Proteins are the most important molecules in the cell since they take care of most of the biological functions which resemble life. To ensure that everything is working properly the cell has a rigorous control system to monitor the proper function of its proteins and sends old or dysfunctional proteins for degradation. Unfortunately, this system sometimes fails and the once so vital proteins start to misbehave or to accumulate and in the worst case scenario these undesired processes cause the death of their host. One example is Amyotrophic Lateral Sclerosis (ALS); a progressive and always fatal neurodegenerative disorder that is proposed to derive from accumulation of aberrant proteins. Over 140 mutations in the human gene encoding the cytosolic homodimeric enzyme Cu/Zn-Superoxide Dismutase (SOD1) are linked to ALS. The key event in SOD1 associated ALS seems to be the pathological formation of toxic protein aggregates as a result of initially unfolded or partly structured SOD1-mutants. Here, we have compared the folding behaviour of a set of ALS associated SOD1 mutants. Based on our findings we propose that SOD1 mediated ALS can be triggered by a decrease in protein stability but also by mutations which reduce the net charge of the protein. Both findings are in good agreement with the hypothesis for protein aggregation. SOD1 has also been found to be able to interact with mitochondrial membranes and SOD1 inclusions have been detected in the inter-membrane space of mitochondria originating from the spinal cord. The obvious question then arose; does the misfolding and aggregation of SOD1 involve erroneous interactions with membranes? Here, we could show that there is an electrostatically driven interaction between the reduced apo SOD1 protein including ALS associated SOD1-mutants and charged lipid membrane surfaces. This association process changes the secondary structures of these mutants in a way quite different from the situation found in membrane free aqueous environment. However, the result show that mutants interact with charged lipid vesicles to lesser extent than wildtype SOD1. This opposes the correlation between decreased SOD1 stability and disease progression. We therefore suggest that the observed interaction is not a primary cause in the ALS mechanism.

Amyotrophic lateral sclerosis (ALS) is a devastating disease that affects people in their late mid-life, with fatal outcome usually within a few years. The progressive degeneration of neurons responsible for muscle movement (motor neurons) throughout the central nervous system (CNS) leads to muscle wasting and paralysis, and eventually affects respiratory function. Most cases have no familial background (sporadic) whereas about 10% of cases have relatives affected by the disease. A substantial number of familial cases are caused by mutations in the gene encoding superoxide dismutase-1 (SOD1). Since the initial discovery of this relationship about 17 years ago, numerous workers have tried to identify the pathogenicity of mutant SOD1 but without any final agreement or consensus regarding mechanism. The experiments in this thesis have been aimed at finding common pathogenic mechanisms by analyzing transgenic mouse models expressing mutant SOD1s with widely different properties.     Mitochondrial pathology and dysfunction have been reported in both ALS patients and murine models. We used density gradient ultracentrifugation for comparison of mitochondrial partitioning of SOD1 in our transgenic models. It was found that models with high levels of mutant protein, overloaded mitochondria with high levels of SOD1-protein whereas models with wild type-like levels of mutant protein did not. No significant association of the truncation mutant G127X with mitochondria was found. Thus, if mitochondrial dysfunction and pathology are fundamental for ALS pathogenesis this is unlikely to be caused by physical association of mutant SOD1 with mitochondria.     Density gradient ultracentrifugation was used to study SOD1 inclusions in tissues from an ALS patient with a mutant SOD1 (G127X). We found large amounts in the ventral horns of the spinal cord but also in the liver and kidney, although at lower levels. This showed that such signs of the disease can also be found outside the CNS.     This method was used further to characterize SOD1 inclusions with regard to the properties of mutant SOD1 and the presence of other proteins. The inclusions were found to be complex detergent-sensitive structures with mutant SOD1 reduced at disulfide C57-C146 being the major inclusion protein, constituting at least 50% of the protein content. Ten co-aggregating proteins were isolated, some of which were already known to be present in cellular inclusions. Of great interest was the presence of several proteins that normally reside in the endoplasmic reticulum (ER), which is in accordance with recent data suggesting that the unfolded protein response (UPR) has a role in ALS.     To obtain unbiased information on the pathogenesis of mutant SOD1, we performed a total proteome study on spinal cords from ALS transgenic mice. By multivariate analysis of the 1,800 protein spots detected, 420 (23%) were found to significantly contribute to the difference between transgenic and control mice. From 53 proteins finally identified, we found pathways such as mitochondrial function, oxidative stress, and protein degradation to be affected by the disease. We also identified a previously uncharacterized covalent SOD1 dimer.    In conclusion, the work described in this thesis suggests that mutant SOD1 affects the function of mitochondria, but not mainly through direct accumulation of SOD1 protein. It also suggests that SOD1 inclusions, present in both the CNS and peripheral tissues, mainly consist of SOD1 but they also trap proteins involved in the UPR. This might be deleterious as motor neurons, unable to renew themselves, are dependent on proper protein folding and degradation.

