Soils represent an essential habitat for a wide diversity of invertebrates. Among these organisms, termites are considered ecosystem engineers for their impact on nutrient cycling and soil functioning, stemming from their wide feeding habits. For instance, soil/litter-feeding termites are one of the groups that incorporate the most organic matter from the soil. Thus, the high abundance and diversity of this group of termites does not only indicate its ecological success, but also its high value for the ecosystem. TheApicotermitinae subfamily (family Termitidae) is a highly diverse group of soil-feeding termites widespread in Afro- and Neotropical regions. However, due to taxonomic difficulties in soldierless groups and poor sampling of species living deep in the soil, it is also one of the most understudied subfamilies. In this thesis, I addressed phylogenetic, anatomical, and ecological aspects of the Apicotermitinae, with the ultimate goal of explaining their high diversity and ecological success. In the first axis, we investigated the phylogenetic relationships of Apicotermitinae species by using de novo mitochondrial genomes. African taxa with soldiers form several basal branches. We confirmed the monophyly of Asian and neotropical lineages, resulting from two independent dispersal events from tropical Africa, and established the relationships of the main lineages in the subfamily. The relationships among and within some genera remain unresolved, probably revealing an explosive radiation. Some genera appear as polyphyletic, showing the need for further taxonomic revision. In the second axis, we described for the first time in detail the anatomical structures of the gizzard. Thepulvillar belt bears a highly diverse, sclerotized and autofluorescent structures. These structures and the ornamentation patterns remain limited to African species since the Neotropical species do not present such specialized structures. Consequently, these ornamentations are proposed as a new complementary taxonomic tool, which can prove useful in the future revisions of genera with phylogenetic incongruences. In the third axis, we characterized the content of crop-gizzard and inferred the isotopic niche of species. Slight variations in the food content suggested that Apicotermitinae can be considered as a single feeding group, with mineralized soil as a primary source. Variations in the crop-gizzard volume can be related to food-collecting behavior. The neotropical species exhibited the broadest isotopic spaces, indicating flexibility to explore large organic matter humification gradients. The broad overlap of isotopic niches and co-occurrence with other termites suggests that this group could be affected by spatial segregation. Finally, in an evolutionary context, it was inferred that the Neotropical soldierless taxa underwent an explosive radiation during the early-middle Miocene. While the reduction of sclerotized structures in the gizzard is associated with the dispersal towards the Neotropics, the African soldierless species developed a pulvillar armature. All of these results provide an overview of the understanding of Apicotermitinae and open up new perspectives on the evolutionary and functional aspects of associations in favor of diet as an agent of their success. / Les sols représentent un habitat essentiel pour une grande diversité d'invertébrés. Parmi ceux-ci, les termitessont considérés comme des ingénieurs de l'écosystème, pour leur impact sur le cycle des nutriments ainsique sur le fonctionnement du sol. Ce rôle écologique majeur est du fait entre autres de la grande diversitéde leur régime alimentaire. Les termites qui se nourrissent de sol et/ou de litière constituent l'un des groupesqui incorpore le plus de matière organique depuis le sol. Ainsi, l'abondance et la diversité élevées de cegroupe de termites indiquent non seulement leur succès écologique, mais aussi leur forte valeur pour lesécosystèmes. La sous-famille des Apicotermitinae (Termitidae) constitue un groupe de termites humivorestrès abondant et diversifié dans les régions afrotropicale et néotropicale. Cette sous-famille est pourtant peuétudiée, du fait des difficultés taxonomiques rencontrées pour les termites sans soldats, mais aussi desdifficultés d’échantillonnage associées aux espèces propres aux sols plus profonds. Le présent travail dethèse se veut être une étude de la diversité des Apicotermitinae selon trois axes, à savoir phylogénétique,anatomique et écologique, dans le but d'expliquer les tenants et aboutissants de la grande diversité desApicotermitinae, ainsi que les raisons de leur succès écologique.Dans le premier axe de la thèse, nous avons étudié les relations phylogénétiques entres les différentesespèces d’Apicotermitinae à l’aide de génome mitochondriaux assemblé de novo. Les taxa africains avecsoldats forment plusieurs branches basales. Nous avons confirmé la monophylie des lignées asiatique etnéotropicale, qui résultent de deux colonisations indépendantes au départ de l'Afrique, et établi les relationsentre les principales lignées de la sous-famille. Les relations, entre ou au sein, de certains genres sont malrésolues et correspondraient à une explosion radiative. Plusieurs genres apparaissent commepolyphylétiques, montrant le besoin d’une révision taxonomique future.Dans la deuxième partie de la thèse, nous avons décrit pour la première fois en détail les différentesstructures