The goal of this project is the development of artificial ion channels that can be actuated by light and thus controlled efficiently. Our artificial system is composed of two regions: the gate and the body part. The gate part is based on light-responsive azo groups while the body part is formed by calix[4]resorcinarene. Key of controlling mechanism is the conformational change between cis and trans isomers, which is translated into movement of the gate. Light-gated artificial ion channels are aimed at eliminating of the stochastic mechanism of artificial ion channels. Such a reversible photocontrol should be a powerful tool for using artificial ion channels as the basis for the development of new pharmaceuticals and drug delivery systems, as photoswitches, and in the field of microfluidics.

Patch clamping is considered the gold standard in measuring the bioelectrical activity of a cell. It is used to detect and measure ion transport through ion channels located throughout a cell membrane. Ion movement is crucial to cell viability and cell-to-cell communication. Pharmaceutical companies increasingly target ion channels because of their significance in disease and to help design better targeted drugs. However, the traditional method of patch clamping is cumbersome and is being replaced by planar high throughput screening (HTS) systems. These systems are reaching their limits due to materials and cost of processing; cell handling methods and small varieties of applicable cell types are also issues to be addressed. In this work, the core components of a new kind of planar patch clamping device have been designed and developed, after analysis of currently available HTS systems. This design approaches patch clamping using polymers to overcome some of the limitations in current systems, specifically cell handling and positioning, by using a simple modification technique to provide distinct attractive areas for cell binding. This uniquely allows the culture of both single cells and cell networks to increase the range of cell types that can be measured and circumvents challenges from using suction to pull cells onto measurement holes. The components of the design are a 10 x 10 array of small holes drilled in a polymer then aligned modifications for precise cell placement are added and a planar electrode array for individual addressing of each cell. A study of methods to produce a leak-tight seal required between microfluidic chambers was done. Cell adhesion parameters for the modification techniques were established. The principle viability of this approach was confirmed using the modification technique to culture cells over holes and measure their resistance using a rig developed for this work.

Astrocytes are the most abundant non-neuronal cells in vertebrate brains. Although Drosophila melanogaster has fewer astrocytic cells relative to neuronal and other glial cell populations, they, like vertebrate astrocytes, are located in synaptic regions, organized into exclusive, minimally-overlapping domains, and play developmental roles in synaptogenesis. But, do Drosophila astrocytes have parallel roles in the regulation of synaptic signaling? Preliminary electron microscopic (EM) data indicates that astrocytic processes are located at a greater distance, on average, from Drosophila synapses than they are from vertebrate synapses, thus raising questions about their capacity to alter synaptic signals. Do astrocytic cells and processes occupy stereotyped synaptic regions across repeating segmental structures and across individuals? In the studies presented here, we have addressed these questions directly in the ventral nerve cord (VNC) of the third-instar larva. We collected the first whole-cell patch-clamp recordings from Drosophila astrocytes. These indicate that intrinsic membrane properties, such as low membrane resistance, high capacitance, a hyperpolarized resting potential relative to neurons, a passive current-voltage relationship, coupling to other astrocytic cells, and an absence of voltage-gated currents, are shared between astrocytes of highly divergent species. Next, we optogenetically activated of a group of glutamatergic pre-motor neurons and showed that astrocytes respond with a glutamate transporter current that is mediated by Eaat1, and that acute, pharmacological and chronic, genetic blockades of this transporter have subsequent effects on the decay of post-synaptic motor neuron currents. Then, we used three-dimensional EM to locate the pre-motor glutamatergic neurons that were activated in the physiological study and measured the distance from each presynaptic site to the nearest astrocytic process. We found that these distances vary 100-fold even along a single neurite and that these structures are rarely in direct contact, but that no synapse is positioned greater than one micron from an astrocytic process. Thus, it is in this anatomical configuration that the regulation of post-synaptic currents by Eaat1 occurs. Finally, we generated a library of single, fluorescently-labeled astrocytes that were co-labeled with fiduciary landmarks, and used this library to compare the placement of astrocyte cell bodies and arbors across VNC segments and individuals. We found substantial variation in the gross shape, size, and territory covered by astrocytes, and conclude that their neuropil domains are not reliably stereotyped. Given the consistent placement of neuronal connectome elements, this indicates that signals of a specific synapse are not regulated by a designated astrocyte. Together, these findings reveal new functional parallels between Drosophila and vertebrate astrocytes. These findings argue for the relevance and applicability of mechanistic discovery in Drosophila astrocytes, and set the stage for further inquiry into the genetic determinants of astrocyte morphology and physiology.

