Augusto Matraga: um hérói além do bem e do mal - uma perpectiva trágico-ntológica

The present dissertation analyses the philosophy of Friedrich Nietzsche (1844-1900) and the short story written by João Guimarães Rosa (1908-1967), A hora e vez de Augusto Matraga (1946) seeking to point out the possibility of the philosophic application of some Nietzschean ideas to enlarge the aesthetic value of the short story. It has been especially aimed at applying the concept of the noble superior being described by Nietzsche, the ubermensch, to the hero s ontological nature, Augusto Matraga. Nietzsche s postulates of the will to power, the elements suggested of the trial between Dionysus and Apollo and the ascetic ideal, will be especially relevant to this work, which intends to establish until what point the hero can be conceived as a good ascetic person, in so far as his noble ontological nature trespass the Christian morality, bringing him closer to the man who, according to Nietzsche, is beyond good and evil. Some Heideggerian concepts will also be relevant in order to reinforce the idea that the judgment created about Matraga is just an appearance that does not contemplate his essential Being, contributing to veil his real tragic-ontological valour, what implies that the aesthetic power of the short has been under valued

ALPHA2 Adrenergic and High Affinity Serotonergic Receptor Changes in the Brain Stem of Streptozotocin induced Diabetic Rats

The brain stems (13S) of streptozotocin (STZ)-diabetic rats were studied lo see the changes in neurotransmitter content and their receptor regulation. The norepinephrine (NE) content determined in the diabetic brain stems did ^ control. an E showed la while PI turnover hri content increased significantly compared N^r eNveFa o the recep significant increase. The alpha2 adrenergic receptor IneP utisoulinntreat d ratsetheNE contentt dec^ sled was significantly reduced during diabetes. in versedcto reanorm sed ulcrea e tK reatment the state. while EPI content remained increased as in die diabetic B,, for a]pha2 adrenergic receptors slw^nificantly while Unlabelled clonidine inhibited [31-I]NE binding in BS of control, diabetic and insulin treated ulations bindi diabetic rats showed that alpha2 adrenergicre^ punks cojnidiabetic animal the ligand bound sites with Hill slopes significantly away from unity. weaker to the low affinity site than in controls. Insulin treatment reversed[ this allumbmn to control levels. The displacement analysis using (-)-epinephrine age in control and diabetic animals revealed two populations of receptor affinidtyo=tat ss. In control animals, when GTP analogue added with epinephrine, the curve nagnlde caofnfitnroit yS model; but in the diabetic BS this effect `not aobserved. In bintact oth the diabetic data thus showlthat the effects of monovalent cations on affinity alphaz adrenergic receptors have a reduced affinity v due in stem ialtered Itscppeomson(5- regulation. The serotonin (5-HT) coat hydroxy) tryptophan (5-HTP) showed an increase and its breakdown metabolite (5-hydroxy) indoleacetic acid (5-I{IAA) showed a significant decrease. This showed that in serotonergic which l nerves there is a disturbance in both synthetic and breankduomwnbers pretma'med ana increased 5-HT. The high affinity serotonin receptor um ese serotonerg decrease in the receptor affinity. The insulin ^treatmentsturtiy showsha decreased serotonergic receptor kinetic parameters to control level. receptor function. These changes in adrenergic and serotonergic receptor function were suggested to be important in insulin function during STZ diabetes.

Identifizierung und funktionelle Charakterisierung neuer A-Kinase-Ankerproteine (AKAPs) im Modellorganismus C. elegans

