This diploma thesis deals with Formula Student Impact Attenuator design. The aim of the thesis is to select suitable material and put it to the test of deformation. From results to suggest dimensions of Impact Attenuator and then perform its test. Next aim of the thesis is to perform FEM analysis of Anti-Intrusion plate and supporting rack.

Thesis (MScEng)--University of Stellenbosch, 2002. / ENGLISH ABSTRACT: The objective of this study is the assessment of the usefulness of an industrial machine vision system for laboratory studies of especially flotation froths and other foams. With this in mind it was attempted to meet three objectives. The first objective is the adjudication of the machine vision system's ability through image analysis to differentiate between characteristics of two-phase and three-phase systems. Secondly, through the use of image analysis the effect of varying surfactant concentration, as well as the addition of quartz particles on the foam/froth stability was quantified and thirdly it was attempted to define the nature of foam decay as stochastic or deterministic. While not as important in the industrial setting, the last objective is quite important when a laboratory system is concerned, even more so owing to the difficulty of quantifying the collapse or decay of foams and froths. Two experimental set-ups were used to attain above-mentioned objectives. A Leeds flotation cell was used for the flotation of the quartz with cetyltrimethylammonium bromide (CTAB) and octadecylamine (ODA) and as well as for the characterisation of the CTAB and ODA foam. A glass column was used to investigate the CTAB foam column decay. In both set-ups the foam/froth structures were monitored with an ELMO charged coupled device (CCD). The CCD was connected to a personal computer equipped with frame grabber, which captured and digitised the images. The following conclusions can be drawn from the experimental results: The machine vision system can differentiate accurately between surfactant groups of different concentration and thus groups with unique characteristics. 11 The stability of foam is highly correlated with all other foam variables extracted from the image analysis system. In particular, the movement of the foam/froth in the flotation cell has a substantial influence on the stability values obtained for the foam/froth. III As far as the experimental system was concerned, small quartz particles (- 75 urn) stabilised the froth, while coarser particles (75-106 urn, 106-150 urn and 150+ urn) destabilised the froth. IV Likewise, the decay of CTAB foam in a column was non-linear and deterministic. When observed manually, the decay of long life foams follow the same trend as short-life foams and short-life emulsions in that a specific decay constant can be calculated for a specific surfactant system. V The machine vision system is a viable alternative for current stability measurements. Firstly, no data processing need to be done to calculate a indirect stability parameter as it computes a direct stability parameter and secondly it also computes the bubble size distribution of the sample. The following recommendations are made: The experimental design of the system needs to be altered to minimise the effect the experimental environment has on results. The possibility of a different camera set-up must be investigated. The focus of the camera on the foam surface needs to be accurate as the computation of the stability value depends on the quality of the image obtained. Il The four factors that affect the stability of a flotation froth, namely solid concentration, particle size, particle type and particle hydrophobicity should be investigated in the same study. There exists many conflicting studies on this subject as many studies for different particles with a variety of sizes, hydrophobicity and concentration are performed. One study should be conducted where all four these factors are investigated under the same experimental conditions. III The method with which the image analysis system deal with fine froth need to be addressed. In laboratory systems a typical system investigated will show a dense foam with small bubble distributions as in the case of CTAB. The extent to which the grouping of fine froth as one bubble influence the instability values obtained needs to be investigated . / AFRIKAANSE OPSOMMING: Die doelwit van hierdie studie is die evaluering van die toepassing van 'n industriële masjienvisiesisteem in 'n laboratorium studie van hoofsaaklik flottasie skuim. Die doel van hierdie toepassing is drie-ledig. Die eerste doelwit is om die onderskeidingsvermoë van die masjienvisiesisteem tussen groepe met verskillende eienskappe te evalueer deur intydse beeld verwerking. Tweedens is die invloed van oppervlakmiddelkonsentrasie veranderinge asook die byvoeging van kwarts partikels op die stabiliteit van die skuim ondersoek. Derdens is die aard van die skuim verval ondersoek om vas te stelof die proses deterministies of stochasties is. Alhoewel laasgenoemde doelwit nie so belangrik is in die industriële toepassing nie, is dit van uiterse belang in die laboratorium studies, veral as gevolg van die probleme wat ondervind word in die kwantifisering van skuimverval. Twee eksperimentele opstellings is gebruik om bogenoemde doelwitte te bereik. 