Avaliação do potencial cancerigénico de microcistinas (cianotoxinas)

Tese de doutoramento em Farmácia (Toxicologia), apresentada à Faculdade de Farmácia da Universidade de Lisboa, 2009.

An evolutionary model explaining the Neolithic transition from egalitarianism to leadership and despotism.

The Neolithic was marked by a transition from small and relatively egalitarian groups, to much larger groups with increased stratification. But the dynamics of this remain poorly understood. It is hard to see how despotism can arise without coercion, yet coercion could not easily have occurred in an egalitarian setting. Using a quanti- tative model of evolution in a patch-structured population, we demonstrate that the interaction between demographic and ecological factors can overcome this conundrum. We model the co-evolution of individual preferences for hierarchy alongside the degree of despotism of leaders, and the dispersal preferences of followers. We show that voluntary leadership without coercion can evolve in small groups, when leaders help to solve coordination problems related to resource production. An example is coordinating construction of an irrigation system. Our model predicts that the transition to larger despotic groups will then occur when: 1. surplus resources lead to demographic expansion of groups, removing the viability of an acephalous niche in the same area and so locking individuals into hierarchy; 2. high dispersal costs limit followers' ability to escape a despot. Empirical evidence suggests that these conditions were likely met for the first time during the subsistence intensification of the Neolithic.

The interplay between tectonics, sediment dynamics and gateways evolution in the Danube system from the Pannonian Basin to the western Black Sea

Understanding the natural evolution of a river–delta–sea system is important to develop a strong scientific basis for efficient integrated management plans. The distribution of sediment fluxes is linked with the natural connection between sediment source areas situated in uplifting mountain chains and deposition in plains, deltas and, ultimately, in the capturing oceans and seas. The Danube River–western Black Sea is one of the most active European systems in terms of sediment re-distribution that poses significant societal challenges. We aim to derive the tectonic and sedimentological background of human-induced changes in this system and discuss their interplay. This is obtained by analysing the tectonic and associated vertical movements, the evolution of relevant basins and the key events affecting sediment routing and deposition. The analysis of the main source and sink areas is focused in particular on the Miocene evolution of the Carpatho-Balkanides, Dinarides and their sedimentary basins including the western Black Sea. The vertical movements of mountains chains created the main moments of basin connectivity observed in the Danube system. Their timing and effects are observed in sediments deposited in the vicinity of gateways, such as the transition between the Pannonian/Transylvanian and Dacian basins and between the Dacian Basin and western Black Sea. The results demonstrate the importance of understanding threshold conditions driving rapid basins connectivity changes superposed over the longer time scale of tectonic-induced vertical movements associated with background erosion and sedimentation. The spatial and temporal scale of such processes is contrastingly different and challenging. The long-term patterns interact with recent or anthropogenic induced modifications in the natural system and may result in rapid changes at threshold conditions that can be quantified and predicted. Their understanding is critical because of frequent occurrence during orogenic evolution, as commonly observed in the Mediterranean area and discussed elsewhere.

The pianosa contourite depositional system (northern Tyrrhenian sea): drift morphology and plio-quaternary stratigraphic evolution

The Pianosa Contourite Depositional System (CDS) is located in the Corsica Trough (Northern Tyrrhenian Sea), a confined basin dominated by mass transport and contour currents in the eastern flank and by turbidity currents in the western flank. The morphologic and stratigraphic characterisation of the Pianosa CDS is based on multibeam bathymetry, seismic reflection data (multi-channel high resolution mini GI gun, single-channel sparker and CHIRP), sediment cores and ADCP data. The Pianosa CDS is located at shallow to intermediate water depths (170 to 850 m water depth) and is formed under the influence of the Levantine Intermediate Water (LIW). It is 120 km long, has a maximum width of 10 km and is composed of different types of muddy sediment drifts: plastered drift, separated mounded drift, sigmoid drift and multicrested drift. The reduced tectonic activity in the Corsica Trough since the early Pliocene permits to recover a sedimentary record of the contourite depositional system that is only influenced by climate fluctuations. Contourites started to develop in the Middle-Late Pliocene, but their growth was enhanced since the Middle Pleistocene Transition (0.7–0.9 Ma). Although the general circulation of the LIW, flowing northwards in the Corsica Trough, remained active all along the history of the system, contourite drift formation changed, controlled by sediment influx and bottom current velocity. During periods of sea level fall, fast bottom currents often eroded the drift crest in the middle and upper slope. At that time the proximity of the coast to the shelf edge favoured the formation of bioclastic sand deposits winnowed by bottom currents. Higher sediment accumulation of mud in the drifts occurred during periods of fast bottom currents and high sediment availability (i.e. high activity of turbidity currents), coincident with periods of sea level low-stands. Condensed sections were formed during sea level high-stands, when bottom currents were more sluggish and the turbidite system was disconnected, resulting in a lower sediment influx.

