes han desarrollado concepciones específicas sobre el ser humano, su relación con la naturaleza, la salud y la enfermedad. Este proceso resultó en un sistema de salud complejo, donde intervienen expertos locales como son los curanderos-adivinos y los parteros y que se basa en un amplio uso y conocimiento de la flora y fauna nativas. A parte de estos conocimientos especializados, la medicina tradicional andina se expresa también en las prácticas terapéuticas diarias de la población, transmitidas y re-inventadas de generación en generación en el contexto de la familia y de la comunidad. La introducción del sistema de salud formal en el área de estudio en los años 90, representado por una posta de salud, impactó las comunidades locales principalmente por el acceso a vacunas – y la consecuente erradicación de algunas enfermedades - y a métodos contraceptivos. Sin embargo, la biomedicina es todavía de acceso muy limitado además de, según la población local, no tener la capacidad para curar ciertas enfermedades locales como el “susto,” el “mal viento” o la “colerina”. Como en otras áreas del conocimiento ecológico local, se observa una pérdida general de los conocimientos vinculados a la medicina tradicional andina debido a varios factores socioeconómicos, el más impactante siendo el proceso masivo de migración de las zonas rurales a los centros urbanos y al trópico boliviano. Sin embargo, estos conocimientos todavía se mantienen vigentes y son altamente valorizados por la población local, como un elemento del mantenimiento de sus formas de vida y de relacionamiento con la “Madre Tierra” (Pachamama). Las familias de Waka Playa integran la medicina tradicional y el sistema de salud formal en sus estrategias de vida, en diferentes grados según factores como la migración, la educación, el sexo y la edad. El uso de la flora medicinal local también es diferenciado según la localización de la vivienda principal de las familias y la consecuente concentración geográfica de sus actividades en las diferentes zonas de producción agroecológicas. Por ultimo, el interés personal influye sobre el grado y el tipo de conocimiento de cada familia. La medicina tradicional juega un rol fundamental en el bien-estar de las familias campesinas andinas y por ello constituye un potencial importante para un desarrollo sostenible. En esta perspectiva, la revalorización de los saberes locales y la construcción de puentes de diálogo con el sistema de salud local son sumamente importantes. Este punto de vista tiene perspectivas prometedoras en el contexto político actual boliviano, que apoya la medicina tradicional en el marco del reconocimiento de las identidades indígenas, en miras a un desarrollo endógeno.

Medicina y Filosofía : ¿Dos disciplinas o un único arte cognoscitivo?

El pensamiento médico y el ejercicio de la medicina plantean un cúmulo de problemas filosóficos que no pueden reducirse a la parcela de lo ético. La medicina está atravesada por cuestiones filosóficas que la definen y la constituyen como ciencia. Se trata de cuestiones sobre las cuales gira y se desarrolla el entero ejercicio médico y la medicina teórica. La filosofía y la medicina no deben ser entendidas como dos cuerpos de conocimiento autónomo e independiente, sino que en orden a alcanzar una ciencia más madura y humana, ambas deben embarcarse en un proyecto común. Paul Karl Feyerabend nos ofrece una primera aproximación a esta tesis.