Study of multi-component systems in polybenzimidazole membrane formation and their impact on membrane performance

Dissertação para obtenção do Grau de Mestre em Engenharia Química e Bioquímica

Synthesis and characterization of sugar containing hydrophilic and hydrophobic polymers with potential application in medicine

Dissertation toobtaina Master of Science degree in Bioorganics

Contribution to drug discovery and development for tauopathies using yeast as a model

This work aimed to contribute to drug discovery and development (DDD) for tauopathies, while expanding our knowledge on this group of neurodegenerative disorders, including Alzheimer’s disease (AD). Using yeast, a recognized model for neurodegeneration studies, useful models were produced for the study of tau interaction with beta-amyloid (Aβ), both AD hallmark proteins. The characterization of these models suggests that these proteins co-localize and that Aβ1-42, which is toxic to yeast, is involved in tau40 phosphorylation (Ser396/404) via the GSK-3β yeast orthologue, whereas tau seems to facilitate Aβ1-42 oligomerization. The mapping of tau’s interactome in yeast, achieved with a tau toxicity enhancer screen using the yeast deletion collection, provided a novel framework, composed of 31 genes, to identify new mechanisms associated with tau pathology, as well as to identify new drug targets or biomarkers. This genomic screen also allowed to select the yeast strain mir1Δ-tau40 for development of a new GPSD2TM drug discovery screening system. A library of unique 138 marine bacteria extracts, obtained from the Mid-Atlantic Ridge hydrothermal vents, was screened with mir1Δ-tau40. Three extracts were identified as suppressors of tau toxicity and constitute good starting points for DDD programs. mir1Δ strain was sensitive to tau toxicity, relating tau pathology with mitochondrial function. SLC25A3, the human homologue of MIR1, codes for the mitochondrial phosphate carrier protein (PiC). Resorting to iRNA, SLC25A3 expression was silenced in human neuroglioma cells, as a first step towards the engineering of a neural model for replicating the results obtained in yeast. This model is essential to understand the mechanisms of tau toxicity at the mitochondrial level and to validate PiC as a relevant drug target. The set of DDD tools here presented will foster the development of innovative and efficacious therapies, urgently needed to cope with tau-related disorders of high human and social-economic impact.

Algorithms and tools for in silico design of cell factories

PhD thesis in Bioengineering

The vulnerability to pollution and hydrochemical variation of eleven springs (catchments) in the karst lowlands of the west of Ireland

The vulnerability to pollution and hydrochemical variation of groundwater in the mid-west karstic lowlands of Ireland were investigated from October 1992 to September 1993, as part of an EU STRIDE project at Sligo Regional Technical College. Eleven springs were studied in the three local authority areas of Co. Galway, Co. Mayo, and Co. Roscommon. Nine of the springs drain locally or regionally important karstic aquifers and two drain locally important sand and gravel aquifers. The maximum average daily discharge of any of the springs was 16,000 m3/day. Determination of the vulnerability of groundwater to pollution relies heavily on an examination of subsoil deposits in an area since they can act as a protecting or filtering layer over groundwater. Within aquifers/spring catchments, chemical reactions such as adsorption, solution-precipitation or acid-base reactions occur and modify the hydrochemistry of groundwater (Lloyd and Heathcote, 1985). The hydrochemical processes) that predominate depend cm the mineralogy of the aquifer, the hydrogeological environment, the overlying subsoils, and the history of groundwater movement. The aim of this MSc research thesis was to investigate the hydrochemical variation of spring outflow and to assess the relationship between these variations and the intrinsic vulnerability of the springs and their catchments. If such a relationship can be quantified, then it is hoped that the hydrochemical variation of a spring may indicate the vulnerability of a spring catchment without the need for determining it by field mapping. Such a method would be invaluable to any of the three local authorities since they would be able to prioritise sources that are most at risk from pollution, using simple techniques of chemical sampling, and statistical analysis. For each spring a detailed geological, hydrogeological and hydrochemical study was carried out. Individual catchment areas were determined with a water balance/budget and groundwater tracing. The subsoils geology for each spring catchment were mapped at the 1:10,560 scale and digitised to the 1:25,000 scale with AutoCad™ and Arclnfo™. The vulnerability of each spring was determined using the Geological Survey's vulnerability guidelines. Field measurements and laboratory based chemistry analyses of the springs were undertaken by personnel from both the EPA Regional Laboratory in Castlebar, Co. Mayo, and the Environment Section of Roscommon Co. Council. Electrical conductivity and temperature (°C) were sampled fortnightly, in the field, using a WTW microprocessor conductivity meter. A percentage (%) vulnerability was applied to each spring in order to indicate the areal extent of the four main classes of vulnerability (Extreme, High, Moderate, and Low) which occurred within the confines of each spring catchment. Hydrochemical variation for the springs were presented as the coefficient of variation of electrical conductivity. The results of this study show that a clear relationship exists between the degree of vulnerability of each catchment area as defined by the subsoil cover and the coefficient of variation of EC, with the coefficient of variation increasing as the vulnerability increases. The coefficient of variation of electrical conductivity is considered to be a parameter that gives a good general reflection of the degree of vulnerability occurring in a spring catchment in Ireland's karstic lowlands.