anatomiques du gésier. La ceinture pulvillaire porte des structures très diverses, sclérifiées etauto-fluorescentes. Ces ornementations sont proposées comme un nouvel outil taxonomiquecomplémentaire, qui serait utile pour les révisions requises dans les genres problématiques susmentionnés.Ces structures et les motifs d'ornementation restent toutefois limités aux espèces africaines, les espècesnéotropicales ne présentant pas de structures spécialisées.Concernant le troisième axe de la thèse, nous avons caractérisé le contenu du jabot-gésier et estimé la nicheisotopique de différentes espèces. Les légères variations du contenu suggèrent que les Apicotermitinaepartagent sensiblement le même régime alimentaire, en ingérant une quantité importante de sol minéralisé.Les variations du volume du jabot-gésier peuvent être liées au comportement de collecte des aliments. Lesespèces néotropicales montrent une étendue des niches isotopiques plus large, ce qui indique la possibilitéd'exploiter plusieurs niveaux d’humification de la matière organique. Le large chevauchement des nichesisotopiques et la co-occurrence avec d'autres termites suggèrent que les espèces de ce groupe tendraient àse ségréger spatialement.Enfin, dans un contexte évolutif, il a été estimé que le groupe des espèces néotropicales sans soldats a subiune explosion radiative au début du Miocène moyen. Alors que la réduction des structures fortementsclérifiées dans le gésier est associée à la dispersion vers les Néo-tropiques, les espèces africaines sanssoldats ont développé une armature pulvillaire. L'ensemble de ces résultats donne un aperçu de lacompréhension des Apicotermitinae et ouvre de nouvelles perspectives quant à des aspects évolutifs etfonctionnels des associations en faveur du régime alimentaire en tant qu'agent de leur succès. / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

Previously, we described prostate cancer (PCa) detection (83% sensitivity; 67% specificity) in seminal plasma by CE-MS/MS. Moreover, advanced disease was distinguished from organ-confined tumors with 80% sensitivity and 82% specificity. The discovered biomarkers were naturally occurring fragments of larger seminal proteins, predominantly semenogelin 1 and 2, representing endpoints of the ejaculate liquefaction. Here we identified proteases putatively involved in PCa specific protein cleavage, and examined gene expression and tissue protein levels, jointly with cell localization in normal prostate (nP), benign prostate hyperplasia (BPH), seminal vesicles and PCa using qPCR, Western blotting and confocal laser scanning microscopy. We found differential gene expression of chymase (CMA1), matrix metalloproteinases (MMP3, MMP7), and upregulation of MMP14 and tissue inhibitors (TIMP1 and TIMP2) in BPH. In contrast tissue protein levels of MMP14 were downregulated in PCa. MMP3/TIMP1 and MMP7/TIMP1 ratios were decreased in BPH. In seminal vesicles, we found low-level expression of most proteases and, interestingly, we also detected TIMP1 and low levels of TIMP2. We conclude that MMP3 and MMP7 activity is different in PCa compared to BPH due to fine regulation by their inhibitor TIMP1. Our findings support the concept of seminal plasma biomarkers as non-invasive tool for PCa detection and risk stratification.

Poultry litter ash (PLA) is a co-product from manure-to-energy systems that originated in response to increased poultry litter (PL) volumes generated in concentrated poultry production regions. Investigating PLA as a crop fertilizer is an alternative solution to balancing poultry and crop regional nutrient cycling in the Commonwealth of Virginia. As the expanding world population places pressure on the poultry industry to meet consumption demands, increased PL production presents an obstacle to identify alternative uses for increased volumes. Currently, Virginia produces 44 million broilers with PL produced predominately in the Shenandoah Valley and Eastern Shore. Likewise, a growing world population places pressure on crop production areas and subsequently finite natural resources used for crop fertilization. Poultry litter ash is an alternative phosphorus (P) and potassium (K) source enhancing transportation logistics, repurposing PL nutrients, and offers dual purpose as a fertilizer and an energy source when compared to PL. Three PLA products [(fluidized bed bulk (FB Bulk), fluidized bed fly (FB Fly), and combustion Mix (CMix)], two manufactured co-products [(granulated poultry litter ash (GPLA), and ash coated urea (ACU)] were evaluated as P, K, and N sources for corn (Zea Mays L.) production in comparison to industry fertilizers [(PL, triple superphosphate (TSP), muriate of potash (KCL), and urea). A comprehensive examination of elemental composition, P speciation, P and K solubility, improved functionality into granulized forms, and field testing were conducted to discern PLA potential as an alternative fertilizer source. Poultry litter ash products were evaluated by total elemental analysis, backscatter-electron dispersive (BSED) microscopy, and X-ray absorption near edge structure (XANES) spectroscopy. Poultry litter ash elemental concentrations were highly variable ranging from 50.6 to 102.0 g P kg -1 and 62.6 to 120.0 g K kg -1 and were comparatively higher than PL concentrations. Phosphorus structures that provided and controlled P solubility were Ca and Ca-Mg-phosphate compounds. Spectroscopy