On the application of optimal control techniques in lossy fieldbuses

The performance of real-time networks is under continuous improvement as a result of several trends in the digital world. However, these tendencies not only cause improvements, but also exacerbates a series of unideal aspects of real-time networks such as communication latency, jitter of the latency and packet drop rate. This Thesis focuses on the communication errors that appear on such realtime networks, from the point-of-view of automatic control. Specifically, it investigates the effects of packet drops in automatic control over fieldbuses, as well as the architectures and optimal techniques for their compensation. Firstly, a new approach to address the problems that rise in virtue of such packet drops, is proposed. This novel approach is based on the simultaneous transmission of several values in a single message. Such messages can be from sensor to controller, in which case they are comprised of several past sensor readings, or from controller to actuator in which case they are comprised of estimates of several future control values. A series of tests reveal the advantages of this approach. The above-explained approach is then expanded as to accommodate the techniques of contemporary optimal control. However, unlike the aforementioned approach, that deliberately does not send certain messages in order to make a more efficient use of network resources; in the second case, the techniques are used to reduce the effects of packet losses. After these two approaches that are based on data aggregation, it is also studied the optimal control in packet dropping fieldbuses, using generalized actuator output functions. This study ends with the development of a new optimal controller, as well as the function, among the generalized functions that dictate the actuator’s behaviour in the absence of a new control message, that leads to the optimal performance. The Thesis also presents a different line of research, related with the output oscillations that take place as a consequence of the use of classic co-design techniques of networked control. The proposed algorithm has the goal of allowing the execution of such classical co-design algorithms without causing an output oscillation that increases the value of the cost function. Such increases may, under certain circumstances, negate the advantages of the application of the classical co-design techniques. A yet another line of research, investigated algorithms, more efficient than contemporary ones, to generate task execution sequences that guarantee that at least a given number of activated jobs will be executed out of every set composed by a predetermined number of contiguous activations. This algorithm may, in the future, be applied to the generation of message transmission patterns in the above-mentioned techniques for the efficient use of network resources. The proposed task generation algorithm is better than its predecessors in the sense that it is capable of scheduling systems that cannot be scheduled by its predecessor algorithms. The Thesis also presents a mechanism that allows to perform multi-path routing in wireless sensor networks, while ensuring that no value will be counted in duplicate. Thereby, this technique improves the performance of wireless sensor networks, rendering them more suitable for control applications. As mentioned before, this Thesis is centered around techniques for the improvement of performance of distributed control systems in which several elements are connected through a fieldbus that may be subject to packet drops. The first three approaches are directly related to this topic, with the first two approaching the problem from an architectural standpoint, whereas the third one does so from more theoretical grounds. The fourth approach ensures that the approaches to this and similar problems that can be found in the literature that try to achieve goals similar to objectives of this Thesis, can do so without causing other problems that may invalidate the solutions in question. Then, the thesis presents an approach to the problem dealt with in it, which is centered in the efficient generation of the transmission patterns that are used in the aforementioned approaches.

Studies of epithelial response to streptozotocin-induced hyperglycemia

A hiperglicemia é a principal característica da diabetes mellitus (DM), uma das causas de morte que mais cresce em Portugal, e cujas complicações a longo prazo são mais debilitantes e mais sobrecarregam os sistemas de saúde. No entanto, os mecanismos subjacentes à resposta fisiológica de alguns tecidos à hiperglicemia não estão completamente esclarecidos. Este estudo teve como objetivo avaliar de que forma o tempo de exposição à hiperglicemia afeta dois tecidos epiteliais, o endotélio e as glândulas salivares, que são frequentemente associados a complicações da DM, utilizando um modelo animal. Adicionalmente, procurou-se encontrar novos biomarcadores para avaliar a suscetibilidade a complicações orais em diabéticos, analisando as modificações pós-traducionais (PTMs) da família de proteínas mais representativa na saliva humana: proteínas ricas em prolina (PRPs). A disfunção vascular está na origem de várias complicações da diabetes. Neste sentido, observou-se uma progressiva disfunção endotelial com o aumento do tempo de exposição à hiperglicemia, que resulta do aumento de danos no endotélio e da diminuição da capacidade de mobilização de células progenitoras. Simultaneamente, o aumento observado na atividade da via de ativação do sistema de complemento mediada por lectinas (MBL), sugere um envolvimento do sistema de imunidade inata na patogénese da disfunção vascular. Outra complicação comum da DM é o desenvolvimento de doenças orais, nomeadamente as relacionadas com a redução da secreção salivar. Na análise às glândulas submandibulares, observou-se uma resposta inicial à hiperglicemia com fortes variações na expressão de proteínas, mas a longo prazo, estas variações foram atenuadas, sugerindo um mecanismo de adaptação à hiperglicemia crónica. Adicionalmente, as proteínas relacionadas com a secreção, como as anexinas, apresentaram-se sobre-expressas, enquanto as calicreinas e proteínas metabólicas estavam sub-expressas. Estas variações sugerem que, apesar de uma diminuição da capacidade de regeneração, as células tentam superar a perda de tecido por meio do aumento da secreção, embora sem êxito. O comprometimento funcional das glândulas salivares tem consequências na composição e funções da saliva. Analisando as PTMs das PRPs salivares humanas, observou-se um aumento da frequência de péptidos com ciclização de resíduos N-terminais de glutamina a piroglutamato, o que confere uma resistência à atividade proteolítica que, por sua vez, se encontra aumentada em diabéticos. Assim, a presença de péptidos com N-piroglutamato poderá ser um potencial biomarcador da suscetibilidade a complicações orais em diabéticos.