Background: Since constrictive pericarditis is most often idiopathic and the pathophysiology remains largely unknown, both the diagnosis and the treatment can be challenging. However, by definition, inflammatory processes are central to this disease process. Amniotic membrane patches have been shown to possess anti-inflammatory properties and are believed to be immune privileged. Due to these properties, amniotic membrane patches were applied intraoperatively in a complicated patient presenting with constrictive pericarditis. Case presentation: A patient with a history of multiple cardiac surgeries presented with marked fatigue, worsening dyspnea and sinus tachycardia. He was found to have constrictive physiology during cardiac catheterization, with cardiac MRI demonstrating hepatic vein dilatation, atrial enlargement and ventricular narrowing. After amniotic membrane patch treatment and pericardiectomy, post-operative cardiac MRI failed to demonstrate any appreciable pericardial effusion or inflammation, with no increased T2 signal that would suggest edema. Conclusions: Given the positive results seen in this complex patient, we suggest continued research into the beneficial properties of amniotic membrane patches in cardiac surgery.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Ecological restorations have become increasingly important as humans have made irreversible changes to ecosystems; biogeochemical cycle alterations and land use changes have led to degraded conditions (Vitousek et al. 1997a). While some modifications to the environment are unavoidable, society is beginning to realize that changes are necessary. While some ecosystem functions may be beyond repair (e.g., biodiversity loss and extinction of species), it is important that the problem is faced head on and those functions that are still salvageable be restored. These environmental repairs can be done through ecological restoration. I looked at two different ecosystems, prairie streams and agricultural streams, that have undergone restoration to help functions. Patch-burn grazing (PBG) is increasingly used as a management practice on the few remaining tallgrass prairie parcels in an effort to simulate effects of large ungulate grazers. Yet, little is known about potential impacts and recovery of aquatic habitats from this management approach. My objective was to assess the influence of PBG with cattle on prairie streams and build on previous research at this site. I hypothesized that cattle grazing would negatively impact water quality and reduce stream biotic integrity, but riparian fencing would mitigate these impacts. We also assessed stream recovery for two years following the removal of ungulates from the study sites and hypothesized that biological and chemical effects would reverse. Six headwater streams (two controls, two PBG with 10 m fenced riparian zones, and two PBG with unfenced riparian zones) on Osage Prairie, Missouri, were sampled over seven years (2009-2015) encompassing pre-PBG (2 years), PBG (3 years), and post-PBG (2 years) periods. Macroinvertebrates and water chemistry were sampled monthly. Nitrate (NO3-) concentrations increased in the fenced and unfenced watersheds compared to the control watersheds (p = 0.015 and p < 0.0001, respectively) and between the fenced and unfenced watersheds (p = 0.001) during the PBG period relative to pre-PBG. Total phosphorus (TP) increased in the fenced and unfenced watersheds after grazing began compared to the control (both p < 0.0001) but did not differ between the fenced and unfenced watersheds (p = 0.187). Relative Chironomidae biomass and abundance increased in the unfenced watersheds in response to grazing (p = 0.008 and p < 0.001, respectively). No differences were observed in relative Chironomidae biomass and abundance, NO3-, and TP from pre-PBG to post-PBG indicating the recovery of these metrics. Results suggest that the negative effects of PBG on prairie streams can be somewhat mitigated by riparian fencing. Additionally, these streams are relatively resilient to PBG and recovery can take place during a rest cycle as brief as two years. Approximately one billion dollars is spent annually on restorations of degraded stream reaches in the United States. However, few projects are monitored upon completion, or monitoring focuses on a single parameter. Other than modifications to physical attributes of streams, the influence of restoration projects on ecosystem processes remains largely unknown. We sampled eleven Midwestern streams that had undergone habitat restorations from 3-15 years prior to sampling. Restoration techniques included in-stream habitat enhancements, bank stabilization, and riparian restoration. We predicted that gross primary production (GPP) would be lower in restored streams due to decreased nutrient inputs, and that respiration would be greater due to increased litter inputs from restored riparian areas. We also hypothesized that the restored streams would have greater invertebrate richness, abundance, and biomass, as well as high densities of intolerant taxa such as Ephemeroptera, Plecoptera, and Trichoptera (EPT taxa) due to higher water quality and better habitat. Restored reaches and unrestored upstream reaches were sampled for physical characteristics, water chemistry, benthic algal biomass, whole-stream metabolism, and macroinvertebrate communities. GPP in restored sites was marginally higher than unrestored sites (t5 = 2.53, p = 0.05), despite no differences in PO4-3, NO3-, or NH4+ concentrations. Three restored sites were autotrophic (P/R > 1), while four unrestored sites were heterotrophic. Total macroinvertebrate biomass was marginally higher in the restored sites compared to unrestored sites (t10 = 1.94; p = 0.08). EPT biomass was also marginally higher in restored sites than unrestored sites (t10 = 1.91; p = 0.09) but no difference was observed in EPT abundance. Results suggest that stream habitat restorations enhance some, but not all ecosystem processes and marginally enhance macroinvertebrate communities.

