New synthetic methodologies from biorenewable resources

Tese de doutoramento, Farmácia (Química Farmacêutica e Terapêutica), Universidade de Lisboa, Faculdade de Farmácia, 2014

Human biomonitoring : biomarkers, individual susceptibility, and nutrigenetics

Tese de doutoramento, Biologia (Genética), Universidade de Lisboa, Faculdade de Ciências, 2015

Avaliação do potencial clínico de novos ligandos para o receptor estrogénico em diagnóstico e terapia de tumores da mama

Tese de doutoramento, Química (Química Inorgânica), Universidade de Lisboa, Faculdade de Ciências, 2015

Taking environmental engineering out of the laboratory

Environmental engineering is a core component of most construction and surveying undergraduate courses. It is generally accepted that students on these courses should have an understanding of thermal comfort, heat transfer, condensation, lighting, noise transmission and acoustics. Experiments are essential in developing students’ awareness and understanding of the underlying physical concepts which drive environmental engineering solutions. Traditionally these experiments have been conducted by students working in small groups in laboratories. However, increasing student numbers and, in particular, the growth in part time study, have placed significant additional demands on limited laboratory resources. The availability of reasonably priced, simple, hand-held equipment has made it possible for students to conduct experiments outside the confines of the laboratory. Furthermore, various professional software packages (some of which are freely available online) enable the resultant data to be further developed and analysed in conjunction with the conventional textbook approach. This paper examines these alternative approaches to the traditional laboratory experiment. An assessment is provided of the types of experiment which are both possible and appropriate, and the efficacy of these approaches is considered.

Detection of MegaTAL-induced HIV pol Mutation By Droplet Digital PCR

Thesis (Master's)--University of Washington, 2015

Corrections

Summary form only given, as follows. In Vol. 12, no. 3 (Summer 2007), page 9, bottom of the left column, in 'Computer Architecture and Amdahl??s Law' by Gene Amdahl, the claim about invalidating Amdahl??s Law in 1988 came from a team at Sandia National Laboratories, and not Los Alamos. The correct text should read: "Several years later I was informed of a proof that Amdahl's Law was invalidated by someone at Sandia National Laboratories, where a number of computers interconnected as an Ncube by communication lines, but with each computer also connected to I/O devices for loading the operating system, initial data, and results." On page 20 of the same issue, in the second sentence of the diagram explanation note by Justin Rattner, the percentage figures for the sequential and the system coordination parts of the workload were interchanged. The correct version of this sentence should read: "assuming a fixed sized problem, Amdahl speculated that most programs would require at least 10% of the computation to be sequential (only one instruction executing at a time), with overhead due to interprocessor coordination averaging 25%."

Technology transfer in Britain: the case of monoclonal antibodies

In 1975 two Cambridge scientists published a short article in Nature which announced the discovery of monoclonal antibodies. The article concluded ‘Such cultures could be valuable for medical and industrial use’. The interest which developed by the end of the decade in the industrial and financial possibilities of the new prospects opening up in biotechnology was to throw the apparent ‘failure’ to follow‐up the potentialities of this discovery into a public prominence rarely achieved by scientific discoveries. By the time Mrs Thatcher came to power it had become a scandal, another example of Britain's apparent inability to exploit effectively the brilliance of its scientific base. It was to explore both the process of scientific discovery and the conditions in Cambridge which nurtured it, and the issues which this particular discovery raised in the area of technology transfer (and the changes of policy that ensued), that the Wellcome Trust's History of Twentieth Century Medicine Group and the Institute of Contemporary British History organised this special witness seminar. It was held at the Wellcome Trust in London on 24 September 1993. The seminar was chaired by Sir Christopher Booth and introduced by Dr Robert Bud of the Science Museum. Those participating included the two authors of the Nature article, Dr César Milstein and Dr Georges Köhler, who received a Nobel Prize for their research, Dr Basil Bard (National Research Development Corporation [NRDC] 1950–74), Sir James Gowans (Secretary of the Medical Research Council [MRC] 1977–87), Sir John Gray (Secretary of the MRC 1968–77), John Newell (BBC World Service science correspondent 1969–79), Dr David Owen (MRC), and Dr David Secher (Laboratory of Molecular Biology [LMB], Cambridge). There were also contributions from Dr Ita Askonas (former head of immunology at the National Institute for Medical Research), Dr John Galloway (former member of MRC headquarters staff), Dr David Tyrrell (former Director, MRC Common Cold Unit), Professor Miles Weatherall (head of Therapeutic Research Division, Wellcome Research Laboratories 1967–75), Dr Guil Winchester (post‐doctoral fellow, Wellcome Institute for the History of Medicine), and Dr Peter Williams (former Director of the Wellcome Trust). The organisers would like to thank the Wellcome Trust for hosting and sponsoring the seminar. We would like to dedicate this publication to the memory of Georges Köhler, who sadly died in April 1995 before this could appear.

