This thesis, themed "Development of antioxidant capacity during the lactic fermentation of vegetables" was elaborated between 2014 and 2016 at the Faculty of Horticulture in Lednice. The aim of the study was to evaluate the development of antioxidant capacity during the lactic fermentation of selected vegetable species. The theoretical part includes characteristics of vegetables, health-promoting substances, antioxidants and lactic fermentation. In the practical part were based samples of cabbage material and during lactic fermentation were observed changes in the basic parameters and the development of antioxidant capacity. Higher values of antioxidant capacity were observed for fermented cabbage compared with unleavened and also values among representatives of red cabbage varieties were higher than for white ones. Based on the measurement results were evaluated that the lactic fermentation of vegetable material, especially cabbage, is one of the most promising methods to preserve and uplift its antioxidant capacity.

L’objectif de cette thèse était d'étudier les propriétés physico-chimiques de réseaux interpénétrés formés par auto-assemblage de copolymères à blocs amphiphiles stimulables. La première partie du travail a consisté à élaborer des hydrogels interpénétrés IPSAN (InterPenetrated Self-Assembled Network) à partir d’un simple mélange de copolymères triblocs associatifs. Ainsi, l’IPSAN correspond à une combinaison des deux réseaux polymères. Le premier réseau est formé par un copolymère tribloc à base de poly(oxyde d'éthylène) (tPOE) porteur de blocs hydrophobes polymérisables sous UV. Le deuxième réseau est constitué d’un copolyélectrolyte tribloc pH sensible à base de poly(acide acrylique) (TH50).L’influence des concentrations en copolymères et du pH sur la structure et les propriétés mécaniques des hydrogels IPSAN a été systématiquement étudiée. Dans une deuxième partie, nous nous sommes intéressés à l'influence de la formation d'un réseau transitoire sur la séparation de phases dans des mélanges de tPOE et de polymères linéaires en solution aqueuse. Cette étude a mis en évidence une augmentation de l'incompatibilité entre les deux polymères induite par l’association du copolymère tribloc. Ce système a permis l'élaboration d'hydrogels macroporeux photo-réticulables.Enfin, la diffusion de polymères linéaires et de particules solides micrométriques dans des réseaux de copolymères pH-sensibles a été étudiée par des techniques de recouvrement de fluorescence après photobleaching (FRAP) et de diffusion dynamique de la lumière. La formation du réseau transitoire ralentit le mouvement de traceurs linéaires de petite taille de la même manière que dans le cas d'hydrogels covalents. Le contrôle de la structure et de la dynamique du réseau par le pH est un levier puissant pour contrôler la diffusion dans ces hydrogels. / The objective of this thesis was to investigate the physical chemical properties of interpenetrating networks formed by self-association of responsive amphiphilic block copolymers. The first part of the work was to develop IPSAN hydrogels(InterPenetrated Self-Assembled Network) simply by mixing two triblock copolymers. Thus, the IPSAN corresponds to a combination of the two polymer networks. The first network is formed by a triblock copolymer based on poly(ethylene oxide) (tPEO) bearing UV-cross-linkable hydrophobic blocks. The second network consists of a pH-sensitive triblock polyelectrolyte based on poly(acrylic acid) (TH50). The influence of the concentration of the copolymers and of the pH on the structure and the mechanical properties of the IPSAN has been systematically studied. In the second part we investigated the influence ofthe formation of a transient network on the phase separation in mixtures of tPEO and linear polymers in aqueous solution. This study revealed an increase of the incompatibility between the two polymers induced by the association of the triblock copolymer. This system enabled the elaboration of photo-cross-linked macroporous hydrogels. Finally, the diffusion of linear polymers and solid particles in pH-sensitive networks was studied by fluorescence recovery after photobleaching (FRAP)and dynamic light scattering. The formation of a transient network restricts the movement of small linear tracers in the same way as a covalent hydrogel would. The control of the structure and dynamics of the network by the pH is a powerful tool to control the diffusion in this hydrogels.

