Fort d’une croissance annuelle de l’ordre de 6%, le secteur des matériaux composites est actuellement en pleine expansion et se doit de répondre aux exigences d’un marché en constante évolution. Dans le même temps, la raréfaction des ressources pétrolières et l’augmentation de la conscience environnementale, conduisent à une demande croissante en matériaux bio-composites. Le remplacement des fibres synthétiques (fibre de verre en particulier) par des fibres naturelles engendre un intérêt certain dont les motivations principales sont la réduction de l’impact environnemental, la diminution des coûts et l’obtention d’un matériau plus léger à volume égal. Néanmoins, la faible compatibilité existante entre les fibres de cellulose hydrophiles et les matrices polymères hydrophobes, est un des inconvénients majeurs qui nuit au bon développement de ces matériaux. L’objectif de cette thèse est de développer une alternative aux fibres de verre pour l’élaboration de matériaux composites à matrice thermoplastique. Pour ce faire, l’intégration de filaments de cellulose (FC), fournis par Kruger notre partenaire industriel, a été étudiée. En plus d’apporter un côté « vert » au matériau final, les FC permettent de réduire le poids des composites par rapport à leurs homologues synthétiques. Néanmoins, la faible compatibilité entre les filaments polaires et la matrice apolaire ainsi que la grande capacité d’absorption d’eau des FC nous a conduit à développer différentes stratégies de modification chimique des FC, afin d’en accroitre le caractère hydrophobe. Ces modifications ont permis de renforcer les matériaux composites grâce à l’amélioration de l’adhésion entre les FC et la matrice, le tout en minimisant la perte de résistance mécanique causée par l’absorption d’eau. Les résultats obtenus après acétylation, alkylation et encollage sont décrits dans ce manuscrit. / Over the past two decades, the increase of environmental concerns and shortage of petroleum resources have provoked a growing interest in the use of natural fibers as an alternative to synthetic fibers for the reinforcement of composites. Natural fibers possess desirable specific properties including biodegradability, renewability and low-cost. In addition, they have densities much lower than synthetic fibers, which makes them interesting for different applications ranging from automotive parts to packaging. Despite their benefits, certain drawbacks such as incompatibility with the hydrophobic polymer matrix, a tendency to form aggregates during processing and a poor resistance to moisture absorption, reduce the potential of these fibers to be used as a reinforcement of hydrophobic thermoplastic matrices.This thesis aims to substitute glass fibers by cellulose fibers for their use in fiber-reinforced composites. Reinforcement of LDPE composites with cellulose filaments (CF), supplied by our industrial partner Kruger, was studied. CF appear to an interesting alternative to glass fibers because they possess desirable specific properties including biodegradability, low density, high tensile strength and modulus as well as providing a low-cost and renewable option. However, the weak interfacial adhesion between CF and LDPE, and the high moisture absorption of CF led us to carry out several chemical modifications of CF in order to increase their hydrophobicity. Modified CF-composites exhibit higher mechanical properties and lower water uptake than unmodified CF-composites. Results obtained from acetylation, alkylation and paper sizing are described in this manuscript.

Denna studie har genomförts i form av en undersökning med 25 respondenter. Arbetet harsyftat till att fackmän inom belysningsbranschen samt privatpersoner och intressenter ska fåen indikation på hur människan uppfattar ljuset från fasdimrade LED-ljuskällor i privata hem.Studien har studerat två frågeställningar gällande människans upplevelse av flimmer blandannat hur känslig människan är för flimmer hos en ljusreglerad LED-ljuskälla innan detuppfattas som obekvämt. Vetenskaplig litteratur har använts för att få en förståelse och djuparekunskap kring problematiken gällande människans hälsopåverkan vid exponering av flimmer,samt vilka riktlinjer och möjliga tillvägagångssätt som finns för att lösa denna problematik. Föratt ta reda på hur väl människan uppfattar flimmer samt hur upplevelsen av obekvämlighetenpåverkas har en enkät tagits fram till studien. Undersökningen konstaterar att människoruppfattar flimmer i olika mängd, trots frekvenser över 90Hertz (Hz). Genom data från digitalamätningar och inhämtad empiri från arbetets undersökningsstudie kan man konstatera att detinte enbart är frekvensnivån som påverkar människans uppfattning av flimmer, utan ävenljuskällans andel flimmerprocent. Ett samband mellan upplevd obekvämlighet och flimmer kananas, men inte fastslås som ett tydligt resultat. Arbetet avgränsades genom att undersökningenenbart utfördes under en kortare period med ett färre antal respondenter. / This study has been carried out in the form of a survey with 25 respondents. The work has aimedto give professionals and individuals an indication of how people perceive the light from phasedimmedLED light sources in private homes. The study has investigated two issues regardingthe human experience of flicker, and how sensitive human beings are to flicker from a phasedimmedLED light source before it will be perceived as inconvenient. Scientific literature hasbeen reviewed to gain an understanding and deeper knowledge about the impact of flicker onhuman health, and which guidelines and possible approaches there are to solve the problem. Asurvey has been developed to find out if flicker is perceived by humans, and how the perceptionof inconvenience is experienced. The study found out that people perceive flicker differently,despite measured frequencies above 90 Hertz (Hz). Through data from digital measurementsand acquired empirical evidence from the study’s results, it is not only the frequency level thataffects humans perception of flicker, but also the percentage of light emitted from the lightsource. A correlation between perceived inconvenience and flicker can be noted, but notdetermined as a clear result. The work was limited by only investigating a small population ofrespondents over a short period of time.

