Producció de material docent per a ciència de materials mitjançant microscòpia electrònica de rastreig i làser confocal

Aquest projecte s'ha realitzat al Servei de Microscòpia de la Universitat Autònoma de Barcelona, i ha tingut una durada de dos anys (2006-2008). La finalitat d’aquest projecte ha estat l’elaboració de material didàctic basat en la captació d’imatges i l’edició de recursos pedagògics de suport digital aplicats a la ciència de materials. Es pretén millorar així la qualitat docent de les pràctiques de diverses assignatures dels ensenyaments de Física i d’Enginyeria de Materials utilitzant tècniques d’anàlisi actuals com són la Microscòpia Electrònica de Rastreig (MER) i la Microscòpia Optica (MO). Amb aquest projecte es vol fomentar també el treball interdisciplinari en equip entre professionals (docents i tècnics superiors de recerca) i acostar la teoria de les assignatures a la realització pràctica, facilitant el suport digital necessari per aconseguir un màxim aprofitament a les aules. Les imatges de MER i MO ajudaran als alumnes a familiaritzar-se amb el món de la recerca i la indústria.

Creació de materials per a l'aprenentatge autònom i guiat de la interpretació de conferències a nivell inicial i intermedi

El desenvolupament del projecte ens ha permès de crear materials per a l´aprenentatge autònom de la interpretació de conferències així com d´un material d´acompanyament i suport del procés d´aprenentatge no presencial. Aquests materials s´integren en les assignatures d´interpretació del Grau en Traducció i Interpretació i serveixen perquè els estudiants continuïn fent un treball guiat fora de l´aula. Es pot considerar també un material de suport a les classes presencials. Els materials desenvolupats cobreixen les cinc llengües dels nostres estudis (català, castellà, anglès, alemany i francès) i estan organitzats en dos blocs diferenciats: itineraris d´aprenentatge i banc de discursos. Aquesta divisió permet a l´estudiant treballar seguint dues opcions diferenciades: a) a partir d´una seqüenciació guiada de tasques; b) abordant directament la interpretació d´un discurs oral. El projecte s'ha dut a terme entre les universitats Pompeu Fabra i Autònoma de Barcelona, durant el període 2007- 2009.

Cellular immune responses to HCV core increase and HCV RNA levels decrease during successful antiretroviral therapy.

BACKGROUND: Hepatitis C virus (HCV) infection is a major cause of morbidity in HIV infected individuals. Coinfection with HIV is associated with diminished HCV-specific immune responses and higher HCV RNA levels. AIMS: To investigate whether long-term combination antiretroviral therapy (cART) restores HCV-specific T cell responses and improves the control of HCV replication. METHODS: T cell responses were evaluated longitudinally in 80 HIV/HCV coinfected individuals by ex vivo interferon-gamma-ELISpot responses to HCV core peptides, that predominantly stimulate CD4(+) T cells. HCV RNA levels were assessed by real-time PCR in 114 individuals. RESULTS: The proportion of individuals with detectable T cell responses to HCV core peptides was 19% before starting cART, 24% in the first year on cART and increased significantly to 45% and 49% after 33 and 70 months on cART (p=0.001). HCV-specific immune responses increased in individuals with chronic (+31%) and spontaneously cleared HCV infection (+30%). Median HCV RNA levels before starting cART were 6.5 log(10) IU/ml. During long-term cART, median HCV-RNA levels slightly decreased compared to pre-cART levels (-0.3 log10 IU/ml, p=0.02). CONCLUSIONS: Successful cART is associated with increasing cellular immune responses to HCV core peptides and with a slight long-term decrease in HCV RNA levels. These findings are in line with the favourable clinical effects of cART on the natural history of hepatitis C and with the current recommendation to start cART earlier in HCV/HIV coinfected individuals.

Cellular pathology of hilar neurons in Ammon's horn sclerosis.

