This paper concerns the solution of plate bending problems in domains composed of rectangles. Domain decomposition (DD) is the basic tool used for both the parallelization of the conjugate gradient method and the construction of efficient parallel preconditioners. A so-called Dirich- let DD preconditioner for systems of linear equations arising from the fi- nite element approximation by non-conforming Adini elements is derived. It is based on the non-overlapping DD, a multilevel preconditioner for the Schur-complement and a fast, almost direct solution method for the Dirichlet problem in rectangular domains based on fast Fourier transform. Making use of Xu's theory of the auxiliary space method we construct an optimal preconditioner for plate problems discretized by conforming Bogner-Fox-Schmidt rectangles. Results of numerical experiments carried out on a multiprocessor sys- tem are given. For the test problems considered the number of iterations is bounded independent of the mesh sizes and independent of the number of subdomains. The resulting parallel preconditioned conjugate gradient method requiresO(h^-2 ln h^-1 ln epsilon^-11) arithmetical operations per processor in order to solve the finite element equations with the relative accuracy epsilon.

The following paper has been dedicated to the verb like, which is one of the verbs in the English language that can take either a to-infinitive or an -ing participle as a complement. The purpose of the paper is to examine if there are any differences in distribution and meaning between the two complements. The focus also lies on the different verbs occurring as complements, and the contrast between the verbs occurring as to-infinitives and as -ing participles. There are many theories which have been proposed on the subject that lie as a basis for the investigation. The analysis was carried out by means of an investigation of sentences taken from the COBUILDDIRECT corpus, and includes both spoken and written British and American English. The outcome of the analysis has demonstrated that there is usually agreement between the theories and the results; however, there is not always a difference of meaning between the two complements. Moreover, the analysis suggests that there is a difference of verbs occurring as to-infinitives and -ing participles; the would like to construction represents a fixed expression and often occurs together with performative verbs. Finally, the conclusion has been drawn that there is a small but visible difference between the occurrences of the spoken and the written subcorpora when discussing both meaning and verbs occurring as complements. Since the to-infinitive complement is more common than the -ing participle in newspapers, books, and spoken English, the difference includes both detached and involved style as well as a regional difference between British and American English.

Chez les vertébrés, le foie est un des organes majeurs du métabolisme en étant le siège de la régulation de différentes voies du métabolisme qui contrôlent l’homéostasie du glucose et des lipides. En se basant sur des travaux de recherche récents suggérant que le système de conjugaison de l'ubiquitine est engagé en réponse à différents signaux métaboliques, nous avons réalisé une analyse protéomique globale dans le but d’identifier des protéines ubiquitylées dans le foie de souris soumises á un protocole de jeûne – re-nourrissage. Parmi les 117 protéines différemment ubiquitylé sur le jeûne ou le re-nourrissage, nous avons identifié complément 3 (C3) dans le foie de souris réalimentées. Nous avons observé qu'un produit d'activation de C3, C3a, est ubiquitylé dans les hépatocytes primaires traités avec les médias riches en nutriments. Ainsi, nous proposons que l'ubiquitination de C3 joue un rôle dans la régulation des fonctions inflammatoires ou métaboliques de C3 dans le foie. / In vertebrates, the liver has developed to be a major metabolic organ able to control glucose and lipid homeostasis. It activates or inhibits specific pathways in a regulated manner, depending on the metabolic state of our organism. Based on the emerging experimental evidence suggesting that the ubiquitin conjugation system is engaged in response to different metabolic cues, we conducted a global proteomic analysis to identify metabolic pathways modified by ubiquitylation. To this end, we used livers of mice subjected to a fasting – refeeding protocol. Amongst the 117 proteins differentially ubiquitylated upon fasting or refeeding conditions, we identified complement 3 (C3) to be ubiquitinated in livers of refed mice. We observed that an activation product of C3, C3a, is ubiquitylated in primary hepatocytes treated with nutrient-rich media. Thus, we suggest that the ubiquitylation of C3 plays a role in the regulation of inflammatory or metabolic functions of C3 in the liver.