confirmed Ca structures as predominately monetite (dicalcium phosphate anhydrous; CaHPO4; log K ̊ 0.30) and brushite (dicalcium phosphate dihydrate; CaHPO4.2H20; 0.63 log K ̊ ) species that were supported by BSED and elemental stoichiometric ratios (Ca:P; 1.12 to 1.71:1). Additionally, GPLA acidified from FB Fly had higher brushite and monetite percentages described by spectra models, translating into a more soluble Ca-phosphate species when compared to FB Fly original P species. Granulated poultry litter acidulation trials successfully identified a desired granulation point of 29% (14.5 g acid to 50 g PLA) phosphoric acid (75% H3PO4) acidulation. Acidulation dose response relationships created simple linear regression (SLR) equations that sufficiently (R2 > 0.80) described changes in total measurable P and water soluble P, pH, and exothermic reaction temperatures to increasing H3PO4 acidulation. Solubility tests included: sequential extraction, particle size effect on solubility, carbon effect on water soluble P, and Mehlich-1 extraction of PLA sources that confirmed decreased P solubility. A majority PLA P was found in bound plant unavailable fractions (87.7 to 97.7% P of total P). Granulated poultry litter ash had improved P plant available P of 36.0% P of total P. Carbon (C) effects on PLA P were examined by ashing PLA samples in a muffle furnace at 550 ̊C. Differences in total carbon content negatively impacted FB Bulk and CMix total P (1.30 and 4.56 g P kg -1); however, muffle furnace temperatures increased FB Fly total P by 6.74 g P kg -1. All fertilizer products were investigated under field conditions in separate P, K and N corn studies across Virginia coastal plain soils to determine fertilizer effects on corn plant parameters [(most mature leaf (V6), corn ear leaf (R1), and grain (R6)]. Poultry litter P treatments, averaged over rate, recorded highest yield in both years. At eight of nine field sites, FB Bulk resulted in numerically or significantly higher Mehlich-1 concentrations than other P sources post-harvest. Although Mehlich-1 P increased, yield and plant parameters did not; which leads to the conclusion that PLA sources increased soil residual P that did not translate into immediate plant availability recorded within a growing season. Across plant efficacy parameters examined, PLA K is a comparable nutrient source and improved plant parameters when compared to control. Eighteen out of twenty-one plant parameters examined found similar ACU and urea effects on N concentrations. Therefore, ACU is a comparable N source to urea. When compared to industry fertilizer sources, we concluded that PLA is a slowly available P source, decreased P availability negatively affected early plant growth, K is a comparable nutrient source and improved plant parameters compared to control, and ACU effectively provided N to maintain sufficient corn growth. In conclusion, PLA co-products serve as a densified nutrient source that may provide plant available nutrients if processed to aid in nutrient distribution to grain producing areas. / Doctor of Philosophy / Poultry litter ash (PLA) is a co-product from manure-to-energy systems that originated in response to increased poultry litter (PL) volumes generated in concentrated poultry production regions. Investigating PLA as an alternative crop fertilizer is essential to balancing poultry and crop regional nutrient cycling in the Commonwealth of Virginia. As the expanding world population places pressure on the poultry industry to meet consumption demands, heightened PL production presents an obstacle to identify alternative uses for increased volumes. Currently, Virginia produces 44,683,904 broilers with PL produced predominately in the Shenandoah Valley and Eastern Shore. Likewise, a growing world population places pressure on crop production areas and subsequently finite natural resources used for fertilization vital to maintaining crop yields. Poultry litter ash, a co-product from manure-to-energy systems, is an alternative phosphorus (P) and potassium (K) source enhancing transportation logistics, repurposing PL nutrients, and offers dual purpose as a fertilizer and an energy source when compared to PL. In this dissertation, three PLA products [(fluidized bed bulk (FB Bulk), fluidized bed fly (FB Fly), and combustion Mix (CMix)], two manufactured co-products [(granulated poultry litter ash (GPLA), and ash coated urea (ACU)] were evaluated as P, K, and N source for corn (Zea Mays L.) production in comparison to industry fertilizers (PL, triple superphosphate (TSP), muriate of potash (KCL), and urea). Each of the following chapters provides a comprehensive examination of the following topics: elemental composition, P speciation, P and K solubility, improved functionality into granulized forms, and field testing designed to provide parameters to conclude PLA potential as an alternative P, K and N source. In the second chapter, PLA products were evaluated by total elemental analysis, backscatter-electron dispersive (BSED) microscopy, and X-ray absorption near edge structure (XANES) spectroscopy. Poultry litter ash elemental concentrations are highly variable and are comparatively higher than PL concentrations. Phosphorus structure and species identified Ca as the primary element controlling P structure and subsequent solubility. The third component of this dissertation is granulation trials investigating