O objetivo deste trabalho foi estudar o comportamento mecânico de cilindros de compósito polimérico reforçado com fibras. Para isso, foram produzidos cilindros com extremidades abertas reforçados com fibra de basalto e fibra de vidro, utilizando a técnica de enrolamento filamentar (filament winding). Estes cilindros foram submetidos a ensaio hidrostático com carregamento circunferencial, ensaio de ruptura de anel (split disk test) e ensaio de resistência ao cisalhamento interlaminar (ILSS). Uma placa do compósito de fibra de basalto foi produzida por enrolamento filamentar, para caracterização por ensaio de resistência à tração. Todos cilindros submetidos ao ensaio hidrostático apresentaram fratura localizada em uma faixa de altura do cilindro, com extensas delaminações das camadas circunferenciais. Os compósitos epóxi/fibra de basalto superaram ou igualaram os de compósito epóxi/fibra de vidro nas comparações entre resultados dos valores das propriedades mecânicas avaliadas, nas porcentagens: resistência à tração aparente de ruptura de anel em 45% e 43% em resistência específica; ILSS, em 11%; resistência/tensão de membrana de ruptura no ensaio hidrostático, em 55%. / The aim of this work was to study the mechanical behavior of fiber reinforced polymer composite cylinders. For this purpose, cylinders reinforced with basalt and glass fibers were produced, with open-ended geometry, using filament winding technique. These cylinders were submitted to hydrostatic test under circunferential loading, split disk (ring segment) test and interlaminar shear strength (ILSS). A basalt fiber composite plate was produced by filament winding for characterization by tensile strength test. All cylinders submitted to hydrostatic test presented fracture located in the height range of the cylinder, with extensive delamination of the circumferential layers. The epoxy/basalt fiber composites overcame or equated the epoxy/glass fiber composites in comparisons between results of the mechanical properties, tensile strength in split disk, in 45% and 43% in specific strength; ILSS in 11%; membrane tensile strength in the hydrostatic test, in 55%.