In dieser Arbeit sollten neue Interaktionspartner der regulatorischen Untereinheit (R-UE) der Proteinkinase A (PKA) und des Modellorganismus C. elegans identifiziert und funktionell charakterisiert werden. Im Gegensatz zu Säugern (vier Isoformen), exprimiert der Nematode nur eine PKA-R-Isoform. Mittels in silico Analysen und so genannten „Pulldown“ Experimenten, wurde insbesondere nach A Kinase Ankerproteinen (AKAP) in C. elegans gesucht. Aus in silico Recherchen resultiert das rgs5 Protein als mögliches Funktionshomolog des humanen AKAP10. Rgs5 enthält eine potenzielle, amphipathische Helix (AS 421-446, SwissProt ID A9Z1K0), die in Peptide-SPOT-Arrays (durchgeführt im Biotechnologie Zentrum in Oslo, AG Prof. K. Taskén) eine Bindung an RI und RII-UE zeigt. Eine ähnliche Lokalisation von rgs5 und hAKAP10 in der Zelle, sowie vergleichende BRET² Studien, weisen auf eine mögliche Funktionshomologie zwischen AKAP10 und rgs5 hin. Die hier durchgeführten Analysen deuten darauf hin, dass es sich bei rgs5 um ein neues, klassisches AKAP mit „RII bindender Domäne“ Motiv im Modellorganismus C. elegans handelt. Basierend auf so genannten „pulldown“ Versuchen können, neben „klassischen“ AKAPs (Interaktion über amphipathische Helices), auch Interaktionspartner ohne typische Helixmotive gefunden werden. Dazu gehört auch RACK1, ein multifunktionales Protein mit 7 WD40 Domänen, das ubiquitär exprimiert wird und bereits mehr als 70 Interaktionspartner in unterschiedlichsten Signalwegen komplexiert (Adams et al., 2011). Durch BRET² Interaktionsstudien und Oberflächenplasmonresonanz (SPR) Analysen konnten hRI und kin2 als spezifische Interaktionspartner von RACK1 verifiziert werden. Untersuchungen zur Identifikation der Interaktionsflächen der beiden Proteine RACK1 und hRI zeigten im BRET² System, dass RACK1 über die WD40 Domänen 1-2 und 6-7 interagiert. Die Analyse unterschiedlicher hRI-Deletionsmutanten deutet auf die DD-Domäne im N-Terminus und zusätzlich auf eine potenzielle BH3 Domäne im C-Terminus des Proteins als Interaktionsfläche mit RACK1 hin. Die Koexpression von hRI BH3 und RACK1 zeigt einen auffälligen ein Phänotyp in Cos7 Zellen. Dieser zeichnet sich unter anderem durch eine Degradation des Zellkerns, DNA Kondensation und eine starke Vakuolisierung aus, was beides als Anzeichen für einen programmierten Zelltod interpretiert werden könnte. Erste Untersuchungen zum Mechanismus des ausgelösten Zelltods deuten auf eine Caspase unabhängige Apoptose (Paraptose) hin und einen bislang unbekannten Funktionsmechanismus der PKA hin.

Optimization-Based Robotic Manipulation for Safe Interaction with Human Operators

This thesis investigates a method for human-robot interaction (HRI) in order to uphold productivity of industrial robots like minimization of the shortest operation time, while ensuring human safety like collision avoidance. For solving such problems an online motion planning approach for robotic manipulators with HRI has been proposed. The approach is based on model predictive control (MPC) with embedded mixed integer programming. The planning strategies of the robotic manipulators mainly considered in the thesis are directly performed in the workspace for easy obstacle representation. The non-convex optimization problem is approximated by a mixed-integer program (MIP). It is further effectively reformulated such that the number of binary variables and the number of feasible integer solutions are drastically decreased. Safety-relevant regions, which are potentially occupied by the human operators, can be generated online by a proposed method based on hidden Markov models. In contrast to previous approaches, which derive predictions based on probability density functions in the form of single points, such as most likely or expected human positions, the proposed method computes safety-relevant subsets of the workspace as a region which is possibly occupied by the human at future instances of time. The method is further enhanced by combining reachability analysis to increase the prediction accuracy. These safety-relevant regions can subsequently serve as safety constraints when the motion is planned by optimization. This way one arrives at motion plans that are safe, i.e. plans that avoid collision with a probability not less than a predefined threshold. The developed methods have been successfully applied to a developed demonstrator, where an industrial robot works in the same space as a human operator. The task of the industrial robot is to drive its end-effector according to a nominal sequence of grippingmotion-releasing operations while no collision with a human arm occurs.