'n Leeds flottasiesel is gebruik vir die flottasie van kwarts met CTAB en ODA asook vir die karakterisering van CTAB en ODA skuimeienskappe. 'n Glaskolom is gebruik vir die ondersoek van die aard van skuimverval. In beide opstellings is die beelde gemonitor deur 'n ELMO CCD. Die CCD is verbind met 'n rekenaar wat toegerus is met 'n raam-vanger wat die beelde vang en digitiseer. Die volgende gevolgtrekkings kan uit die eksperimentele resultate gemaak word: Die masjienvisiesisteem differensieer akkuraat tussen oppervlakmiddel groepe van verskillende konsentrasies en dus ook tussen groepe met unieke eienskappe. 11 Die skuim stabiliteit korreleer met die ander skuim eienskappe naamlik gemiddelde area, gemiddelde omtrek, aantal borrels, fynheid van die beeld (SNE), beweging van die skuim en kleur van die skuim. Veral die beweging van die skuim in die flottasiesel het 'n beduidende invloed op die skuimstabiliteit. 111 In die genoemde eksperimentele sisteem IS gevind dat klein kwartspartikels (-75 urn) skuim stabiliseer terwyl groter partikels (75-106 urn, 106-150 urn en 150+ urn) skuim destabiliseer. IV Die verval van CTAB skuim in 'n kolom is nie-lineêr en deterministies. In die geval waar die verval van die CTAB met die hand gemeet is, is gevind dat die verval van 'n lang-lewe skuim dieselfde tendens as kort-lewe skuim en kort-lewe emulsies volg. 'n Spesifieke verval konstante kan bereken word vir 'n spesifieke oppervlakmiddel sisteem. v Die masjienvisiesisteem IS 'n werkbare alternatief in die meting van skuimstabiliteit. Anders as in bestaande stabiliteitsmetings waar 'n indirekte stabiliteitsparameter bereken moet word, bereken die sisteem 'n direkte stabiliteitsparameter asook die borrelverspreiding van die skuim monster. Die volgende aanbevelings kan gemaak word: Die eksperimentele ontwerp van die sisteem moet aangepas word om sodoende die invloed van eksperimentele kondisies op die resultate te verminder. Die moontlikheid van 'n alternatiewe kamera opstelling moet ook ondersoek word, aangesien die verkrygde stabiliteitswaardes afhanklik is van die kwaliteit van die skuimbeelde. 11 Daar is talle teenstrydige studies ten opsigte van die vier faktore wat die stabiliteit van flotasie skuim beinvloed naamlik erts tipe, partikel grootte, partikel hidrofobisiteit en vastestof konsentrasie. Hierdie faktore moet gelyktydig in 'n studie ondersoek word onder dieselfde eksperimentele kondisies. 111 Die masjienvisiesisteem groepeer fyn skuim gedeeltes saam as een borrel. Hierdie wyse van fyn skuim hantering moet aangespreek word aangesien 'n tipiese oppervlakmiddel sisteem in die laboratorium, soos in die geval van CTAB, digte skuim met 'n fyn borrelverspreiding toon. Die mate waarin die onvermoë van die sisteem om fyn borrels te herken die stabiliteitswaardes beïnvloed, moet ondersoek word.

Atherosclerosis is a disease characterized by cholesterol-rich plaques within the intima of medium and large arteries. Cholesterol deposition is thought to occur by infiltration of low-density lipoprotein (LDL) into lesions followed by uptake into macrophages, generating lipid-loaded “foam cells.” Foam cells can also be generated in vitro by treatment of macrophages with LDL or oxidized LDL (oxLDL). The purpose of the current investigation was to determine the contribution of selective cholesteryl ester (CE) uptake versus whole-particle uptake during LDL-induced foam cell formation in cultured macrophages. Murine bone marrow-derived macrophages (BMMs) exhibited significant cholesterol accumulation when treated with LDL as indicated by quantification of cellular cholesterol and visualization of Oil Red-O-stained neutral lipid droplets. Uptake of LDL cholesterol was determined by measuring uptake of 3H and 125I into BMMs during treatment with [3H]CE/125I-LDL. [3H]CE uptake was linearly related to the LDL concentration at the concentrations used and was much larger than 125I uptake, indicating that the majority of LDL-cholesterol was acquired by nonsaturable, selective CE uptake. This pathway was demonstrated to be independent of whole-particle uptake by showing that inhibition of actin polymerization blocked LDL particle uptake but not selective CE uptake. Analysis by thin-layer chromatography (TLC) indicated that following uptake, [3H]CE was rapidly hydrolyzed into [3H]cholesterol by cells and largely effluxed into the culture medium. In contrast to LDL, studies of [3H]CE/125I-oxLDL uptake demonstrated that CE was acquired from oxLDL by whole-particle uptake with little or no selective CE uptake. Using a series of ten different [3H]CE/125I-oxLDLs oxidized for 0-24 hours, selective [3H]CE uptake was shown to be progressively impaired by LDL oxidation, while 125I-LDL particle uptake was increased as expected. Interestingly, the impairment of selective CE uptake occurred very early in LDL oxidation and this minimally oxidized LDL induced significantly less cholesterol accumulation in BMMs compared to native LDL. Together, these results demonstrate that selective CE uptake is the primary mode of cholesterol uptake from LDL but not oxidized LDL, a finding that has important implications for cholesterol metabolism in atherosclerotic lesions. Future studies seek to identify the molecular components that participate in the macrophage selective CE uptake mechanism.

