The two-component system of a novel copper resistant operon of Marinobacter hydrocarbonoclasticus

Dissertation for the Master’s Degree in Structural and Functional Biochemistry

Structural characterization of the urea-unfolded state of Colicin Immunity Protein Im7 and Im9

Dissertação para obtenção do Grau de Mestre em Bioquímica Estrutural e Funcional

Use of leaves for drinking water

The use of leaves for drinking water is a widespread tool-use behavior among chimpanzees. Although this tool-use behavior is widely described as the use of leaf sponges, it can actually be found in three different forms: leaf sponges, leaf-folding, and leaf spoons. Among the chimpanzee community of Bossou, we can observe all three forms, albeit in different frequencies. Here I describe the longitudinal record of manufacture and use of leaf tools for drinking water, highlighting the learning process underlying the acquisition of the skill. The degree of laterality evident in both immature and mature performers is also presented here. The use of leaves for drinking water emerges at the age of 1.5 years old, but the manufacture of leaf tools only starts at 3.5 years of age. Infants and juveniles were observed to use drinking tools that had been discarded by other individuals after use. Concerning handedness in general, the chimpanzees are ambidextrous, with some individuals biased to one side.

Modulation of α-synuclein aggregation and toxicity

Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina

Geospatial assessment of pest-induced forest damage through the use of UAV-bases NIR imaging and Gi-technology

Dissertation submitted in partial fulfillment of the requirements for the Degree of Master of Science in Geospatial Technologies.

Characterization of novel heme-containing sensor proteins from Geobacter sulfurreducens

The focus of this Thesis was the study of the sensor domains of two heme-containing methyl-accepting chemotaxis proteins (MCP) from Geobacter sulfurreducens: GSU0582 and GSU0935. These domains contain one c-type heme, form swapped dimers with a PAS-like fold and are the first examples of a new class of heme sensors. NMR spectroscopy was used to assign the heme and polypeptide signals in both sensors, as a first step to probe conformational changes in the vicinity of the hemes. However, the presence of two conformations in solution impaired the confident assignment of the polypeptide signals. To understand how conformational changes and swapped dimerization mechanism can effectively modulate the function of the two sensor domains and their signal transduction process, the sensor domains folding and stability were studied by circular dichroism and UV-visible spectroscopy. The results showed differences in the thermodynamic stability of the sensors, with GSU0582 displaying higher structural stability. These studies also demonstrated that the heme moiety undergoes conformational changes matching those occurring at the global protein structure and that the content of intrinsically disordered segments within these proteins (25% for GSU0935; 13% for GSU0582) correlates with the stability differences observed. The thermodynamic and kinetic properties of the sensor domains were determined at different pH and ionic strength by visible spectroscopy and stopped-flow techniques. Despite the remarkably similar spectroscopic and structural features of the two sensor domains, the results showed that their properties are quite distinct. Sensor domain GSU0935 displayed more negative reduction potentials and smaller reduction rate constants, which were more affected by pH and ionic strength. The available structures were used to rationalize these differences. Overall, the results described in this Thesis indicate that the two G. sulfurreducens MCP sensor domains are designed to function in different working potential ranges, allowing this bacterium to trigger an adequate cellular response in distinct anoxic subsurface environments.

Land cover mapping analysis and urban growth modeling using remote sensing techniques