Les moteurs moléculaires permettent le transport actif de molécules et d’organelles le long de filaments polaires du cytosquelette de la cellule eucaryote. Les filaments peuvent s’organiser en réseaux parallèles, antiparallèles, ou désordonnés. Malgré son importance, l’influence de l’architecture du cytosquelette sur le transport de cargos par des moteurs moléculaires est souvent ignorée. Dans ce travail de thèse, nous avons utilisé des patrons surfaciques de nucléation pour contrôler la géométrie de polymérisation de filaments d’actine. Cette approche permet de reproduire in vitro les trois types de réseaux qui sont observés in vivo. En adsorbant des moteurs moléculaires purifiés à la surface de nano-billes fluorescentes (diamètre 200 nm), nous avons étudié le transport de ces cargos dans des réseaux antiparallèles émergeant de deux lignes de nucléation parallèles. Nous avons observé que les billes recouvertes de moteurs processifs HMM-V (Heavy Mero-Myosine V) génèrent des mouvements dirigés en direction du milieu du réseau où la polarité moyenne des filaments est nulle et où les billes s’accumulent. De plus, la distribution de positions des billes à l’état stationnaire peut être déduite des profils de vitesse et de diffu-sion des billes, indiquant que le transport actif suit un processus de convection-diffusion. Les billes recouvertes de moteurs non-processifs HMM-II (Heavy Mero-Myosine II) dé-montrent des comportements similaires. Cependant, bien que le gradient de vitesse des billes HMM-II soit plus important que pour les billes HMM-V au centre du réseau, le coeffi-cient de diffusion l’est bien davantage. Le centrage de ces billes est ainsi moins précis qu’avec la HMM V. A notre surprise, nous avons observé que la précision du centrage ne dépend pas de l’espacement entre les deux lignes de nucléation, donc de la taille du sys-tème. Le comportement des billes est décrit par un modèle à trois états dans lequel la bille sonde localement la polarité nette du réseau en se détachant et en se rattachant fré-quemment aux filaments. Une séquence stochastique de déplacements dirigés dans un état attaché aux filaments et d’explorations diffusives dans un état détaché, conduit à une marche aléatoire biaisée avec un coefficient de diffusion et une vitesse effectifs. Notre description physique indique que la précision du positionnement des billes dépend du gra-dient de polarité nette du réseau de filaments ainsi que de la distance moyenne de par-cours des billes dans l’état attaché à l’actine. Nos résultats démontrent ainsi le rôle clef que joue l’architecture du réseau de filaments sur les propriétés du transport. Dans la cellule, les moteurs moléculaires permettent également la réorganisation des filaments du cytosquelette. En ajoutant nos moteurs moléculaires en solution, nous avons observé qu’un réseau de filaments d’actine parallèles, polymérisés in vitro à partir d’une ligne ou d’un disque de nucléation, peut former spontanément des faisceaux oscil-lants. Ces oscillations ressemblent aux battements du flagelle du spermatozoïde. En parti-culier, le système génère des ondes de déformation transverses se propageant de la base à l’apex du faisceau oscillant à une vitesse de 0,5 µm/s. Au cours du temps, les faisceaux d’actine s’épaississent, ce qui conduit à un ralentissement de l’oscillation avec une période qui croît de 25 s à 40 s. De plus, nous avons observé que des faisceaux d’actine voisins sont capables de se synchroniser, comme c’est le cas entre les deux flagelles de l’algue Chlamydomonas. Notre système acto-myosine minimal permet donc d’imiter le battement de flagelles, bien que la nature des moteurs moléculaires et des filaments en jeu soit com-plètement différente. Ainsi, ce système fournit un nouvel outil pour étudier les propriétés physiques génériques du battement flagellaire. / Molecular motors navigate the cytoskeleton to position vesicles and organelles at specific locations in the cell. Cytoskeletal filaments assemble into parallel, antiparallel or disordered networks, providing a complex environment that constrains active transport properties. Using surface micro-patterns of nucleation-promoting factors to control the geometry of actin polymerization, we studied in vitro the interplay between the actin-network architec-ture and cargo transport by small myosin assemblies. With two parallel nucleation lines, we produced an antiparallel network of overlapping filaments. We found that 200nm beads coated with processive myosin V motors displayed directed movements towards the mid-line of the pattern, where the net polarity of the actin network was null, and accumulated there. The bead distribution was dictated by the spatial profiles of bead velocity and diffu-sion coefficient, indicating that a diffusion-drift process was at work. Interestingly, beads coated with skeletal heavy mero-myosin II motors showed a similar behavior. However, although velocity gradients were sharper with myosin II, the much larger bead diffusion observed with this non-processive motor resulted in less precise positioning. Strikingly, bead positioning did not depend on the spacing between the nucleation lines. Our observa-tions are well described by a three-state model of bead transport, in which active beads locally sense the net polarity of the filament network by frequently detaching from and re-attaching to the filaments. A stochastic sequence of processive runs and diffusive search-es results in a biased random walk with an effective drift velocity and diffusion coefficient. Positioning relies on spatial gradients of the net actin polarity, as well as on the run length of the cargo in the attached state. Altogether, our results on a minimal acto-myosin system demonstrate the key role played by the actin-network architecture on motor transport. Molecular motors can also deform and reorganize the cytoskeleton. Adding heavy mero-myosin II or V in bulk, we observed that parallel networks of actin filaments emerg-ing from a nucleation line or disk can self-assemble into bundles that beat periodically like the flagellum of the spermatozoid. In a preliminary analysis, we observed waves of defor-mation travelling from the base to the tip of the bundle at a speed of 0,5 µm/s. As time went by, the bundles grew thicker, resulting in an increase of the beating period (range: 25-40 s). In addition, neighboring actin ‘flagella’ were able to synchronize, as observed in vivo for instance with the the two flagella of the algae Chlamydomonas. Our minimal acto-myosin system thus mimicked key properties of microtubule-based flagellar beating, de-spite the different nature of the motors and cytoskeletal filaments involved. This system thus provides a new tool to study the generic physical properties of flagellar beating.