Apicobasal polarity is essential for epithelia formation and maintenance. Cell junctions, namely the zonula adherens in Drosophila melanogaster, are the morphological landmarks that define and distinguish the apical from the basal surface. This resulting compartmentalisation is key for the cell and consequently the epithelia. To maintain proper junctions, cells make use of several protein complexes and their interactions. Among these complexes, the Crumbs (Crb) network stands out. Mutations in Crumbs (crb11A22) lead to zonula adherens collapse, consequent loss of apical surface and disaggregation of the epithelia. However, the mechanisms behind this are not known and havenʼt been addressed using modern techniques such as live imaging. Several things came out of the dataset obtained from the FRAP experiments. Firstly, protein kinetics are better described when a double exponential fit curve is used, which raises the possibility that two cell processes might be involved in the recovery observed for the different markers. Another finding was the fact that the kinetics of some polarised protein markers is not the same in every region of the embryo. Distinct areas of the embryo with different morphogenetic activity levels show different kinetics for the same compartment marker. That was the case with SpiderGFP (whole plasma membrane marker) and SASVenus (apical plasma membrane marker) where τ2 was lower in the posterior region of the embryo which is characterised by intense cell movements resulting from convergence extension. DE-CadGFP (zonula adherens marker) and lacGFP (basolateral marker) behaved similarly in the whole embryo. This indicates that convergence extension shows different trafficking needs for the apical surface. In crb11A22, SpiderGFP kinetic spatial differences were not observed. τ2 in the anterior (low level of morphogenesis) is affected and similar to wild type τ2 levels in the posterior. This could pinpoint the fact that the epithelia disaggregation is a result of trafficking failure of apical components. Live imaging of DE-CadGFP in crb11A22 background revealed initial disaggregation in the anterior part of the embryo, which strengthens the idea that Crb is required for adherens junction stabilisation and maintenance.

In this thesis the need for nutrients for rabbits was described. Anthocyanins were described as biological substances contained in wheat with non-traditional coloring and their possible antioxidant effects. The second part of the review is devoted to bio-chemical parameters of animal blood. The practical part was aimed at the study of the influence of feed mixture with increased content of anthocyanins. In the experiment, a wheat species PS Karkulka with purple pericarp was used and Vanessa was used as control wheat species. Both of these made 15% of the feed diet. 18 rabbit females of which 9 were in the experimental group and 9 in the control group were included in the experiment. They were fed a complete pelleted mixture. In the experiment, the weight of the animals, the average weight gains, the feed consump-tion, the feed conversion ratio were monitored. After slaughter, slaughter yield, bio-chemical blood analysis and antioxidant activity were monitored. The results show that the differences in average live weight were statistically in-significant, only at week 8 the differences in live weight was statistically significantly higher in the control group. Average weight gains were evaluated as statistically insig-nificant. Statistically significantly higher average feed consumption was found in the control group. The carcass weight was significantly higher in the control group. There were no statistically significant differences in biochemical parameters and blood anti-xodation activity.

Cellular biophysics deals with, among other things, transport processes within cells. This thesis presents two studies where mathematical models have been used to explain how two of these processes occur. Cellular membranes separate cells from their exterior environment and also divide a cell into several subcellular regions. Since the 1970s lateral diffusion in these membranes has been studied, one the most important experimental techniques in these studies is fluorescence recovery after photobleach (FRAP). A mathematical model developed in this thesis describes how dopamine 1 receptors (D1R) diffuse in a neuronal dendritic membrane. Analytical and numerical methods have been used to solve the partial differential equations that are expressed in the model. The choice of method depends mostly on the complexity of the geometry in the model. Calcium ions (Ca2+) are known to be involved in several intracellular signaling mechanisms. One interesting concept within this field is a signaling microdomain where the inositol 1,4,5-triphosphate receptor (IP3R) in the endoplasmic reticulum (ER) membrane physically interacts with plasma membrane proteins. This microdomain has been shown to cause the intracellular Ca2+ level to oscillate. The second model in this thesis describes a signaling network involving both ER membrane bound and plasma membrane Ca2+ channels and pumps, among them store-operated Ca2+ (SOC) channels. A MATLAB® toolbox was developed to implement the signaling networks and simulate its properties. This model was also implemented using Virtual cell. The results show a high resemblance between the mathematical model and FRAP data in the D1R study. The model shows a distinct difference in recovery characteristics of simulated FRAP experiments on whole dendrites and dendritic spines, due to differences in geometry. The model can also explain trapping of D1R in dendritic spines. The results of the Ca2+ signaling model show that stimulation of IP3R can cause Ca2+ oscillations in the same frequency range as has been seen in experiments. The removing of SOC channels from the model can alter the characteristics as well as qualitative appearance of Ca2+ oscillations. / Cellulär biofysik behandlar bland annat transportprocesser i celler. I denna avhandling presenteras två studier där matematiska modeller har använts för att förklara hur två av dess processer uppkommer. Cellmembran separerar celler från deras yttre miljö och delar även upp en cell i flera subcellulära regioner. Sedan 1970-talet har lateral diffusion i dessa membran studerats, en av de viktigaste experimentella metoderna i dessa studier är fluorescence recovery after photobleach (FRAP). En matematisk modell utvecklad i denna avhandling beskriver hur dopamin 1-receptorer (D1R) diffunderar i en neural dendrits membran. Analytiska och numeriska metoder har använts för att lösa de partiella differentialekvationer som uttrycks i modellen. Valet av metod beror främst på komplexiteten hos geometrin i modellen. Kalciumjoner (Ca2+) är kända för att ingå i flera intracellulära signalmekanismer. Ett intressant koncept inom detta fält är en signalerande mikrodomän där inositol 1,4,5-trifosfatreceptorn (IP3R) i endoplasmatiska nätverksmembranet (ER-membranet) fysiskt interagerar med proteiner i plasmamembranet. Denna mikrodomän har visats vara orsak till oscillationer i den intracellulära Ca2+-nivån. Den andra modellen i denna avhandling beskriver ett signalerande nätverk där både Ca2+-kanaler och pumpar bundna i ER-membranet och i plasmamembranet, däribland store-operated Ca2+(SOC)-kanaler, ingår. Ett MATLAB®-verktyg utvecklades för att implementera signalnätverket och simulera dess egenskaper. Denna modell implementerades även i Virtual cell. Resultaten visar en stark likhet mellan den matematiska modellen och FRAP-datat i D1R-studien. Modellen visar en distinkt skillnad i återhämtningsegenskaper hos simulerade FRAP-experiment på hela dendriter och dendritiska spines, beroende på skillnader i geometri. Modellen kan även förklara infångning av D1R i dendritiska spines. Resultaten från Ca2+-signaleringmodellen visar att stimulering av IP3R kan orsaka Ca2+-oscillationer inom samma frekvensområde som tidigare setts i experiment. Att ta bort SOC-kanaler från modellen kan ändra karaktär hos, såväl som den kvalitativa uppkomsten av Ca2+-oscillationer. / QC 20101111