APP RERMS enriched matrix as an adhesion substrate for neuritic outgrowth

Specific domains can determine protein structural functional relationships. For the Alzheimer’s Amyloid Precursor Protein (APP) several domains have been described, both in its intracellular and extracellular fragments. Many functions have been attributed to APP including an important role in cell adhesion and cell to cell recognition. This places APP at key biological responses, including synaptic transmission. To fulfil these functions, extracellular domains take on added significance. The APP extracellular domain RERMS is in fact a likely candidate to be involved in the aforementioned physiological processes. A multidisciplinary approach was employed to address the role of RERMS. The peptide RERMS was crosslinked to PEG (Polyethylene glycol) and the reaction validated by FTIR (Fourier transform infrared spectrometry). FTIR proved to be the most efficient at validating this reaction because it requires only a drop of sample, and it gives information about the reactions occurred in a mixture. The data obtained consist in an infrared spectra of the sample, where peaks positions give information about the structure of the molecules, and the intensity of peaks is related to the concentration of the molecules. Subsequently substrates of PEG impregnated with RERMS were prepared and SH-SY5Y (human neuroblastoma cell line) cells were plated and differentiated on the latter. Several morphological alterations were clearly evident. The RERMS peptide provoked cells to take on a flatter appearance and the cytoskeletal architecture changed, with the appearance of stress fibres, a clear indicator of actin reorganization. Given that focal adhesions play a key role in determining cellular structure the latter were directly investigated. Focal adhesion kinase (FAK) is one of the most highly expressed proteins in the CNS (central nervous system) during development. It has been described to be crucial for radial migration of neurons. FAK can be localized in growth cones and mediated the response to attractive and repulsive cues during migration. One of the mechanisms by which FAK becomes active is by auto phosphorylation at tyrosine 397. It became clearly evident that in the presence of the RERMS peptide pFAK staining at focal adhesions intensified and more focal adhesions became apparent. Furthermore speckled structures in the nucleus, putatively corresponding to increased expression activity, also increased with RERMS. Taken together these results indicate that the RERMS domain in APP plays a critical role in determining cellular physiological responses. Here is suggested a model by which RERMS domain is recognized by integrins and mediate intracellular responses involving FAK, talin, actin filaments and vinculin. This mechanism probably is responsible for mediating cell adhesion and neurite outgrowth on neurons.

Immune function, gene expression, blood indices and performance in transition dairy cows affected by diet and inflammation

The transition period is associated with the peak incidence of production problems, metabolic disorders and infectious diseases in dairy cows (Drackley, 1999). During this time the cow’s immune system seems to be weakened; it is apparent that metabolic challenges associated with the onset of lactation are factors capable of affecting immune function. However, the reasons for this state are not entirely clear (Goff, 2006). The negative energy balance associated with parturition leads to extensive mobilization of fatty acids stored in adipose tissue, thus, causing marked elevations in blood non-esterified fatty acids (NEFA) and B-hydroxybutyrate (BHBA) concentrations (Drackley et al., 2001). Prepartal level of dietary energy can potentially affect adipose tissue deposition and, thus, the amount of NEFA released into blood and available for metabolism in liver (Drackley et al., 2005). The current feeding practices for pregnant non-lactating cows has been called into question because increasing amounts of moderate-to-high energy diets (i.e. those more similar to lactation diets in the content of energy) during the last 3 wk postpartum have largely failed to overcome peripartal health problems, excessive body condition loss after calving, or declining fertility (Beever, 2006). Current prepartal feeding practices can lead to elevated intakes of energy, which can increase fat deposition in the viscera and upon parturition lead to compromised liver metabolism (Beever, 2006, Drackley et al., 2005). Our general hypothesis was that overfeeding dietary energy during the dry period, accompanied by the metabolic challenges associated with the onset of lactation would render the cow’s immune function less responsive early postpartum. The chapters in this dissertation evaluated neutrophil function, metabolic and inflammation indices and gene expression affected by the plane of dietary energy prepartum and an early post-partum inflammatory challenge in dairy cows. The diet effect in this experiment was transcendental during the transition period and potentially during the entire lactation. Changes in energy balance were observed and provided a good model to study the challenges associated with the onset of lactation. Overall the LPS model provided a consistent response representing an inflammation incident; however the changes in metabolic indices were sudden and hard to detect in most of the cases during the days following the challenge. In general overfeeding dietary energy during the dry period resulted in a less responsive immune function during the early postpartum. In other words, controlling the dietary energy prepartum has more benefits for the dairy cow during transition.