I det här utvecklas en reproducerbar polymerisationsmetod för att uppnå en stabil produktion av poly(ε-caprolakton-co-p-dioxanon) (PCLDX), skala upp filamenttillverkningen för att producera 1.75 mm långa filament och optimera 3D-utskriftsprocessen för att tillverka ställningar/anordningar för mjukvävnadsteknik. PCLDX, med högre nedbrytningshastighet och bättre flexibilitet jämfört med poly(ε-caprolactone) (PCL), syntetiserades på ett reproducerbart sätt genom sampolymerisering. Den syntetiserade PCLDX uppvisade önskvärd sammansättning (85 mol% CL : 15 mol% DX), molmassa (cirka 40 kg∙mol-1), dispersitet (cirka 1.8) och relativt låg smältpunkt (cirka 45 ℃). För att tillverka tredimensionella matriser av PCLDX utformades och optimerades två processer, filamenttillverkning och 3D printning. För filamenttillverkningsprocessen användes låg extruderingstemperatur (65 och 80 ℃) och låg extruderingshastighet (100 cm∙min-1) för att spara energi och minimera nedbrytningen. PCLDX-filament med en jämn diameter på 1.75 mm tillverkades genom att använda en passande partikelstorlek (diameter på 3-4 mm) och en kylmetod (blandning av vatten och torris, 0 ℃). De erhållna filamenten uppvisade lägre Youngs modul (25 % lägre än PCL), PCLDX batch oberoende termiska egenskaper, god ytkvalitet och printbarhet. Den termiska nedbrytningen av PCLDX under processen var försumbar och molmassan var nästintill oförändrad. Processen har skalats upp för att producera stora mängder PCLDX-filament, vars produktivitet nådde upp till 140 g∙h-1. Tredimensionella matriser tillverkades genom att printa önskad design genom manuell matning och låg printhastighet (5 mm/s). En isplatta användes för att kyla ner maskinen under printningen för att undvika bucklingproblem. Det optimerade printprotokollet genererade ingen termisk nedbrytning av polymeren, påverkade inte polymerens molmassa eller dispersitet. De producerade matriserna hade samma termiska egenskaper oavsett polymerbatch och god ytkvalitet. Det optimerade printprotokollet användes också framgångsrikt för att skriva ut komplicerade prototyper, t.ex. menisk och knäprotes för potentiella biomedicinska tillämpningar. / In this project, we develop a reproducible polymerization method to achieve stable production of poly(ε-caprolactone-co-p-dioxanone) (PCLDX), scale-up the filament fabrication to produce 1.75 mm filaments and optimize 3D printing process to manufacture scaffolds/devices for soft tissue engineering. PCLDX, with a higher degradation rate and better pliability compared to poly(ε-caprolactone) (PCL), was successfully synthesized by reproducible copolymerization of ε-caprolactone (CL) and p-dioxanone (DX). The synthesized PCLDX exhibited a polymer composition of 85 mol% CL : 15 mol% DX, molar mass around 40 kg∙mol-1, dispersity around 1.8, and relatively low melting point around 45 ℃. From PCLDX particles to final scaffolds, two processes, including filament fabrication and scaffold manufacturing, were designed and optimized. For the filament fabrication process, low extrusion temperature (65 and 80 ℃) and low extrusion speed (100 cm∙min-1) were applied to save energy and minimize degradation. PCLDX filaments with an even diameter of 1.75 mm were fabricated using suitable particle sizes (diameter of 3-4 mm) and a cooling method (mixture of water and dry ice, 0℃). The obtained filaments exhibited lower young’s modulus (25% lower than PCL), consistent thermal properties, good surface quality, and printability. The thermal degradation of PCLDX during the process was negligible, and the molar mass was kept almost unchanged. The process has been scaled up to produce high amounts of PCLDX filaments, whose productivity rate reached up to 140 g∙h-1. For the scaffold manufacturing process, porous scaffolds were manufactured by feeding manually and printing slowly (5 mm/s). The printability was assessed and validated using produced PCL/PCLDX filaments and commercial PCL filaments. The optimized printing protocol maintained the molar mass and dispersity of the material. The produced scaffolds possessed consistent thermal properties independent on polymer batches and good surface quality. The optimized printing protocol was also successfully applied to print complicated prototypes, such as meniscus and knee prosthesis for potential biomedical applications.