phosphoric acid effects on granulizing and increasing total and water soluble P. Our results identified 29% (14.5 g acid to 50 g PLA) phosphoric acid acidulation for desired granule size. The third dissertation component examines PLA solubility. The results demonstrated PLA decreased P water solubility when compared to industry fertilizer sources. Granulated poultry litter ash demonstrated improved P plant availability due to the granulation process. The final and fourth dissertation components investigated PLA sources under field conditions in separate P, K and N corn studies across Virginia coastal plain soils to determine fertilizer effects on corn plant parameters. Minority of plant parameters tested revealed P control yielded numerically higher P concentrations than PLA P sources tested. Poultry litter P treatments, averaged over rate, recorded highest yield in both years. At eight of nine field sites, FB Bulk resulted in numerically or significantly higher Mehlich-1 concentrations than other P sources post-harvest. Although Mehlich-1 P concentrations increased, yield and plant parameters did not; which leads to the conclusion that PLA sources increased soil residual P that did not translate into immediate plant availability recorded within a growing season. Across plant efficacy parameters examined, PLA K is a comparable nutrient source and improved plant parameters when compared to controls. The majority of plant parameters examined found similar ACU and urea effects on N concentrations. Therefore, ACU is a comparable N source to urea. When compared to industry fertilizer sources, field results concluded that PLA is a slowly available P source, decreased P availability negatively affected early plant growth, K is a comparable nutrient source and improve plant parameters compared to control, ACU effectively provides N to maintain sufficient corn growth. In conclusion, PLA co-products serve as a densified nutrient source that may provide plant available nutrients if processed to aid in nutrient distribution to grain producing areas.

Denna rapport behandlar genomförandet av ett examensarbete på högskolenivå inom maskinteknik vid Högskolan i Borås. Examensarbetet har utförts hos SP Sveriges Tekniska Forskningsinstitut AB, enheterna SP Safety – Mechanical Research i Borås och Göteborg samt Food and Bioscience i Göteborg. Den största delen av arbetet har utförts vid sektionen Mechanical Research Göteborg. Målet med examensarbetet var att kombinera analys av experimentell bilddata från konfokalmikroskopi och mekanisk lastdata från en dragcell som gradvis deformerar ett polyuretanskum med modellering av skummets mekaniska egenskaper med hjälp av finita elementmetoder (FEM). Syftet var att bestämma elasticitetsmodul och Poissons tal. En viktig del av projektet var också att säkerställa hög repeterbarhet och möjliggöra vidareutveckling av metodiken genom att skapa rutiner för hur de olika delmomenten i arbetscykeln bäst utförs. Polyuretanskum, liksom många andra mjuka heterogena biomaterial saknar i dagsläget uppmätta eller beräknade mekaniska egenskaper. Därför finns potential för att den framtagna metodiken kommer till användning för att bestämma materialparametrar och analysera beteenden för fler av dessa material. Genom att bestämma materialparametrarna är det sedan möjligt att¬ utföra hållfasthetsberäkningar på sådana material, och korrelera materialparametrarna till processparametrarna vid tillverkningen för att optimera materialets egenskaper. Studien började med att ett prov av polyuretanskum placerades i en dragcell där det utsattes för en kraft så att det gradvis deformerades. Med hjälp av ett konfokalmikroskop kan hela deformationsprocessen följas i hög upplösning. De framtagna bildserierna analyserades sedan med hjälp av DaVis, en mjukvara som genomför så kallad digital image correlation-analys, med vars hjälp lokala förskjutningar kunde bestämmas. För att kunna utföra FEM-beräkningar delades materialstrukturen in i elementnät med hjälp av den fritt tillgängliga programvaran OOF2. Elementnät och förskjutningsdata importerades sedan till Matlab och insticksmodulen CalFEM. Med hjälp av CalFEM konstruerades en materialmodell med elasticitetsmodul och Poissons tal som inparametrar. Valideringskriterium användes för att säkerställa korrektheten i finita elementanalyserna. Elasticitetsmodulen bestämdes till 4.6 MPa och Poissons tal till 0.33 ± 0.06. Med tillgängliga data kunde inte modellen användas för att uppskatta båda parametrarna samtidigt. Poissons tal bestämdes genom manuell analys av bildserierna. Metodiken kan förbättras och vidareutvecklas genom att analysera fler provbitar för att ta hänsyn till lokala fluktuationer i materialstrukturen, samt avbilda provet i tredimensioner. Tredimensionell avbildning skulle också möjliggöra konstruktion av en tredimensionell beräkningsmodell av materialet. / This bachelor thesis deals with the implementation of a degree in mechanical engineering at the University of Borås. The thesis work has been conducted at SP Technical Research Institute of Sweden AB at the departments SP Safety – Mechanical Research in Borås and Gothenburg and Food and Bioscience in Gothenburg. The major part of the work has been done at the Mechanical Research department in Gothenburg. The aim of the thesis work was to combine analysis of experimental image data from confocal laser scanning microscopy and mechanical load data from