This research investigates the decomposition and burning behaviours of polyurethane foams experimentally and compares the experimental results obtained with the numerical results from the pyrolysis model of Fire Dynamics Simulator, Version 5 (FDS 5). Based on the comparison of model and experimental heat release rates, the accuracy of the pyrolysis model is quantified. In total, this research tested seven polyurethane foams consisting of three non-fire retardant (NFR) and four fire retardant (FR) foams. According to the simultaneous differential scanning calorimetry and thermogravimetric analysis (SDT) experiments, the decomposition behaviour of polyurethane foams under nitrogen environment is represented by two pyrolysis reactions. The first reaction consists of foam decomposition into melts and gases while the second reaction consists of the decomposition of the remaining melts into gases. The kinetic properties which govern the rate of decomposition are the activation energy (E), pre-exponential factor (A), reaction order (n) and heat of reaction (Δhr). Using graphical techniques, E, A and n of the first and second reactions are determined from the thermogravimetric analysis (TGA) results. Through analysing the differential scanning calorimetry (DSC) results, Δhr is determined from the changes in heat flow and sample mass. The thermophysical properties govern the heat transfer through material and these are the thermal conductivity (λ) and specific heat (cp) which are measured experimentally at ambient temperature through the Hot Disk method. Through the Sample Feeding Vertical Cone, the decomposition and melting behaviours of polyurethane foams in a vertical orientation are investigated and the foams tested can be categorised into those which produce melts only after ignition and those which produce melts and char after ignition. The 1-dimensional burning behaviour of foams is obtained from the cone calorimeter experiments. The NFR foams show a change from plateau burning behaviour at low heat flux to two stage burning behaviour at high heat flux while the FR foams consistently show two stage burning behaviour. The combustion property governs the amount of heat released when fuel combusts and this is the effective heat of combustion (Δhc,eff) which is determined from the heat released and mass consumed in the cone experiment. The 1-dimensional burning behaviour is simulated using the pyrolysis model of FDS 5 and two different modelling approaches are considered. The direct method uses the material properties determined experimentally as FDS 5 inputs while the refined method uses the genetic algorithm of Gpyro to refine the kinetic properties which are later used as FDS 5 inputs. The heat release rate of the model and experiment are compared through linear regression analysis which quantifies the accuracy of both methods. The accuracy is defined as the percentage of data points within the boundary of acceptance which is bounded by 25 % of the greatest experimental heat release rate. This assessment method places greater emphasis on the accuracy of developed burning phases and lesser emphasis on the accuracy of initial growth and final decay. The accuracy of the direct method is found to be 56 % while the refined method with estimated kinetic properties achieves a higher accuracy of 75 %. The 2-dimensional burning behaviours are investigated in the foam slab experiments for two different slab thicknesses, 120 and 100 mm. The opposed-flow spread of 120 mm slab is more intense and rapid while for the 100 mm slab, the flame spread is less intense and slow. FDS 5 is used to simulate the experimental results but when the material properties either developed experimentally or refined by Gpyro are used as inputs, the model fails to produce flame spread. This is because FDS 5 does not yet have the features which address the dynamics of foam melting and the reactive nature of the flame. In order to produce flame spread in the model, E of the reactions have been reduced to increase the decomposition rate.