Denna rapport detaljerar ett examensarbete där användbarheten på en lokalt skapad 3D-skrivare förbättrats i syfte av att tillåta skrivarens användning av studenter på KTH Södertälje. Skrivarens har ursprungligen byggts för att agera som ett pågående projekt som kan återupptas som examensarbete, varav detta är det tredje. Skrivaren i fråga är en FFF-skrivare i ”box” stil med utskriftsvolymen 400 x 400 x 375 mm. Vid projektstart är skrivaren i nominellt användbart skick men saknar dokumentation, processrutiner och användbara slicerprofiler. Skrivaren besitter även hårdvaran för att tillåta utskrifter med två material men denna funktion har aldrig implementerats. Framtagningen av slicerprofiler har tagit upp majoriteten av arbetet och har utförts genom iterativ testning där inställningar modifieras och testas, vartefter positiva ändringar implementeras för att gradvis eliminera defekter och minska haveririsk. Projektet har levererat en full slicerprofil som tillåter pålitliga utskrifter med två olika PLA-filament samt en användarmanual till skrivaren. Utöver användarverktygen så detaljeras även de reparationsarbeten och hårdvaruändringar som gjorts. Profil, manual, rådata och filer för utskrivna komponenter är tillgängliga i en projektmapp som kan hittas från en länk i början av kapitel 4. / This report details a thesis project for getting a 3D-printer internally built at KTH Södertälje into a state where it can be used by other students for unrelated projects. The printer acts as an ongoing project that can be used for thesis work, with this project in particular being the third in line. The printer in question is an FFF-printer with the ”box” configuration and a print volume of 400 x 400 x 375 mm. At project start, the printer is nominally in working condition but lacks any documentation for proper handling and slicer settings. Additionally, its multi-material function has never been successfully implemented. Adjustment of slicer settings is done using iterative testing, wherein settings are modified and tested, with positive changes being integrated and negative changes discarded with the intent of removing defects and minimizing risk of print failure. The project has delivered a slicer profile for Cura capable of reliably printing with two different PLA filaments as well as a user manual. Hardware upgrades and repair work has also been detailed. The profile, user manual, raw data and object files for upgrades and spares can be found through a link in the beginning of chapter 4.

Hypertrophic cardiomyopathy (HCM) is a primary disease of the myocardium. 4-11% of HCM is caused by mutations in cardiac troponin T (cTnT) and 65% of them are within the tropomyosin (TM)-binding TNT1 domain. Two of the known mutational hotspots within TNT1 are in the N and C-terminal domains. Unlike the N-terminal domain; no high-resolution structure exists for the highly conserved C-terminal domain limiting both our ability to understand the functional role of this extended domain in myofilament activation and molecular mechanism(s) of HCM. The Δ160E mutation is an in-frame deletion of a glutamic acid residue at position 160 of cTnT. This TNT1 C-terminal mutation is associated with an especially poor prognosis. The Δ160E mutation is located in a putative "hinge region" immediately adjacent to the unstructured flexible linker connecting the TM-binding TNT1 domain to the Ca²⁺-sensitive TNT2 domain. Unwinding of this α-helical hinge may provide the flexibility necessary for thin filament function. Previous regulated in vitro motility assay (R-IVM) data showed mutation-induced impairment of weak actomyosin binding. Thus, we hypothesized that the Δ160E mutation repositions the flexible linker which impairs weak electrostatic binding and ultimately leads to severe cardiac remodeling. The goal of our studies is two-fold: 1) to gain high-resolution insight into the position of the cTnT linker with respect to the C-terminus of TM, and 2) to identify Δ160E-induced positional changes using Fluorescence Resonance Energy Transfer (FRET) in a fully reconstituted thin filament. To this end, residues in the middle and distal regions of the cTnT linker were sequentially cysteine-substituted (A168C, A177C, A192C and S198C) and labeled with the energy donor IAEDANS. The energy acceptor, DABMI was attached to cysteine 190 (C190) in the C-terminal region of TM and FRET measurements were obtained in the presence and absence of Ca²⁺ and myosin subfragment 1 (S1). An all-atom thin filament model in the Ca²⁺–on state was employed to predict the pathogenic effects of the Δ160E mutation on the structure and the dynamics of the cTnT linker region. Our data suggest that the linker domain runs alongside the C-terminus of TM and is differentially repositioned by calcium, myosin and the Δ160E mutation. The Δ160E mutation moves the linker closer to the C-terminus of TM. The in silico model supported this finding and demonstrated a mutation-induced decrease in linker flexibility. Moreover, the model predicted a pathogenic change in the orientation of the middle region of the linker and in the position of the Ca²⁺-sensitive TNT2 domain and the TM-binding TNT1 domain in response to Δ160E mutation. Collectively, our findings suggest that the Δ160E mutation-induced changes in the structure, position and dynamics of the linker region cause steric blocking of weak myosin binding sites on actin and subsequent impairment of contraction and disruption of sarcomeric integrity. These studies, for the first time, provided information regarding the role of the extended linker in both myofilament activation and disease.