In addition to functionally affected neuronal signaling pathways, altered axonal, dendritic, and synaptic morphology may contribute to hippocampal hyperexcitability in chronic mesial temporal lobe epilepsies (MTLE). The sclerotic hippocampus in Ammon's horn sclerosis (AHS)-associated MTLE, which shows segmental neuronal cell loss, axonal reorganization, and astrogliosis, would appear particularly susceptible to such changes. To characterize the cellular hippocampal pathology in MTLE, we have analyzed hilar neurons in surgical hippocampus specimens from patients with MTLE. Anatomically well-preserved hippocampal specimens from patients with AHS (n = 44) and from patients with focal temporal lesions (non-AHS; n = 20) were studied using confocal laser scanning microscopy (CFLSM) and electron microscopy (EM). Hippocampal samples from three tumor patients without chronic epilepsies and autopsy samples were used as controls. Using intracellular Lucifer Yellow injection and CFLSM, spiny pyramidal, multipolar, and mossy cells as well as non-spiny multipolar neurons have been identified as major hilar cell types in controls and lesion-associated MTLE specimens. In contrast, none of the hilar neurons from AHS specimens displayed a morphology reminiscent of mossy cells. In AHS, a major portion of the pyramidal and multipolar neurons showed extensive dendritic ramification and periodic nodular swellings of dendritic shafts. EM analysis confirmed the altered cellular morphology, with an accumulation of cytoskeletal filaments and increased numbers of mitochondria as the most prominent findings. To characterize cytoskeletal alterations in hilar neurons further, immunohistochemical reactions for neurofilament proteins (NFP), microtubule-associated proteins, and tau were performed. This analysis specifically identified large and atypical hilar neurons with an accumulation of low weight NFP. Our data demonstrate striking structural alterations in hilar neurons of patients with AHS compared with controls and non-sclerotic MTLE specimens. Such changes may develop during cellular reorganization in the epileptogenic hippocampus and are likely to contribute to the pathogenesis or maintenance of temporal lobe epilepsy.

Estudi del procés de tall per a la millora dels esbocats en materials AHSS

En aquest projecte es presenta un estudi sobre punxonats i esbocats per a materials d’Alt límit elàstic (Acers AHSS; Advanced High Strength Steels), amb l’objectiu d’aconseguir saber quin és el millor procés de tall per aconseguir esbocats correctes. Utilitzant la premsa hidràulica de la Fundació Centre Tecnològic de Manresa (CTM) es realitza el punxonat i els esbocats a tots els materials per tal d’avaluar el seu comportament alhora de ser esbocats. A partir d’aquests resultats obtindrem les gràfiques de ràtio d’esbocat per a cadascun dels materials en els diferents sentits de esbocat. Com a mètode alternatiu del procés de tall, es realitzen unes mostres amb tall per aigua. S’observarà si els esbocats trenquen o no trenquen i en quines condicions ho fan. Una vegada realitzats aquests assajos, es procedirà a realitzar l’estudi de microdureses amb un indentador Vickers per tal de comprovar l’afectació de l’operació de tall en la microestructura del material. Es realitzarà també algun perfil de dureses utilitzant el Nanoindentador. Per finalitzar el projecte, s’inclourà un apartat de conclusions i un estudi mediambiental produït com a conseqüència de l’elaboració del projecte, així com un pressupost total d’aquest.

Materials per a una edició crítica i comentada del París e Viana català

Edició crítica del París e Viana català que té com a testimoni de base l'incunable de Girona de 1495, que es conserva a la Biblioteca Reial de Copenhaguen. El treball conté una anàlisi comparativa de les variants de l'edició de Girona i de les de l'edició de Barcelona. Es desmenteix la hipòtesi de Cátedra (1986: 37-38), segons la qual el Curial e Güelfa podria dependre del París e Viana i s'argumenta la inconveniència de classificar aquesta novel·la cavalleresca com a ficció sentimental. Les notes permeten resseguir els motius literaris comuns que constitueixen el relat

Extracellular serine-proteinases isolated from Streptomyces alboniger: Partial characterization and effect of aprotinin on cellular structure

