Characterization of PPP1 interacting proteins in male reproduction

A fosforilação reversível de proteínas é um importante mecanismo de controlo em eucariotas. A fosfoproteína fosfatase 1 (PPP1) é uma fosfatase de serina/treonina envolvida em vários processos celulares. Existem três isoformas da subunidade catalítica (α/CA, δ/β/CB e γ/CC) com pequenas diferenças nos terminais amino e carboxílico. O gene PPP1CC sofre ainda splicing alternativo para produzir duas isoformas, a PPP1CC1 ubíqua e a PPP1CC2 enriquecida em testículo e específica de esperma. A localização e especificidade de substratos da PPP1 está dependente da formação de complexos oligoméricos com proteínas que interagem com a PPP1 (PIPs). O objetivo principal desta tese foi estudar novas PIPs, específicas de testículo e esperma, a fim de melhor caracterizar o papel desta fosfatase e dos respetivos complexos na reprodução em mamíferos. Com este fim, estudou-se a presença, localização e possíveis funções de uma PIP previamente conhecida, PPP1R2, e de duas novas PIPs, PPP1R2P3 e Tctex1d4. PPP1R2 e PPP1R2P3 estão presentes em esperma humano colocalizando com a PPP1CC2, na cabeça e na cauda. A hipótese é que as holoenzimas localizadas na cabeça terão um papel na reação acrossómica, enquanto que as holoenzimas presentes no axonema são relevantes para o controlo da motilidade flagelar. De seguida foram estudados os pseudogenes da PPP1R2, em termos de história evolutiva e de possíveis funções. Na espécie humana, a PPP1R2 tem 10 pseudogenes, 7 deles específicos de primatas. Estudos de bioinformática e dados de expressão mostram que os PPP1R2P1/P3/P9 são os pseudogenes com maior probabilidade de serem transcritos e traduzidos. Também identificámos o PPP1R2P9 em esperma humano e mostrámos que alguns pseudogenes poderão estar associados a estados fisiopatológicos. Isto indica que o processo de evolução poderá estar ligado á formação de novos genes ou ao controlo do mRNA da PPP1R2. A sobre-expressão da PPP1R2 ou PPP1R2P3 em testículo de ratinho também foi realizada, para caracterizar os mecanismos envolvidas na função dos complexos PPP1R2/PPP1R2P3-PPP1CC2 na espermatogénese e fisiologia dos espermatozoides. A dineína de cadeia leve, Tctex1d4, foi encontrada como interagindo com a PPP1C e como estando presente em testículo de ratinho e em esperma humano. Demonstrámos que a Tctex1d4 e a PPP1 colocalizam no centro organizador de microtúbulos e nos microtúbulos e que o motivo de ligação à PPP1 presente na Tctex1d4 parece ser importante para manter a PPP1 no centro organizador de microtúbulos e/ou para disromper ou atrasar o seu movimento ao longo dos microtúbulos emergentes. Estes resultados abrem novos caminhos para os possíveis papéis do complexo Tctex1d4-PPP1 na dinâmica dos microtúbulos, motilidade do esperma, reação acrossómica e na regulação da barreira hemato-testicular, provavelmente, através da via de sinalização do TGFß. A análise do motivo de ligação à PPP1 mostra que este é altamente conservado entre os mamíferos, com exceção das Pikas, sugerindo que esta perda aconteceu antes da radiação das Pikas, há 6-20 milhões de anos atrás. Através de um rastreio por mutações demonstrámos que a capacidade da Tctex1d4 se ligar à PPP1 é mantida nas Pikas, embora o motivo de ligação à PPP1 esteja disrompido. Este estudo abre portas para novas descobertas na área da reprodução mostrando o papel da PPP1CC2 na espermatogénese e fisiologia do esperma.