This study aims to present and evaluate a new method for measuring the distribution of snow within built-up environments by differencing elevations collected by an Airborne Laser Scanner (ALS) before, and during peak snow accumulation. Few efforts have been made to study the distribution of snow within built-up environments due to the false assumption that high-intensity rainfall is the main contributor to peak yearly runoff rates. Traditional techniques for measuring snow are often difficult to replicate in built-up environments due to incompatibility of methods and barriers such as buildings, roads and private property. Light Detection and Ranging (LiDAR) technology, specifically ALSs, have previously been used to characterize the distribution of snow under forest canopy, and in remote mountain environments. This study investigates and assesses the utility of high resolution, non-intrusive ALS data for estimating the depth and distribution of snow within the town of Uxbridge, Ontario. ALS flights for this study were completed before the onset of snow accumulation, as well as near peak snow accumulation for the winters of 2010 and 2011. Pre and post snow accumulation ALS measured elevations were differenced to estimate the depth of the snowpack across the entire study area at a resolution of 0.5 m. Ground measurements of snow depth were also completed within 24 hours of each of the winter flights. The LiDAR-estimated and ground-measured snow depths were compared using Spearman's rank correlation coefficient as well as Mean Absolute Error (MAE) and Root Mean Squared Error (RMSE). Results from this thesis show that: 1) Snow depths estimated by differencing elevations from two ALS flights show a MAE of 3 cm and an RMSE of 10 cm when compared to ground-measured snow depths. (2) There is a strong, statistically significant relationship (ρ = 0:82, p < 0:001) between LiDAR-estimated and ground-measured snow depths. (3) An average bias of -3 cm was found for the entire dataset showing an underestimation in the LiDAR-estimated snow depths most likely caused by the effects of low lying vegetation on the fall ALS measurements. The results presented in this study demonstrate that ALSs are capable of providing high spatial resolution snow depth estimates within built-up environments. Furthermore, snow depth measurements made using an ALS can be used to increase the current body of knowledge on the distribution and re-distribution of snow within built-up environments. Snow distributions measured by an ALS could also be used for future development and verification of urban hydrological models.