A absorção de nitrogênio, fósforo, potássio, cálcio, magnésio, enxôfre e silício pela cana de açúcar, Co 419, e o seu crescimento em função da idade

This paper describes the data obtained for the growth of sugar cane, Variety Co 419, and the amount and rate of absorption of nitrogen, phosphorus, potassium, calcium, magnesium, sulfur, and silicon, according to the age of the plant, in the soil and climate conditions of the state of S. Paulo, Brazil. An experiment was installed in the Estação Experimental de Cana de Açúcar "Dr. José Vizioli", at Piracicaba, state of S. Paulo, Brazil, and the soil "tèrra-roxa misturada" presented the following composition: Sand (more than 0,2 mm)........................................................................ 8.40 % Fine sand (from 0,2 to less than 0,02 mm)................................................. 24.90 % Silt (from 0,02 to less than 0,002 mm)...................................................... 16.40 % Clay (form 0,002 mm and less)................................................................ 50.20 % pH 10 g of soil and 25 ml of distilled water)..................................................... 5.20 %C (g of carbon per 100 g of soil)................................................................. 1.00 %N (g of nitrogen per 100 g of soil)............................................................... 0.15 P0(4)-³ (me. per 100 g of soil, soluble in 0,05 normal H2SO4) ............................... 0.06 K+ (exchangeable, me. per 100 g of soil)....... 0.18 Ca+² (exchangeable, me. per 100 g of soil)...... 2.00 Mg+² (exchangeable, me. per 100 g of soil)...... 0.66 The monthly rainfall and mean temperature from January 1956 to August 1957 are presented in Table 1, in Portuguese. The experiment consisted of 3 replications of the treatments: without fertilizer and with fertilizer (40 Kg of N, from ammonium sulfate; 100 Kg of P(2)0(5) from superphosphate and 40 Kg K2 O, from potassium chloride). Four complete stools (stalks and leaves) were harvested from each treatment, and the plants separated in stalks and leaves, weighed, dried and analysed every month from 6 up to 15 months of age. The data obtained for fresh and dry matter production are presented in table 2, and in figure land 2, in Portuguese. The curves for fresh and dry matter production showed that fertilized and no fertilized sugar cane with 6 months of age presents only 5% of its total weight at 15 months of age. The most intense period of growth in this experiment is located, between 8 and 12 months of age, that is between December 1956 and April 1957. The dry matter production of sugar cane with 8 and 12 months of age was, respectively, 12,5% and 87,5% of the total weight at 15 months of age. The growth of sugar cane in relation to its age follows a sigmoid curve, according to the figures 1, 2 and 3. The increase of dry matter production promoted by using fertilizer was 62,5% when sugar cane was 15 months of age. The concentration of the elements (tables 4 and 5 in Portuguese) present a general trend of decreasing as the cane grows older. In the stalks this is true for all elements studied in this experiment. But in the leaves, somme elements, like sulfur and silicon, appears to increase with the increasing of age. Others, like calcium and magnesium do not show large variations, and finally a third group, formed by nitrogen, phosphorus and potassium seems to decrease at the beginning and later presents a light increasing. The concentration of the elements was higher in the leaves than in the stalks from 6 up to 15 months of age. There were some exceptions. Potassium, magnesium and sulfur were higher in the stalks than in the leaves from 6 up to 8 or 9 months of age. After 9 months, the leaves presented more potassium, magnesium and sulfur than the stalks. The percentage of nitrogen in the leaves was lower in the plants that received fertilizer than in the plants without fertilizer with 6, 7, 8, 10, 11 and 13 months of age. This can be explained by "dilution effect". The uptake of elements by 4 stools (stalks and leaves) of sugar cane according to the plant age is showed in table 6, in Portuguese. The absorption of all studied elements, nitrogen, phosphorus, potassium, calcium, magnesium, sulfur and silicon, was higher in plants that received fertilizer. The trend of uptake of nitrogen and potassium is similar to the trend of production of dry matter, that is, the maximum absorption of those two nutrients occurs between 9 and 13 months of age. Finaly, the maxima amounts of elements absorbed by 4 stools (stalks and leaves) of sugar cane plants that received fertilizer are condensed in the following table: Element Maximum absorption in grams Age of the plants in months Nitrogen (N) 81.0 14 Phosphorus (P) 6.8 15 Potassium (K) 81.5 15 Calcium (Ca) 19.2 15 Magnesium (Mg) 13.9 13 Sulfur (S) 9.3 15 Silicon (Si) 61.8 15 It is very interesting to note the low absorption of phosphorus even with 100 kg of P2O5 per hectare, aplied as superphosphate. The uptake of phosphorus was lower than calcium, magnesium and sulfur. Also, it is noteworthy the large amount of silicon absorbed by sugar cane.