Numerical and combinatorial applications of generalized Appell polynomials

This thesis studies properties and applications of different generalized Appell polynomials in the framework of Clifford analysis. As an example of 3D-quasi-conformal mappings realized by generalized Appell polynomials, an analogue of the complex Joukowski transformation of order two is introduced. The consideration of a Pascal n-simplex with hypercomplex entries allows stressing the combinatorial relevance of hypercomplex Appell polynomials. The concept of totally regular variables and its relation to generalized Appell polynomials leads to the construction of new bases for the space of homogeneous holomorphic polynomials whose elements are all isomorphic to the integer powers of the complex variable. For this reason, such polynomials are called pseudo-complex powers (PCP). Different variants of them are subject of a detailed investigation. Special attention is paid to the numerical aspects of PCP. An efficient algorithm based on complex arithmetic is proposed for their implementation. In this context a brief survey on numerical methods for inverting Vandermonde matrices is presented and a modified algorithm is proposed which illustrates advantages of a special type of PCP. Finally, combinatorial applications of generalized Appell polynomials are emphasized. The explicit expression of the coefficients of a particular type of Appell polynomials and their relation to a Pascal simplex with hypercomplex entries are derived. The comparison of two types of 3D Appell polynomials leads to the detection of new trigonometric summation formulas and combinatorial identities of Riordan-Sofo type characterized by their expression in terms of central binomial coefficients.

Oxidation and glycoxidation of phosphatidylserine and their effect in immune response : a lipidomic approach

