Aptasensores impedimétricos para la detección de trombina

En el presente trabajo se ha desarrollado el primer aptasensor (biosensor de aptámero) en nuestro grupo de investigación, Grup de Sensors i Biosensors de la Universidad Autònoma de Barcelona. En concreto se han desarrollado dos aptasensores para la detección de la proteína trombina, uno basado en la inmovilización de aptámeros por adsorción física, y otro basado en la inmovilización de aptámeros por enlace covalente mediante la reacción EDAC-NHS. El aptasensor utiliza la afinidad específica de la cadena de DNA (aptámero) por la proteína con la que interacciona. Los cambios de carga y estéricos del complejo aptámero proteína alteran la capacidad y la resistencia de transferencia interfacial de electrones en la superficie del electrodo. El principio de detección se basa en la detección de cambios de estas propiedades de interfase del electrodo con el marcador redox [Fe(CN)6]3- / [Fe(CN)6]4-, utilizando mediciones de Espectroscopia Electroquímica de Impedancia. El aptasensor basado en adsorción física del aptámero mostró una respuesta lineal a trombina en el rango de 7.5 a 75 pM y un límite de detección de 5pM, después de optimizar todas las condiciones experimentales. Posteriormente se estudió la especificidad del sistema respecto proteínas potencialmente interferentes presentes en suero sanguíneo, obteniendo cierta interferencia por parte de fibrinógeno e inmunoglobulina G, pero no por parte de albúmina. El sensor demostró ser regenerable mediante la ruptura del complejo formado entre el aptámero y la trombina con una solución de NaCl 2.0 M, aumento de la temperatura y agitación. El segundo aptasensor, basado en enlace covalente del aptámero mostró una respuesta lineal a trombina y limite de detección mejor que el anterior sensor; de 2.5 a 100 pM y 1.5 pM respectivamente. Aunque cabe destacar que este aptasensor está siendo optimizado actualmente.

Irisin is expressed and produced by human muscle and adipose tissue in association with obesity and insulin resistance.

CONTEXT Recently irisin (encoded by Fndc5 gene) has been reported to stimulate browning and uncoupling protein 1 expression in sc adipose tissue of mice. OBJECTIVE The objective of the study was to investigate FNDC5 gene expression in human muscle and adipose tissue and circulating irisin according to obesity, insulin sensitivity, and type 2 diabetes. DESIGN, PATIENTS, AND MAIN OUTCOME MEASURE Adipose tissue FNDC5 gene expression and circulating irisin (ELISA) were analyzed in 2 different cohorts (n = 125 and n = 76); muscle FNDC5 expression was also evaluated in a subcohort of 34 subjects. In vitro studies in human preadipocytes and adipocytes and in induced browning of 3T3-L1 cells (by means of retinoblastoma 1 silencing) were also performed. RESULTS In both sc and visceral adipose tissue, FNDC5 gene expression decreased significantly in association with obesity and was positively associated with brown adipose tissue markers, lipogenic, insulin pathway-related, mitochondrial, and alternative macrophage gene markers and negatively associated with LEP, TNFα, and FSP27 (a known repressor of brown genes). Circulating irisin and irisin levels in adipose tissue were significantly associated with FNDC5 gene expression in adipose tissue. In muscle, the FNDC5 gene was 200-fold more expressed than in adipose tissue, and its expression was associated with body mass index, PGC1α, and other mitochondrial genes. In obese participants, FNDC5 gene expression in muscle was significantly decreased in association with type 2 diabetes. Interestingly, muscle FNDC5 gene expression was significantly associated with FNDC5 and UCP1 gene expression in visceral adipose tissue. In men, circulating irisin levels were negatively associated with obesity and insulin resistance. Irisin was secreted from human adipocytes into the media, and the induction of browning in 3T3-L1 cells led to increased secreted irisin levels. CONCLUSIONS Decreased circulating irisin concentration and FNDC5 gene expression in adipose tissue and muscle from obese and type 2 diabetic subjects suggests a loss of brown-like characteristics and a potential target for therapy.

Epidemiological and clinical complexity of amoxicillin-clavulanate-resistant Escherichia coli.