La immigració xinesa a Catalunya ha viscut un augment exponencial en els darrers anys: mentre que al 1998 només hi havia 2.440 xinesos censats, al 2011 es va arribar als 47.493, una xifra que continua augmentant. L’arribada d’aquests nous ciutadans xinesos, juntament amb persones d’altres orígens, ha motivat l’aparició de dos nous perfils professionals: els intèrprets als serveis públics i els mediadors interculturals, que comparteixen la tasca de facilitar la comunicació entre els ciutadans que desconeixen les llengües oficials i el personal dels serveis públics de la societat receptora. La nostra tesi s’emmarca en aquest context i pretén unir dues línies de recerca que, fins a data d’avui, han avançat per separat: l’estudi de la interpretació als serveis públics (ISP) i l’estudi d’un col·lectiu immigrat, com és el cas dels xinesos. Per això, partim d’un marc teòric que tant inclou una panoràmica de la immigració xinesa a Catalunya i dels estudis que s’han fet sobre comunicació intercultural amb els xinesos, com una revisió àmplia de l’estat de la qüestió de la recerca en ISP a nivell general. El nostre objectiu principal és analitzar els tipus de resposta que s’han donat als problemes de comunicació amb el col·lectiu xinès als serveis públics a Catalunya, és a dir, analitzar la ISP i la mediació intercultural en aquests àmbits. D’aquest objectiu principal es desprenen d’altres objectius secundaris, tots amb el denominador comú d’intentar definir una mica millor el perfil professional de l’intèrpret als serveis públics i les especificitats de la comunicació mediada amb el col·lectiu xinès. Per assolir aquests objectius, ens hem basat en un enfocament mixt: qualitatiu i quantitatiu. D’una banda, hem realitzat entrevistes en profunditat a intèrprets i mediadors per al col·lectiu xinès i a representants d’entitats que contracten a aquests professionals; les hem transcrit i les hem analitzat mitjançant l’anàlisi qualitativa de continguts i l’anàlisi del discurs. De l’altra, hem distribuït qüestionaris en xinès a usuaris xinesos dels serveis públics, d’on hem extret unes dades que ens han permès una aproximació quantitativa a la nostra realitat d’estudi. Gràcies a la diversitat de dades obtingudes hem pogut entendre les diferents percepcions que diferents agents tenen de la ISP i de la mediació intercultural, així com també, afirmar que són pràctiques que sovint comparteixen molts punts en comú. D’entre les conclusions més destacades, hem subratllat la necessitat d’una millor definició del perfil professional i d’una formació coherent amb aquesta definició. Essent conscients del moment de crisi econòmica actual, prescindir o reduir els serveis d’ISP no hauria de ser ser la solució, sinó més aviat reestructurar-los basant-nos en l’experiència adquirida, per tal d’aproximar-nos al model integrador de solucions que ja fa uns anys que és una realitat als països més avançats en la ISP. Pel que fa a la comunicació amb el col·lectiu xinès, hem constat que existeixen certes especificitats: p. ex., la diversitat de variants lingüístiques, que caldria tenir-se en compte en la contractació d’intèrprets, o certs trets de la comunicació intercultural (p. ex., el llenguatge no verbal), que s’haurien de treballar en la formació. De fet, la formació apareix com un leitmotiv al llarg de les conclusions i, sobretot, quan tenim presents les dificultats que intèrprets i mediadors expliquen trobar-se en l’exercici de la seva pràctica professional. Finalment, la tesi conclou assenyalant noves línies d’investigació que s’obren a partir de la nostra recerca, d’entre les quals cal destacar l’estudi d’altres modalitats de comunicació mediada als serveis públics (p. ex., interpretació telefònica o traducció escrita de materials d’acollida) o l’anàlisi d’interaccions reals per tal d’aprofundir en els aspectes més relacionats amb la comunicació intercultural. / Chinese immigration in Catalonia has experienced a sharp increase in recent years: while in 1998 there were only 2,440 registered Chinese citizens, in 2011 they had already reached 47,493 –a number that keeps increasing. The presence of these new Chinese citizens, as well as of people from other origins, has motivated the emergence of two new professionals: public service interpreters and intercultural mediators. They both share the responsibility of enabling communication between citizens who do not know the official languages and public service providers of the host country. The present dissertation is set in this background and aims to combine two research lines that have advanced separately up to date: research into Public Service Interpreting (PSI) and the study of a specific immigrant community, as the Chinese may be. Therefore, the dissertation departs from a theoretical framework that includes an overview of Chinese immigration in Catalonia and of research in intercultural communication with the Chinese, as well as a wide revision of the state of the art of PSI in general. The main objective is to analyse the kind of responses that have been put forward to overcome the communication barriers with the Chinese in public services in Catalonia; i.e. to analyse PSI and intercultural mediation. This main objective is the mainstay of other secondary objectives, which to better describe the professional profile of public service interpreters and the specificities of mediated communication with the Chinese. To reach these objectives, the thesis bases on a mixed method: qualitative and quantitative. On the one hand, a sample of interpreters and mediators for the Chinese and a sample of coordinators in charge of the provision of PSI or intercultural mediation were interviewed. These informal interviews were transcribed and examined by means of qualitative content analysis and discourse analysis. On the other hand, written surveys in Chinese were distributed to a sample of Chinese users of public services. The data collected by means of the surveys has provided a quantitative approach to the object of study. The variety of data obtained reflects the different perceptions different agents have of PSI and intercultural mediation and pinpoints the convergences and divergences between the two practices. Among the main conclusions, the need for a better definition of the professional profile and for coherent training according to this definition is stressed. Conscious of the current economic crisis, reduction or even omission of PSI services should not be the solution, but to restructure them basing on the experienced gained in these fields, in order to approach the comprehensive model which is already a reality in the countries where PSI is most developed. Regarding the communication with the Chinese, certain specificities have been detected: for instance, the different linguistic varieties, that should be taken into account when contracting interpreters, or certain aspects of intercultural communication (e.g., non-verbal communication), that should be included in training courses. In fact, training appears as a leitmotiv throughout the conclusions and especially when thinking of the difficulties that interpreters and mediators have to face in their daily practice. Finally, the thesis concludes pointing out new research lines that arise from the research developed; for instance, the study of new modalities of mediated communication in public services (e.g. telephone interpreting and written translation of “welcome materials”) or the analysis of real interactions in order to shed light onto issues related to intercultural communication