Während der Gewebeentwicklung beeinflusst die Anpassung der mechanischen Spannung bei Zell-zu-Zell-Kontakten das Gewebewachstum, die Musterbildung und die Morphogenese. Die Erzeugung und Kontrolle der mechanischen Spannung hängt von Komponenten des Zytoske- letts wie dem Aktomyosin und den Mikrotubuli-Netzwerken ab. Die Bildung von Komparti- mentgrenzen ist ein wichtiger Entwicklungsprozess, der auf der Anpassung mechanischer Spannungen beruht. Kompartimentgrenzen sind Abstammungsbeschränkungen, die Zellen mit unterschiedlichen Funktionen und Identitäten innerhalb von Geweben trennen. Zellverbindun- gen entlang der Kompartimentgrenzen sind häufig durch eine Anreicherung von filamentösen (F-) Aktin sowie nicht-muskulären Myosin II (Myosin II) Motorprotein und erhöhter mechani- scher Spannung gekennzeichnet. Die Mechanismen, durch die F-Aktin und Myosin II an die- sen Verbindungsstellen angereichert werden, sind jedoch kaum verstanden. Hier zeigen wir, dass an der sich bildenden anteroposterioren Kompartimentgrenze der Puppenepidermis von Drosophila melanogaster F-Aktin und Myosin II vorübergehend angereichert werden. Die An- reicherung von F-Aktin scheint nicht von mechanischer Spannung abzuhängen. Die Fluores- zenzerholung nach Photobleichversuchen (Fluorescence recovery after photobleaching, FRAP) weist eher darauf hin, dass Myosin II vorzugsweise an Zellübergängen entlang der Komparti- mentgrenze stabilisiert wird. Darüber hinaus zeigen wir unter Verwendung einer photokonver- tierbaren Form von Myosin II, dass Myosin II vorzugsweise aus einem zytosolischen Pool an Zellverbindungen entlang der Kompartimentgrenze rekrutiert wird. Um die Rolle des Mikro- tubuli-Netzwerks bei der Bildung von Kompartimentgrenzen zu testen, haben wir außerdem dessen Organisation in der Puppenepidermis charakterisiert. Wir zeigen, dass sich Mikrotubuli und das Mikrotubuli-Minus-Ende-bindende Protein Patronin in einem Streifen anteriorer Zellen entlang der Kompartimentgrenze ansammeln. Interessanterweise haben die Zellen in diesem Streifen, im Vergleich zu anderen Zellen in der Epidermis, eine unterschiedliche Form. Zusammengefasst enthüllen unsere Daten Unterschiede in der Organisation der Mikrotubuli, die mit Kompartimentgrenzen verbunden sind, und zeigen, dass die Anreicherung von Myosin II entlang der Kompartimentgrenze der Puppen-Abdominalepidermis sowohl eine bevorzugte Stabilisierung als auch eine Rekrutierung beinhaltet.