Diagnosis socioambiental y presupuesto para el Parque Nacional Los Quetzales (Costa Rica)

Se realiza una diagnosis socioambiental y un presupuesto para el Plan de Manejo del Parque Nacional Los Quetzales (PNLQ). El PNLQ ocupa una superficie de 4117 ha. Presenta un régimen hí­drico elevado favorecido por la topografí­a quebrada que permite la presencia de fajas de condensación. Estas condiciones favorecen la presencia del bosque nublado que como se ha observado es el hábitat de innumerables especies, algunas de las cuales presentan gran valor ecológico. El presupuesto para el Plan de Manejo refleja cómo los gastos superan en una proporción importante a los ingresos, por lo que se recomienda centrar los esfuerzos en la búsqueda de formas de financiación (pago por servicios ambientales, convenios, co-manejo y donaciones). También se hace precisa una aportación anual del Gobierno Central para la ejecución del Plan.

Molecular approaches to malaria and Babesisosis diagnosis

The development of additional methods for detecting and identifuing Babesia and Plasmodium infections may be useful in disease monitoring, management and control efforts. To preliminarily evaluate sunthetic peptide-based serodiagnosis, a hydrophilic sequence (DDESEFDKEK)was selected from published BabR gene of B. bovis. Immunization of rabbits and cattle with the hemocyanin-conjugated peptide elicited antibody responses that specifically detected both P. falciparum and B. bovis antigens by immunofluorescence and Western blots. Using a dot-ELISA with this peptide, antisera from immunized and naturally-infected cattle, and immunized rodents, were specifically detected. Reactivity was weak and correlated with peptide immunization or infection. DNA-based detection using repetitive DNA was species-specific in dot-blot formats for B. bovis DNA, and in both dot-blot and in situ formats for P. falciparum; a streamlined enzymelinked synthetic DNA assay for P. falciparum detected 30 parasites/mm(cúbicos) from patient blood using either colorimetric (2-15 h color development) or chemiluminescent detection (0.5-6-min. exposures). Serodiagnostic and DNA hybridization methods may be complementary in the respective detection of both chronic and acute infections. However, recent improvements in the polymerase chain reaction (PCR) make feasible a more sensitive and uniform approach to the diagnosis of these and other infectious disease complexes, with appropriate primers and processing methods. An analysis of ribosomal DNA genes of Plasmodium and Toxoplasma identified Apicomplexa-conserved sequence regions. Specific and distinctive PCR profiles were obtained for primers spanning the internal transcribed spacer locus for each of several Plasmodium and Babesia species.