In this work, prestressed elasticity problem as a model of the so-called glacial isostatic adjustment (GIA) process is studied. The model problem is described by a set of partial differential equations (PDE) and discretized with a mixed finite element (FE) formulation. In the presence of prestress the so-constructed system of equations is non-symmetric and indefinite. Moreover, the resulting system of equations is of the saddle point form. We focus on a robust and efficient block lower-triangular preconditioning method, where the lower diagonal block is and approximation of the so-called Schur complement. The Schur complement is approximated by the so-called element-wise Schur complement. The element-wise Schur complement is constructed by assembling exact local Schur complements on the cell elements and distributing the resulting local matrices to the global preconditioner matrix. We analyse the properties of the element-wise Schur complement for the symmetric indefinite system matrix and provide proof of its quality. We show that the spectral radius of the element-wise Schur complement is bounded by the exact Schur complement and that the quality of the approximation is not affected by the domain shape. The diagonal blocks of the lower-triangular preconditioner are combined with inner iterative schemes accelerated by (numerically) optimal and robust algebraic multigrid (AMG) preconditioner. We observe that on distributed memory systems, the top pivot block of the preconditioner is not scaling satisfactorily. The implementation of the methods is further studied using a general profiling tool, designed for clusters. For nonsymmetric matrices we use the theory of Generalized Locally Toeplitz (GLT) matrices and show the spectral behavior of the element-wise Schur complement, compared to the exact Schur complement. Moreover, we use the properties of the GLT matrices to construct a more efficient AMG preconditioner. Numerical experiments show that the so-constructed methods are robust and optimal.

The work described in this thesis is an experimental study into the application of Electron Paramagnetic Resonance (EPR) Spectroscopy for the study of biological systems. Using a variety of methods of site-directed spin-labelling (SDSL), this thesis aims to explore long range structure in an assortment of recombinant and native proteins, and complexes thereof. The work described in this thesis covers all aspects of the work, from experimental design, molecular biology and cloning, protein expression and purification, as well as functional characterisation, and finally EPR distance measurements, data analysis and interpretation. Challenges and pitfalls will also be addressed. Chapters 1 and 2 introduce EPR spectroscopy, and its application in the study of long range structure in biological systems. The experimental techniques employed throughout this thesis are also introduced. Chapter 3 details an investigation into the complement C3b:factor H complex. This chapter addresses the challenges associated with the SDSL of cysteine rich proteins. Utilising hidden cysteine residues in native proteins for spin-labelling purposes will also be addressed. Chapter 4 looks at the interactions of the human myosin regulatory light chain (RLC) with cardiac myosin binding protein C (cMyBP-C). Optimisation of expression and purification protocols will be the focus, as well as addressing issues with protein solubility and spin labelling efficiencies. Chapter 5 explores the development of new methods of SDSL, for the specific labelling of cysteine rich proteins. The ability of Escherichia coli to read through the amber stop codon will be exploited for the incorporation of unnatural amino acids for labelling purposes, and novel spin labels, specific for labelling cysteine pairs tested in several model systems. Furthermore, native paramagnetic centres in recombinant proteins will be explored as potential labelling sites.