Engineering the transition from non-living to living matter

Re-creating and understanding the origin of life represents one of the major challenges facing the scientific community. We will never know exactly how life started on planet Earth, however, we can reconstruct the most likely chemical pathways that could have contributed to the formation of the first living systems. Traditionally, prebiotic chemistry has investigated the formation of modern life’s precursors and their self-organisation under very specific conditions thought to be ‘plausible’. So far, this approach has failed to produce a living system from the bottom-up. In the work presented herein, two different approaches are employed to explore the transition from inanimate to living matter. The development of microfluidic technology during the last decades has changed the way traditional chemical and biological experiments are performed. Microfluidics allows the handling of low volumes of reagents with very precise control. The use of micro-droplets generated within microfluidic devices is of particular interest to the field of Origins of Life and Artificial Life. Whilst many efforts have been made aiming to construct cell-like compartments from modern biological constituents, these are usually very difficult to handle. However, microdroplets can be easily generated and manipulated at kHz rates, making it suitable for high-throughput experimentation and analysis of compartmentalised chemical reactions. Therefore, we decided to develop a microfluidic device capable of manipulating microdroplets in such a way that they could be efficiently mixed, split and sorted within iterative cycles. Since no microfluidic technology had been developed before in the Cronin Group, the first chapter of this thesis describes the soft lithographic methods and techniques developed to fabricate microfluidic devices. Also, special attention is placed on the generation of water-in-oil microdroplets, and the subsequent modules required for the manipulation of the droplets such as: droplet fusers, splitters, sorters and single/multi-layer micromechanical valves. Whilst the first part of this thesis describes the development of a microfluidic platform to assist chemical evolution, finding a compatible set of chemical building blocks capable of reacting to form complex molecules with endowed replicating or catalytic activity was challenging. Abstract 10 Hence, the second part of this thesis focuses on potential chemistry that will ultimately possess the properties mentioned above. A special focus is placed on the formation of peptide bonds from unactivated amino acids, despite being one of the greatest challenges in prebiotic chemistry. As opposed to classic prebiotic experiments, in which a specific set of conditions is studied to fit a particular hypothesis, we took a different approach: we explored the effects of several parameters at once on a model polymerisation reaction, without constraints on hypotheses on the nature of optimum conditions or plausibility. This was facilitated by development of a new high-throughput automated platform, allowing the exploration of a much larger number of parameters. This led us to discover that peptide bond formation is less challenging than previously imagined. Having established the right set of conditions under which peptide bond formation was enhanced, we then explored the co-oligomerisation between different amino acids, aiming for the formation of heteropeptides with different structure or function. Finally, we studied the effect of various environmental conditions (rate of evaporation, presence of salts or minerals) in the final product distribution of our oligomeric products.