Esse estudo teve como objetivo comparar duas formas de manejo de cultivos de banana em relação ao seu potencial de abrigar espécies florestais. Nós assumimos que o registro de espécies florestais utilizando um elemento da matriz mostra que a espécie não evita a matriz totalmente e pode ser capaz de cruzá-la, indicando a importância dessas estruturas para a manutenção de conectividade funcional entre as áreas de hábitat. Foram amostrados 30 cultivos incluindo áreas sob manejo convencional e agroflorestal. Em cada unidade amostral foram realizadas transecções para acessar a proporção de aves florestais, generalistas e de áreas abertas. Variáveis descritoras da estrutura interna e da estrutura da paisagem do entorno dos cultivos foram consideradas para verificar se a avifauna responde à características intrínsecas dos cultivos ou ao contexto de paisagem em que os cultivos estão inseridos. A proporção de espécies florestais foi maior em cultivos agroflorestais (F28;0,001=19,29; p<0,0004), enquanto a proporção de generalistas não apresentou diferença significativa (F28;0,001=0,22; p<0,22) e a de espécies de áreas abertas foi maior em cultivos convencionais (F28;0,001=8,92; p<0,006). Os dois conjuntos de variáveis considerados, isto é, a estrutura interna do cultivo e do entorno deste foram importantes para predizer a proporção de espécies florestais. Embora tenhamos observado o efeito da paisagem na proporção de espécies florestais, salientamos aqui que sistemas convencionais, mesmo em contextos de paisagem similares à sistemas agroflorestais, não apresentaram a mesma proporção de espécies florestais. Esse resultado indica um diferencial de cultivos agroflorestais de banana para a ocorrência de espécies florestais. Em nível de paisagem, a proporção de florestas e consequentemente a diminuição do isolamento entre manchas é o responsável pela presença potencial de espécies em fragmentos. No entanto, em nível de mancha, mesmo esta apresentando um contexto de paisagem favorável para que seja ocupada por espécies de áreas fonte, o mecanismo que influencia a presença das espécies é a elevada estrutura interna dos bananais. / This study aimed to compare two management of banana crops in relation to its potential to host forest bird species. We assume that the record of forest bird species using a matrix element shows that the species does not entirely avoid this area and may be able to cross it, indicating the importance of these structures for the maintenance of functional connectivity between areas of habitat. We sampled 30 areas including crops under conventional and agroforestry management. In each sampling unit we used transects to access the composition and abundance of bird species. Descriptive variables of the internal structure and the structure of the surrounding landscape of crops were considered to determine whether the bird responds to the intrinsic characteristics of the crops or the landscape context in which crops are inserted. The proportion of forest species was higher in agroforestry crops (F28, 0,001 = 19.29, p <0.0004), while the proportion of generalists showed no significant difference (F28, 0,001 = 0.22, p <0.22) and species of open areas was greater in conventional crops (F28, 0,001 = 8.92, p <0.006). The two sets of variables considered, i.e. the internal structure of the crop and of surroundings, were important to predict the proportion of forest birds species. Although we observed the effect of landscape in the proportion of forest species, we note here that conventional systems, even in contexts similar to the landscape agroforestry systems, did not show the same proportion of forest species. This result indicates a upper value of agroforestry crops for the occurrence of forest bird species. At landscape level, the proportion of forests and consequently the lower isolation between patches is responsible for the potential presence of species in fragments. However, at patch level, even it is presenting a landscape context favorable to be occupied by a species of source areas, the mechanism that influences the presence of the species is the high internal structure of the banana plantations.