a tensile cell that gradually deforms a polyurethane foam with modelling of the mechanical properties of the foam using finite element methods (FEM). The purpose was to determine Young’s modulus and Poisson's ratio. A crucial part of the project was also to facilitate a high degree of repeatability and further development of the method through establishing routines and best practices for how to implement different parts of the method. There is currently a lack of measured or calculated properties for polyurethane foams, as is the case also for many other soft heterogeneous biomaterials. This implies that the developed method has potential use for determining material parameters and analyzing behavior also for other materials of this type. Determining the material parameters facilitates strength calculations on these materials and makes it possible to correlate material parameters to process parameters during manufacturing to optimize material performance. The polyurethane foam was placed in a tensile cell, exposed to a force and slowly, gradually deformed. Using a confocal microscope, the entire deformation process can be observed at high resolution. The obtained image series were then analyzed using DaVis, a software that can perform so called digital image correlation analysis where local displacements could be determined. In order to perform the finite element calculations, the material structure was divided into an element mesh using the software OOF2. The element mesh and displacement data were then imported to Matlab and the plugin module CalFEM. Using CalFEM, a material model involving Young’s modulus and Poisson’s ratio was created. Young’s modulus was determined to be 4.6 MPa and Poisson’s ratio 0.33 ± 0.06. Using the available data, the model was insufficient to determine both parameters simultaneously. Therefore, Poisson’s ratio was determined through manual analysis of the image series. The method can be improved and further developed mainly by analyzing several samples to account for local fluctuations in the material structure and by using three-dimensional imaging methods. The latter would also open up for creating a three-dimensional model of the material.

L’objectif de cette étude a été de formuler et caractériser des émulsions simples huile/eau d’intérêt pharmaceutique stabilisées par de la β-lactoglobuline (β-lg), de la gomme arabique (GA), de la gomme xanthane (GX) et des mélanges β-lg:GA et β-lg:GX. Les concentrations massiques totales des dispersions de biopolymères étaient de 1 % et ont été augmentées à 2,5 % si les émulsions formulées n’étaient pas stables. Le mélange β-lg:GA a été réalisé à pH 4,2 afin de permettre la formation de complexes par interactions électrostatiques attractives entre la β-lg et la GA. Deux ratios β-lg:GA ont été étudiés : 2:1 et 1:2. Enfin, le mélange β-lg:GX a été effectué à pH 7, où les deux biopolymères étant chargés négativement ne se complexent pas et à un ratio de 1:1. Une étude de stabilité des émulsions a été menée sur 6 mois. Les stabilités obtenues ont pu être classées par ordre croissant : GA 2,5 % < β-lg:GA 2,5 % < β-lg 2,5 % < GX 1 % = β-lg:GX 1 %. Plusieurs mécanismes de stabilisation ont été mis en évidence grâce à l’étude des propriétés interfaciales des biopolymères, à l’étude des propriétés rhéologiques des émulsions et à des observations au microscope confocal à balayage laser des émulsions après marquage des biopolymères à la fluorescence. La β-lg et la GA sont toutes deux capables de s’adsorber à l’interface des globules huileux alors que la GX augmente la viscosité de la phase continue. L’association β-lg:GA conduit à la formation d’une double couche interfaciale stabilisante. Enfin, l’association β-lg:GX combine les mécanismes de stabilisation de la protéine, par adsorption interfaciale et de la gomme, par augmentation de la viscosité de la phase continue. / The main objective of this study was to formulate and characterize oil-in-water simple emulsions of pharmaceutical interest stabilized by β-lactoglobulin (β-lg), gum arabic (GA), xanthan gum (XG), and mixtures of β-lg:GA and β-lg:XG. The total biopolymer final concentration in the dispersions was 1 (w/w) % and could be raised to 2.5 (w/w) % if the formulated emulsions were not stable. β-lg:GA mixing was performed at pH 4.2 to allow attractive electrostatic interactions between the two biopolymers and thus the formation of complexes. Two protein:polysaccharide ratios were investigated: 2:1 and 1:2. Conversely, β8lg:XG mixing was performed at pH 7, where both biopolymers are negatively charged, in order to avoid the complex formation, and with a 1:1 ratio. A stability study was conducted for emulsions over a 6-month period. The obtained stabilities could be classified increasingly: GA 2.5 % < β-lg:GA 2.5 % < β-lg 2.5 % < XG 1 % = β-lg:XG 1 %. Several stabilization mechanisms were evidenced by the study of the biopolymer interfacial properties, the study of emulsion rheology and by confocal laser scanning microscopy observations with labeled fluorescent biopolymers. β-lg and GA were both able to adsorb at the interface of oil globule. XG enhanced the continuous phase viscosity. β-lg:GA mixing led to the formation of a stabilizing interfacial double layer. Finally, β-lg:XG association combined the stabilization mechanisms of both biopolymers, respectively: interfacial adsorption and enhancement of the continuous phase viscosity.