Two hundred twelve patients with colonization/infection due to amoxicillin-clavulanate (AMC)-resistant Escherichia coli were studied. OXA-1- and inhibitor-resistant TEM (IRT)-producing strains were associated with urinary tract infections, while OXA-1 producers and chromosomal AmpC hyperproducers were associated with bacteremic infections. AMC resistance in E. coli is a complex phenomenon with heterogeneous clinical implications.

És la diversitat sembrada una eina de resistència enfront a les males herbes en cultius farratgers?

El creixement de males herbes és un problema habitual als camps de cultiu de plantes farratgeres. Tradicionalment, molts agricultors han utilitzat tot tipus d’estratègies basades en l’energia exògena per evitar la invasió d’espècies no desitjades als monocultius. No obstant, diversos estudis han demostrat que no és la tàctica més adequada per fer front a aquesta situació. A més a més, s’han realitzat diverses anàlisis per tal de determinar la resistència dels policultius davant la invasibilitat i, al mateix temps, augmentar la productivitat, obtenint resultats més satisfactoris als policultius.

Mecanismos de deformación en aceros inoxidables austeníticos metaestables

La deformación plástica puede inducir a la transformación de la austenita a martensita en los aceros inoxidables austeníticos metaestables. Para analizar este hecho, el inoxidable austenítico metaestable grado AISI 301 LN fue estudiado en dos condiciones diferentes: recocido y laminado en frío. En el primer caso, el acero era completamente austenítico, mientras que después de la laminación presentaba un importante porcentaje de α’-martensita. Se evaluó el cambio de fase cuando el acero es sometido a ensayos monotónicos y cíclicos, así como cuando ha sido modificada la superficie mediante el granallado o se han realizado tratamientos térmicos de reversión. Se utilizaron diferentes técnicas de caracterización microestructural para detectar y cuantificar la martensita, como microscopía óptica, difracción de rayos-X (DRX) y difracción de electrones retrodispersados (EBSD); como también de caracterización mecánica para evaluar el comportamiento de los aceros, trabajo esencial de fractura (TEF), conformabilidad, fatiga de alto número de ciclos (HCF) y nanoindentación. Los resultados mostraron un incremento en la resistencia mecánica del acero laminado en comparación al acero recocido; este hecho está relacionado con la presencia de martensita originada por la laminación. Sin embargo, en términos de deformación y endurecimiento el acero recocido presenta un mejor desempeño como consecuencia del elevado porcentaje de fase austenítica. Así mismo, revertir la martensita de laminación a austenita y refinar la austenita presente permite obtener un acero con una propiedades mecánicas similares a cuando esta en la condición laminado.

Selected ABCB1, ABCB4 and ABCC2 Polymorphisms Do Not Enhance the Risk of Drug-Induced Hepatotoxicity in a Spanish Cohort.

BACKGROUND AND AIMS Flawed ABC transporter functions may contribute to increased risk of drug-induced liver injury (DILI). We aimed to analyse the influence of genetic variations in ABC transporters on the risk of DILI development and clinical presentations in a large Spanish DILI cohort. METHODS A total of ten polymorphisms in ABCB1 (1236T>C, 2677G>T,A, 3435T>C), ABCB4 (1954A>G) and ABCC2 (-1774G>del, -1549A>G, -24C>T, 1249G>A, 3972C>T and 4544G>A) were genotyped using Taqman 5' allelic discrimination assays or sequencing in 141 Spanish DILI patients and 161 controls. The influence of specific genotypes, alleles and haplotypes on the risk of DILI development and clinical presentations was analysed. RESULTS None of the individual polymorphisms or haplotypes was found to be associated with DILI development. Carriers homozygous for the ABCC2 -1774del allele were however only found in DILI patients. Hence, this genotype could potentially be associated with increased risk, though its low frequency in our Spanish cohort prevented a final conclusion. Furthermore, carriers homozygous for the ABCC2 -1774G/-1549A/-24T/1249G/3972T/4544G haplotype were found to have a higher propensity for total bilirubin elevations when developing DILI. CONCLUSIONS Our findings do not support a role for the analysed polymorphisms in the ABCB1, ABCB4 and ABCC2 transporter genes in DILI development in Spanish patients. The ABCC2 -1774deldel genotype was however restricted to DILI cases and could potentially contribute to enhanced DILI susceptibility.

