On-line diagnostics of transformer winding insulation failures, by Park`s vector approach

This paper presents a non-invasive approach for diagnosing winding insulation failures in three-phase transformers, which is based on the on-line monitoring of the primary and secondary current Park's Vector. Experimental and simulated results demonstrate the effectiveness of the proposed technique, for detecting winding inter-turn insulation faults in operating three-phase transformers.

Incipient turn-to-turn winding fault diagnosis of power transformers by the on-load exciting current Extended Park`s Vector Approach

This paper presents the application of the on-load exciting current Extended Park's Vector Approach for diagnosing incipient turn-to-turn winding faults in operating power transformers. Experimental and simulated test results demonstrate the effectiveness of the proposed technique, which is based on the spectral analysis of the AC component of the on-load exciting current Park's Vector modulus.

Transformers on-load exciting current Park`s vector approach as a tool for winding faults diagnostics

This paper presents the development of a new approach for diagnosing the occurrence of inter-turn short-circuits in the windings of three-phase transformers, which is based on the on-line monitoring of the on-load exciting current Park's Vector patterns. Experimental and simulated results demonstrate the effectiveness of the proposed technique for detecting winding inter-turn insulation faults in operating three-phase transformers.

Diagnóstico de avarias em transformadores trifásicos em serviço através da análise do Vector de Park das correntes de excitação em carga

Devido ao seu custo e importância estratégica, os transformadores de potência constituem um componente vital dos sistemas de produção, transmissão e distribuição de energia eléctrica. A sua fiabilidade constitui assim um factor crucial no funcionamento dos sistemas eléctricos de energia. Não admira, pois, que sobre estes equipamentos recaiam grandes preocupações relativamente à sua manutenção e, consequentemente, ao desenvolvimento de métodos capazes de fornecerem um diagnóstico completo e fiável do seu estado de funcionamento. Para o efeito, torna-se porém indispensável possuir um conhecimento detalhado acerca das avarias susceptíveis de ocorrerem nos transformadores, bem como dos mecanismos específicos que lhe estão subjacentes.

Estimating bottom properties with a vector sensor array during MakaiEx 2005

Nowadays, vector sensors which measure both acoustic pressure and particle velocity begin to be available in underwater acoustic systems, normally configured as vector sensor arrays (VSA). The spatial filtering capabilities of a VSA can be used, with advantage over traditional pressure only hydrophone arrays, for estimating acoustic field directionality as well as arrival times and spectral content, which could open up the possibility for its use in bottom properties' estimation. An additional motivation for this work is to test the possibility of using high frequency probe signals (say above 2 kHz) for reducing size and cost of actual sub bottom profilers and current geoacoustic inversion methods. This work studies the bottom related structure of the VSA acquired signals, regarding the emitted signal waveform, frequency band and source-receiver geometry in order to estimate bottom properties, specially bottom reflection coefficient characteristics. Such a system was used during the Makai 2005 experiment, off Kauai I., Hawai (USA) to receive precoded signals in a broad frequency band from 8 up to 14 kHz. The agreement between the observed and the modelled acoustic data is discussed and preliminary results on the bottom reflection estimation are presented.

Detection of trasformer intermittent winding faults by the on-load exciting current Park`s vector approach

This paper presents the application of the on-load exciting current Park's Vector Approach for diagnosing permanent and intermittent turn-to-turn winding faults in operating power transformers. First, an experimental investigation of the behaviour of the transformer under the occurrence of both permanent and intermittent winding faults is presented. Finally, experimental test results demonstrate the effectiveness of the proposed diagnostic technique, which is based on the on-line monitoring of the on-load exciting current Park's Vector patterns.

Source localization with vector sensor array during the makai experiment

Vector sensors measure both the acoustic pressure and the three components of particle velocity. Because of this, a vector sensor array (VSA) has the advantage of being able to provide substantially higher directivity with a much smaller aperture than an array of traditional scalar (pressure only) hydrophones. Although several, most of them theoretic, works were published from early nineties, only in the last years due to improvements and availability of vector sensor technology, the interest on field experiments with VSA increased in the scientific community. During the Makai Experiment, that took place off the coast of Kauai I., Hawaii, in September 2005, real data were collected with a 4 element vertical VSA. These data will be discussed in the present paper. The acoustic signals were emitted from a near source (low frequency ship noise) and two high frequency controlled acoustic sources located within a range of 2km from the VSA. The advantages of the VSA over traditional scalar hydrophone arrays in source localization will be addressed using conventional beamforming.

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.

Synthesis and evaluation of polymer nanoparticles for delivery in RPE cells

Ocular pathologies are among the most debilitating medical conditions affecting all segments of the population. Traditional treatment options are often ineffective, and gene therapy has the potential to become an alternative approach for the treatment of several pathologies. Methacrylate polymers have been described as highly biocompatible and are successfully used in medical applications. Due to their cationic nature, these polymers can be used to form polyplexes with DNA for its delivery. This work aims to study the potential of PDMAEMA (poly(2-(N,N’-dimethylamino)ethyl methacrylate)) as a non viral gene delivery system to the retina. The first part of this work aimed to study the potential for gene delivery of a previously synthesized PDMAEMA polymer of high molecular weight (354kDa). In the second part, we synthesized by RAFT a PDMAEMA with a lower molecular weight (103.3kDa) and similarly, evaluated its ability to act as a gene delivery vehicle. PDMAEMA/DNA polyplexes were prepared at 5, 7.5, 10, 12.5 and 20 nitrogen/phosphorous (N/P) ratio for the 354kDa PDMAEMA and at 5 and 7.5 for the 103.3kDa PDMAEMA. Dynamic light scattering and zeta potential measurements confirmed the nanosize and positive charge of polyplexes for all ratios and for both polymers. Both high and low Mw PDMAEMA were able to efficiently complex and protect DNA from DNase I degradation. Their cytotoxicity was evaluated using a non-retinal cell line (HEK293) and a retinal pigment epithelium (RPE) cell line (D407). We have found that cytotoxicity of the free polymer is concentration and time dependent, as expected, and negligible for all the concentrations of the PDMAEMA-DNA polyplexes. Furthermore, for the concentrations to be used in vivo, the 354kDa PDMAEMA showed no signs of inflammation upon injection in the intravitreal space of C57BL/6 mice. The transfection efficiency, as evaluated by fluorescence microscopy and flow cytometry, showed that the D407 retinal cells were transfected by polyplexes of both high and low Mw PDMAEMA, but with varied efficiency, which was dependent on the N/P ratio. Althogether, these results suggest that PDMAEMA is a feasible candidate for non-viral gene delivery to the retina, and this work constitutes the basis of further studies to elucidate the bottleneck in transfection and further optimization of the material.

Development of non-viral vectors for gene therapy for pathologies of the retina

Tese de doutoramento, Ciências Biomédicas, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, 2015