Brazilian savanna forest : conservation, medicinal reservoir and bioprospecting

This study aimed to analyse the Brazilian savanna forest from a Legal Reserve (LR) area from a perspective of conservation, reservoir of organic carbon and medicinal biomass for a prospective use of native medicinal plants. An ethnobotanical and ethnopharmacological survey was carried out close to a community settled in the rural area in the south of Tocantins, being selected 9 of the most cited species (cajuí- Anacardium othonianum; inharé-Brosimum gaudichaudii; jatobá-Hymenaeae courbaril; jenipapo-Genipa americana, aroeira-Myracrodruon urundeuva; negramina-Siparuna guianensis; barbatimão- Stryphnodendron obovatum; assa peixe-Vernonia brasiliana, embaúba-Cecropia pachystachya). Crude foliar extracts were subjected to a preliminary phytochemical prospection and triage of secondary metabolites with antimicrobial activity of potential interest in health and familiar agriculture. Phenolic compounds, terpenes and flavonoids were detected in the extracts of most species, which suggests the presence of antimicrobial, antioxidant and anti-insect activities. It was evident the need to better know the LR as a reservoir of medicinal biomass in an area under ecological tension where 35% (610ha) of the property is LR and should be protected by law. Therefore, a forest inventory of live woody species was performed using the allometric or indirect method. This identified a rare remnant of Semidecidual Seasonal Forest amidst the largest world savannah, the Cerrado biome. An analysis of the forest average productivity per basal area (m².ha), aerial live biomass (ton.ha-1) and carbon stock was carried out. The forest fragment was considered relatively rich in species and diversity, although showing signs of disturbance and dominance by a few species. Its horizontal structure suggests biotic regeneration conditions. It is an important reservoir of medicinal plants. Of the families (57.5%) presenting medicinal species, 19 from a total of 33 are represented in the area and contain 44% (27) of the total species (61) and 63% (432) of the total individuals catalogued. Medicinal species have ecological importance for the equilibrium of the local flora and represent 80% of the 10 species with higher Importance Value Index (IVI): Tetragastris altissima, Chrysophyllum marginatum, Oenocarpus distichus, Sclerolobium paniculatum, Simarouba versicolor, Alibertia macrophylla, Siparuna guianensis, Maprounea guianensis, Licania parvifolia e Physocalymma scaberrimum. Medicinal productivity was high for this type of phytophysionomy: 183,2 ton. ha-1 of biomass and 91,51 ton. ha-1 of carbon representing 66% of the total biomass and carbon of this Cerrado forest. From this stage S. guianensis (Siparunaceae) was selected for performing bioassays in order to verify its biological activity against microorganisms of health and agricultural relevance. This is a native aromatic medicinal plant recommended as priority for conservation, with local popular medicinal validation and availability of medicinal feedstock (3300 Kg.ha-1), with the foliar fraction giving 38Kg/ha of crude extract and 5L/ha of essential oil. Foliar crude extracts and essential oil were obtained and tested in vitro using a disk diffusion bioassay. Different concentrations of these natural products were tested against gram-positive bacteria (Staphylococcus aureus ATCC 29213), gram-negative bacteria (Escherichia coli ATCC 25922 and ATCC 35218; Pseudomonas aeruginosa ATCC 10145) and fungi (Candida albicans ATCC 6258 e Fusarium oxysporum). The essential oil inhibited the growth of S. aureus in its crude concentration (380μg.mL-1), as well as diluted to half (190μg.mL-1) and a quarter strength (95μg.mL-1). It’s likely that such action is due to sesquiterpenes major components, such as bisabolol and bisabolene (10.35%), measured by gas chromatography (GC-MS, GC-FID). Extracts did not exhibit any antimicrobial activity against the microorganisms tested. The native medicinal plants prospective market is an alternative that favours the conservation of biodiversity while generating benefits for the development of sustainable family productive activities within local ecosystems instead of the current inappropriate uses. This strengthens conservation policies of Legal Reserve in rural settlements and is in agreement with public policy on global warming and climate changes.