Laser assisted directional solidification of zirconia-based eutectics

Directionally solidified zirconia-based eutectic (DSE) fibres were obtained using the laser floating zone (LFZ) method. Two systems were investigated: zirconia-barium zirconate and zirconia-mullite. The purpose was to take advantage of zirconia properties, particularly as an ionic conductor and a mechanical rein-forcement phase. The influence of processing conditions in the structural and microstructural characteristics and their consequences on the electrical and mechanical behaviour were the focus of this thesis. The novel zirconia-barium zirconate eutectic materials were developed in order to combine oxygen ionic conduction through zirconia with protonic conduction from barium zirconate, promoting mixed ionic conduction behaviour. The mi-crostructure of the fibres comprises two alternated regions: bands having coarser zirconia-rich microstructure; and inter-band regions changing from a homogeneous coupled eutectic, at the lowest pulling rate, to columnar colony microstructure, for the faster grown fibres. The bands inter-distance increases with the growth rate and, at 300 mm/h, zirconia dendrites develop enclosed in a fine-interpenetrated network of 50 vol.% ZrO2-50 vol.% BaZrO3. Both phases display contiguity without interphase boundaries, according to impedance spec-troscopy data. Yttria-rich compositions were considered in order to promote the yttrium incorporation in both phases, as revealed by Raman spectroscopy and corroborated by the elemental chemical analysis in energy dispersive spectros-copy. This is a mandatory condition to attain simultaneous contribution to the mixed ionic conduction. Such results are supported by impedance spectrosco-py measurements, which clearly disclose an increase of total ionic conduction for lower temperatures in wet/reduction atmospheres (activation energies of 35 kJ/mol in N2+H2 and 48 kJ/mol in air, in the range of 320-500 ºC) compared to the dry/oxidizing conditions (attaining values close to 90 kJ/mol, above 500 ºC). At high temperatures, the proton incorporation into the barium zirconate is un-favourable, so oxygen ion conduction through zirconia prevails, in dry and oxi-dizing environments, reaching a maximum of 1.3x10-2 S/cm in dry air, at ~1000 ºC. The ionic conduction of zirconia was alternatively combined with another high temperature oxygen ion conductor, as mullite, in order to obtain a broad elec-trolytic domain. The growth rate has a huge influence in the amount of phases and microstructure of the directionally solidified zirconia-mullite fibres. Their microstructure changes from planar coupled eutectic to dendritic eutectic mor-phology, when the growth rate rises from 1 to 500 mm/h, along with an incre-ment of tetragonal zirconia content. Furthermore, high growth rates lead to the development of Al-Si-Y glassy phase, and thus less mullite amount, which is found to considerably reduce the total ionic conduction of as-grown fibres. The reduction of the glassy phase content after annealing (10h; 1400 ºC) promotes an increase of the total ionic conduction (≥0.01 S/cm at 1370 °C), raising the mullite and tetragonal zirconia contents and leading to microstructural differ-ences, namely the distribution and size of the zirconia constituent. This has important consequences in conductivity by improving the percolation pathways. A notable increase in hardness is observed from 11.3 GPa for the 10 mm/h pulled fibre to 21.2 GPa for the fibre grown at 500 mm/h. The ultra-fine eutectic morphology of the 500 mm/h fibres results in a maximum value of 534 MPa for room temperature bending strength, which decreases to about one-fourth of this value at high temperature testing (1400 ºC) due to the soft nature of the glassy-matrix.

IB-DFE receiver techniques for CP-assisted block transmission within DS-CDMA and MC-CDMA systems

MC-CDMA (MultiCarrier Code Division Multiple Access), currently regarded as a promissing multiple access scheme for broadband communications, is known to combine the advantages of an OFDM-based (Orthogonal Frequency Division Multiplexing), CP-assisted (Cyclic Prefix) block transmission with those of CDMA systems. Recently, it was recognised that DS-CDMA (Direct Sequence) implementations can also take advantage of the beneficts of the CP-assisted block transmission approach, therefore enabling an efficient use of FFT-based (Fast Fourier Transform), chip level FDE (Frequency- Domain Equalisation) techniques. In this paper we consider the use of IB-DFE (Iterative Block Decision Feedback Equalisation) FDE techniques within both CP-assisted MC-CDMA systems with frequency-domain spreading and DS-CDMA systems. Our simulation results show that an IB-DFE receiver with moderate complexity is suitable in both cases, with excellent performances that can be close to the single-code matched filter bound (especially for the CP-assisted DSCDMA alternative), even with full code usage.

A frequency-domain receiver for asynchronous systems employing CP-assisted DS-CDMA schemes

In this paper we consider the uplink transmission within CP-assisted (Cyclic Pre¯x) DS-CDMA (Direct Sequence Code Division Multiple Access) systems and we present a frequency-domain MUD (MultiUser Detection) receiver with iterative estimation and compensation of residual frequency errors. The proposed receiver is suitable for broadband wireless systems, with performances that can be close to the single-user MFB (Matched Filter Bound), even for fully loaded systems and/or in the presence of strong interfering signals. The receiver is powerful enough for typical asynchronous scenarios, requiring only a coarse synchronization between users.