Lyophilisation of influenza, rabies and Marburg lentiviral pseudotype viruses for the development and distribution of a neutralisation-assay based diagnostic kit

Pseudotype viruses (PVs) are chimeric, replication-deficient virions that mimic wild-type virus entry mechanisms and can be safely employed in neutralisation assays, bypassing the need for high biosafety requirements and performing comparably to established serological assays. However, PV supernatant necessitates -80°C long-term storage and cold-chain maintenance during transport, which limits the scope of dissemination and application throughout resource-limited laboratories. We therefore investigated the effects of lyophilisation on influenza, rabies and Marburg PV stability, with a view to developing a pseudotype virus neutralisation assay (PVNA) based kit suitable for affordable global distribution. Infectivity of each PV was calculated after lyophilisation and immediate reconstitution, as well as subsequent to incubation of freeze-dried pellets at varying temperatures, humidities and timepoints. Integrity of glycoprotein structure following treatment was also assessed by employing lyophilised PVs in downstream PVNAs. In the presence of 0.5M sucrose-PBS cryoprotectant, each freeze-dried pseudotype was stably stored for 4 weeks at up to 37°C and could be neutralised to the same potency as unlyophilised PVs when employed in PVNAs. These results confirm the viability of a freeze-dried PVNA-based kit, which could significantly facilitate low-cost serology for a wide portfolio of emerging infectious viruses.

Accessing Planktothrix species diversity and associated toxins using quantitative real-time PCR in natural waters

Tese de Doutoramento em Biologia apresentada à Faculdade de Ciências da Universidade do Porto, 2015.

Supercritical carbon dioxide extraction of bioactive compounds from microalgae and volatile oils from aromatic plants

A discussion of the most interesting results obtained in our laboratories, during the supercritical CO(2) extraction of bioactive compounds from microalgae and volatile oils from aromatic plants, was carried out. Concerning the microalgae, the studies on Botryococcus braunii and Chlorella vulgaris were selected. Hydrocarbons from the first microalgae, which are mainly linear alkadienes (C(23)-C(31)) with an odd number of carbon atoms, were selectively extracted at 313 K increasing the pressure up to 30.0 MPa. These hydrocarbons are easily extracted at this pressure, since they are located outside the cellular walls. The extraction of carotenoids, mainly canthaxanthin and astaxanthin, from C. vulgaris is more difficult. The extraction yield of these components at 313 K and 35.0 MPa increased with the degree of crushing of the microalga, since they are not extracellular. On the other hand, for the extraction of volatile oils from aromatic plants, studies on Mentha pulegium and Satureja montana L were chosen. For the first aromatic plant, the composition of the volatile and essential oils was similar, the main components being the pulegone and menthone. However, this volatile oil contained small amounts of waxes, which content decreased with decreasing particle size of the plant matrix. For S. montana L it was also observed that both oils have a similar composition, the main components being carvacrol and thymol. The main difference is the relative amount of thymoquinone, which content can be 15 times higher in volatile oil. This oxygenated monoterpene has important biological activities. Moreover, experimental studies on anticholinesterase activity of supercritical extracts of S. montana were also carried out. The supercritical nonvolatile fraction, which presented the highest content of the protocatechuic, vanilic, chlorogenic and (+)-catechin acids, is the most promising inhibitor of the enzyme butyrylcholinesterase. In contrast, the Soxhlet acetone extract did not affect the activity of this enzyme at the concentrations tested. (C) 2011 Elsevier B.V. All rights reserved.