Research and teaching: a holy or an unholy alliance?

Much of the self-image of the Western university hangs on the idea that research and teaching are intimately connected. The central axiom here is that research and teaching are mutually supportive of each other. An institution lacking such a set of relationships between research and teaching falls short of what it means to be a university. This set of beliefs raises certain questions: Is it the case that the presence of such a mutually supportive set of relationships between research and teaching is a necessary condition of the fulfilment of the idea of the university? (A conceptual question). And is it true that, in practice today, such a mutually supportive set of relationships between research and teaching characterises universities? (An empirical question). In my talk, I want to explore these matters in a critical vein. I shall suggest that: a) In practice today, such a mutually supportive set of relationships between research and teaching is in jeopardy. Far from supporting each other, very often research and teaching contend against each other. Research and teaching are becoming two separate ideologies, with their own interest structures. b) Historically, the supposed tight link between research and teaching is both of recent origin and far from universally achieved in universities. Institutional separateness between research and teaching is and has been evident, both across institutions and even across departments in the same institution. c) Conceptually, research and teaching are different activities: each is complex and neither is reducible to the other. In theory, therefore, research and teaching may be said to constitute a holy alliance but in practice, we see more of an unholy alliance. If, then, in an ideal world, a positive relationship between research and teaching is still a worthwhile goal, how might it be construed and worked for? Seeing research and teaching as two discrete and unified sets of activity is now inadequate. Much better is a construal of research and teaching as themselves complexes, as intermingling pools of activity helping to form the liquid university that is emerging today. On this view, research and teaching are fluid spaces, ever on the move, taking up new shapes, and themselves dividing and reforming, as the university reworks its own destiny in modern society. On such a perspective, working out a productive relationship between research and teaching is a complex project. This is an alliance that is neither holy nor unholy. It is an uneasy alliance, with temporary accommodations and continuous new possibilities.

Skin permeation and metabolism of di(2-ethylhexyl) phthalate (DEHP).

Phthalates are suspected to be endocrine disruptors. Di(2-ethylhexyl) phthalate (DEHP) is assumed to have low dermal absorption; however, previous in vitro skin permeation studies have shown large permeation differences. Our aims were to determine DEHP permeation parameters and assess extent of skin DEHP metabolism among workers highly exposed to these lipophilic, low volatile substances. Surgically removed skin from patients undergoing abdominoplasty was immediately dermatomed (800 μm) and mounted on flow-through diffusion cells (1.77 cm(2)) operating at 32°C with cell culture media (aqueous solution) as the reservoir liquid. The cells were dosed either with neat DEHP or emulsified in aqueous solution (166 μg/ml). Samples were analysed by HPLC-MS/MS. DEHP permeated human viable skin only as the metabolite MEHP (100%) after 8h of exposure. Human skin was able to further oxidize MEHP to 5-oxo-MEHP. Neat DEHP applied to the skin hardly permeated skin while the aqueous solution readily permeated skin measured in both cases as concentration of MEHP in the receptor liquid. DEHP pass through human skin, detected as MEHP only when emulsified in aqueous solution, and to a far lesser degree when applied neat to the skin. Using results from older in vitro skin permeation studies with non-viable skin may underestimate skin exposures. Our results are in overall agreement with newer phthalate skin permeation studies.

NS2 protein of hepatitis C virus interacts with structural and non-structural proteins towards virus assembly.