Le moussage du polypropylène (PP) et du polystyrène (PS) par le scCO2 est étudié de manière systématique avec l’accent sur la relation entre le procédé, la microstructure et les propriétés mécaniques. La première partie de cette thèse porte sur l’amélioration de la résistance au choc du PP par cristallisation induite par le scCO2. Dans un deuxième temps, le PP fortement orienté avec des structures shish-kebab et sphérolite est utilisé pour étudier l’influence de la structure cristalline sur la nucléation et la croissance des cellules pendant le procédé de moussage sous le scCO2. Ensuite, les influences des conditions de moussage ainsi que celles des paramètres structuraux de la mousse de PS ont été étudiées. Les mousses de PS à cellules isotropes ont été comparées avec celles de PS à cellules orientées. Enfin, un procédé de moussage discontinu en deux étapes est développé afin de produire des mousses de PS à cellules bi-modales en utilisant le scCO2 en tant qu’agent de moussage. Cette structure unique de petites et grandes cellules réparties de manière homogène au travers de l’ensemble du volume de la mousse confère à cette dernière des propriétés particulières. / In this work, scCO2 induced foaming of PP or PS is systematically studied with emphasis on the relationship between process, microstructure and mechanical properties. The first part of the thesis deals with the toughening of iPP by scCO2 induced crystallization for the fine separation of rigid crystalline domains and soft amorphous ones in the polymer matrix. The highly oriented iPP with “shish-kebab” and “spherulite” are then used for CO2 foaming to investigate the effect of crystalline structure on the formation of cell nucleation and growth. In addition, the effect of the foaming conditions and the cell structural parameters of PS foams on the mechanical properties are studied systematically. PS foams with isotropic cell morphology and oriented cell one are prepared. Finally, a two-step depressurization batch process is developed to produce bi-modal cell structure PS foams by using scCO2 as the blowing agent. This unique cell structure with both small and large cells homogenously distributed throughout the entire volume of the foam sample might have particular properties.

La flottation ionique est un procédé de séparation permettant d'extraire et de concentrer des ions dans une mousse formée à l'aide d'un tensioactif. Pour les systèmes classiques, la forte interaction entre les ions et le tensioactif collecteur entraîne généralement la formation de précipités. Une fois la mousse collapsée, le résidu solide récupéré nécessite un traitement particulier pour permettre sa valorisation ou son conditionnement. Afin de remédier à cet inconvénient, ce travail propose d'utiliser comme collecteur un tensioactif carboxylique polyéthoxylé, l'AKYPO® RO 90 VG, ayant la particularité de former des complexes ion-tensioactifs solubles, et ce également avec des ions multichargés. Ce travail présente une étude poussée des mécanismes fondamentaux qui régissent l'extraction d'ions par formation de mousses. Dans une première partie, les propriétés tensioactives et acide base du collecteur en solution sont étudiées en combinant nombre de techniques indépendantes que sont le dosage pHmétrique, la tensiométrie et la diffusion aux petits angles. L'évolution de ces propriétés en présence de différents sels de nitrates (Nd, Eu, Ca, Sr, Cu, Li, Na, Cs) couplée à des mesures électrophorétiques donneront une première approche de la sélectivité. Enfin, l'ensemble de ces données associées à une étude de la formation des complexes tensioactif/ion permettra d'établir la spéciation du système Nd/AKYPO® en fonction du pH. Dans une seconde partie, l'analyse de la conductimétrie et de la diffusion de neutron aux petits angles permettront d'établir les paramètres régissant la formation et stabilité des mousses de flottation. Le pH et la nature des ions ajoutés, leur nombre de charge ainsi que leur nature chimique, apparaissent ainsi comme les paramètres majeurs gérant l'humidité et l'épaisseur de film des mousses. La dernière partie est dédiée à la compréhension des expériences d'extraction/séparation ionique par flottation en s'appuyant sur l'ensemble des résultats obtenus précédemment. Il est montré que la flottation du néodyme est fortement liée à sa spéciation, pouvant conduire à sa dés-extraction ou sa flottation sous forme particulaire. Il est également montré que le néodyme induit un phénomène de déplétion des ions monovalents dans la mousse. Cette spécificité ionique permet d'envisager le système étudié pour la séparation d'ions par le procédé de flottation. / Ion foam flotation allows to concentrate ions in a foam phase formed by a soap. For classical systems, the strong interaction between ions and surfactant generally leads to the formation of precipitates and of froth. When the froth collapses, the solid residue thus recovered requires a recycling or conversion. In order to remedy this, the present work uses as collector a polyethoxylated carboxylic surfactant, AKYPO® RO 