In dieser Doktorarbeit wird die Hypothese geprüft, ob Osteozyten einen direkten Einfluss auf die Knocheneigenschaften in ihrer unmittelbaren Umgebung haben. Der zentrale Experimentieransatz ist dabei die Korrelation der Organisation des Osteozytennetzwerks mit den Mineraleigenschaften des Knochens auf der Submikrometerebene. Es wird gezeigt, dass bereits die anfängliche Ausrichtung der Osteoblasten entscheidend für die Synthese von hoch ausgerichtetem Knochenmaterial ist. Die dabei entstehenden Osteozytennetzwerke sind so organisiert, dass die Osteozyten und ihre Zellfortsätze jeweils einen möglichst kleinen Abstand zum Knochenmineral haben. Deshalb wird vermutet, dass genau diese Netzwerkorganisation mitentscheidend ist, wie gut die Zellen das Mineral in ih-rer Umgebung beeinflussen können. Messungen der Knochenmineraleigenschaften auf Submikrometerebene mit Röntgenkleinwinkelstreuung bestätigen diese Vermutung. Dabei wird deutlich, dass Knochenmaterial in der Nähe der Osteozyten durch andere Mineraleigenschaften geprägt ist. Um zu klären, wie Osteozyten Mineral in ihrer direkten Umgebung verändern können, werden Mechanismen der passiven Mineralherauslösung aus der mineralisierten Oberfläche des Osteozytennetzwerks untersucht. Es wird gezeigt, dass kalziumarme ionische Lösungen unter physiologischen Bedingungen große Mengen von Kalzium-Ionen aus dem Knochen lösen und diese dann durch die Osteozytennetzwerkstrukturen diffundieren können. Zum Abschluss wurde medullärer Knochen von Hühnern als ein Modellsystem für rasanten Knochenumbau untersucht. Dieser spezielle Knochentyp dient den Hennen als labiles Kalziumreservoir und ermöglicht dadurch die tägliche Eierschalenproduktion. Experimente am medullären Knochen-material zeigen insbesondere die Bedeutung von weniger stabilen Mineralstrukturen die benötigt werden um den Knochen an den schnellen, sich wiederholenden Knochenauf- sowie Abbau optimal anzupassen. / This thesis aims to test the hypothesis whether osteocytes have a direct influence on bone material properties in their vicinity. In this regard, the concomitant ana-lysis of osteocyte network organization and bone ultrastructural properties on the submicron level is the central approach to answer this question. In this work, it is shown that already initial cell-cell alignment during the process of bone formation is crucial for the synthesis of highly organized bone. Furthermore it is proposed that the occurrence of highly ordered osteocyte networks visualized with confocal laser scanning microscopy (CLSM) has a strong impact on the ability of osteocytes to directly influence bone material properties. These highly organized networks are another consequence of initial cell-cell alignment and are found to be arranged such as to feature short mineral cell distances. Examination of sub-micron mineral properties with scanning small angle x-ray scattering (sSAXS) shows that bone material in the direct vicinity of osteocytes and their cell proc-esses shows different mineral properties compared to bone further away in the depth of the tissue. Moreover, mechanisms of passive mineral extraction from the mineralized surface of the osteocyte network, due to the treatment with calcium poor ionic solutions, are investigated. It is shown that this chemical process occurring under physiological conditions leads not only to the dissolution of considerable amounts of calcium, but also to efficient diffusion of these ions through the osteocyte network structures. Finally, medullary bone which is intended as a labile calcium source for daily egg shell formation in hens is used as a model system for rapid bone turnover rates. This bone type in particular indicates the importance of uniquely adapted, less stable mineral structures to fit the requirements for rapid bone resorption as well as reformation.