The rapid growth of big cities has been noticed since 1950s when the majority of world population turned to live in urban areas rather than villages, seeking better job opportunities and higher quality of services and lifestyle circumstances. This demographic transition from rural to urban is expected to have a continuous increase. Governments, especially in less developed countries, are going to face more challenges in different sectors, raising the essence of understanding the spatial pattern of the growth for an effective urban planning. The study aimed to detect, analyse and model the urban growth in Greater Cairo Region (GCR) as one of the fast growing mega cities in the world using remote sensing data. Knowing the current and estimated urbanization situation in GCR will help decision makers in Egypt to adjust their plans and develop new ones. These plans should focus on resources reallocation to overcome the problems arising in the future and to achieve a sustainable development of urban areas, especially after the high percentage of illegal settlements which took place in the last decades. The study focused on a period of 30 years; from 1984 to 2014, and the major transitions to urban were modelled to predict the future scenarios in 2025. Three satellite images of different time stamps (1984, 2003 and 2014) were classified using Support Vector Machines (SVM) classifier, then the land cover changes were detected by applying a high level mapping technique. Later the results were analyzed for higher accurate estimations of the urban growth in the future in 2025 using Land Change Modeler (LCM) embedded in IDRISI software. Moreover, the spatial and temporal urban growth patterns were analyzed using statistical metrics developed in FRAGSTATS software. The study resulted in an overall classification accuracy of 96%, 97.3% and 96.3% for 1984, 2003 and 2014’s map, respectively. Between 1984 and 2003, 19 179 hectares of vegetation and 21 417 hectares of desert changed to urban, while from 2003 to 2014, the transitions to urban from both land cover classes were found to be 16 486 and 31 045 hectares, respectively. The model results indicated that 14% of the vegetation and 4% of the desert in 2014 will turn into urban in 2025, representing 16 512 and 24 687 hectares, respectively.

Analysis of temporal and spatial variations of forest. A case of study in notheastern Armenia

The forest has a crucial ecological role and the continuous forest loss can cause colossal effects on the environment. As Armenia is one of the low forest covered countries in the world, this problem is more critical. Continuous forest disturbances mainly caused by illegal logging started from the early 1990s had a huge damage on the forest ecosystem by decreasing the forest productivity and making more areas vulnerable to erosion. Another aspect of the Armenian forest is the lack of continuous monitoring and absence of accurate estimation of the level of cuts in some years. In order to have insight about the forest and the disturbances in the long period of time we used Landsat TM/ETM + images. Google Earth Engine JavaScript API was used, which is an online tool enabling the access and analysis of a great amount of satellite imagery. To overcome the data availability problem caused by the gap in the Landsat series in 1988- 1998, extensive cloud cover in the study area and the missing scan lines, we used pixel based compositing for the temporal window of leaf on vegetation (June-late September). Subsequently, pixel based linear regression analyses were performed. Vegetation indices derived from the 10 biannual composites for the years 1984-2014 were used for trend analysis. In order to derive the disturbances only in forests, forest cover layer was aggregated and the original composites were masked. It has been found, that around 23% of forests were disturbed during the study period.

Analyzing Geospatial patterns of syrian refugee flows in southeastern Turkey by use of remote sensing and complementary data

Crisis-affected communities and global organizations for international aid are becoming increasingly digital as consequence geotechnology popularity. Humanitarian sector changed in profound ways by adopting new technical approach to obtain information from area with difficult geographical or political access. Since 2011, turkey is hosting a growing number of Syrian refugees along southeastern region. Turkish policy of hosting them in camps and the difficulty created by governors to international aid group expeditions to get information, made such international organizations to investigate and adopt other approach in order to obtain information needed. They intensified its remote sensing approach. However, the majority of studies used very high-resolution satellite imagery (VHRSI). The study area is extensive and the temporal resolution of VHRSI is low, besides it is infeasible only using these sensors as unique approach for the whole area. The focus of this research, aims to investigate the potentialities of mid-resolution imagery (here only Landsat) to obtain information from region in crisis (here, southeastern Turkey) through a new web-based platform called Google Earth Engine (GEE). Hereby it is also intended to verify GEE currently reliability once the Application Programming Interface (API) is still in beta version. The finds here shows that the basic functions are trustworthy. Results pointed out that Landsat can recognize change in the spectral resolution clearly only for the first settlement. The ongoing modifications vary for each case. Overall, Landsat demonstrated high limitations, but need more investigations and may be used, with restriction, as a support of VHRSI.