The player and video game interplay in the gameplay experience construct

Among the many discussions and studies related to video games, one of the most recurrent, widely debated and important relates to the experience of playing video games. The gameplay experience – as appropriated in this study – is the result of the interplay between two essential elements: a video game and a player. Existing studies have explored the resulting experience of video game playing from the perspective of the video game or the player, but none appear to equally balance both of these elements. The study presented here contributes to the ongoing debate with a gameplay experience model. The proposed model, which looks to equally balance the video game and the player elements, considers the gameplay experience to be both an interactive experience (related to the process of playing the video game) and an emotional experience (related to the outcome of playing the video game). The mutual influence of these two experiences during video game play ultimately defines the gameplay experience. To this gameplay experience contributes several dimensions, related to both the video game and player: the video game includes a mechanics, interface and narrative dimension; the player includes a motivations, expectations and background dimension. Also, the gameplay experience is initially defined by a gameplay situation, conditioned by an ambient in which gameplay takes place and a platform on which the video game is played. In order to initially validate the proposed model and attempt to show a relationship among the multiple model dimensions, a multi-case study was carried out using two different video games and player samples. In one study, results show significant correlations between multiple model dimensions, and evidence that video game related changes influence player motivations as well as player visual behavior. In specific player related analysis, results show that while players may be different in terms of background and expectations regarding the game, their motivation to play are not necessarily different, even if their performance in the game is weak. While further validation is necessary, this model not only contributes to the gameplay experience debate, but also demonstrates in a given context how player and video game dimensions evolve during video game play.

Planeamento do território: metodologias, actores e participação

O argumento que compagina esta investigação sustenta que, no contexto actual de crescente complexidade dos problemas territoriais e de densidade de actores com interesses contraditórios e conflituantes, o desenvolvimento de metodologias de planeamento territorial promotoras da participação dos actores pode ser particularmente útil, possibilitando a construção de compromissos entre actores, alinhando as suas motivações e mobilizando os seus meios, gerando novas formas de “acção colectiva” em torno dos objectivos de planeamento. Procurou-se, assim: i) perceber o que é território, objecto da actividade do planeamento; ii) estabelecer um conceito de planeamento do território; iii) produzir um quadro de referência sobre planeamento territorial, com particular enfoque para o papel dos actores e participação e iv) desenvolver uma análise crítica de um conjunto de experiências de planeamento territorial, à luz do anterior quadro de referência. Do ponto de vista empírico, ambicionava-se examinar um conjunto de experiências de planeamento territorial onde a participação dos actores tivesse sido particularmente relevante, em diferentes contextos quanto ao entendimento dos objectivos de planeamento e sua concretização, com a particularidade de em ambas ter havido um forte envolvimento do investigador num duplo papel: investigador/cidadão e investigador/promotor do planeamento do território. Durante o trajecto de investigação percorrido, revisitaram-se as principais referências sobre a problemática do planeamento do território e suas metodologias, com enfoque particular nos actores e formas de participação. Assume-se a conceptualização do planeamento do território como uma actividade desenvolvida por um promotor público de planeamento (Estado), o qual, num determinado contexto e em defesa do interesse colectivo, define objectivos para um determinado objecto de planeamento (território, temática e escala). Para os cumprir, mobiliza os meios e actores necessários, produzindo um quadro de referência e identificando as acções, os palcos e os métodos para os alcançar. Identificaram-se seis famílias de metodologias de planeamento do território. Discutiu-se o conceito de actor em planeamento e propôs-se uma tipologia de actores, consoante meios, motivações e palcos. Em seguida, produziu-se uma conceptualização e tipologia de formas de participação dos actores, que vai para além da tradicional participação dos cidadãos. Concluiu-se com a sugestão de princípios de orientação metodológica para o planeamento territorial e participação dos actores. Os dados da análise empírica efectuada permitiram concluir que o papel dos diferentes actores e a forma como foram tidos em conta os seus meios disponíveis e as suas motivações foram determinantes nos resultados alcançados nos dois contextos analisados. Num conjunto de experiências, o envolvimento dos actores visou fortalecer o quadro social e institucional de apoio, o fomento à criação de plataformas de diálogo e colaboração, de valorização de diferentes formas de conhecimento (técnico-científico e “local”) e a mobilização para a construção de políticas ou para a validação do processo de decisão. Noutro conjunto de experiências, o entendimento da participação dos actores foi desvalorizado, produzido de forma burocrática, sem o devido reconhecimento das suas motivações e valorização dos meios disponíveis, sem um adequado envolvimento, o que conduziu a situações de elevada conflitualidade, fragilização e descredibilização do exercício de planeamento e a resultados que contrariam os objectivos inicialmente definidos. Neste contexto, parece pois confirmar-se a hipótese de que as metodologias de planeamento onde a participação dos actores é considerada de forma explícita, coerente e consequente, condicionam de forma clara o envolvimento e alinhamento dos actores e determinam os resultados do planeamento do território.