[pt] Neste trabalho, foi realizada uma revisão crítica do tema na gramática de modelo tradicional, especificamente nas gramáticas de Cunha e Cintra (2001) e de Rocha Lima (2007). Através dessas análises, buscamos comprovar que o modo como o conceito de complementação verbal, bem como as noções de termo complementar e acessório e de transitividade, vêm sendo apresentados tradicionalmente em gramáticas e livros didáticos tem demonstrado algumas lacunas e incoerências que vem causando dificuldades ao ensino/aprendizagem da língua materna. Costuma-se relacionar a transitividade verbal à ideia de completude ou não da informação transmitida pelo verbo, sem levar em consideração a circunstância e o sentido nos quais o verbo está sendo empregado, tampouco a natureza do(s) elemento(s) a ele ligado(s). Observa-se, portanto, que aquilo que se apresenta aos estudantes como a gramática da língua portuguesa é um conjunto de regras que nem sempre leva em consideração a realidade dos fatos linguísticos. Com o propósito de encontrar um caminho de análise que ajudasse a solucionar o problema, foram examinados alguns trabalhos mais recentes que fogem em parte ou totalmente à orientação tradicional, como a Moderna Gramática Portuguesa de Bechara (edição a partir de 1999), a Gramática de usos do português de Moura Neves (2000), a dissertação de Mestrado de Maria Eliana Duarte Alves de Brito (1986) e a Gramática de Valências de Busse e Vilela (1986). Partindo de uma orientação funcionalista e tomando como base os princípios da Gramática de Valências, este trabalho tem por objetivo demonstrar que, no estudo das orações, é preciso que se levem em consideração dois pólos de análise: a sintaxe e a semântica. Sendo o verbo o elemento central da oração, é ele que determina o número de lugares-vazios, além das propriedades morfo-sintáticas e semânticas dos actantes que realizam esses lugares-vazios. Desse modo, a distinção entre objeto direto e objeto indireto da gramática tradicional mostra-se insuficiente, sendo necessário distinguir um número maior de tipos de actantes. Além disso, foi possível detectar, através da análise do verbo como elemento central, quais termos devem ser considerados como actantes (complementos) ou como circunstantes (adjuntos). / [en] This work carries a critical review of the traditional model of grammar, specifically the Grammars by Cunha e Cintra (2001) and Rocha Lima (2007). Through these analysis, we seek to prove that the manner how the concept of verb complementation, as well as the notions of complementary and accessory terms and of transitivity, have been traditionally presented in grammar and didactic books present a number of gaps and incoherencies that have been causing difficulties in the teaching/learning of our mother tongue [Portuguese]. Usually verbal transitivity is related to the idea if there is or not a completion of the information transmitted by the verb, without considering the circumstances and meaning in which the verb is being used or the nature of the element(s) connected to it. We can thus state that which is being presented to students as the grammar of the Portuguese language is a set of rules that often doesn’t take into consideration the reality of linguistic facts. Aiming to find an analytical way that could help us to solve the problem, we studied some more recent works that partially or totally escape from the traditional orientation, such as Moderna Gramática Portuguesa by Bechara (the 1999 edition onwards), Gramática de usos do português by Moura Neves (2000), the Master’s thesis by Maria Eliana Duarte Alves de Brito (1986) and Gramática de Valências by Busse e Vilela (1986). Departing from a functionalist orientation and taking as basis the principle of dependency grammar, the goal of this work is to demonstrate that two poles of analysis must be taken into consideration in the study of phrases: syntax and semantic. Being the verb the central element of a phrase, it’s the verb that determines the number of empty-spaces, besides of the morphosyntatic and semantic properties of the actants that convey these empty-spaces. In this way, the distinction between direct and indirect object of traditional grammar comes up as insufficient, being thus necessary to identify a greater number of types of actant. Besides, we were able to distinguish, through the analysis of the verb as central element, which terms must be considered actants (complements) or circumstantial (adjuncts).

Streptococcus pneumoniae é um importante agente causador de pneumonia, meningite e septicemia. O alto custo e a cobertura limitada da vacina conjugada atual reforçam a necessidade de se desenvolver uma vacina mais abrangente e acessível. A proteína de superfície de pneumococo A (PspA) é imunogênica e protetora em modelos animais; porém, devido à sua diversidade - há 6 clados e 3 famílias de PspA - uma vacina baseada em PspA deverá incluir fragmentos das duas famílias prevalentes (1 e 2). Neste estudo, foram produzidos fragmentos contendo a região N-terminal de PspA das famílias 1 e 2, e proteínas híbridas - contendo fusões destes fragmentos. Os anticorpos gerados contra os híbridos reconheceram PspAs nativas das duas famílias, foram capazes de se ligar a bactérias íntegras, e de aumentar a deposição de complemento em sua superfície. Finalmente, a imunização de camundongos com os híbridos foi capaz de proteger contra desafio com pneumococos contendo PspAs diversas, mostrando que estes seriam candidatos promissores na composição de uma vacina anti-pneumocócica. / Streptococcus pneumoniae is an important cause of pneumonia, meningitis and septicaemia. The high cost and limited coverage of the available conjugate vaccine reinforce the need for cost effective strategies, with broader coverage. Pneumococcal Surface Protein A (PspA) is immunogenic and protective in animal models; however, due to its diversity - there are six clades and threee families of PspA - a PspA based vaccine should include fragments of each major family (1 and 2). In the present study, we have produced fragments of the N-terminal region of PspAs families 1 and 2, and hybrid proteins - containing fusions of these fragments. Sera made against the hybrids induced antibodies that recognized PspAs from both families; these sera were also able to bind pneumococcal strains bearing diverse PspAs, and to increase complement deposition on their surface. Finally, immunization of mice with PspA hybrids was protective against challenge with pneumococci bearing diverse PspAs, showing that these hybrids should constitute promising candidates in an anti-pneumococcal vaccine.