<p dir="ltr">Spatial heterogeneity is ubiquitous across life and the universe; the same is true for phase-separating pharmaceutical formulations, cells, and tissues. To interrogate these spatially-varying complicated samples, simple analysis techniques such as fluorescence recovery after photobleaching (FRAP) can provide information on molecular transport. Conventional FRAP approaches localize analysis to small spots, which may not be representative of trends across the full field of view.</p><p dir="ltr">Taking advantage of strategies used for structures illumination, an approach has been developed to use patterned illumination in combination with FRAP for probing large fields of view while representatively sampling. Patterned illumination is used to establish a concentration gradient across a sample by irreversibly photobleaching fluorophores, such as with the simple comb pattern photobleach presented in Chapters 1 and 4. Patterned photobleaching allows spatial Fourier-domain analysis of multiple spatial harmonics simultaneously. In the spatial FT-domain the real-space photobleach signal is integrated into puncta, greatly increasing the signal to noise ratio compared to conventional point-bleach FRAP. The order of the spatial harmonic is directly related to the length-scale of translational diffusion measured, with a series of harmonics accessing diffusion over many length scales in a single experiment. Measurements of diffusion at multiple length scales informs on the diffusion mechanism by sensitively reporting on deviations away from normal diffusion.</p><p dir="ltr">Complementing the physical hardware for inducing patterned illumination, this dissertation introduces novel algorithms for reconstructing spatially-resolved diffusion maps in heterogeneous materials by combining Fourier domain analysis with patterned photobleaching. FT-FRAP is introduced in Chapter 1 for interrogating phase-separating samples using beam-scanning instrumentation for comb-bleach illumination. This analysis allowed disentangling separate contributions to diffusion from normal bulk diffusion and an interfacial exchange mechanism only available due to multi-harmonic analysis. The introduction of a dot-array bleach pattern using widefield microscopy is presented in Chapter 2 for high-throughput detection of mobility in simple binary systems as well as for segmentation in phase-separating pharmaceutical formulations. The analysis becomes more complicated as more components are added to the system such as a surfactant. Introduced in chapter 3, FT-FRAP with dot-array photobleaching was shown to be useful for characterizing diffusion of phase-separating micro-domain smaller than a single pixel of the camera. Supported by simulations, a biexponential fitting model was developed for quantification of diffusion by multiple species simultaneously. Chapter 4 introduces imaging inside of 3D particles comprised of an active pharmaceutical ingredient (API) in microencapsulated agglomerates which exhibited strong interfacial exchange. Multi-photon excited fluorescence enabled imaging a small focal volume within the particles.</p>

Brucella abortus is a zoonotic agent that primarily infects cattle and causes brucellosis. B. abortus strain RB51 is a live, attenuated vaccine licensed for cattle. However, there is no available vaccine to prevent human brucellosis. Outer membrane vesicles have been tested as potential vaccines to prevent diseases caused by bacterial species. OMV are constantly released from Gram-negative bacteria. They are comprised principally of the outer membrane components and periplasmic proteins from the bacterial cell envelope. The research in this thesis examined the adjuvant property of non-replicative, metabolically active irradiated strain RB51 and the protective ability of OMV derived from strain RB51. Irradiated B. abortus strain RB51 was assessed for its ability to act as an adjuvant to induce protection against malaria. It was found that irradiated B. abortus strain RB51 administered along with fasciclin related adhesive protein (FRAP) to mice induced a protective immune response and a significant decrease in parasitemia after challenge with Plasmodium berghei. Strain RB51 and strain RB51 over-producing Cu/Zn superoxide dismutase (Cu/Zn SOD) were used to produce OMV. Western blotting and SDS-PAGE gel staining confirmed the presence of OMV and the over-production of Cu/Zn SOD. OMV were delivered to mice using an intraperitoneal route and, in some cases, with aluminum hydroxide adjuvant. The immune response was assessed by antibody isotyping with respect to OMV and measuring splenic clearance (i.e. protection) from a B. abortus strain 2308 challenge. The results demonstrate that OMV from B. abortus strain RB51 or strain RB51 over producing Cu/Zn SOD produced a Th1 polarized immune response as measured by specific OMV antibodies and cytokines but no statistically significant protection was observed. / Master of Science

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal.

No description available.