Excitonic behaviour in polymeric semiconductors : the effect of morphology and composition in heterostructures

La compréhension des interrelations entre la microstructure et les processus électroniques dans les polymères semi-conducteurs est d’une importance primordiale pour leur utilisation dans des hétérostructures volumiques. Dans cette thèse de doctorat, deux systémes diffèrents sont étudiés ; chacun de ces systèmes représente une approche diffèrente pour optimiser les matériaux en termes de leur microstructure et de leur capacité à se mettre en ordre au niveau moléculaire. Dans le premier système, j’ai effectué une analyse complète des principes de fonctionnement d’une cellule photovoltaïque hybride à base des nanocristaux d’oxyde de zinc (ZnO) et du poly (3-hexylthiophène) (P3HT) par absorption photoinduite en régime quasi-stationnaire (PIA) et la spectroscopie PIA en pompage modulé dépendant de la fréquence. L’interface entre le donneur (le polymère P3HT) et l’accepteur (les nanoparticules de ZnO), où la génération de charges se produit, joue un rôle important dans la performance des cellules photovoltaïques hybrides. Pour améliorer le mécanisme de génération de charges du P3H: ZnO, il est indispensable de modifier l’interface entre ses constituants. Nous avons démontré que la modification d’interface moléculaire avec cis-bis (4, 40 - dicarboxy-2, 20bipyridine) ruthénium (II) (N3-dye) et a-Sexithiophen-2 yl-phosphonique (6TP) a améliorée le photocourant et la performance dans les cellules P3HT: ZnO. Le 6TP et le N3 s’attachent à l’interface du ZnO, en augmentant ainsi l’aire effective de la surface donneur :accepteur, ce qui contribue à une séparation de charge accrue. De plus, le 6TP et le N3 réduisent la densité de pièges dans le ZnO, ce qui réduit le taux de recombinaison des paires de charges. Dans la deuxième partie, jai introduit une matrice hôte polymérique de polystyréne à masse molaire ulra-élevée, qui se comporte comme un solide pour piéger et protéger une solution de poly [2-méthoxy, 5- (2´-éthyl-hexoxy) -1,4-phénylènevinylène- PPV] (MEHPPV) pour utilisation dans des dispositifs optoèlectroniques quantiques. Des travaux antérieurs ont montré que MEH-PPV en solution subit une transition de conformation, d’une conformation enroulé à haute température (phase bleue) à une conformation de chaîne étendue à basse température (phase rouge). La conformation de la chaîne étendue de la solution MEH-PPV favorise les caractéristiques nécessaires à l’amélioration des dispositifs optoélectroniques quantiques, mais la solution ne peut pas être incorporées dans le dispositif. J’ai démontré que la caractéristique de la phase rouge du MEH-PPV en solution se maintient dans une matrice hôte polymérique de polystyrène transformé de masse molaire très élevée, qui se comporte comme un solide (gel de MEH-PPV/UHMW PS), par le biais de la spectroscopie de photoluminescence (PL) dépendant de la température (de 290K à 80 K). La phase rouge du gel MEH-PPV/UHMW PS se manifeste par des largeurs de raie étroites et une intensité augmentée de la transition 0-0 de la progression vibronique dans le spectre de PL ainsi qu’un petit décalage de Stokes entre la PL et le spectre d’absorption à basse température. Ces approches démontrent que la manipulation de la microstructure et des propriétés électroniques des polymères semi-conducteurs ont un impact direct sur la performance de dispositifs pour leurs développements technologiques continus.

New insights into the transport processes controlling the sulfate-methane-transition-zone near methane vents

Over the past years, several studies have raised concerns about the possible interactions between methane hydrate decomposition and external change. To carry out such an investigation, it is essential to characterize the baseline dynamics of gas hydrate systems related to natural geological and sedimentary processes. This is usually treated through the analysis of sulfate-reduction coupled to anaerobic oxidation of methane (AOM). Here, we model sulfate reduction coupled with AOM as a two-dimensional (2D) problem including, advective and diffusive transport. This is applied to a case study from a deep-water site off Nigeria’s coast where lateral methane advection through turbidite layers was suspected. We show by analyzing the acquired data in combination with computational modeling that a two-dimensional approach is able to accurately describe the recent past dynamics of such a complex natural system. Our results show that the sulfate-methane-transition-zone (SMTZ) is not a vertical barrier for dissolved sulfate and methane. We also show that such a modeling is able to assess short timescale variations in the order of decades to centuries.