Metabolic signature of lung cancer: a metabolomic study of human tissues and biofluids

This thesis reports the application of metabolomics to human tissues and biofluids (blood plasma and urine) to unveil the metabolic signature of primary lung cancer. In Chapter 1, a brief introduction on lung cancer epidemiology and pathogenesis, together with a review of the main metabolic dysregulations known to be associated with cancer, is presented. The metabolomics approach is also described, addressing the analytical and statistical methods employed, as well as the current state of the art on its application to clinical lung cancer studies. Chapter 2 provides the experimental details of this work, in regard to the subjects enrolled, sample collection and analysis, and data processing. In Chapter 3, the metabolic characterization of intact lung tissues (from 56 patients) by proton High Resolution Magic Angle Spinning (HRMAS) Nuclear Magnetic Resonance (NMR) spectroscopy is described. After careful assessment of acquisition conditions and thorough spectral assignment (over 50 metabolites identified), the metabolic profiles of tumour and adjacent control tissues were compared through multivariate analysis. The two tissue classes could be discriminated with 97% accuracy, with 13 metabolites significantly accounting for this discrimination: glucose and acetate (depleted in tumours), together with lactate, alanine, glutamate, GSH, taurine, creatine, phosphocholine, glycerophosphocholine, phosphoethanolamine, uracil nucleotides and peptides (increased in tumours). Some of these variations corroborated typical features of cancer metabolism (e.g., upregulated glycolysis and glutaminolysis), while others suggested less known pathways (e.g., antioxidant protection, protein degradation) to play important roles. Another major and novel finding described in this chapter was the dependence of this metabolic signature on tumour histological subtype. While main alterations in adenocarcinomas (AdC) related to phospholipid and protein metabolisms, squamous cell carcinomas (SqCC) were found to have stronger glycolytic and glutaminolytic profiles, making it possible to build a valid classification model to discriminate these two subtypes. Chapter 4 reports the NMR metabolomic study of blood plasma from over 100 patients and near 100 healthy controls, the multivariate model built having afforded a classification rate of 87%. The two groups were found to differ significantly in the levels of lactate, pyruvate, acetoacetate, LDL+VLDL lipoproteins and glycoproteins (increased in patients), together with glutamine, histidine, valine, methanol, HDL lipoproteins and two unassigned compounds (decreased in patients). Interestingly, these variations were detected from initial disease stages and the magnitude of some of them depended on the histological type, although not allowing AdC vs. SqCC discrimination. Moreover, it is shown in this chapter that age mismatch between control and cancer groups could not be ruled out as a possible confounding factor, and exploratory external validation afforded a classification rate of 85%. The NMR profiling of urine from lung cancer patients and healthy controls is presented in Chapter 5. Compared to plasma, the classification model built with urinary profiles resulted in a superior classification rate (97%). After careful assessment of possible bias from gender, age and smoking habits, a set of 19 metabolites was proposed to be cancer-related (out of which 3 were unknowns and 6 were partially identified as N-acetylated metabolites). As for plasma, these variations were detected regardless of disease stage and showed some dependency on histological subtype, the AdC vs. SqCC model built showing modest predictive power. In addition, preliminary external validation of the urine-based classification model afforded 100% sensitivity and 90% specificity, which are exciting results in terms of potential for future clinical application. Chapter 6 describes the analysis of urine from a subset of patients by a different profiling technique, namely, Ultra-Performance Liquid Chromatography coupled to Mass Spectrometry (UPLC-MS). Although the identification of discriminant metabolites was very limited, multivariate models showed high classification rate and predictive power, thus reinforcing the value of urine in the context of lung cancer diagnosis. Finally, the main conclusions of this thesis are presented in Chapter 7, highlighting the potential of integrated metabolomics of tissues and biofluids to improve current understanding of lung cancer altered metabolism and to reveal new marker profiles with diagnostic value.