Immunotherapy reduces allergen-mediated CD66b expression and myeloperoxidase levels on human neutrophils from allergic patients.

CD66b is a member of the carcinoembryonic antigen family, which mediates the adhesion between neutrophils and to endothelial cells. Allergen-specific immunotherapy is widely used to treat allergic diseases, and the molecular mechanisms underlying this therapy are poorly understood. The present work was undertaken to analyze A) the in vitro effect of allergens and immunotherapy on cell-surface CD66b expression of neutrophils from patients with allergic asthma and rhinitis and B) the in vivo effect of immunotherapy on cell-surface CD66b expression of neutrophils from nasal lavage fluid during the spring season. Myeloperoxidase expression and activity was also analyzed in nasal lavage fluid as a general marker of neutrophil activation. RESULTS CD66b cell-surface expression is upregulated in vitro in response to allergens, and significantly reduced by immunotherapy (p<0.001). Myeloperoxidase activity in nasal lavage fluid was also significantly reduced by immunotherapy, as were the neutrophil cell-surface expression of CD66b and myeloperoxidase (p<0.001). Interestingly, CD66b expression was higher in neutrophils from nasal lavage fluid than those from peripheral blood, and immunotherapy reduced the number of CD66+MPO+ cells in nasal lavage fluid. Thus, immunotherapy positive effects might, at least in part, be mediated by the negative regulation of the CD66b and myeloperoxidase activity in human neutrophils.

España en Europa - Europa en España (1986-2006)

Aquesta publicació té per objectiu oferir un balanç dels primers vint anys des de l'adhesió d'Espanya a l'Europa comunitària en termes de «europeïtzació», una formulació que és objecte de creixent atenció per part dels estudiosos del procés d'integració europea. Per això s'analitzen tant la incidència d'Espanya a la Unió Europea com la d'aquesta en el sistema institucional, els processos polítics i les polítiques públiques espanyoles. El resultat reflecteix el grau creixent de simbiosi que, amb alguns matisos, s'ha anat generant entre ambdós nivells al llarg d'aquests anys.

La Unión Europea en el mundo: Entre el poder de transformación y los límites de la Constitución

La construcción europea se ha enfrentado en los últimos años a un triple desafío: la mayor, y más desafiante (valga la redundancia) adhesión de nuevos miembros, pasando de Quince a Veinticinco; la profundización del proceso político, económico y social a través de la adopción del Tratado por el que establece una Constitución para Europa (términos altamente simbólicos1) y; finalmente, la crisis transatlántica, que ha explotado en torno a la guerra contra Irak, y que ha desencadenado, a su vez, una crisis entre los miembros de la Unión en torno a su actuación internacional. Estas páginas van a abordar, justamente, el tema de la acción exterior de la Unión, sin olvidar que la misma, como veremos, es un producto de factores estructurales (lo que nos permitirá hablar de una política exterior europea estructural) pero, también, un producto de factores coyunturales (la evolución del contexto internacional determinado, en buena medida, por la política de Estados Unidos). Tres temas van a ser desarrollados en estas páginas: el poder de “transformación” que la Unión tiene en la escena internacional; el balance de la política exterior de la Unión bajo el marco legal del Tratado de la Unión Europea y, finalmente, una breve reflexión en torno al tratamiento que la acción exterior merece en el Tratado Constitucional (en proceso de ratificación).

Disruption of GIP/GIPR axis in human adipose tissue is linked to obesity and insulin resistance.