Introduction : La chronicité de la rhinosinusite, sa résistance aux antibiotiques, et ses exacerbations aiguës laissent croire que les biofilms sont impliqués dans la rhinosinusite chronique. Objectifs : Nous avons évalué la capacité des bactéries Pseudomonas aeruginosa, staphylocoques à coagulase négative et Staphylococcus aureus à former des biofilms par un essai in vitro, et si cette capacité de formation a un lien avec l’évolution de la maladie. Nous avons évalué in vitro l’effet de la moxifloxacine, un antibiotique utilisé dans le traitement de la rhinosinusite chronique sur des biofilms matures de Staphylococcus aureus. Méthodes : Trent et une souches bactériennes ont été isolées de 19 patients atteints de rhinosinusite chronique et qui ont subit au moins une chirurgie endoscopique des sinus. L’évolution de la maladie a été notée comme "bonne" ou "mauvaise" selon l’évaluation du clinicien. La production de biofilm a été évaluée grâce à la coloration au crystal violet. Nous avons évalué la viabilité du biofilm après traitement avec la moxifloxacine. Ces résultats ont été confirmés en microscopie confocale à balayage laser et par la coloration au LIVE/DEAD BacLight. Résultat et Conclusion : Vingt deux des 31 souches ont produit un biofilm. La production d’un biofilm plus importante chez Pseudomonas aeruginosa et Staphylococcus aureus était associée à une mauvaise évolution. Ceci suggère un rôle du biofilm dans la pathogenèse de la rhinosinusite chronique. Le traitement avec la moxifloxacine, à une concentration de 1000X la concentration minimale inhibitrice réduit le nombre des bactéries viables de 2 à 2.5 log. Ces concentrations (100 µg/ml - 200 µg/ml) sont faciles à atteindre dans des solutions topiques. Les résultats de notre étude suggèrent que l’utilisation de concentrations supérieure à la concentration minimale inhibitrice sous forme topique peut ouvrir des voies de recherche sur de nouveaux traitements qui peuvent être bénéfiques pour les patients atteints de forme sévère de rhinosinusite chronique surtout après une chirurgie endoscopique des sinus. / Introduction: The role of biofilms in chronic diseases is increasingly recognized. Chronic rhinosinusitis, with its chronic indolent course, resistance to antibiotics, and acute exacerbations, has an evolution that parallels that of other biofilm-related diseases. Objectives: 1-To develop an in vitro method to assess the biofilm formation capacity. 2- To determine whether biofilm-forming capacity of bacteria demonstrated in chronic rhinosinusitis has an impact on persistence of the disease following endoscopic sinus surgery. 3- To determine the in vitro activity of moxifloxacin against Staphyylococcus aureus in biofilm form. Method: Thirty-one bacterial strains recovered from 19 patients with chronic rhinosinusitis at least one year post-endoscopic sinus surgery. Evolution of disease was assessed by questionnaire and endoscopy as favorable or unfavorable. The bacteria were cultured on a 96-well culture plaque and a semi-quantitative method using crystal violet to quantify biofilm production was used. Confirmation of the effect of the antimicrobial agents on viability was performed with confocal laser microscopy, using a LIVE/DEAD BacLight staining. Results: Twenty-two of 31 samples produced a biofilm thicker or equal to the positive control. Biofilm formation was associated with a poor evolution for Pseudomonas aeruginosa and Staphylococcus aureus, but not for coagulase-negative staphylococci. Biofilm treated with moxifloxacin at 1000X (0.1mg/ml – 0.2 mg/ml) gave a 2 to 2.5 log reduction in number of viable bacteria. Conclusion: We have shown that Crystal violet method is able to detect biofilm formation. There is a correlation between in vitro biofilm production by Pseudomonas aeruginosa and Staphylococcus aureus and unfavorable evolution after endoscopic sinus surgery, suggesting a role for biofilm in chronic rhinosinusitis. Increased concentrations of moxifloxacin, easily attainable in topical solutions have a potential role in the management of biofilm infections.

A conventional membrane bioreactor (MBR) and two moving bed bioreactors coupled with ultrafiltration membrane filtration were operated for close to six months to investigate biological nutrient removal and potential fouling inducing parameter mitigation. Unique to one of the moving bed membrane bioreactors (MBMBR) was a newly designed media that incorporated a hydrodynamic exterior carrier with a highly porous interior packing. Preliminary investigation indicates that nitrogen compounds were superiorly removed in the two MBMBRs when compared with the MBR. This is a result of denitrification processes occurring in anoxic micro-zones found within the depths of the biofilm affixed to media. Fouling propensity was found to be increased by over four times in the MBMBR systems as compared to the MBR. Mixed liquor, permeate and filtrate analysis, membrane fibre examination and permeability tests indicated that colloidal organic carbon, as well as soluble microbial products were the dominant fouling inducing compounds. / Manuscript format / The Natural Sciences and Engineering Research Council of Canada

Sphaerodoridae is a group of benthic marine worms (Annelida) characterized by the presence of spherical tubercles covering their whole surface. They are commonly considered as belonging to Phyllodocida although sistergroup relationships are still far from being understood. Primary homology assessments of their morphological features are lacking, hindering the appraisal of evolutionary relationships between taxa. Therefore, our detailed morphological investigation focuses on different Sphaerodoridae as well as on other members of Phyllodocida using an integrative approach combining scanning electron microscopy (SEM) as well as immunohistochemistry with standard neuronal (anti-5-HT) and muscular (phalloidin-rhodamine) markers and subsequent CLSM analysis of whole mounts and sections. Furthermore, we provide histological (HES) and light microscopical data to shed light on the structures and hypothetical function of sphaerodorid key morphological features. We provide fundamental details into the sphaerodorid morphology supporting a Phyllodocida ancestry of these enigmatic worms. However, the muscular arrangement and the presence of an axial muscular pharynx is similar to conditions observed in other members of the Errantia too. Furthermore, nervous system and muscle staining as well as SEM and histological observations of different types of tubercles indicate a homology of the so called microtubercles, present in the long-bodied sphaerodorids, to the dorsal cirri of other Errantia. The macrotubercles seem to represent a sphaerodorid autapomorphy based on our investigations. Therefore, our results allow comparisons concerning morphological patterns between Sphaerodoridae and other Phyllodocida and constitute a starting point for further comparative investigations to reveal the evolution of the remarkable Sphaerodoridae.