A formação em rastreio e intervenções breves do álcool muda as atitudes dos profissionais de saúde ocupacional? : estudo quasi experimental exploratório para uma estratégia para o meio laboral

RESUMO - Estudos anteriores nos Cuidados de Saúde Primários (CSP) apontam para um incremento da Deteção Precoce (DP) e Intervenções Breves (IB) em pacientes com Problemas Ligados ao Álcool (PLA) após formação apropriada dos profissionais de saúde (com melhoria das suas atitudes). Este estudo quasi experimental, exploratório, é pioneiro no âmbito da Saúde Ocupacional (SO) para a implementação de Rastreio/DP e IB nos PLA. O objetivo principal foi avaliar se uma formação sobre Rastreio/DP e IB pode melhorar as atitudes dos profissionais de SO ao lidar com os PLA em Meio Laboral (PLAML). Foi aplicado um questionário em duas partes: Q1/pré-formação (variáveis sociodemográficas, profissionais, competências em alcoologia, experiência pessoal com álcool/familiares com PLA, dificuldades em lidar com trabalhadores com PLA, AUDIT-C e SAAPPQ) e Q2/pós-formação (avaliação da formação e das atitudes SAAPPQ - Adequação, Legitimidade, Motivação, Autoestima e Satisfação). Os resultados na amostra (N=39) revelaram um aumento sobretudo na Adequação e Satisfação. No entanto, naqueles com níveis baixos das atitudes pré-formação constou-se melhoria das atitudes à excepção da Autoestima e foram encontradas algumas associações com as características do perfil inicial dos participantes. Sugerem-se estudos posteriores para identificação de pacotes formativos mais efetivos e propostas para um Plano integrador de RIB para PLAML

Design of bio-inspired ionic liquids for protein stabilisation

Ionic Liquids (ILs) consist in organic salts that are liquid at/or near room temperature. Since ILs are entirely composed of ions, the formation of ion pairs is expected to be one essential feature for describing solvation in ILs. In recent years, protein - ionic liquid (P-IL) interactions have been the subject of intensive studies mainly because of their capability to promote folding/unfolding of proteins. However, the ion pairs and their lifetimes in ILs in P-IL thematic is dismissed, since the action of ILs is therefore the result of a subtle equilibrium between anion-cation interaction, ion-solvent and ion-protein interaction. The work developed in this thesis innovates in this thematic, once the design of ILs for protein stabilisation was bio-inspired in the high concentration of organic charged metabolites found in cell milieu. Although this perception is overlooked, those combined concentrations have been estimated to be ~300 mM among the macromolecules at concentrations exceeding 300 g/L (macromolecular crowding) and transient ion-pair can naturally occur with a potential specific biological role. Hence the main objective of this work is to develop new bio-ILs with a detectable ion-pair and understand its effects on protein structure and stability, under crowding environment, using advanced NMR techniques and calorimetric techniques. The choline-glutamate ([Ch][Glu]) IL was synthesized and characterized. The ion-pair was detected in water solutions using mainly the selective NOE NMR technique. Through the same technique, it was possible to detect a similar ion-pair promotion under synthetic and natural crowding environments. Using NMR spectroscopy (protein diffusion, HSQC experiments, and hydrogen-deuterium exchange) and differential scanning calorimetry (DSC), the model protein GB1 (production and purification in isotopic enrichment media) it was studied in the presence of [Ch][Glu] under macromolecular crowding conditions (PEG, BSA, lysozyme). Under dilute condition, it is possible to assert that the [Ch][Glu] induces a preferential hydration by weak and non-specific interactions, which leads to a significant stabilisation. On the other hand, under crowding environment, the [Ch][Glu] ion pair is promoted, destabilising the protein by favourable weak hydrophobic interactions , which disrupt the hydration layer of the protein. However, this capability can mitigates the effect of protein crowders. Overall, this work explored the ion-pair existence and its consequences on proteins in conditions similar to cell milieu. In this way, the charged metabolites found in cell can be understood as key for protein stabilisation.