Diferencias entre la regla de exclusión probatoria nacional y la regla de exclusión del Derecho Internacional Penal: análisis de tres casos de injerencias en comunicaciones privadas.

Teniendo en cuenta tres casos dentro del contexto del conflicto armado donde la jurisdicción penal colombiana estudia la admisión o exclusión de medios probatorios producidos a partir de injerencias de comunicaciones, encontramos que no siempre se aplica la regla de exclusión cuando no cuentan con una orden judicial previa. Ante este problema, buscamos como se ha solucionado, teniendo como referente la el Derecho Internacional Humanitario, el Derecho Internacional de los Derechos Humanos. Lo anterior nos da base para entender la regla de exclusión de que se ha venido desarrollando dentro del Derecho Internacional Penal. Si bien estas reglas nos sirven para solucionar estos tres casos, terminan siendo contradictorios a lo que ha venido desarrollando la Corte Constitucional. De esta manera, si bien esta Corte consideró que la regla de Estatuto de Roma, va en concordancia con la nuestra regla de exclusión nacional, tal afirmación es errónea.

Producció i caracterització de variants de la ribonucleasa pancreàtica humana dissenyades per a adquirir propietats citotòxiques

Amb la finalitat d'aprofundir en les bases moleculars de la citotoxicitat de les ribonucleases pancreàtiques, es van construir variants derivades de l'HP-RNasa seguint dues estratègies. En la primera, es van generar variants de l'enzim resistents a l'acció de l'inhibidor proteic de les ribonucleases (hRI), substituint residus implicats en la interfície de contacte entre la ribonucleasa i l'hRI. En la segona, es va addicionar el motiu RGD en regions de superfície de la proteïna implicades en la formació del complex amb l'hRI, a fi de promoure la seva interacció amb la membrana plasmàtica de les cèl·lules i a la vegada disminuir l'afinitat de les variants per l'hRI. Es va comprovar que només les variants portadores de substitucions múltiples adquirien la capacitat de resistència a l'hRI. L'estudi del percentatge d'inhibició de la síntesi proteica en cèl·lules incubades amb cadascuna de les variants va mostrar que només dues de les variants construïdes havien adquirit propietats citotòxiques. La citotoxicitat més elevada la va presentar una variant que no era resistent a l'hRI, amb valors que eren només entre 5 i 15 vegades inferiors als de l'onconasa. Aquest resultat demostrà que la sensibilitat a l'hRI no és necessàriament un paràmetre limitant per a la citotoxicitat de les ribonucleases. Cap de les variants que incorporava un motiu RGD presentà citotoxicitat, evidenciant que aquest motiu no és efectiu a fi de dotar les ribonucleases pancreàtiques de propietats citotòxiques. Es van estudiar les bases moleculars de la citotoxicitat de la variant més citotòxica. En primer lloc, l'anàlisi de la internalització per marcatge radioactiu d'aquesta variant en relació amb l'onconasa i amb altres variants de l'HP-RNasa no citotòxiques, va posar en evidència que només l'onconasa era internalitzada eficientment. Es descartava així la possibilitat que l'acció citotòxica de l'enzim estudiat fos conseqüència d'una major eficiència d'endocitosi. També es va comprovar que l'addició del motiu RGD no era capaç de promoure la internalització de les proteïnes amb més eficàcia. Per microscòpia confocal de fluorescència, les variants humanes només es van començar a detectar a l'interior de la cèl·lula a partir de les 24 h d'incubació. Totes les variants generades van presentar una eficiència catalítica superior al 50 % de l'activitat de la seva proteïna parental, PM5, indicant que probablement l'estructura del centre actiu no havia estat afectada de manera dràstica per les substitucions introduïdes. No obstant, en tots els casos es va produir una disminució en la termoestabilitat respecte a PM5. Aquest resultat indicà que la correlació descrita a la bibliografia entre l'increment de termoestabilitat i l'increment de citotoxicitat per les ribonucleases no sempre es compleix. Per microscòpia confocal es va comprovar que tant la proteïna més citotòxica, com una variant no citotòxica resistent a l'hRI, així com la proteïna parental, seguien la via de degradació lisosomal. Aquesta ruta de trànsit no va ser afectada per l'addició de drogues que alteren les vies de trànsit retrògrad (monensina i brefeldina A), però sí per l'addició de la bafilomicina A1, una droga que neutralitza el pH endosomal i que va actuar alentint el trànsit de les proteïnes als lisosomes. D'acord amb aquests resultats, els valors de citotoxicitat de les variants es van incrementar de manera significativa només en presència de bafilomicina A1, suggerint que les ribonucleases transloquen al citoplasma a partir d'algun punt de la via de trànsit endosomal. Es va comprovar que l'acció de la variant més citotòxica era deguda a que l'addició d'un segon motiu de tres Arg en PE5 dota a aquesta proteïna amb un senyal de transport nuclear. La fracció d'enzim que aconsegueix translocar al citoplasma a partir d'algun punt de la via endosomal previ als lisosomes, és conduït ràpidament al nucli de la cèl·lula per mitjà del mecanisme clàssic de transport actiu. Per la seva afinitat amb l'rRNA, l'enzim es concentra en el nuclèol, on probablement duu a terme la seva activitat catalítica. La interacció d'aquesta variant amb els receptors nucleocitoplasmàtics, les importines, impediria per altra banda el bloqueig de l'enzim per part de l'hRI. Els resultats obtinguts presenten una nova estratègia de disseny de ribonucleases citotòxiques, basada en l'addició de segments NLS a fi de promoure el transport nuclear dels enzims. Aquesta estratègia podria permetre superar limitacions que fins al moment han estat descrites com a limitants de la citotoxicitat de les ribonucleases pancreàtiques, com la sensibilitat a l'hRI o la baixa eficiència d'internalització.