Streptomyces alboniger ATCC 12461 grown in brain heart infusion (BHI) medium produced two extracellular serine-proteinases, denoted SP I and SP II, which were purified by ammonium sulfate precipitation and aprotinin-agarose affinity chromatography. SP I was purified 88,9-fold and SP II 66,7- fold, with 33.4% and 10.4% yield, respectively. The optimum pH for the proteinases activity, using a-N-p-tosyl-L-arginine-methyl ester (TAME) as substrate, was 9-10 and the optimum temperature was 37ºC. The proteolytic activity of SP I and SP II was inhibited by aprotinin and SP I was partially inhibited by leupeptin, both serine-proteinase inhibitors. S. alboniger growth in BHI-liquid medium decreased when 5 mg/ml, 10 mg/ml of aprotinin was used, being completely inhibited with 20 mg/ml and 40 mg/ml. At the ultrastructural level, aprotinin-treated S. alboniger cells showed swelling of the bacterial body and condensation of the genetic material, probably related to the inhibition of its growth.

Cellular variability of RpoS expression underlies subpopulation activation of an integrative and conjugative element.

Conjugative transfer of the integrative and conjugative element ICEclc in the bacterium Pseudomonas knackmussii is the consequence of a bistable decision taken in some 3% of cells in a population during stationary phase. Here we study the possible control exerted by the stationary phase sigma factor RpoS on the bistability decision. The gene for RpoS in P. knackmussii B13 was characterized, and a loss-of-function mutant was produced and complemented. We found that, in absence of RpoS, ICEclc transfer rates and activation of two key ICEclc promoters (P(int) and P(inR)) decrease significantly in cells during stationary phase. Microarray and gene reporter analysis indicated that the most direct effect of RpoS is on P(inR), whereas one of the gene products from the P(inR)-controlled operon (InrR) transmits activation to P(int) and other ICEclc core genes. Addition of a second rpoS copy under control of its native promoter resulted in an increase of the proportion of cells expressing the P(int) and P(inR) promoters to 18%. Strains in which rpoS was replaced by an rpoS-mcherry fusion showed high mCherry fluorescence of individual cells that had activated P(int) and P(inR), whereas a double-copy rpoS-mcherry-containing strain displayed twice as much mCherry fluorescence. This suggested that high RpoS levels are a prerequisite for an individual cell to activate P(inR) and thus ICEclc transfer. Double promoter-reporter fusions confirmed that expression of P(inR) is dominated by extrinsic noise, such as being the result of cellular variability in RpoS. In contrast, expression from P(int) is dominated by intrinsic noise, indicating it is specific to the ICEclc transmission cascade. Our results demonstrate how stochastic noise levels of global transcription factors can be transduced to a precise signaling cascade in a subpopulation of cells leading to ICE activation.

Role of glutathione in macrophage activation: effect of cellular glutathione depletion on nitrite production and leishmanicidal activity.

We have examined the effects of two agents depleting the intracellular pool of glutathione (GSH) on macrophage activation induced by IFN-gamma + LPS, as measured by nitrite production and leishmanicidal activity. Diethylmaleate (DEM), which depletes intracellular GSH by conjugation via a reaction catalyzed by the GSH-S-transferase, strongly inhibited nitrite secretion and leishmanicidal activity when added before or at the time of addition of IFN-gamma + LPS; this inhibition was progressively lost when addition of DEM was delayed up to 10 hr. A close correlation was observed between levels of intracellular soluble GSH during activation and nitrite secretion. Inhibition was partially reversed by the addition of glutathione ethyl ester (GSH-Et). Buthionine sulfoximine (BSO), a specific inhibitor of gamma-glutamylcysteine synthetase, also inhibited macrophage activation, although to a lesser extent than DEM despite a more pronounced soluble GSH depletion. This inhibition was completely reversed by the addition of GSH-Et. DEM and BSO did not alter cell viability or PMA-triggered O2- production by activated macrophages, suggesting that the inhibitory effects observed on nitrite secretion and leishmanicidal activity were not related to a general impairment of macrophage function. DEM and BSO treatment reduced iNOS specific activity and iNOS protein in cytosolic extracts. DEM also decreased iNOS mRNA expression while BSO had no effect. Although commonly used as a GSH-depleting agent, DEM may have additional effects because it can also act as a sulhydryl reagent; BSO, on the other hand, which depletes GSH by enzymatic inhibition, has no effect on protein-bound GSH. Our results suggest that both soluble and protein-bound GSH may be important for the induction of NO synthase in IFN-gamma + LPS-activated macrophages.