The durability of concrete structures is affected by a number of factors such as environmental exposure, electrochemical reactions, mechanical loading, impact damage and others. Of all of these, corrosion of the reinforcement is probably the main cause for the deterioration of steel reinforced concrete (RC) structures. Corrosion management is becoming increasingly necessary as a result of the growing number of ageing infrastructure assets (e.g. bridges, tunnels etc.) and the increased requirement for unplanned maintenance in order to keep these structures operational throughout their design life (and commonly, beyond). The main RC repair, refurbishment and rehabilitation approaches generally employed can be broadly categorised under a) conventional, b) surface treatments, c) electrochemical treatments and d) design solutions. The overarching aim of this research was to identify the key corrosion management techniques and undertake empirical investigations focused on full-scale RC structures to investigate their long-term performance. To achieve this, individual research packages were identified from the above broad five approaches for repair, replacement and rehabilitation. These were 1) Patch repairs and incipient anodes, 2) Impressed Current Cathodic Protection, 3) Galvanic Cathodic Protection and 4) Hydrophobic treatments. The selection of the above research packages was based on past and present use by the construction industry to repair, refurbish and rehabilitate RC structures. Their contributions may be broadly categorised as i) Investigations on how specific treatments and materials perform, ii) Investigations on the effectiveness of existing methods of measurements and developing alternatives, iii) Changes to the existing theory of corrosion initiation and arrest and iv) Changes to management framework strategies. The key findings from each research package can be summarised as follows: Macrocell activity appears to be a consequence rather than a cause of incipient anode formation in repaired concrete structures, as has previously been presented; ICCP has persistent protective effects even after interruption of the protective current; Discrete galvanic anodes installed in the parent concrete surrounding the patch repair are a feasible alternative to galvanic anodes embedded within the patch repairs of RC structures; Silanes may have a residual hydrophobic effect even after 20 years of service.

Une brève stimulation du Noyau Sous-thalamique (STN) dans la tranche des ganglions de la base induit une réponse polysynaptique de longue durée dans ses cibles : Substantia Nigra Pars reticulata (SNr), GP (Globus Pallidus). Cette réponse consiste en un courant NMDA suivi d'un barrage de courant excitateurs de type AMPA qui génère des bouffées de potentiels d'action. La même stimulation dans des branches provenant de souris déplétée en dopamine génère aussi des réponses complexes de plus longue durée. Leur seuil de déclenchement étant 2 à 3 fois plus faibles et ne peuvent dans certains cas se réverbérer. Afin de comprendre les mécanismes sous-sous-jacents dans le STN, nous avons étudié la réponse de ces neurones. en contrôle de courtes réponses sont enregistrées tandis qu'en conditions "parkinsoniennes" cette réponse est considérablement augmentée. Nous proposons qu'un réseau glutamatergique est présent dans le STN et est sous le contrôle négatif des récepteurs dopaminergiques / Single pulse stimulation of the Subthalamic Nucleus (STN) induces a polysynaptic response in the Basal Ganglia Slice in its targets : Substantia Nigra Pars Reticulata (SNr), Globus Pallidus (GP). This response consists of a slow NMDA current with superimposed AMPA transients which generate burst of action potential. The same stimulation in slices depleted in dopamine generates complex response of longer duration. The threshold is also lowered (2-3 times) and in some cases this response reverberates. Theses bursts of synaptic activities could be generated spontaneously. In order to understand the mechanisms, we recorded the evoked response in the STN. In control, only short responses are recorded whereas in "Parkinsonian" conditions, the polysynaptic response is increased. We propose that a polysynaptic network of dopaminegic receptors

Interstitial cystitis (IC)/painful bladder syndrome (PBS) is a functional visceral disorder characterized by increased bladder activity and chronic pelvic pain in the absence of a pathobiological condition. Enhanced sensory transduction of peripheral bladder afferents is hypothesized to contribute to the pain and mechanical hypersensitivity of IC/PBS patients. The aim of this thesis is to test the hypothesis that purinergic receptors, including ionotropic P2X and metabotropic P2Y, are important for sensory transmission in bladder afferent neurons and may be involved in bladder hypersensitivity after bladder tissue insults. Electrophysiological, single cell RT-PCR and immunohistochemistry techniques were performed in bladder afferent neurons from naïve and bladder inflamed mice to test the hypothesis. In Chapter 2, I characterized the distribution and function of P2X receptors in thoracolumbar (TL) and lumbosacral (LS) dorsal root ganglia (DRG) neurons innervating the urinary bladder, and found that LS and TL bladder neurons have differential purinergic signaling and distinct membrane electrical properties. In Chapter 3, I examined the sensitization of bladder afferent neurons and the plasticity of P2X receptor function in a mouse model of chemical induced bladder inflammation. P2X-mediated signals in LS and TL bladder neurons after bladder inflammation were enhanced compared with those in saline-treated controls, suggesting the importance of P2X in bladder hypersensitivity associated with cystitis. In Chapter 4, the modulation of P2Y on P2X function and the co-localization of P2Y and P2X were examined in bladder sensory neurons. It has been found that P2Y2 receptor enhances bladder sensory neuron excitability and facilitates the response of homomeric P2X2 receptor to the purinergic agonist (ATP). The present study provides evidence that LS and TL mouse bladder sensory neurons exhibit distinct P2X signaling, and the function of P2X receptors could be facilitated during bladder inflammation and modulated by activation of P2Y2 receptor, indicating an involvement of P2X and P2Y2 receptors as mechano- and chemosensors in bladder sensory transmission under normal conditions and in bladder hypersensitivity associated with inflammation.