Precision placement of fertiliser for optimising the early nutrition of vegetable crops: a review of the implications for the yield and quality of crops, and their nutrient use efficiency

The research outlined in this paper highlights the importance of the early nutrition of vegetable crops, and its long-term effects on their subsequent growth and development. Results are also presented to demonstrate how the nutrient supply during the establishment stages of young seedlings and transplants can be enhanced by targeting fertiliser to a zone close to their developing roots. Three different precision fertiliser placement techniques are compared for this purpose: starter, band or side-injected fertiliser. The use of each of these methods consistently produced the same (or greater) yields at lower application rates than those from conventional broadcast applications, increasing the apparent recovery of N, P and K, and the overall efficiency of nutrient use, while reducing the levels of residual nutrients in the soil. Starter fertilisers also advanced the maturity of some crops, and enhanced produce quality by increasing the proportions of the larger and/or more desirable marketable grades. The benefits of the different placement techniques are illustrated with selected examples from research at Warwick HRI using different vegetable crops, including lettuce, onion and carrot.

Utanförskapets ”frustrerade ungdom” : Talet om de ”problematiska” förorterna i svensk kvällspress under augusti- september månad 2009

Uppsatsen är en innehållsanalytisk studie av kvällstidningarna Expressen och Aftonbladet under augusti- september 2009. Syftet är att undersöka hur s.k. ”invandrartäta områden”/förorter framställs i kvällstidningarna under nämnda period samt att ge exempel på hur detta kan analyseras ur ett sociologiskt perspektiv. Urvalet av artiklar tillåter sig avgränsas genom att se till vad som skrivs om de förortsområden som under aktuell period tillåtit sig förknippas med ”problem”. Frågeställningarna vill således belysa vad som under aktuell period skrivs om dessa områden: hur kan dessa kan budskap tolkas, och hur kan de tolkas med sociologiska teorier? ”Invandrartäta områden” har länge varit föremål för negativ stereotypisering bl.a. i media. Varför uppsatsen använder sig av teorier som kan förklara hur mediediskursen kan vara möjlig, och i vilket Elias teori om figurationer bildar en yttre ram. Analysen av resultatet uppvisar att flertalet artiklar ger utrymme åt reproduktion av ”moralisk differentiering” och ”illusorisk korrelation”, och med detta en förstärkning av ”common sense” uppfattningar hos läsaren. En uppfattning där förorten och dess invånare alltjämt tillskrivs rollen som de ”Andra”. Emellertid är inte endast sådana artiklar häri förekomna. En rimlig hypotes är dock att; hur artiklarna påverkar läsaren är avhängigt dennes befintliga attityd, samt hur artiklarna är utformade, och där det ges mer eller mindre utrymme för attitydförändringar.

Modeling the heterogeneous ice and gas coma of Comet 103P/Hartley 2