Understanding and modelling hydrological and soil erosion processes in burnt forest catchments

Forest fires implications in overland flow and soil erosion have been researched for several years. Therefore, is widely known that fires enhance hydrological and geomorphological activity worldwide as also in Mediterranean areas. Soil burn severity has been widely used to describe the impacts of fire on soils, and has being recognized as a decisive factor controlling post-fire erosion rates. However, there is no unique definition of the term and the relationship between soil burn severity and post-fire hydrological and erosion response has not yet been fully established. Few studies have assessed post-fire erosion over multiple years, and the authors are aware of none which assess runoff. Small amount of studies concerning pre-fire management practices were also found. In the case of soil erosion models, the Revised Universal Soil Loss Equation (RUSLE) and the revised Morgan–Morgan–Finney (MMF) are well-known models, but not much information is available as regards their suitability in predicting post-fire soil erosion in forest soils. The lack of information is even more pronounced as regards post-fire rehabilitation treatments. The aim of the thesis was to perform an extensive research under the post fire hydrologic and erosive response subject. By understanding the effect of burn severity in ecosystems and its implications regarding post fire hydrological and erosive responses worldwide. Test the effect of different pre-fire land management practices (unplowed, downslope plowed and contour plowed) and time-since-fire, in the post fire hydrological and erosive response, between the two most common land uses in Portugal (pine and eucalypt). Assess the performance of two widely-known erosion models (RUSLE and Revised MMF), to predict soil erosion rates during first year following two wildfires of distinctive burn severity. Furthermore, to apply these two models considering different post-fire rehabilitation treatments in an area severely affected by fire. Improve model estimations of post-fire runoff and erosion rates in two different land uses (pine and eucalypt) using the revised MMF. To assess these improvements by comparing estimations and measurements of runoff and erosion, in two recently burned sites, as also with their post fire rehabilitation treatments. Model modifications involved: (1) focusing on intra-annual changes in parameters to incorporate seasonal differences in runoff and erosion; and (2) inclusion of soil water repellency in runoff predictions. Additionally, validate these improvements with the application of the model to other pine and eucalypt sites in Central Portugal. The review and meta-analysis showed that fire occurrence had a significant effect on the hydrological and erosive response. However, this effect was only significantly higher with increasing soil burn severity for inter-rill erosion, and not for runoff. This study furthermore highlighted the incoherencies between existing burn severity classifications, and proposed an unambiguous classification. In the case of the erosion plots with natural rainfall, land use factor affected annual runoff while land management affected both annual runoff and erosion amounts significantly. Time-since-fire had an important effect in erosion amounts among unplowed sites, while for eucalypt sites time affected both annual runoff and erosion amounts. At all studied sites runoff coefficients increase over the four years of monitoring. In the other hand, sediment concentration in the runoff, recorded a decrease during the same period. Reasons for divergence from the classic post-fire recovery model were also explored. Short fire recurrence intervals and forest management practices are viewed as the main reasons for the observed severe and continuing soil degradation. The revised MMF model presented reasonable accuracy in the predictions while the RUSLE clearly overestimated the observed erosion rates. After improvements: the revised model was able to predict first-year post-fire plot-scale runoff and erosion rates for both forest types, these predictions were improved both by the seasonal changes in the model parameters; and by considering the effect of soil water repellency on the runoff, individual seasonal predictions were considered accurate, and the inclusion of the soil water repellency in the model also improved the model at this base. The revised MMF model proved capable of providing a simple set of criteria for management decisions about runoff and erosion mitigation measures in burned areas. The erosion predictions at the validation sites attested both to the robustness of the model and of the calibration parameters, suggesting a potential wider application.

Pathways of oxidative degradation of wine

The production of color/flavor compounds in wine is the result of different interrelated mechanism reactions. Among these, the oxidation phenomenon and the Maillard reaction stands out with particular relevance due to their large impact on the sensory quality of wines and consequently on the product shelflife. The aim of this thesis is to achieve a global vision of wine degradation mechanisms. The identification of mediators’ reactions involved in oxidative browning and aromatic degradation will be attempted based on different detectors. Two approaches are implemented in this work: a “non-target” approach by which relevant analytical tools will be used to merge the information of cyclic voltammetry and Diode-Array (DAD) detectors, allowing a broader overview of the system and the note of interesting compounds, and a “target” approach by which the identification and quantification of the different compounds related to the wine degradation process will be performed using different detectors (HPLC-UV/Vis, LC-MS, GC-MS, and FID). Two different patterns of degradation will be used in this study: wines generated by O2 and temperature perturbations, and synthetic solutions with relevant wine constituents for mechanisms validation. Results clearly demonstrate a “convolution” of chemical mechanisms. The presence of oxygen combined with temperature had a synergistic effect on the formation of several key odorant compounds.The results of this work could be translated to the wine-making and wine-storage environment from the modelling of the analysed compounds.