Atypical hemolytic uremic syndrome (aHUS) is a rare renal disorder characterized by thrombotic microangiopathy, thrombocytopenia, and acute kidney injury. Its pathogenesis has been attributed to a ‘triggering' event that leads to dysregulation of the complement cascade at the level of the endothelial cell surface. Consistent with this understanding of the disease, mutations in complement genes have been definitively implicated in aHUS. However, the existence of other genetic contributors is supported by two observations. First, in ~50% of cases, disease-causing variants are not identified in complement genes, and second, disease penetrance is typically incomplete and highly variable. To test this hypothesis, we identified pathways established to have crosstalk with the complement cascade, focusing initially on the coagulation pathway. Using targeted genomic enrichment and massively parallel sequencing we screened 36 European-American patients with sporadic aHUS patients for genetic variants in 85 complement and coagulation genes, identifying deleterious rare variants in several coagulation genes. The most frequently mutated coagulation gene in our study cohort was PLG, which encodes a zymogen of plasmin and plays key role in fibrinolysis. These results implicate the coagulation pathway in the pathogenesis of aHUS. Based on this outcome, we developed a clinical genetic testing panel to screen disease-related genes in a group of ultra-rare complement-mediated diseases that includes, in addition to aHUS, thrombotic thrombocytopenic purpura (TTP), C3 glomerulonephritis (C3GN) and dense deposit disease (DDD) patients. Data from 193 patients validate the usage of this panel in clinical practice and also provide confirmatory insight into the pathogeneses of these diseases. Specifically, we found that in aHUS and TTP patients, variants were frequently identified in complement regulator genes, while in C3GN and DDD patients, variants were additionally found in C3 convertase genes. To understand variability in disease penetrance, we completed targeted genetic screening in two aHUS families grossly discordant for disease penetrance, identifying in one family a co-segregating Factor X-deficiency variant (F10 p.Glu142Lys) that abrogated the effect of the complement mutation. Functional studies of the F10 p.Glu142Lys variant show that it decreases Factor X activity predicting to a hypo-coagulable state and further illustrating the importance of complement-coagulation crosstalk in exacerbating, but also mitigating the aHUS phenotype. In our final studies, we have sought to complete a comprehensive analysis for other potentially related pathways by using bioinformatics to identify candidate pathways coupled with whole exome sequencing. Preliminary data from 43 aHUS patients and 300 controls suggest that pathways for dermatan and heparan sulfate synthesis, which are relevant to the formation of the extra-cellular matrix and cell surface adhesion, may be implicated in the aHUS.

För att elever ska utveckla kunskap om matematiska principer behöver de ha kunskap om tals additiva del-helhetsrelationer och relationen mellan addition och subtraktion. Addition och subtraktion är varandras invers och det är viktigt att eleverna lär sig att addition och subtraktion inte är åtskilda räknesätt. En välstuderad matematisk princip som beskriver den här relationen är inverse principle, vilket skrivs som 𝑎+ 𝑏− 𝑏= 𝑎 eller 𝑎− 𝑏+ 𝑏= 𝑎. Däremot har få studier riktat in sig på den matematiska principen complement principle, vilket innebär sambandet mellan 𝑎+ 𝑏= 𝑐 och 𝑐− 𝑏= 𝑎. Därför är syftet med litteraturstudien att utifrån matematikdidaktisk forskning belysa elevers förståelse för matematiska principer genom addition och subtraktion. Syftet besvaras genom frågorna: Hur kan förståelse för addition och subtraktion underlätta för elever när de utvecklar kunskap om inverse principle och complement principle samt vilken betydelse har konkret material för elevers förståelse för inverse principle och complement principle.  Genom en systematisk informationssökning som utgår från tidigare forskning har internationellt vetenskapligt material samlats in. Litteraturstudien har inriktats mot elever i skolans lägre årskurser. Resultatet visar att förståelse för de matematiska principerna grundas i elevers förståelse för del-helhetsrelationer och hur de förstår relationen mellan addition och subtraktion. Resultatet visar även att begreppet complement principle är komplext men med hjälp av konkret material kan elever lättare att ta till sig och använda principen. Elever verkar förstå de matematiska principerna lättare när de får det beskrivet med konkret material. Det finns en möjlighet att elever utvecklar förståelse för complement principle genom sin förståelse för inverse principle.