Growing experimental evidence indicates that, in addition to the physical virion components, the non-structural proteins of hepatitis C virus (HCV) are intimately involved in orchestrating morphogenesis. Since it is dispensable for HCV RNA replication, the non-structural viral protein NS2 is suggested to play a central role in HCV particle assembly. However, despite genetic evidences, we have almost no understanding about NS2 protein-protein interactions and their role in the production of infectious particles. Here, we used co-immunoprecipitation and/or fluorescence resonance energy transfer with fluorescence lifetime imaging microscopy analyses to study the interactions between NS2 and the viroporin p7 and the HCV glycoprotein E2. In addition, we used alanine scanning insertion mutagenesis as well as other mutations in the context of an infectious virus to investigate the functional role of NS2 in HCV assembly. Finally, the subcellular localization of NS2 and several mutants was analyzed by confocal microscopy. Our data demonstrate molecular interactions between NS2 and p7 and E2. Furthermore, we show that, in the context of an infectious virus, NS2 accumulates over time in endoplasmic reticulum-derived dotted structures and colocalizes with both the envelope glycoproteins and components of the replication complex in close proximity to the HCV core protein and lipid droplets, a location that has been shown to be essential for virus assembly. We show that NS2 transmembrane region is crucial for both E2 interaction and subcellular localization. Moreover, specific mutations in core, envelope proteins, p7 and NS5A reported to abolish viral assembly changed the subcellular localization of NS2 protein. Together, these observations indicate that NS2 protein attracts the envelope proteins at the assembly site and it crosstalks with non-structural proteins for virus assembly.

Physical and Sensory Disability Strategy and Action Plan 2011-2015 - Summary of the feedback and findings from the public consultation (PDF 248KB)

On 27 January 2011 the Department of Health, Social Services and Public Safety (DHSSPS) launched a three month public consultation for a new draft Physical and Sensory Disability Strategy and Action Plan (2011-2015). åÊ The aim of the consultation was to provide the opportunity for a range of different stakeholders (public authorities and organisations, individuals including persons with disabilities and community and voluntary organisations) from across Northern Ireland to give feedback on the suggested priorities and challenges detailed in the document. The Department recognised the need for a new Disability Strategy and Action Plan not least to address new and developing challenges and opportunities. These include: åÊ • Obligations taken by the UK and NI in signing and ratifying the UN Convention on the Rights of Persons with Disabilities; åÊ • New innovations and models of care, support and treatment available within health and social care; åÊ • The current demographic trends and financial constraints being faced by everyone. åÊ åÊ åÊ

Sand fly evolution and its relationship to Leishmania transmission

The evolutionary relationships of sand flies and Leishmania are discussed in this report, which draws distinctions between co-association, co-evolution and co-speciation (or co-cladogenesis). Examples focus on Phlebotomus vectors of Le. infantum and Le. major in the Mediterranean subregion.

Care and Case Management for Older People in Ireland

This study provides, for the first time, a systematic review of how health and social services are currently being delivered to older people with the purpose of identifying models of best practice within a Care Management framework. The research allows the voices of a wide range of health and social service providers to comment individually about how care is currently delivered in Ireland. In addition, the research provides them with an opportunity to express how they feel about Care and Case Management as a model for the planning, co-ordination and delivery of services and its feasibility in an Irish context Download the Report here

Performance of OptiMAL® in the diagnosis of Plasmodium vivax and Plasmodium falciparum infections in a malaria referral center in Colombia

Alternative, non-microscopic methods for the diagnosis of malaria have recently become available. Among these, rapid dipstick methods stand out. One such test, OptiMAL®, is based on the immunochromatographic detection of Plasmodium lactate dehydrogenase (pLDH) and has the capacity to detect and distinguish infections caused by P. falciparum and Plasmodium sp. This capacity is particularly important in countries where different species of Plasmodium co-exist. In this study we evaluated the performance of OptiMAL® in an urban referral center for malaria diagnosis. Two sets of patients were included: one (n = 112) having predetermined infections with P. falciparum or P. vivax and individuals with negative blood smears; and another consisting of all eligible consecutive patients (n = 80) consulting for diagnosis at the referral center during one month. The overall diagnostic efficiency of OptiMAL® for both sets of patients was 96.9%. Efficiency was higher for P. vivax (98.1%) than for P. falciparum (94.9%). These results corroborate the diagnostic utility of OptiMAL® in settings where P. vivax and P. falciparum co-exist and support its implementation where microscopic diagnosis is unavailable and in circumstances that exceed the capacity of the local microscopic diagnosis facility.