Tropomyosins(Tm) are a group of proteins that regulate the actin filaments in both muscle and non-muscle cells. In mammalian cells four Tm species are found: α-Tm (fast) encoded by α-Tm /TPM1 gene, β-Tm, encoded by β-Tm/ TPM2 gene, α-Tm (slow) encoded by γTm gene/ TPM3 and δ-Tm encoded by δTm / TPM4gene. Mutations in Tm are linked to many cardiac and skeletal diseases like hypertrophic cardiac myopathy (TPM1 and TPM2), familial cardiac myopathy (TPM1) and skeletal diseases like nemaline myopathy (TPM2 and TPM3) along with other sarcomere proteins. The hypothesis on which this study is based is, the isoform composition in both muscle and non-muscle cells adapts in response to disease and physiological changes. A significant part of that adaptation is changes in the thin filament protein isoforms expressed and the post translational modifications of these proteins. In this study Tpm3.12st isoform of γTm and other striated muscle tropomyosin isoforms (Tpm1 and Tpm2) and a non-muscle Tmp4 were characterised using a variety of techniques. The aim was to enhance our understanding of the role of tropomyosin interactions in regards to its efficiency of actin binding capacity as well as its effect on actin polymerisation. Human tropomyosin 3 (Tpm3.12st) was expressed in E. coli to produce recombinant protein with three N-terminal sequence variants (Met, MM and (M)ASM). The proteins were characterised for their binding affinity with actin as this isoform has not been well characterised so far. Its properties are compared with other striated muscle tropomyosin Tpm1.1st and Tpm2.2st and non-muscle Tpm4.1cy. The proteins were purified through ion exchange chromatography and the purity was checked by using SDS-PAGE and UV spectrometry. The molecular weights of the recombinant proteins produced were confirmed by mass spectrometry. Cosedimentation assays were performed for their actin binding affinity using ultracentrifugation. The variant of Tpm3.12st with AS N-terminal extension was found to have similar actin affinity to Tpm1.1st in the range of 0.1-0.8 μM (half saturation). However the variants with Met and MM N-termini bound to actin weakly with high half saturation concentration of ~ 6 μM and ~8 μM tropomyosin respectively. Measurement of actin polymerisation kinetics showed it is affected in presence of tropomyosin. From this study it is shown that tropomyosin accelerates the initiation step in actin polymerisation with varying differences within the isoforms in contrast to several previous studies. There have been very few studies of the effect of tropomyosin on actin polymerisation in the last two decades. This work shows that tropomyosin isoforms have a large and variable role in controlling actin polymerisation and understanding tropomyosin function will need further investigation in this area. This study also developed an ELISA screening method using monoclonal antibodies for identification and quantification of Tpm3.12st which was tested against all the four tropomyosin isoforms. None of the twelve antibodies studied showed reactivity only with Tpm3.12st. From the data analysed it is deduced the amino acid residues in the region of 24-43 shows the prospect of designing a monoclonal antibody specific to Tpm3.12st isoform. Accurate quantification of tropomyosin isoforms is key to understanding their function and the effects of modulation of isoform composition in health and disease. A reverse phase liquid chromatography method was developed which is compatible with the analysis of the thin filament proteome using top-down mass spectrometry. Reverse phase liquid chromatography (RPLC) is one of the most popular methods used in mass spectrometry analysis where proteins are separated based on their hydrophobicity. The RPLC method developed in this study gives an efficient separation of major thin filament proteins along with small soluble proteins that is compatible to use for top down mass spectrometry for identification and quantification of proteins, PTMs and isoform composition. With a minimum amount of 2 mg of tissue using chicken and mouse heart and skeletal muscle samples a buffer system was optimized to extract thin filament proteins. With the optimized RPLC method actin, tropomyosin and troponin complex subunits (TnC, TnI and TnI) were successfully separated and the proteins were identified using SDS-page by comparison with the previous research results. This novel method of extraction and the optimised RPLC method will provide a “bird’s eye view” of thin filament proteome providing information of PTMs of all the proteins together within one single extraction, reducing the time for analysis and the sample size. This has the potential to give insight into tissue, muscle and heart adaptations that could act as a prognostic indicator.

