Fungal communities in archives: assessment strategies and impact on paper conservation and human health

The main results presented in this PhD Dissertation have been published in interna-tional journals included in the Science Citation Index (SCI)

Blocking tumor exosome release using nanovectorization systems

Exosomes are small membrane vesicles secreted by most cell types, either normal or malignant and are found in most body fluids such as saliva, plasma and breast milk. In the past decade, the interest in these vesicles has been growing more and more since it was found that besides their beneficial functions such as the removal of cellular debris and unnecessary proteins during cell maturation process, they can also interact with other cells and transfer information between them, thus helping diseases like cancer to progress. The present work intended to use gold nanoparticles as vehicles for gene silencing in an attempt to reduce the tumor-derived exosome secretion, regulated by Rab27a protein, and also aimed to compare the exosome secretion between two breast cell lines, MCF7 and MDA. Changes in RAB27A gene expression were measured by Real-time Quantitative PCR and it was revealed a decreased in RAB27A gene expression, as expected. Exosomes were isolated and purified by two different methods, ultracentrifugation and the commercial kit ExoQuick™ Solution, and further characterized using Western Blot analysis. ExoQuick™ Solution was proven to be the most efficient method for exosome isolation and it was revealed that MDA cells secrete more exosomes. Furthermore, the isolated MCF7-derived exosomes were placed together with a normal bronchial/tracheal epithelial cell line (BTEC) for an additional assay, which aimed to observe the uptake of exosomes by other cells and the exosomes’ capability of promoting cell-cell communication. This observation was made based on alterations in the expression levels of c-Myc and miR-21 genes and the fact that they both have an increased expression in BTEC cells incubated with tumor-derived exosomes when compared to control cells (without incubation with the exosomes) lead us to the conclusion that the exosome uptake and exchange of information between the exosomes and the normal cells did occurred.

Novel antibiotic-loaded orthopaedic bone cements: insights on drug release profiles and biocompatibility

Acrylic bone cement (BC) is widely used as an anchor of artificial joints. Bacterial infection due to biofilm formation and inflammation are common and difficult to treat problems associated with commercial available BC formulations. Research on novel BC compositions is urgently needed. The main objective of this thesis was to develop a new biocompatible antibiotic-loaded BC with improved release profile. To achieve that aim several additives were incorporated, as an antibiotic (levofloxacin) to combat bacterial growth, an anti-inflammatory drug (diclofenac) to decrease the inflammatory process and two well-known and broadly used biopolymers, alginate and chitosan in order to increase matrix porosity, and in this way to intensify the amount of released drug. Novel BC formulations were tested in order to find the most suitable one that had potential to proceed to clinical application. Numerous tests were conducted as: a) evaluation of drug release profiles in different biomimetic media, b) mechanical and surface studies, c) microbiological activity testing against Staphylococcus aureus and d) in vitro biocompatibility assays (fibroblasts and osteoblasts). In general, the addition of biopolymers increased drug release, didn’t compromised BC mechanical properties and increased BC hydrophilicity. Microbiological testing revealed that Lev[BC]Chi was the only matrix that reduced significantly biofilm formation. On the contrary, alginate and diclofenac loading into BC seemed to increase biofilm growth. Biocompatibility studies showed some decrease in cell viability, in particularly on osteoblasts, mainly due to the high amounts of released drugs. In conclusion, the present work has shown that the matrix with more potential to proceed in further investigations was Lev[BC]Chi. Other conditions (namely additives and drugs concentrations) should be evaluated with the other tested BC matrices before being discharged.

Development of paper transistor with memory effect

This work will discuss the use of different paper membranes as both the substrate and dielectric for field-effect memory transistors. Three different nanofibrillated cellulose membranes (NFC) were used as the dielectric layer of the memory transistors (NFC), one with no additives, one with an added polymer PAE and one with added HCl. Gallium indium zinc oxide (GIZO) was used as the device’s semiconductor and gallium aluminium zinc oxide (GAZO) was used as the gate electrode. Fourier transform infrared spectroscopy (FTIR) was used to access the water content of the paper membranes before and after vacuum. It was found that the devices recovered their water too quickly for a difference to be noticeable in FTIR. The transistor’s electrical performance tests yielded a maximum ION/IOFF ratio of around 3,52x105 and a maximum subthreshold swing of 0,804 V/decade. The retention time of the dielectric charge that grants the transistor its memory capabilities was accessed by the measurement of the drain current periodically during 144 days. During this period the mean drain current did not lower, leaving the retention time of the device indeterminate. These results were compared with similar devices revealing these devices to be at the top tier of the state-of-the-art.

A arte ao serviço do império e das colónias: o contributo de alguns programas expositivos e museológicos para o discurso de legitimação territorial

Artigo escrito no âmbito da realização de tese de doutoramento em História da Arte financiado por bolsa FCT (SFRH/BD/63763/2009)