Non-metallic inclusions in steels are generally considered to be detrimental for mechanical properties. However, it has been recognized that certain inclusions, such as Ti-oxide and TiN, can serve as potent nucleation sites for the formation of intragranular ferrite (IGF) in low-alloy steels. The formation of IGF could improve the toughness of the coarse grained heat affected zone (CGHAZ) of weld metals. Thus, the present thesis mainly focuses on the effect of size of nucleation sites on the IGF formation. Quantitative studies on the composition, size distribution and nucleation probability for each size of the inclusions as well as the area fraction, starting temperature and morphology of an IGF have been carried out. In the present work, the Ti-oxide and TiN powders were mixed with metallic powders. The mixed powders were heated up to the liquid state and cooled with a slow cooling rate of 3.6 ºC/min. These as-cast steels with Ti-oxide and TiN additions were used to simulate the IGF formation in the CGHAZ of weld metals. Specifically, the inclusion and microstructure characteristics in as-cast steels have been investigated. The results show that the nucleant inclusion was identified as a TiOx+MnS phase in steels with Ti2O3 additions and as a TiN+Mn-Al-Si-Ti-O+MnS phase in steels with TiN additions. In addition, the TiOx and TiN phases are detected to be the effective nucleation sites for IGF formation. It is clearly shown that an increased inclusion size leads to an increased probability of IGF nucleation. This probability of IGF nucleation for each inclusion size of the TiOx+MnS inclusions is clearly higher than that of the complex TiN+Mn-Al-Si-Ti-O+MnS inclusions. In addition, the area fraction of IGF in the steels with Ti2O3 additions is larger than that of the steels with TiN additions. This result agrees with the predicted tendency of the probability of IGF nucleation for each inclusion size in the steels with Ti2O3 and TiN additions. In order to predict the effective inclusion size for IGF formation, the critical diameters of the TiO, TiN and VN inclusions, which acted as the nucleation sites of IGF formation, were also calculated based on the classical nucleation theory. The critical diameters of TiO, TiN and VN inclusions for IGF formation were found to be 0.192, 0.355 and 0.810 μm in the present steels. The calculation results were found to be in agreement with the experiment data of an effective inclusion size. Moreover, the effects of the S, Mn and C contents on the critical diameters of inclusions were also calculated. It was found that the critical diameter of the TiO, TiN and VN inclusions increases with an increased content of Mn or C. However, the S content doesn’t have a direct effect on the critical diameter of the inclusions for IGF formation. The probability of IGF nucleation for each inclusion size slightly decreases in the steel containing a higher S content. This fact is due to that an increased amount of MnS precipitation covers the nucleant inclusion surface. In the as-cast experiment, it was noted that an IGF can be formed in steels with Ti2O3 and TiN additions with a cooling rate of 3.6 ºC/min. In order to control the microstructure characteristics, such as the area fraction and the morphology of an IGF, and to investigate the starting temperature of IGF and grain boundary ferrite (GBF) formation, the dynamic transformation behavior of IGF and GBF was studied in-situ by a high temperature confocal laser scanning microscope (CLSM). Furthermore, the chemical compositions of the inclusions and the morphology of IGF after the in-situ observations were investigated by using scanning electron microscopy (SEM), electron backscatter diffraction (EBSD) and electron probe microanalysis (EPMA) which equipped wavelength dispersive spectrometer (WDS). The results show that the area fraction of IGF is larger in the steels with Ti2O3 additions compared to the steels with TiN additions, after the same thermal cycle has been imposed. This is due to that the TiOx phase provides more potent nucleation sites for IGF than the TiN phase does. Also, the area fraction of IGF in the steels is highest after at an intermediate cooling rate of 70 ºC/min, since the competing phase transformations are avoided. This fact has been detected by using a hybrid methodology in combination with CLSM and differential scanning calorimetry (DSC). In addition, it is noted that the morphology of an IGF is refined with an increased cooling rate. / <p>QC 20150325</p>