90 VG, which forms soluble ion/surfactant complexes, even with multi-charge ions. This work presents a detailed study of the fundamental mechanisms that govern the extraction of ions by foaming. In the first part, surface activity and acid/base properties of the surfactant in solution are determined by combining numerous independant techniques which are pHmetric dosage, tensiometry and small angle scattering. The evolution of these properties in the presence of different nitrate salts (Nd, Eu, Ca, Sr, Cu, Li, Na, Cs) coupled with electrophoretic measurements give a first approach to selectivity. Finally, all of these data combined with a study of the formation of surfactant/ion complexes allow us to determine the speciation of Nd/AKYPO® system as a function of pH. In the second part, the analysis of the foam by conductivity and neutron scattering provides information on the wetness and foam film thickness, parameters governing foam stability. The pH and the nature of the added ions, their number of charge and also their chemical nature thus appear to be major parameters that governed wetness and foam film thickness. The last part is devoted to the understanding of the ion extraction/separation experiments by flotation based on all previous results. It is shown that the flotation of neodymium is strongly related to its speciation, which could lead to its re-extraction or its flotation in precipitated form. It is shown that, neodymium induces a phenomenon of mono-charge ion depletion in the foam. This ionic specificity allows to consider the studied system for ion separation by the flotation process.

O objetivo deste projeto é verificar a possibilidade da substituição dos produtos PVPP (Polivinilpolipirolidona) e APG (Alginato de Propileno Glicol), estabilizantes empregados na fabricação de cervejas, pela goma Oenogum, analisando parâmetros físico-químicos, composição química e atributos sensoriais da bebida. O PVPP é utilizado como estabilizante coloidal da cerveja, porém é de alto custo. O APG é estabilizante da espuma de cervejas, principalmente aquelas com baixo teor de proteínas, aquelas que recebem na sua fabricação adjuntos não maltados. Em vinhos, o Oenogum diminui adstringência, inibe o turvamento e previne o depósito de compostos polifenólicos, contribuindo para a produção de vinhos de qualidade superior. O Oenogum cria uma película ao redor dos taninos e dos polifenóis, deixando-os inativos, sem, no entanto, afetar os compostos voláteis do vinho e, assim sendo, não afetando o equilíbrio organoléptico da bebida. Será produzida cerveja elaborada com 55% de malte e 45% de maltose de milho em pó (cerveja com malte e adjuntos não maltados), com a adição de 0, 5 ou 10 g/hL da goma Oenogum e a cerveja denominada como padrão comercial com PVPP e APG em substituição à goma Oenogum para efeito de comparação. As cervejas serão analisadas quanto a parâmetros físico-químicos (extrato, fermentabilidade, pH, cor, amargor, estabilidade de espuma e turbidez), composição química (acidez, etanol, ésteres, cetonas, aldeídos, álcoois superiores, metanol e polifenóis) e atributos sensoriais (impressão global, cor, odor e sabor) da bebida. Espera-se que a goma Oenogum possa promover estabilização coloidal e de espuma na cerveja produzida, sem, no entanto, alterar negativamente a composição química e parâmetros físico-químicos e sensoriais da bebida. / The objective of this project is to verify the possibility of replacing the products PVPP (Polivinilpolipirolidona) and APG (Propylene glycol alginate), stabilizers used in making beer, by gum Oenogum, analyzing physical and chemical parameters, chemical composition and sensory attributes of the drink. The PVPP is used as a colloidal stabilizer in beer, but it is expensive. The APG is the foam stabilizer of beers, especially those with low protein content, those who receive in its manufacture Assistant unmalted. In wine, the astringency Oenogum decreases, inhibits and prevents blurring the deposit of polyphenolic compounds, contributing to the production of superior quality wines. The Oenogum creates a film around the tannins and polyphenols, making them inactive, without, however, affect the volatile compounds of wine and, therefore, not affecting the organoleptic balance of the drink. Will be produced beer made with 55% malt and 45% maltose corn powder (beer with malt and adjuncts unmalted), with the addition of 0, 5 or 10 g / hL Oenogum gum and beer named as standard commercial PVPP and APG to replace gum Oenogum for comparison. The beers will be analyzed for physico-chemical parameters (extract, fermentability, pH, color, bitterness, foam stability and turbidity), chemical (acidity, ethanol, esters, ketones, aldehydes, higher alcohols, methanol and polyphenols) and attributes sensory (overall impression, color, odor and taste) of drinking. It is expected that the gum can promote Oenogum and colloidal stabilization of beer foam produced without, however, adversely affected the chemical composition and physico-chemical and sensory characteristics of the drink.