Materiais compósitos são conhecidos pela alta resistência mecânica e baixo peso, desempenho superior, resistência à corrosão e baixa densidade. A produção de um material composto possui vários processos com particularidades diferentes. O enrolamento filamentar (Filament Winding) é um processo no qual fibras de reforço contínuas são posicionadas de maneira precisa e de acordo com um padrão predeterminado para compor a forma do componente desejado. As fibras, submetidas à tração, são enroladas continuamente ao redor de um mandril que tem a forma do produto final, em geral utilizando equipamentos automáticos. No final do processo, após a cura da resina, o mandril é geralmente removido. Desta forma, é de fundamental importância que o projetista disponha de recursos computacionais adequados para o cálculo das trajetórias e sequenciamento de posicionamento das fibras. Esse trabalho tem como objetivo o desenvolvimento de procedimentos matemáticos para cálculo de trajetórias geodésicas no processo de \"Filament Winding\" e implementá-los em um ambiente computacional em linguagem de alto nível Java, considerando-se os casos de revestimento circunferencial, helicoidal e polar. São desenvolvidos dois estudos de caso: tubos cônicos e vasos de pressão, e os resultados apresentados e discutidos, validando os procedimentos e ambiente implementado. / Composite materials are well known by the high strength and low weight, superior performance, resistance to the corrosion and low density. The production of a composite material part involves some processes with different requirements. The filament winding process is an automated process in which continuous reinforcement fibers are lay down in prescribed paths on the surface of a mandrel, which is generally removed after the cure of the resin. In such a way, it is fundamental that the designer uses computational resources for the calculation of the paths and sequence of the fibers. In this work is developed the mathematical procedures for calculation of geodesic trajectories in the Filament Winding process and implements them in a computational environment in high level language Java, considering the circumferential, helical and polar strategies. Two case studies are developed successfully: conical pipes and pressure vessels, and the results presented and discussed, validating the procedures and implemented environment.

Essa dissertação trata da modelagem física de filamentos de fibras visando adequá-las a aplicações em equipamentos de enrolamento filamentar - filament winding. No decorrer do trabalho é abordada a criação do subsistema para condução e posicionamento dos fios que compõem as bobinas - rovings. São feitas análises por elementos finitos e realizados testes de bancada com os componentes mecânicos visando validar o subsistema como parte integrante das máquinas de filament winding. É estudada a forma de impregnação dos fios, a tensão e o grau de espalhamento dos fios no decorrer da sua passagem pelo sistema, o qual é composto por um conjunto de polias, um tracionador e um enrolador desses fios. / This dissertation deals with the use of fiber reinforcements, in order to improve its appliance in filament winding machines. Throughout this work it is created a mechanism system to remodel the fiber filaments from rovings to the part that is been produced. Finite element analyses and experimental tests are performed to validate the system as part of the filament winding machines. Its is studied the impregnation characteristics, the tensioning and the spreading of the fibers along the designed system, which is composed by a set of pulleys and tensioners. The system is built and satisfactory results are obtained. The results obtained as well as proposals to improve the system are presented and the results discussed.