Phosphatidylserine (PS) is a member of the class of phospholipids, and is distributed among all cells of mammalians, playing important roles in diverse biological processes, including blood clotting and apoptosis. When externalized, PS is a ligand that is recognized on apoptotic cells. It has been considered that before externalization PS is oxidized and oxPS enhance the recognition by macrophages receptors, however the knowledge about oxidation of PS is still limited. PS, like others phospholipids, has two fatty acyl chains and one polar head group, in this case is the amino acid serine. The modifications in PS structure can occur by oxidation of the unsaturated fatty acyl chains and by glycation of the polar head group, due to free amine group, thus increasing the susceptibility to oxidative events. The main goal of this work was to characterize and identify oxidized and glycoxidized PS, contributing to the knowledge of the biological role of oxidation products of PS, as well as of glycated PS, in immune and inflammatory processes. To achieve this goal, PS standards (1-palmitoyl-2-oleoyl-sn-glycero-3-phospho- L-serine (POPS), 1,2-dipalmitoyl-sn-glycero-3-phospho-L-serine (DPPS), 1- palmitoyl-2-linoleoyl-sn-glycero-3-phospho-L-serine (PLPS) and 1-palmitoyl-2- arachidonoyl-sn-glycero-3-phospho-L-serine (PAPS)) and glycated PS (PAPS and POPS) were induced to oxidize in model systems, using different oxidant reagents: HO• and 2,2'-azobis-2-methyl-propanimidamide dihydrochloride (AAPH) . The detailed structural characterization of the oxidative products was performed by ESI-MS and MS/MS coupled to separation techniques such as off line TLC-MS and on line LC-MS, in order to obtained better characterization of the larger number of PS and glycated PS oxidation products. The results obtained in this work allowed to identify several oxidation products of PS and glycated PS with modifications in unsaturated fatty acyl chain. Also, oxidation products formed due to structural changes in the serine polar head with formation of terminal acetamide, terminal hydroperoxyacetaldehyde.and terminal acetic acid (glycerophosphacetic acid, GPAA) were identified. The mass spectrometric specific fragmentation pathway of each type of oxidation product was determined using different mass spectrometry approaches. Based on the identified fragmentation pathways, targeted lipidomic analysis was performed to detect oxidation products modified in serine polar head in HaCaT cell line treated with AAPH. The GPAA was detected in HaCaT cells treated with AAPH to induce oxidative stress, thus confirming that modifications in PS polar head is possible to occur in biological systems. Furthermore, it was found that glycated PS species are more prone to oxidative modifications when compared with non glycated PS. During oxidation of glycated PS, besides the oxidation in acyl chains, new oxidation products due to oxidation of the glucose moiety were identified, including PS advanced glycation end products (PSAGES). To investigate if UVA oxidative stress exerted changes in the lipidome of melanoma cell lines, particularly in PS profile, a lipidomic analysis was performed. The lipid profile was obtained using HILIC-LC-MS and GC-MS analysis of the total lipid extracts obtained from human melanoma cell line (SKMEL- 28) after UVA irradiation at 0, 2 and 24 hours. The results did not showed significant differences in PS content. At molecular level, only PS (18:0:18:1) decreased at the moment of irradiation. The most significant changes in phospholipids content occurred in phosphatidylcholines (PC) and phosphatidylinositol (PI) classes, with an increase of mono-unsaturated fatty acid (MUFA), similarly as observed for the fatty acid analysis. Overall, these data indicate that the observed membrane lipid changes associated with lipogenesis after UVA exposure may be correlated with malignant transformations associated with cancer development and progression. Despite of UVA radiation is associated with oxidative damage, in this work was not possible observe oxidation phospholipids. The anti/pro-inflammatory properties of the oxidized PLPS (oxPLPS) versus non-oxidized PLPS were tested on LPS stimulated RAW 264.7 macrophages. The modulation of intracellular signaling pathways such as NF-kB and MAPK cascades by oxPLPS and PS was also examined in this study. The results obtained from evaluation of anti/pro-inflammatory properties showed that neither PLPS or oxPLPS species activated the macrophages. Moreover only oxidized PLS were found to significantly inhibit NO production and iNOS and il1β gene transcription induced by LPS. The analysis at molecular level showed that this was the result of the attenuation of LPS-induced c-Jun-N-terminal kinase (JNK) and p65 NF-kB nuclear translocation. Overall these data suggest that oxPLPS, but not native PLPS, mitigates pro-inflammatory signaling in macrophages, contributing to containment of inflammation during apoptotic cell engulfment. The results obtained in this work provides new information on the modifications of PS, facilitating the identification of oxidized species in complex samples, namely under physiopathologic conditions and also contributes to a better understanding of the role of oxPS and PS in the inflammatory response, in the apoptotic process and other biological functions.

Altered regulatory response of Rab7a and Rab9 in MM1 and VV2 subtype of Creutzfeldt-Jakob disease

The present study was undertaken to identify proteins interacting with PrPC that could provide new insights into its physiological functions and pathological role. We performed a target search for lysosomal network protein, Rab7a and Rab9, in frontal cortex and cerebellum of human brain from patients with sCJD-MM1 and sCJD-VV2. The intracellular level of Rab7a was increased significantly, when compared with healthy age-matched control. Interactions of PrPC and Rab7a/Rab9 were further investigated by using confocal laser scanning microscopy. Immunofluorescence results suggested potential interactions of Rab7a and PrPC. siRNA against the Rab7a gene was used to knockdown the expression of Rab7a protein in primary cell culture of cortical neurons from wild type mice. This depleted Rab7a resulted an impairment of PrPC trafficking leading to an accumulation of PrPC in the endocytosis pathway. Furthermore, interactions of Tau and Rab7a were investigated by using western blot analysis and confocal laser scanning microscopy. Cell cultures of cortex of wildtype mice were treated with siRNA-Tau, siRNA-Rab7 and control siRNA followed by immunofluorescence. The results of immunofluorescence suggested potential interaction of Tau and Rab7a. Cells lines treated with siRNA-Tau, the intracellular levels of Rab7a and Rab9 significantly increases and their localization is also modified. When we transfected this cells lines with siRNA-rab7a the accumulation of Tau decreases in cytosolic region and their localization was also modified when compared with control cells. In conclusion, this study may help to understand and characterize the subtype specific disease progression in CJD cases. Furthermore, it could be a step ahead to development of new treatment strategies for diseases subtype specific manner.