CONTEXT Glucose-dependent insulinotropic peptide (GIP) has a central role in glucose homeostasis through its amplification of insulin secretion; however, its physiological role in adipose tissue is unclear. OBJECTIVE Our objective was to define the function of GIP in human adipose tissue in relation to obesity and insulin resistance. DESIGN GIP receptor (GIPR) expression was analyzed in human sc adipose tissue (SAT) and visceral adipose (VAT) from lean and obese subjects in 3 independent cohorts. GIPR expression was associated with anthropometric and biochemical variables. GIP responsiveness on insulin sensitivity was analyzed in human adipocyte cell lines in normoxic and hypoxic environments as well as in adipose-derived stem cells obtained from lean and obese patients. RESULTS GIPR expression was downregulated in SAT from obese patients and correlated negatively with body mass index, waist circumference, systolic blood pressure, and glucose and triglyceride levels. Furthermore, homeostasis model assessment of insulin resistance, glucose, and G protein-coupled receptor kinase 2 (GRK2) emerged as variables strongly associated with GIPR expression in SAT. Glucose uptake studies and insulin signaling in human adipocytes revealed GIP as an insulin-sensitizer incretin. Immunoprecipitation experiments suggested that GIP promotes the interaction of GRK2 with GIPR and decreases the association of GRK2 to insulin receptor substrate 1. These effects of GIP observed under normoxia were lost in human fat cells cultured in hypoxia. In support of this, GIP increased insulin sensitivity in human adipose-derived stem cells from lean patients. GIP also induced GIPR expression, which was concomitant with a downregulation of the incretin-degrading enzyme dipeptidyl peptidase 4. None of the physiological effects of GIP were detected in human fat cells obtained from an obese environment with reduced levels of GIPR. CONCLUSIONS GIP/GIPR signaling is disrupted in insulin-resistant states, such as obesity, and normalizing this function might represent a potential therapy in the treatment of obesity-associated metabolic disorders.

Analysis of genes encoding penicillin-binding proteins in clinical isolates of Acinetobacter baumannii.

There is limited information on the role of penicillin-binding proteins (PBPs) in the resistance of Acinetobacter baumannii to β-lactams. This study presents an analysis of the allelic variations of PBP genes in A. baumannii isolates. Twenty-six A. baumannii clinical isolates (susceptible or resistant to carbapenems) from three teaching hospitals in Spain were included. The antimicrobial susceptibility profile, clonal pattern, and genomic species identification were also evaluated. Based on the six complete genomes of A. baumannii, the PBP genes were identified, and primers were designed for each gene. The nucleotide sequences of the genes identified that encode PBPs and the corresponding amino acid sequences were compared with those of ATCC 17978. Seven PBP genes and one monofunctional transglycosylase (MGT) gene were identified in the six genomes, encoding (i) four high-molecular-mass proteins (two of class A, PBP1a [ponA] and PBP1b [mrcB], and two of class B, PBP2 [pbpA or mrdA] and PBP3 [ftsI]), (ii) three low-molecular-mass proteins (two of type 5, PBP5/6 [dacC] and PBP6b [dacD], and one of type 7 (PBP7/8 [pbpG]), and (iii) a monofunctional enzyme (MtgA [mtgA]). Hot spot mutation regions were observed, although most of the allelic changes found translated into silent mutations. The amino acid consensus sequences corresponding to the PBP genes in the genomes and the clinical isolates were highly conserved. The changes found in amino acid sequences were associated with concrete clonal patterns but were not directly related to susceptibility or resistance to β-lactams. An insertion sequence disrupting the gene encoding PBP6b was identified in an endemic carbapenem-resistant clone in one of the participant hospitals.

Zinc-α2-Glycoprotein Modulates AKT-Dependent Insulin Signaling in Human Adipocytes by Activation of the PP2A Phosphatase.

OBJECTIVE Evidence from mouse models suggests that zinc-α2-glycoprotein (ZAG) is a novel anti-obesity adipokine. In humans, however, data are controversial and its physiological role in adipose tissue (AT) remains unknown. Here we explored the molecular mechanisms by which ZAG regulates carbohydrate metabolism in human adipocytes. METHODS ZAG action on glucose uptake and insulin action was analyzed. β1 and β2-adrenoreceptor (AR) antagonists and siRNA targeting PP2A phosphatase were used to examine the mechanisms by which ZAG modulates insulin sensitivity. Plasma levels of ZAG were measured in a lean patient cohort stratified for HOMA-IR. RESULTS ZAG treatment increased basal glucose uptake, correlating with an increase in GLUT expression, but induced insulin resistance in adipocytes. Pretreatment of adipocytes with propranolol and a specific β1-AR antagonist demonstrated that ZAG effects on basal glucose uptake and GLUT4 expression are mediated via β1-AR, whereas inhibition of insulin action is dependent on β2-AR activation. ZAG treatment correlated with an increase in PP2A activity. Silencing of the PP2A catalytic subunit abrogated the negative effect of ZAG on insulin-stimulated AKT phosphorylation and glucose uptake but not on GLUT4 expression and basal glucose uptake. ZAG circulating levels were unchanged in a lean patient cohort stratified for HOMA-IR. Neither glucose nor insulin was associated with plasma ZAG. CONCLUSIONS ZAG inhibits insulin-induced glucose uptake in human adipocytes by impairing insulin signaling at the level of AKT in a β2-AR- and PP2A-dependent manner.