DE LA TESIS:Desde la aparición de las quinolonas, las resistencias bacterianas a las mismas han evolucionado de forma paralela. Conforme han ido apareciendo nuevas moléculas con mayor capacidad bactericida y mejores parámetros farmacodinámicos y farmacocinéticos, también se han ido identificando nuevos y más sofisticados mecanismos de resistencia, que se han ido adaptando a las necesidades de cada momento. Es por ello que se hace necesaria una nueva metodología para el diseño y la síntesis de nuevos agentes antibacterianos con el objetivo de aumentar su potencia antibacteriana y, al mismo tiempo, disminuir la probabilidad de generar a corto plazo nuevos mecanismos de resistencia. Con este fin, es fundamental conocer todos los mecanismos de resistencia y cuál es, en detalle, su mecanismo de acción, para poder diseñar nuevas moléculas capaces de, manteniendo su capacidad bactericida, eludir dichos mecanismos.En esta tesis nos propusimos como objetivos fundamentales, en primer lugar, investigar las bases moleculares de los mecanismos de resistencia a quinolonas en bacterias Gram-negativas. En concreto, en Escherichia coli, Yersinia enterocolitica y Citrobacter freundii. Por otro lado, se llevaron a cabo cálculos de acoplamiento o "docking" mediante la utilización de programas informáticos con el fin de incrementar nuestros conocimientos respecto al modelo de interacción entra ADN-ADN girasa-quinolona, para de esta manera mejorar nuestra comprensión de los mecanismos de resistencia a fluoroquinolonas asociados a las mutaciones más comúnmente encontradas en el gen gyrA. Finalmente, el último de los objetivos que nos propusimos fue el de diseñar, sintetizar y evaluar diferentes derivados de ciprofloxacino y norfloxacino con el fin de encontrar entre ellos alguno que tuviera la capacidad de interaccionar con la enzima ADN girasa, poseyendo ésta uno o dos cambios aminoacídicos en su subunidad A.

Exoner är de sekvenser i DNA vilka rymmer koden för proteiner i människan och i alla andra organismer. Intronerna, vilka utgör utrymmet mellan exoner, består av ickekodande sekvenser och kontrollelement. Exoner tillhörande en gen måste inte alltid inkluderas i den slutliga mRNA produkten, alternativ splicing tillåter exkludering av vissa sekvenser och gör att en gen kan ha mer än en mRNA produkt, därigenom kan en gen koda för flera olika proteiner. Alternativ splicing är ett fält som snabbt utvecklas och dess relevans för många sjukdomar har blivit uppenbar. Detta arbete går igenom ett flertal av dessa sjukdomar för att sammanställa ny forskning och tydliggöra rollen av alternativ splicing i dem. De sjukdomar som undersökts är cystisk fibros, ärftlig frontotemporal dementia, systemisk lupus erythematosus, aniridi, myotonisk dystrofi, amyotrophic lateral sclerosoch familial dysautonomia. Dessa sjukdomar har involvering av alternativ splicing, de genetiska processerna bakom dem är dock mycket olika och kan visa på de många sätt alternativ splicing kan påverka cell och kroppsfunktion. Målet med arbetet är en översiktlig bild av framstegen som gjorts och vilken forskning som nu bedrivs.

Amyotrofisk Lateral Skleros, ALS, är en neurologisk sjukdom vilken leder till att samtliga kroppens muskler förtvinas och dör. Då sjukdomen saknar bot blir all behandling symptomatisk och individuellt anpassad för varje enskild persons behov. I denna systematiska litteraturstudie har det sökts efter olika sätt att stötta denna patientgrupp då syftet att belysa hur vi som personal kan hjälpa och stötta personer med ALS relaterad dysfagi och andningsproblem till en så bra tillvaro som möjligt skulle belysas.Författarna har funnit att omvårdnaden sällan sätts i fokus. Det är istället de lösningar som tar bort symtomet som fått fokus i flertalet av de artiklar som granskats. Att hjälpa dessa personer till trygga och oberoende människor som kan fortsätta att leva istället för som många av artiklarna visade då det gjordes insatser som ledde till att personerna blev mer bundna till sina anhöriga och sina vårdare.