La millor comprensió del desenvolupament cardíac i el dany cel•lular en els miòcits ajudarà a definir dianes terapèutiques

En el cor embrional, la senyalització de mort cel•lular apoptòtica iniciada per receptors de mort i les caspases executores 3 i 7 exerceixen un paper important durant el desenvolupament cardíac no relacionat amb la mort cel•lular, i posteriorment són silenciats en l’adult, on les vies independents de caspases estan implicades en la mort cel•lular patològica. Resultats previs del nostre grup han contribuït a entendre com es regula i silencia en el cor l’expressió dels gens de la via apoptòtica depenent de caspases durant el desenvolupament; a més, resultats no publicats demostren que les caspases regulen el procés d’expressió de gens en el cor i, contràriament a la maquinària depenent de caspases, TatD, una nucleasa, ’expressa abundantment al cor postnatal. Es desconeixen les funcions de la senyalització apoptòtica durant el desenvolupament cardíac, tot i que són essencials per al desenvolupament, a més, la senyalització independent de caspases implicada al dany cel•lular en els miòcits només es coneix parcialment, el nostre objectiu és contribuir al coneixement d’ambdós fenòmens. Creiem que les caspases podrien processar proteïnes reguladores de l’expressió de gens musculars alterant la seva activitat, mentre que TatD té un paper rellevant en el dany cel•lular però també en la funció cardíaca normal. Volem caracteritzar la contribució de les caspases 3 i 7 en el desenvolupament cardíac, utilitzant models in vivo (estem finalitzant els creuaments necessaris per a disposar dels animals amb el genotip desitjat) i in vitro (pràcticament hem preparat tot el material i hem optimitzat els protocols per a tirar-ho endavant). També volem caracteritzar la funció de TatD durant el desenvolupament i fisiologia del cor i conèixer-ne la seva funció utilitzant models in vitro i in vivo.

Early functional differentiation in the chick embryonic disc: interaction between mechanical activity and extracellular matrix

The mechanical behaviour of ectodermal cells in the area opaca and the supracellular organization of fibronectin in the adjacent extracellular matrix were studied in whole chick blastoderms developing in vitro. The pattern of spontaneous mechanical activity and its modification by immunoglobulins against fibronectin were determined using a real-time image-analysis system. The pattern of fibronectin was studied using immunocytochemical techniques. It was found that the ectodermal cells in the area opaca actively develop a radially oriented contraction, which leads to a distension of the area pellucida from which the embryo develops. Abnormally increased tension resulted in perturbations of gastrulation and neurulation. An optimized mechanical equilibrium within the blastoderm seems to be necessary for normal development. Anti-fibronectin antibodies applied to the basal side of the blastoderm led rapidly and reversibly to an increase of tension in the contracted cells. This observation indicates that modifications of the extracellular matrix can be transmitted to cytoskeletal elements within adjacent cells. The extracellular matrix of the area opaca contains fibronectin arranged in radially oriented fibrils. This orientation corresponds to the direction of migration of the mesodermal cells. Interestingly, the radial pattern of fibronectin is found in the regions where the ectodermal cells are contracted and develop radially oriented forces. This observation suggests that the supracellular assembly of the extracellular materials could be influenced by the mechanical activity of adjacent cells. Possible modulations of the supracellular organization of extracellular matrix by other factors, e.g. diffusible metabolites, is also discussed. The presence of characteristically organized extracellular matrix components, of spatially differentiated cell activities and of reciprocal interactions between them makes the young chick blastoderm an excellent system for physiological studies of the coordinated cellular activities that lead to changes in form, complexity and function.