Random matrix theory eta aplikazioa kaos kuantikoan

[eus] Gradu amaierako lan honetan ausazko matrizeen teoriari, RMT-ri, buruzko sarrera orokor bat egiten da ondoren aplikazio fisiko bat emateko. Teoriaren aplikazioa egiteko Kaos kuantikoa deritzon fisikaren arloa erabiliko da. Lehenik eta behin, RMT-ren kontzeptu batzuk azalduko dira helburutzat lehen auzokideen distantziaren distribuzioaren espresio lortzea izanik. Izan ere, distribuzio honek erakutsiko baititu Kaosak kuantikoki uzten dituen aztarnak. Bigarren kapituluan, aplikazio fisikoa azalduko da. Lehenengo Kaosean RMT nola aplikatzen den ikusiko da, ondoren adibide batzuen bidez argituz, eremu magnetiko batean dagoen hidrogeno atomoa eta billar kuantikoak izenarekin ezagutzen diren sistemak, batik bat.

THERMODYNAMICS AND APPLICATION OF A PAIR OF SYNTHETIC NUCLEOBASES FROM ARTIFICIALLY EXPANDED GENETIC INFORMATION SYSTEM

UV-melting experiments were performed on 9-mer duplexes containing a pair of synthetic nucleobases P·Z, two members of Expanded Genetic Information System (AEGIS), or P, Z containing mismatches. Enthalpy, entropy and free energy change were derived from simulation using two-state transition model. Nearest neighbor thermodynamic parameters of trimers or tetramers containing P·Z pair or P, Z containing mismatches were derived based on known nearest neighbor parameters. Proposed structures based on thermodynamic parameters are discussed. An application using P·Z pair as reverse selection tool of desired nucleic acid secondary structure is described.

Jiao Tong: A Grounded Theory of Chinese International Students' Transition to American Tertiary Education

University students are more globally mobile than ever before, increasingly receiving education outside of their home countries. One significant student exchange pattern is between China and the United States; Chinese students are the largest population of international students in the U.S. (Institute of International Education, 2014). Differences between Chinese and American culture in turn influence higher education praxis in both countries, and students are enculturated into the expectations and practices of their home countries. This implies significant changes for students who must navigate cultural differences, academic expectations, and social norms during the process of transition to a system of higher education outside their home country. Despite the trends in students’ global mobility and implications for international students’ transitions, scholarship about international students does not examine students’ experiences with the transition process to a new country and system of higher education. Related models were developed with American organizations and individuals, making it unlikely that they would be culturally transferable to Chinese international students’ transitions. This study used qualitative methods to deepen the understanding of Chinese international students’ transition processes. Grounded theory methods were used to invite the narratives of 18 Chinese international students at a large public American university, analyze the data, and build a theory that reflects Chinese international students’ experiences transitioning to American university life. Findings of the study show that Chinese international students experience a complex process of transition to study in the United States. Students’ pre-departure experiences, including previous exposure to American culture, family expectations, and language preparation, informed their transition. Upon arrival, students navigate resource seeking to fulfill their practical, emotional, social, intellectual, and ideological needs. As students experienced various positive and discouraging events, they developed responses to the pivotal moments. These behaviors formed patterns in which students sought familiarity or challenge subsequent to certain events. The findings and resulting theory provide a framework through which to better understand the experiences of Chinese international students in the context of American higher education.

Nonlinear development of matrix-converter instabilities

Matrix converters convert a three-phase alternating-current power supply to a power supply of a different peak voltage and frequency, and are an emerging technology in a wide variety of applications. However, they are susceptible to an instability, whose behaviour is examined herein. The desired “steady-state” mode of operation of the matrix converter becomes unstable in a Hopf bifurcation as the output/input voltage transfer ratio, q, is increased through some threshold value, qc. Through weakly nonlinear analysis and direct numerical simulation of an averaged model, we show that this bifurcation is subcritical for typical parameter values, leading to hysteresis in the transition to the oscillatory state: there may thus be undesirable large-amplitude oscillations in the output voltages even when q is below the linear stability threshold value qc.

Challenges and Opportunities in Transforming Laser System Industry to Deliver Integrated Product and Service Offers

Part 4: Transition Towards Product-Service Systems

Multifactorial approach to non-viral gene therapy: development of an efficient system for the retina

Tese de Doutoramento, Ciências Biomédicas, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, 2016