Les cellules myéloïdes jouent un rôle majeur dans le développement et les complications de l’athérosclérose. TREM (Triggering Receptor Expressed on Myeloid cells)-1 est un récepteur, exprimé par les cellules myéloïdes, impliqué dans l’amplification de la réponse inflammatoire en réponse à une stimulation de la voie des TLRs. Notre objectif était d’étudier les conséquences de l’inhibition de TREM-1 sur le développement de l’athérosclérose expérimentale.Matériel et résultats : Dans un modèle de souris chimères Ldlr-/- retransplantées avec une moelle de souris Trem-1+/+ ou Trem-1-/-, nous avons montré que la déficience hématopoïétique en Trem-1 induisait une réduction de 42% de la taille des plaques d’athérosclérose au niveau du sinus aortique après 6 semaines de régime gras et une réduction de 60% après 14 semaines. De plus, la déficience hématopoïétique en Trem-1 induisait un phénotype lésionnel plus stable avec une moindre accumulation macrophagique, un centre nécrotique moins volumineux. Ces résultats ont été confirmés dans un modèle de souris ApoE-/-/Trem1-/-. Parallèlement, l’inhibition pharmacologique de TREM-1 par des injections répétées de peptide LR12 induisait une réduction significative de la taille des plaques au niveau du sinus et de l’aorte thoracique chez la souris ApoE-/- sous un régime riche en matières grasses. En utilisant différentes approches in vitro et in vivo, nous avons montré que l’invalidation génique de Trem-1 ou son inhibition pharmacologique induisait une déviation de la réponse immune vers un profil moins inflammatoire, réduisait le recrutement des monocytes non classiques dans la plaque d’athérosclérose et enfin limitait l’endocytose des lipides oxydés en régulant l’expression du CD36. Ces données de modulation de la réponse immune et de l’endocytose des lipides ont été confirmées sur des macrophages humains. Nous avons enfin montré que TREM-1 est exprimé dans des plaques d’athérosclérose humaines essentiellement par les macrophages spumeux. L’expression de TREM-1 est significativement plus importante dans les plaques athéromateuses par rapport aux plaques fibreuses. Conclusion : Nous avons montré que TREM-1 est impliqué dans le développement de l’athérosclérose à différents stades de la maladie, en modulant la réponse immune systémique et l’endocytose des lipides oxydés. L’inhibition de TREM-1 pourrait constituer une nouvelle cible thérapeutique des maladies cardiovasculaires. / Innate immune responses activated through myeloid cells contribute to the initiation, progression and complications of atherosclerosis in experimental models. However, the critical upstream pathways that link innate immune activation to foam cell formation are still poorly identified.Objectives: We hypothesized that activation of TREM (Triggering Receptor Expressed on Myeloid cells)-1 plays a determinant role in macrophage atherogenic responses. Methods and Results: Ldlr-/- mice reconstituted with bone marrow deficient for Trem-1 (Trem-1-/-) showed a strong reduction of atherosclerotic plaque size in both the aortic sinus and the thoraco-abdominal aorta, and were less inflammatory compared to plaques of Trem-1+/+ chimeric mice. Genetic invalidation of Trem-1 led to alteration of monocyte recruitment into atherosclerotic lesions and inhibited Tlr4-initiated pro-inflammatory macrophage responses. Furthermore, we identified a critical role for Trem-1 in the upregulation of Cd36, thereby promoting the formation of inflammatory foam cells. Genetic invalidation of Trem-1 in Apoe-/-/Trem-1-/- mice or pharmacological blockade of Trem-1 in Apoe-/- mice using LR-12 peptide also significantly reduced the development of atherosclerosis throughout the vascular tree, and lessened plaque inflammation. TREM-1 was expressed in human atherosclerotic lesions mainly in lipid-rich areas, with significantly higher levels of expression in atheromatous compared to fibrous plaques. Conclusion: We identify TREM-1 as a major upstream pro-atherogenic receptor. We propose that TREM-1 activation orchestrates monocyte/macrophage pro-inflammatory responses and foam cell formation through coordinated and combined activation of CD36 and TLR4. Blockade of TREM-1 signaling may constitute an attractive novel and double-hit approach for the treatment of atherosclerosis.

