Simulation of economic benefits achieved by water quality improvement for recreational and professional users, within the SPICOSSA SSA (SPICOSSA System Approach) 11 - Guadiana estuary ecosystem sic

Dissertação de Mestrado, Gestão da Água e da Costa, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2010

A scalable and open source linear positioning system controller

This paper is on the implementation of a dual axis positioning system controller. The system was designed to be used for space-dependent ultrasound signal acquisition problems, such as pressure field mapping. The work developed can be grouped in two main subjects: hardware and software. Each axis includes one stepper motor connected to a driver circuit, which is then connected to a processing unit. The graphical user interface is simple and clear for the user. The system resolution was computed as 127 mu m with an accuracy of 2.44 mu m. Although the target application is ultrasound signal acquisition, the controller can be applied to other devices that has up to four stepper motors. The application was developed as an open source software, thus it can be used or changed to fit different purposes.

Development of a Piggybac based direct reprogramming system for derivation of integration free induced pluripotent stem cells

Induced pluripotent stem cells (iPSc) have great potential for applications in regenerative medicine, disease modeling and basic research. Several methods have been developed for their derivation. The original method of Takahashi and Yamanaka involved the use of retroviral vectors which result in insertional mutagenesis, presence in the genome of potential oncogenes and effects of residual transgene expression on differentiation bias of each particular iPSc line. Other methods have been developed, using different viral vectors (adenovirus and Sendai virus), transient plasmid transfection, mRNA transduction, protein transduction and use of small molecules. However, these methods suffer from low efficiencies; can be extremely labor intensive, or both. An additional method makes use of the piggybac transposon, which has the advantage of inserting its payload into the host genome and being perfectly excised upon re-expression of the transposon transposase. Briefly, a policistronic cassette expressing Oct4, Sox2, Klf4 and C-Myc flanked by piggybac terminal repeats is delivered to the cells along with a plasmid transiently expressing piggybac transposase. Once reprogramming occurs, the cells are re-transfected with transposase and subclones free of tranposon integrations screened for. The procedure is therefore very labor intensive, requiring multiple manipulations and successive rounds of cloning and screening. The original method for reprogramming with the the PiggyBac transposon was created by Woltjen et al in 2009 (schematized here) and describes a process with which it is possible to obtain insert-free iPSc. Insert-free iPSc enables the establishment of better cellular models of iPS and adds a new level of security to the use of these cells in regenerative medicine. Due to the fact that it was based on several low efficiency steps, the overall efficiency of the method is very low (<1%). Moreover, the stochastic transfection, integration, excision and the inexistence of an active way of selection leaves this method in need of extensive characterization and screening of the final clones. In this work we aime to develop a non-integrative iPSc derivation system in which integration and excision of the transgenes can be controlled by simple media manipulations, avoiding labor intensive and potentially mutagenic procedures. To reach our goal we developed a two vector system which is simultaneously delivered to original population of fibroblasts. The first vector, Remo I, carries the reprogramming cassette and GFP under the regulation of a constitutive promoter (CAG). The second vector, Eneas, carries the piggybac transposase associated with an estrogen receptor fragment (ERT2), regulated in a TET-OFF fashion, and its equivalent reverse trans-activator associated with a positive-negative selection cassette under a constitutive promoter. We tested its functionality in HEK 293T cells. The protocol is divided in two the following steps: 1) Obtaining acceptable transfection efficiency into human fibroblasts. 2) Testing the functionality of the construct 3) Determining the ideal concentration of DOX for repressing mPB-ERT2 expression 4) Determining the ideal concentration of TM for transposition into the genome 5) Determining the ideal Windows of no DOX/TM pulse for transposition into the genome 6) 3, 4 and 5) for transposition out of the genome 7) Determination of the ideal concentration of GCV for negative selection We successfully demonstrated that ENEAS behaved as expected in terms of DOX regulation of the expression of mPB-ERT2. We also demonstrated that by delivering the plasmid into 293T HEK cells and manipulating the levels of DOX and TM in the medium, we could obtain puromycin resistant lines. The number of puromycin resistant colonies obtained was significantly higher when DOX as absent, suggesting that the colonies resulted from transposition events. Presence of TM added an extra layer of regulation, albeit weaker. Our PCR analysis, while not a clean as would be desired, suggested that transposition was indeed occurring, although a background level of random integration could not be ruled out. Finally, our attempt to determine whether we could use GVC to select clones that had successfully mobilized PB out of the genome was unsuccessful. Unexpectedly, 293T HEK cells that had been transfected with ENEAS and selected for puromycin resistance were insensitive to GCV.

Medium-term prospects for portuguese agriculture under health check Proposals: a quantitative analysis with the CAPRI modelling system

The challenges of Common Agricultural Policy are driven by internal and external factors, such as the budgetary constraints, the budget reform, the globalization and the world financial crisis. According to this work results, CAP will continue its evolution from a sectorial to a territorial approach, with a slow re-balance of its two pillars. The Portuguese agriculture will slowly adjust itself to the disappearance of prices and markets policy and the reinforcement of rural development policy. As in the past, agriculture will accommodate the reform effects and adjust to a new framework without sudden brakes or disclosers.

Acoustic oceanographic buoy telemetry system: an ‘advanced’ sonobuoy that meets acoustic rapid environmental assessment requirements

The Acoustic Oceanographic Buoy (AOB) Telemetry System has been designed to meet acoustic rapid environmental assessment requirements. It uses a standard institute of Electrical and Electronics Engineers 802.11 wireless local area network (WLAN) to integrate the air radio network (RaN) and a hydrophone array and acoustic source to integrate the underwater acoustic network (AcN). It offers advantages including local data storage, dedicated signal processing, and global positioning system (GPS) timing and localization. The AOB can also be integrated with other similar systems, due to its WLAN transceivers, to form a flexible network and perform on-line high speed data transmissions. The AOB is a reusable system that requires less maintenance and can also work as a salt-water plug-and-play system at sea as it is designed to operate in free drifting mode. The AOB is also suitable for performing distributed digital signal processing tasks due to its digital signal processor facility.