SIC and PIC multiuser detection for prefix-assisted DS-CDMA systems

In this paper we present iterative frequency-domain multiuser detection (MUD) receivers for the uplink transmission of direct sequence code division multiple access systems (DS-CDMA) that combine iterative block decision feedback equalization (IB-DFE) principles with interference cancelation techniques. Both successive interference cancelation (SIC) and parallel interference cancelation (PIC) structures are considered. Our performance results show that the proposed receiver structures have excellent bit error rate (BER) performances, that can be close to the single-user matched filter bound (MFB), even for fully loaded systems and severely time-dispersive channels1.

B-splines neural network assisted PID autotuning

This papers describes an extantion of previous works on the subject of neural network proportional, integral and derivative (PID) autotuning. Basically, neural networks are employed to supply the three PID parameters, according to the integral of time multiplied by the absolute error (ITAE) criterion, to a standard PID controller.

B-splines neural networks assisted PID autotuning

This paper describes previous works (1), (2), on neural network pid autotuning. Basically, neural networks are employed to supply PID parameters, according to the ITAE criterion, to a standard PID controller.

Neural networks assisted PID autotuning

The Proportional, Integral and Derivative (PID) controllers are standard building blocks for industrial automation. Their popularity comes from their rebust performance and also from their functional simplicity.

Neural network assisted autotuning of industrial PID controllers

A recent servey (1) has reported that the majority of industrial loops are controlled by PID-type controllers and many of the PID controllers in operation are poorly tuned. poor PID tuning is due to the lack of a simple and practical tuning method for avarage users, and due to the tedious procedurs involved in the tuning and retuning of PID controllers.

Neural networks assisted PID autotuning

A scheme of automatically tuning the existing industrial PID controllers using neural networks is proposed. The scheme estimates the process critical data on-line in proportional control mode.

Turbo equalization with cancellation of nonlinear distortion effects for CP-Assisted and Zero-Padded MC-CDMA signals

We consider MC-CDMA schemes, with reduced envelope fluctuations. Both CP-assisted (cyclic prefix) and ZP (zero-padded) MC-CDMA schemes are addressed. We develop turbo FDE (frequency-domain equalization) schemes, combined with cancelation of nonlinear distortion effects. The proposed turbo receivers allow significant performance improvements at low and moderate SNR, even when the transmitted signals have reduced envelope fluctuations.

Multiuser Detection for the Uplink of Prefix-Assisted DS-CDMA Systems Employing Multiple Transmit and Receive Antennas

In this paper we consider the uplink transmission within a DS-CDMA system employing CP-assisted (Cylic Prefix) block transmission techniques combined with spatial multiplexing techniques that require multiple antennas at both the transmitter and the receiver. We present an efficient frequency-domain receiver structure with iterative MUD (MultiUser Detection). The performance of the proposed receiver can be close to the single user matched filter bound, even for fully loaded systems and/or severely time-dispersive channels.

Synthesis and computer-assisted design of mitochondrial electron transport-chain inhibitors as antimalarial agents

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

Laccase-assisted approach to graft multifunctional materials of interest: keratin-EC based novel composites and their characterisation

This study focuses on the evaluation of raw keratin as a potential material to develop composites with novel characteristics. Herein, we report a mild and eco-friendly fabrication of in-house extracted feather keratin-based novel enzyme assisted composites consisting of ethyl cellulose (EC) as a backbone material. A range of composites between keratin and EC using different keratin: EC ratios were prepared and characterised. Comparing keratin to the composites, the FT-IR peak at 1,630 cm-1 shifted to a lower wavenumber of 1,610 cm-1 in keratin-EC which typically indicates the involvement of β-sheet structures of the keratin during the graft formation process. SEM analysis revealed that the uniform dispersion of the keratin increases the area of keratin-EC contact which further contributes to the efficient functionality of the resulting composites. In comparison to the pristine keratin and EC, a clear shift in the XRD peaks was also observed at the specific region of 2-Theta values of keratin-g-EC. The thermo- mechanical properties of the composites reached their highest levels in comparison to the keratin which was too fragile to be measured for its mechanical properties. Considerable improvement in the water contact angle and surface tension properties was also recorded.