<p>Epilepsy is comorbid with cognitive and psychiatric dysfunctions. This pathophysiology, associated with hippocampal synaptodendritic structural and functional changes, is exacerbated by prolonged seizures (status epilepticus; SE). We found a correlation between hippocampal dendritic loss and microgliosis after SE, along with hyperactivation of the classical complement pathway (C1q-C3). These paralleled increased seizure frequency and memory deficits in a rat model of SE and acquired epilepsy. C1q leads to C3 cleavage into biologically active fragments C3a and C3b. Evidence suggests that C1q and C3b contribute to synaptic stripping by microglia in the developing brain and neurodegenerative disorders. Thus, we hypothesized that SE-induced C3 activation may alter hippocampal synaptic protein levels thereby promoting memory deficits. </p> <p>To test the hypothesis, different groups of wild type (WT) or C3 deficient (C3KO) mice were injected with pilocarpine (350mg/kg) to induce SE or saline (controls): WT-C, WT-SE, C3KO-C, and C3KOSE. At two weeks after SE, mice were subjected to novel object recognition (NOR) to evaluate recognition memory, and Barnes maze (BM) to measure hippocampal-dependent spatial learning and memory. Following behavioral testing, mice were sacrificed and hippocampi collected at either 2 or 5 weeks after SE to measure changes in C3 protein levels and levels of synaptic proteins including PSD95, Vglut1, and Vgat. As a method of verifying our findings, we used a second model of pilocarpine-induced SE in male Sprague Dawley rats. Starting at 7 days after SE, rats were treated with cobra venom factor (CVF; 100ng/g, i.p.) or vehicle (veh) every third day. On days 14-15 rats were subjected to open field and NOR to measure anxiety and recognition memory. On day 16, rats were sacrificed and hippocampi collected for western blotting.</p> <p>WT and C3KO mice were able to reach stage 4.5-6 seizures after pilocarpine injections. In NOR trial 1, exploration time for both objects was similar in all groups (<i>p</i> > .05). In trial 2, WT-C and C3KO-C mice spent more time exploring the novel object than the familiar one (<i>p</i> < .05) while WT-SE mice explored both objects equally (<i>p</i> > .05). Interestingly, C3KO-SE mice spent more time with the novel object similar to controls (<i>p</i> > .05), suggesting that the deficit in object recognition memory induced by SE was attenuated in C3KO mice. Similarly, veh- and CVF-treated control rats spent more time exploring the novel object during trial 2 (<i>p</i> < .05). The veh-treated SE rats did not show significant preference for the novel object versus familiar (<i>p</i> > .05), whereas the CVF-treated SE rats explored the novel object significantly more than the familiar (p < .05). These findings support that C3 inhibition after SE prevents recognition memory deficits. Furthermore, there was a reduction in synaptic proteins PSD95 and Vgat in the SE-veh group compared to the C-veh group. This difference was not observed in the C-CVF and SE-CVF groups, suggesting that blocking C3 after SE is neuroprotective against hippocampal synaptic loss.</p> <p>Taken together, these findings are the first to show an association between C3 activation and hippocampal and cognitive deficits in two rodent models of SE and acquired TLE. We found that depletion of C3 is sufficient to attenuate SE-induced deficits in NOR-evaluated recognition memory and changes in the levels of an inhibitory synaptic protein. In conclusion, our data suggest that SE-induced complement C3 activation contributes to hippocampal synaptic remodeling and impairments in recognition memory, and that the complement C3 may be a potential therapeutic target for the memory comorbidities associated with SE. Future studies will determine the effect of C3 inhibition on spontaneous recurrent seizures, and whether C3-guided and microglial-dependent phagocytosis is an underlying mechanism for the SE-induced epileptogenic synaptic remodeling.</p>