Large agminated cellular 'plaque-type' blue nevus surrounding the ear: a case and review.

Large or giant cellular blue nevi are usually congenital and represent a challenge for the physician. Close anatomic structures may be altered by the size of the moles. In this article, we report the case of an uncommon large, agminated, cellular blue nevus of the 'plaque type' in a 42-year-old female. Due to the risks of malignant melanoma development on a large or giant blue nevus, we highlight the importance of proper histopathological diagnosis. Furthermore, because of the possibility that the nevus may invade the bone and cerebral tissues, we discuss the indication of a radiological diagnosis. The accurate correlation to clinical and histopathological findings and appropriate multidisciplinary management can save the lives of patients. © 2013 S. Karger AG, Basel.

Funcions de les molècules associades a la mielina i la proteïna priònica cel•lular (PrPc) en neurodegeneració (Malaltia d’Alzheimer) i neuroinflamació (Esclerosi Múltiple)

Les proteïnes associades a la mielina (MAIS), Nogo-A, MAG i OMgp, són molècules que presenten una capacitat inhibitòria molt important per el recreixement axonal i la neuroreparació després de lesió. No obstant des de fa anys les seves funcions han estat ampliades i s’han involucrat en diferents processos degeneratius del sistema nerviós o en processos neuroinflamatoris del sistema nerviós central i el perifèric com ara l'Escleresi Múltiple (MS). La base neurobiològica d’indicadors moleculars que són responsables del dany axonal en MS segueixen sense estar plenament descrits. Recentment s’ha publicat que el mecanisme de senyalització Nogo-A pot regir els primers canvis de la desmielinització immunomediada del sistema nerviós central en el model animal de MS, l’encefalomielitis autoimmune experimental (EAE). De la mateixa forma la proteïna priònica cel•lular és una proteïna que s’ha associat majoritàriament a malalties espongiformes, però que recentment s’ha vinculat (no sense controvèrsia) amb la seva possible relació amb la Malaltia d'Alzheimer (AD), ja que seria capaç de reclutar els oligòmers d’Aβ (ADDLs), els quals correlacionen millor amb el grau de demència, i amb els que interacciona directament, actuant així com un possible mediador de la fosforilació de tau en la malaltia. No obstant, les funcions de les MAIS i de la PrPc en aquests models de la malaltia no estan clarament definits i, per altra banda, es desconeixen els mecanismes de senyalització implicats, no descartant de forma clara el component neural i l’immune.

The influence of hydrocortisone on cellular defence mechanisms of Biomphalaria glabrata

Since the internal defense system of mollusks consists of cellular and humoral mechanisms, we examined the role of hydrocortisone in mollusks defense cells and the influence of this steroid on the development of Schistosoma mansoni in its intermediary host. Hydrocortisone had an immunosuppressive action in Biomphalaria glabrata, as reflected in the reduced number of defense cells and the altered cell physiology. Histopathological analysis showed that hydrocortisone facilitated the intramolluscan development of S. mansoni, by reducing the extent of the inflammatory response, seen as a greater number of viable sporocysts with no surrounding hemocytes.

Role of the sub-cellular localization of RasGAP fragment N2 for its ability to sensitize cancer cells to genotoxin-induced apoptosis.

The specific sensitization of tumor cells to the apoptotic response induced by genotoxins is a promising way of increasing the efficacy of chemotherapies. The RasGAP-derived fragment N2, while not regulating apoptosis in normal cells, potently sensitizes tumor cells to cisplatin- and other genotoxin-induced cell death. Here we show that fragment N2 in living cells is mainly located in the cytoplasm and only minimally associated with specific organelles. The cytoplasmic localization of fragment N2 was required for its cisplatin-sensitization property because targeting it to the mitochondria or the ER abrogated its ability to increase the death of tumor cells in response to cisplatin. These results indicate that fragment N2 requires a spatially constrained cellular location to exert its anti-cancer activity.