Os produtos de modificação de lipoproteínas de baixa densidade (LDL) como a subfração eletronegativa [LDL(-)] desempenham um importante papel na progressão da aterosclerose. O acúmulo massivo de LDL modificada captada por macrófagos resulta em células espumosas que liberam mediadores inflamatórios e contribuem para a aterogênese. O scFv (single chain fragment variable) é um fragmento de anticorpo recombinante que contém o sítio completo de ligação ao antígeno. Diante do papel da LDL(-) na aterogênese e da necessidade de novas intervenções terapêuticas que possam inibir o acúmulo de lipídeos em macrófagos, este trabalho objetivou a expressão do scFv anti-LDL(-) 2C7 em Pichia pastoris, bem como a avaliação do efeito deste fragmento de anticorpo sobre a formação de células espumosas em cultura de macrófagos RAW 264.7. O vetor inicial de expressão pPIgLE apresentava como estratégia de detecção e purificação o fusionamento com a proteína A. No entanto, a alta imunogenicidade da proteína A inviabilizaria o estudo da proteína de fusão em cultura de macrófagos, o que determinou a substituição da estratégia de purificação anterior pela cromatografia com resina de níquel através da inserção de hexahistidina na região C-terminal da proteína. A análise de sequenciamento confirmou a presença da inserção e das regiões determinantes de complementariedade. O cassete de expressão com hexahistidina foi inserido no vetor pPIgLE de P. pastoris e transformado na linhagem SMD1168 (Invitrogen®). Testes preliminares de expressão em pequena escala permitiram a análise entre sete clones diferentes, demonstrando uma banda correspondente ao peso molecular de 28 KDa em SDS-PAGE, confirmado por Western Blot. A separação do scFv 2C7 através de resina de níquel obteve uma proteína pura, conforme foi analisado em SDS-PAGE corado com prata. A afinidade do scFv 2C7 a 9 LDL(-) foi confirmada por Dot Blot. O ensaio de captação de LDL(-) demonstrou que o scFv 2C7 foi eficaz na redução de células espumosas e este efeito foi acompanhado pela diminuição na expressão gênica de CD36, TLR-4 e COX-2. Baseado nestes dados, o scFv 2C7 demonstra uma propriedade importante para uma futura intervenção terapêutica para a aterosclerose. / The modification products of low-density lipoprotein (LDL), as the electronegative subfraction [LDL(-)], play an important role in the progression of atherosclerosis. The massive accumulation of modified LDL uptake by macrophages results in foam cells that release inflammatory mediators and contribute to atherogenesis. The scFv (singlechain fragment variable) is a recombinant antibody fragment that contains the complete site antigen-binding. Considering the role of LDL(-) in atherogenesis and the need for new therapeutic interventions that may inhibit the accumulation of lipids in macrophages, this study aimed the expression of anti-LDL(-) 2C7 scFv in Pichia pastoris and the evaluation of the effect of this recombinant antibody fragment on foam cells formation in cultured RAW 264.7 macrophages. The pPIgLE expression initial vector presented as a strategy for detection and purification the fusion with protein A. However, the high immunogenicity of the protein impairs the study of the fusion protein in cultured macrophages, leading to the replacement of the previous strategy of purification by chromatography with nickel resin by inserting hexahistidine tag at the C-terminus of the protein. The sequence analysis confirmed the presence of insertion and the complementary determining regions. The expression cassete with hexahistidine was inserted into the pPIgLE vector of P. pastoris and transformed in the SMD1168 strain (Invitrogen®). Preliminary tests of expression in small-scale allowed the analysis of seven different clones, showing a band corresponding to the molecular weight of 28KDa on SDS-PAGE, confirmed by Western Blot. The separation of 2C7 scFv by the nickel resin yield a pure protein, as it was shown by SDS-PAGE stained with silver. The affinity of 2C7 scFv was confirmed by Dot Blot. The assay of LDL(-) uptake showed that the 2C7 scFv was effective in reducing foam cells and this effect was determined by the decrease in gene expression of CD36, TLR-4 and COX-2. Based on these data, the 2C7 scFv demonstrates an important property for future therapeutic intervention for atherosclerosis