Metabolomic insights into the intricate gut microbial-host interaction in the development of obesity and type 2 diabetes.

Gut microbiota has recently been proposed as a crucial environmental factor in the development of metabolic diseases such as obesity and type 2 diabetes, mainly due to its contribution in the modulation of several processes including host energy metabolism, gut epithelial permeability, gut peptide hormone secretion, and host inflammatory state. Since the symbiotic interaction between the gut microbiota and the host is essentially reflected in specific metabolic signatures, much expectation is placed on the application of metabolomic approaches to unveil the key mechanisms linking the gut microbiota composition and activity with disease development. The present review aims to summarize the gut microbial-host co-metabolites identified so far by targeted and untargeted metabolomic studies in humans, in association with impaired glucose homeostasis and/or obesity. An alteration of the co-metabolism of bile acids, branched fatty acids, choline, vitamins (i.e., niacin), purines, and phenolic compounds has been associated so far with the obese or diabese phenotype, in respect to healthy controls. Furthermore, anti-diabetic treatments such as metformin and sulfonylurea have been observed to modulate the gut microbiota or at least their metabolic profiles, thereby potentially affecting insulin resistance through indirect mechanisms still unknown. Despite the scarcity of the metabolomic studies currently available on the microbial-host crosstalk, the data-driven results largely confirmed findings independently obtained from in vitro and animal model studies, putting forward the mechanisms underlying the implication of a dysfunctional gut microbiota in the development of metabolic disorders.

Filmar la historia a partir de Peter Weiss

La lectura de La estética de la resistencia nos permite detectar una serie de operaciones que, durante el proceso de escritura, Peter Weiss pone en marcha y queencontraremos reflejadas a su vez en algunos cineastas contemporáneos en la aproximación al tratamiento de la Historia. La novela nos da pie a intentar sacar a la luz sus costuras: el entrecruzamiento de temporalidades como forma de hacer saltarel presente; la recuperación del archivo personal que permitirá el tratamiento de laHistoria en primera persona; la evolución desde la incorporación del registro enbruto a la escenificación de las discusiones entre partes; la descripción exhaustiva deacontecimientos y obras de arte y su continua puesta en relación con la vidacotidiana de los personajes, que configurará lo que Fredric Jameson entiende comouna “pedagogía del subalterno” y que coincide plenamente con la “emancipaciónintelectual” que propone Jacques Rancière.

La fábrica sensible de Fontainhas

Tras el rodaje de Ossos (1997), emplazado en el barrio liboeta de Fontainhas, el realizador Pedro Costa consagraba los dos años siguientes a registrar ese escenario urbano –en proceso de demolición– a partir de una videocámara digital. Lo haría desde el cuarto de Vanda Duarte, vecina y actriz en la anterior película. Este encuentro entre el dispositivo del cineasta y lo real de la balieue portuguesa instituiría, para No quarto da Vanda (2000) y Juventude emmarcha (2006), una instancia de producción estética que da título a este trabajo, toda vez que se la ha bautizado también en él como La fábrica sensible de Fontainhas. Se trata de examinar cómo esa fábrica mancomunada engrenda una resistencia desde la luz, el espacio, el tiempo y el lenguaje, tanto al derrumbe del barrio –durante el primer largometraje– como al realojo de sucomunidad en los edificios blancos de Casal de Boba –en el segundo.