Imazethapyr was recently approved for use in rice, but limited information is available regarding its efficacy, environmental fate or potential red rice resistance. Therefore, experiments were conducted to 1) determine the effect of flooding time, and stage of imazethapyr application in red rice control, 2) assess the acetolactate synthase resistance to imazethapyr on red rice ecotypes, 3) determine the relative photolysis of imazethapyr, and 4) determine the effect of soil and moisture on imazethapyr adsorption and availability. When imazethapyr was applied in sequential application of PRE followed by a POST application, to achieve >95% red rice control, flood needed to be established within 14 DAT when imazethapyr was applied EPOST, and 7 DAT when imazethapyr was applied LPOST. Delaying the flood up to 21 DAT reduced rice grain yield for both EPOST and LPOST application timings. Based on enzymatic activity, the mean I50 values were 1.5, 1.1, 1.5, 1.6, 20.8, and 590.6 mM of imazethapyr, respectively, for LA 5, MS 5, TX 4, ??Cypress??, ??CL-121??, and ??CL-161??. CL-161 was 32 times more resistant than CL-121, and at least 420 times more resistant than the average of the red rice ecotypes and ??Cypress??. Results from the ALS assay showed that red rice ecotypes and Cypress had high susceptibility to imazethapyr when compared with the tolerant CL-121 and the resistant CL-161. Measurable enzymatic tolerance to ALS-inhibiting herbicides has not yet developed in these red rice ecotypes. Imazethapyr quantum yield (fI ) was 0.023 ?? 0.002 while the hydroxyl radical rate constant ( I OH k?? ) was 2.8 ?? 0.44 x 1013 M-1 h-1. These results show that imazethapyr is susceptible to both direct and indirect photolysis. The results also show that imazethapyr photolysis in paddy water will be affected by turbidity due to its impact on the availability of sunlight to drive direct and indirect photolysis reactions. Imazethapyr was more available and more concentrated in sandy soil. With higher amounts of water in soil there was greater amount of imazethapyr in soil solution and a lower concentration of herbicide due to dilution. The double centrifuge method provided a better estimate of plant available herbicide.

Mitochondrial dysfunction plays a pivotal role in the development of aging phenotypes and aging-associated neurodegenerative disorders, such as Alzheimer’s disease (AD), Parkinson’s disease (PD) and Amyotrophic lateral sclerosis (ALS). Strategies that restore mitochondrial dysfunction may rescue the deficits of central metabolism in these disorders and improve cell survival. For example, we found that modulating the mTOR signaling pathway in a tissue culture model of aging-induced mitochondrial DNA mutation enhanced mitochondrial function as evidenced by increased oxygen consumption. Our previous melatonin studies also led us to hypothesize that caloric restriction and the hormone melatonin would reverse brain mitochondrial dysfunction in animal models of AD. Although caloric restriction did not improve mitochondrial function in a transgenic P301L tau model of AD, novel insight into the regulation of F0-F1 ATP synthase activity under CR was gained that may help explain the protective effects of CR in other disease models. In addition, we determined the effects of melatonin treatment on brain mitochondrial cytochrome c oxidase (COX) activity using the transgenic APPSWE mouse model of AD bred to double melatonin receptor (MT1 and MT2) knockout mice. COX activity declined with aging in control mice, but increased with aging in AD mice, most likely as a response to mitochondrial reactive oxygen species (ROS) induced by amyloid-beta generated through APP proteolysis. Both effects were blunted by melatonin treatment. The effects of melatonin were partially dependent on the G-protein coupled melatonin receptors. We also used PD models to identify therapies that restore mitochondrial dysfunction. We showed that overexpression of wild-type alpha synuclein (α-syn) in human neuroblastoma M17 cells resulted in mitochondrial oxygen consumption deficits; similar to the levels observed when PD mutant forms (A30P α-syn, E46K α-syn, and, A53T α-syn) were overexpressed. Mitochondria from cells overexpressing α-syn were more sensitive to a high iron environment, mimicking the physiological conditions in which dopaminergic neurons are found. Diethyl oxaloacetate, succinate, and several amino acids were protective, suggesting the possibility for effective dietary interventions for PD. Lastly, we delineated the level of mitochondrial complex IV activity between gray and white matter in human cervical and lumbar spinal cord, as well as mitochondrial aggregation in the entire neurovascular units (NVU) as a consequence of ALS. At the conclusion of these projects a better understanding of the molecular mechanisms leading to mitochondrial dysfunction in AD, PD, ALS, and aging was gained and promising strategies to delay or reverse these dysfunctions were developed.

Was der Körper vermag, ist auch dem Archiv gegeben: Bewegung sinnreich zu gestalten, „in Beziehung zu setzen“ und über das bloß Präsentische des „Da Seins“ zu erheben. Wie bei der Performance selbst ist aber auch im Archiv nichts selbst-verständlich. Erst die komplexe Struktur – der Umgang mit „dem, was da ist“ – verweist über sich selbst hinaus.