[pt] As dispersões bifásicas (ex. espumas e emulsões), de grande relevância prática na química e na engenharia, são termodinamicamente instáveis devido à diminuição de energia livre no processo de coalescência. Esta limitação é comumente minimizada através de mecanismos de estabilização eletrostática e estérica. Recentemente, há um interesse crescente na estabilização de emulsões e espumas usando partículas sólidas (tipo Pickering), devido à forte adsorção das mesmas nas interfaces para formar uma película sólida. Dependendo do tamanho e do ângulo de contato, a adsorção das partículas pode tornar-se irreversível, melhorando a estabilidade em comparação com sistemas estabilizados por surfactante. Como nanopartículas de molhabilidade intermédia são relativamente raras, o ajuste deste parâmetro é o principal desafio para obter dispersões estáveis. No presente estudo, nanopartículas de sílica (SiNPs) foram modificadas superficialmente com 3-aminopropil trimetoxisilano e hexadeciltrimetoxisilano como agentes sililantes, para se obter três tipos diferentes de nanopartículas híbridas de maior hidrofobicidade, visando a formulação de dispersões bifásicas altamente estáveis usando um método de baixa energia (agitação manual). As SiNPs modificadas foram caracterizadas através de testes qualitativos, infravermelho, analise termogravimétrica, analise elementar e ressonância magnética nuclear de carbono treze e de silício vinte nove no estado sólido para confirmar o enxerto dos grupos funcionais. As propriedades das NPs em relação à sua dispersabilidade e comportamento em interfaces foram avaliadas por espalhamento de luz dinâmica e a tensão superficial crítica de molhabilidade. Finalmente, foram preparadas espumas líquidas e emulsões água-em-óleo (A/O) utilizando as partículas híbridas obtidas e pequenas quantidades de surfactantes, para facilitar o processo de dispersão. Os resultados mostraram que as NPs mais hidrofílicas estabilizam melhor a interface ar/água (espumas), enquanto a estabilização máxima em emulsões é obtida com as NPs mais hidrofóbicas, uma vez que a interação com a fase contínua é otimizada. Além disso, a presença de grupos funcionais na superfície das SiNPs oferece a possibilidade de obter efeitos de sinergia com os surfactantes, controlando a adsorção mista na interface. As aplicações potenciais destes resultados podem abranger diversas áreas, tais como biomedicina, ciência dos materiais, recuperação de petróleo, cosméticos e alimentos. / [en] Biphasic dispersions (e.g. foams and emulsions) are of practical relevance to many chemical and engineering fields. These dispersions are thermodynamically unstable, since their decay results in a decrease of the free energy. Different mechanisms can prevent the coalescence of the dispersion, such as electrostatic and steric stabilization. Recently, there has been an increased interest in the study of stabilization of emulsions and foams using solid particles (Pickering type), since particles can strongly adsorb at interfaces to form a solid film, replacing the incompatible water-oil(ar) interface by the more favorable oil(ar)-particle and particlewater interfaces. Depending on their size and contact angle of selective wetting, particle adsorption can become irreversible, with increased stability of particle-stabilized dispersions compared to surfactant-stabilized systems. Since particles of intermediate wettability by nature are relatively rare, tailoring particle wettability is the major challenge to obtain stable dispersions. In the present study, hybrid silica nanoparticles (SiNPs) were prepared by modification of their surface using 3 aminopropyltrimethoxysilane and hexadecyltrimethoxysilane as silanizing agents, to obtain three different nanoparticles with increased hydrophobicity, aiming to achieve highly stable biphasic dispersions using a low-energy method. The modified SiNPs were characterized using qualitative tests, FTIR, TGA, CHN and NMR (Si) to confirm grafting of functional groups. In addition, nanoparticle properties regarding aqueous dispersions and behavior at interfaces were evaluated by DLS and critical wetting surface tension. Liquid foams and water-in-oil (W/O) emulsions were prepared using the obtained hybrid particles and small amounts of different surfactants, to facilitate the dispersion process. The results showed that while more hydrophilic NPs stabilize better the air/water interface (foams), maximum stabilization in W/O emulsions is achieved with the most hydrophobic NPs, since interaction with continuous phase is optimized. In addition, the presence of different functional groups at the surface of the SiNPs offers the possibility for increased synergy with surfactants, controlling the mixed adsorption at the interface. The potential applications of these results can cover diverse fields such as biomedicine, materials science, oil recovery, cosmetics and food.