The spectacular images of Comet 103P/Hartley 2 recorded by the Medium Resolution Instrument (MRI) and High Resolution Instrument (HRI) on board of the Extrasolar Planet Observation and Deep Impact Extended Investigation (EPOXI) spacecraft, as the Deep Impact extended mission, revealed that its bi-lobed very active nucleus outgasses volatiles heterogeneously. Indeed, CO2 is the primary driver of activity by dragging out chunks of pure ice out of the nucleus from the sub-solar lobe that appear to be the main source of water in Hartley 2's coma by sublimating slowly as they go away from the nucleus. However, water vapor is released by direct sublimation of the nucleus at the waist without any significant amount of either CO2 or icy grains. The coma structure for a comet with such areas of diverse chemistry differs from the usual models where gases are produced in a homogeneous way from the surface. We use the fully kinetic Direct Simulation Monte Carlo model of Tenishev et al. (Tenishev, V.M., Combi, M.R., Davidsson, B. [2008]. Astrophys. J. 685, 659-677; Tenishev, V.M., Combi, M.R., Rubin, M. [2011]. Astrophys. J. 732, 104-120) applied to Comet 103P/Hartley 2 including sublimating icy grains to reproduce the observations made by EPOXI and ground-based measurements. A realistic bi-lobed nucleus with a succession of active areas with different chemistry was included in the model enabling us to study in details the coma of Hartley 2. The different gas production rates from each area were found by fitting the spectra computed using a line-by-line non-LTE radiative transfer model to the HRI observations. The presence of icy grains with long lifetimes, which are pushed anti-sunward by radiation pressure, explains the observed OH asymmetry with enhancement on the night side of the coma.

Mechanisms Underlying the Heterogeneous Sensitivities of Cancer Cells to Proteasome Inhibitors

The mechanisms underlying cellular response to proteasome inhibitors have not been clearly elucidated in solid tumor models. Evidence suggests that the ability of a cell to manage the amount of proteotoxic stress following proteasome inhibition dictates survival. In this study using the FDA-approved proteasome inhibitor bortezomib (Velcade®) in solid tumor cells, we demonstrated that perhaps the most critical response to proteasome inhibition is repression of global protein synthesis by phosphorylation of the eukaryotic initiation factor 2-α subunit (eIF2α). In a panel of 10 distinct human pancreatic cancer cells, we showed marked heterogeneity in the ability of cancer cells to induce eIF2α phosphorylation upon stress (eIF2α-P); lack of inducible eIF2α-P led to excessive accumulation of aggregated proteins, reactive oxygen species, and ultimately cell death. In addition, we examined complementary cytoprotective mechanisms involving the activation of the heat shock response (HSR), and found that induction of heat shock protein 70 kDa (Hsp72) protected against proteasome inhibitor-induced cell death in human bladder cancer cells. Finally, investigation of a novel histone deacetylase 6 (HDAC6)-selective inhibitor suggested that the cytoprotective role of the cytoplasmic histone deacetylase 6 (HDAC6) in response to proteasome inhibition may have been previously overestimated.