Re-evaluation of a reported increased micronucleus frequency in lymphocytes of workers occupationally exposed to formaldehyde

A replicate evaluation of increased micronucleus (MN) frequencies in peripheral lymphocytes of workers occupationally exposed to formaldehyde (FA) was undertaken to verify the observed effect and to determine scoring variability. May–Grünwald–Giemsa-stained slides were obtained from a previously performed cytokinesis-block micronucleus test (CBMNT) with 56 workers in anatomy and pathology laboratories and 85 controls. The first evaluation by one scorer (scorer 1) had led to a highly significant difference between workers and controls (3.96 vs 0.81 MN per 1000 cells). The slides were coded before re-evaluation and the code was broken after the complete re-evaluation of the study. A total of 1000 binucleated cells (BNC) were analysed per subject and the frequency of MN (in ‰) was determined. Slides were distributed equally and randomly between two scorers, so that the scorers had no knowledge of the exposure status. Scorer 2 (32 exposed, 36 controls) measured increased MN frequencies in exposed workers (9.88 vs 6.81). Statistical analysis with the two-sample Wilcoxon test indicated that this difference was not significant (p = 0.17). Scorer 3 (20 exposed, 46 controls) obtained a similar result, but slightly higher values for the comparison of exposed and controls (19.0 vs 12.89; p = 0.089). Combining the results of the two scorers (13.38 vs 10.22), a significant difference between exposed and controls (p = 0.028) was obtained when the stratified Wilcoxon test with the scorers as strata was applied. Interestingly, the re-evaluation of the slides led to clearly higher MN frequencies for exposed and controls compared with the first evaluation. Bland–Altman plots indicated that the agreement between the measurements of the different scorers was very poor, as shown by mean differences of 5.9 between scorer 1 and scorer 2 and 13.0 between scorer 1 and scorer 3. Calculation of the intra-class correlation coefficient (ICC) revealed that all scorer comparisons in this study were far from acceptable for the reliability of this assay. Possible implications for the use of the CBMNT in human biomonitoring studies are discussed.

Development of triplet repeat primed PCR (TP-PCR) technique as a support of molecular test in Machado-Joseph disease

Dissertação de Mestrado, Ciências Biomédicas, 3 de Fevereiro de 2016, Universidade dos Açores.

Chromatographic techniques for the determination of free phenol in foundry resins

Phenol is a toxic compound present in a wide variety of foundry resins. Its quantification is important for the characterization of the resins as well as for the evaluation of free contaminants present in foundry wastes. Two chromatographic methods, liquid chromatography with ultraviolet detection (LC-UV) and gas chromatography with flame ionization detection (GC-FID), for the analysis of free phenol in several foundry resins, after a simple extraction procedure (30 min), were developed. Both chromatographic methods were suitable for the determination of phenol in the studied furanic and phenolic resins, showing good selectivity, accuracy (recovery 99–100%; relative deviations <5%), and precision (coefficients of variation <6%). The used ASTM reference method was only found to be useful in the analysis of phenolic resins, while the LC and GC methods were applicable for all the studied resins. The developed methods reduce the time of analysis from 3.5 hours to about 30 min and can readily be used in routine quality control laboratories.

Remoção de fibrocimento: amianto

Dissertação para obtenção do grau de Mestre em Engenharia Civil Ramo Edificações

Validação do método de deteção de eritropoietina recombinante humana em urina humana por focalização isoelétrica

A eritropoietina (EPO) é uma substância que estimula a produção de eritrócitos, aumentando a oxigenação muscular, sendo segregada de forma natural pelo organismo e excretada na urina em baixas concentrações. Devido às suas propriedades e características, a EPO foi rapidamente introduzida no mundo do desporto, como substância ilícita, proporcionando vantagens no rendimento desportivo. No início de 2000 foi desenvolvido um método de deteção direta de EPO Recombinante (rHuEPO) em urina humana por Lasne, baseado na focalização isoelétrica (IEF) em gel de poliacrilamida, seguido de duplo blote, tendo este sido publicado e validado. Em 2002, a Agência Mundial Antidopagem (AMA) implementou este mesmo método, sendo atualmente um dos métodos oficiais utilizado pelos laboratórios acreditados pela AMA. Desta forma, o ponto de partida para a realização deste trabalho consistiu na necessidade de implementar e validar o método de referência de IEF para a deteção de rHuEPO em urina humana. O trabalho foi realizado no Laboratório de Análises e Dopagem (LAD) do Instituto do Desporto de Portugal (IDP), atual Instituto Português do Desporto e Juventude (IPDJ). O principal objetivo deste trabalho consistiu no estudo/investigação de diferentes parâmetros de validação (especificidade/seletividade; capacidade de identificação; limite de deteção; exatidão e repetibilidade), de acordo com o protocolado no Procedimento Geral interno do Laboratório de Análises de Dopagem de Lisboa (LAD). O referido método de triagem e confirmação revelou possuir características de desempenho conformes com os requisitos aplicáveis, pelo que é considerado validado e apto.