HUNT: launch of a full-length cDNA database from the Helix Research Institute

The Helix Research Institute (HRI) in Japan is releasing 4356 HUman Novel Transcripts and related information in the newly established HUNT database. The institute is a joint research project principally funded by the Japanese Ministry of International Trade and Industry, and the clones were sequenced in the governmental New Energy and Industrial Technology Development Organization (NEDO) Human cDNA Sequencing Project. The HUNT database contains an extensive amount of annotation from advanced analysis and represents an essential bioinformatics contribution towards understanding of the gene function. The HRI human cDNA clones were obtained from full-length enriched cDNA libraries constructed with the oligo-capping method and have resulted in novel full-length cDNA sequences. A large fraction has little similarity to any proteins of known function and to obtain clues about possible function we have developed original analysis procedures. Any putative function deduced here can be validated or refuted by complementary analysis results. The user can also extract information from specific categories like PROSITE patterns, PFAM domains, PSORT localization, transmembrane helices and clones with GENIUS structure assignments. The HUNT database can be accessed at http://www.hri.co.jp/HUNT.

Role of heme regulated eIF2α kinase in pancreatic beta cell function and viability

The eukaryotic translation initiation factor 2 alpha (eIF2α) is part of the initiation complex that drives the initiator amino acid methionine to the ribosome, a crucial step in protein translation. In stress conditions such as virus infection, endoplasmic reticulum (ER) stress, amino acid or heme deficiency eIF2α can be phosphorylated and thereby inhibit global protein synthesis. This adaptive mechanism prevents protein accumulation and consequent cytotoxic effects. Heme-regulated eIF2α kinase (HRI) is a member of the eIF2α kinase family that regulates protein translation in heme deficiency conditions. Although present in all tissues, HRI is predominantly expressed in erythroid cells where it remains inactive in the presence of normal heme concentrations. In response to heme deficiency, HRI is activated and phosphorylates eIF2α decreasing globin synthesis. This mechanism is important to prevent accumulation of heme-free globin chains which cause ER stress and apoptosis. RNA sequencing data from our group showed that in human islets and in primary rat beta cells HRI is the most expressed eIF2α kinase compared to the other family members. Despite its high expression levels, little is known about HRI function in beta cells. The aim of this project is to identify the role of HRI in pancreatic beta cells. This was investigated taking a loss-of-function approach. HRI knock down (KD) by RNA interference induced beta cell apoptosis in basal condition. HRI KD potentiated the apoptotic effects of palmitate or proinflammatory cytokines, two in vitro models for type 2 and type 1 diabetes, respectively. Increased cytokine-induced apoptosis was also observed in HRI-deficient primary rat beta cells. Unexpectedly, we observed a mild increase in eIF2α phosphorylation in HRI-deficient cells. The levels of mRNA or protein expression of C/EBP homologous protein (CHOP) and activating transcription factor 4 (ATF4) were not modified. HRI KD cells have decreased spliced X-box binding protein 1 (XBP1s), an important branch of the ER stress response. However, overexpression of XBP1s by adenovirus in HRI KD cells did not protect from HRI siRNA-induced apoptosis. HRI deficiency decreased phosphorylation of Akt and its downstream targets glycogen synthase kinase 3 (GSK3), forkhead box protein O1 (FOXO1) and Bcl-2-associated death promoter (BAD). Overexpression of a constitutively active form of Akt by adenovirus in HRI-deficient beta cells partially decreased HRI KD-mediated apoptosis. Interestingly, BAD silencing protected from apoptosis caused by HRI deficiency. HRI silencing in beta cells also induced JNK activation. These results suggest an important role of HRI in beta cell survival through modulation of the Akt/BAD pathway. Thus, HRI may be an interesting target to modulate beta cell fate in diabetic conditions.