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Different methods to predict barrier failure in packaging board or paper after converting were investigated. The approach was to compare substrates before and after creasing/folding by applying different barrier tests and to propose a methodology to predict failure in the barrier layer.  Different coatings were used to develop and verify the methodology; a hemicellulose based dispersion barrier coating, a dispersion coated PVOH coating and an extrusion coated PE. Creasing was performed according to standard procedure using recommended creasing geometries. Folding of paper was performed by a gentle creasing with a board backing followed by folding the paper between two metal plates with a well defined distance. The first step in the evaluation was to visually inspect creased/folded substrates by light microscopy to search for coating failures in form of cracks. Both good and bad samples were then tested for grease resistance with a standard test, i.e. TAPPI 454. The TAPPI 454 test showed to be effective to expose barrier failure since oil would penetrate quite fast through the creasing line of cracked samples. Even some samples that appeared to have no cracks in the light microscope showed failure with the grease test. The results showed that only the PE coated samples could sustain a barrier after creasing and folding. This was probably due to a high ductility of the PE-coating combined with a high thickness. The water vapour transmission rate, WVTR, of the samples that passed the TAPPI 454 test was then measured on the samples that endured the grease resistance test. Since PE is a good water vapour barrier, WVTR-measurements were proper for detecting barrier defects. The VWTR of the creased/folded samples was slightly higher for the creased samples than the un-creased references despite the absence of cracks. This was probably due to that the barrier layer got thinner as a result of the strains applied on the coating during the creasing/folding operation.  A methodology to predict barrier failure in barrier coated packaging board and paper after creasing and folding was proposed. Well defined creasing and folding geometries were used in combination with screening for cracks in the barrier layer, first by visual inspection in light microscopy and then by a standard grease resistance test. The samples that passed then screening tests could then be analyzed using more exact but also more time consuming methods such as WVTR. / Olika metoder att förutspå skador i barriärskikt på kartong eller papper efter konvertering undersöktes. Tillvägagångssättet var att jämföra substrat före och efter bigning och vikning genom att tillämpa olika barriärtest och att föreslå en metod för att förutspå defekter i barriärlager. Olika barriärmaterial användes för att utveckla och bekräfta metoden: en hemicellulosa baserad dispersionsbestrykning, en dispersionsbestrykt PVOH barriär och en extruderad PE barriär. Bigandet utfördes enligt standard proceduren och rekomenderade biggeometrier användes. Vikningen av papret utfördes genom varsam bigning med kartong som stöd följt av vikning av pappret genom två metallplattor med ett bestämt avstånd. Utvärderingen började med visuell inspektion av bigade/vikta substrat i ljusmikroskop för att finna barriärdefekter i form av sprickor. Både bra och dåliga prover testades sedan för fettbeständighet med hjälp av ett standardtest, dvs TAPPI 454. TAPPI 454 testet visades sig att vara ett effektivt sätt att identifiera barriärdefekter på grund av att penetration av olja vid biglinjen skedde snabbt på de prov som uppvisade sprickor. Även några av de prov som ej uppvisade sprickor i ljusmikroskop klarade inte av fettbeständighetstest. Resultatet visade att det enda material som kunde bibehålla barriäregenskaper efter bigning och vikning var de PE belagda proven. Detta är antagligen tack vare PE-bestrykningens höga duktulitet och tjocklek. Vattenångspermeabiliteten, WVTR, uppmättes på de prov som uthärdade fettbeständighetstestet. Eftersom PE är en utmärkt vattenångbarriär, var WVTR-mätningar lämpliga för att upptäcka barriärfel. WVTR resultaten för de bigade/vikta proven visade ett något högre värde än de obigade referenserna även om de inte hade sprickor. Det något högre WVTR värdet beror antagligen på att barriärskiktet blev tunnare på grund av töjningen i barriärskiktet under big/vikningen. En metod för att förutspå skador i barriärbestrykt kartong och papper efter bigning och vikning föreslogs. Definierade big- och vikgeometrier användes i kombination av screening av sprickor i barriärskikten, först genom visuell inspektion i ljusmikroskop och sedan ett standarderiserat fettbeständighetstest. Proven som passerar screeningen kan sedan bli analyserade för mer exakta och tidskrävande metoder som WVTR.

Les pathologies intestinales sont des maladies multifactorielles dont l’incidence ne cesse d’augmenter. Les maladies organiques regroupent les maladies inflammatoires chroniques de l'intestin tel que la maladie de Crohn (MC) et les pathologies fonctionnelles digestives comprennent les différentes formes du syndrome de l'intestin irritable (SII). Ces pathologies présentes des caractéristiques communes comme des troubles du transit, des douleurs abdominales, un dysfonctionnement de la barrière intestinale et des modifications de la communication bidirectionnelle de l’axe intestin-cerveau et une altération de la composition du microbiote intestinal (dysbiose). Cette dernière peut être en partie associée à un déficit de production en peptides et protéines antimicrobiennes (PAMs) par les cellules de Paneth. Parmi les cibles thérapeutiques stratégiques, une réduction de la dysbiose dans le but de réduire l’état inflammatoire ou micro-inflammatoire de la muqueuse font aujourd’hui l’objet de différentes études. Ces traitements alternatifs montrent l’efficacité d’un régime nutritionnel adapté, d’un apport en probiotiques ou en prébiotiques sur le microbiote intestinal des patients MC et SII, et sur l’intégrité de leur barrière intestinale. Parmi les matrices alimentaires présentant une composition nutritionnelle intéressante proche du lait maternel humain, le lait d’ânesse (LA) contient également une forte teneur en PAMs (lysozyme). En parallèle, un observatoire économique et social de la filière asine commandité par l’Institut National Âne et Mulet fait le diagnostic d’une activité grandissante autour du LA chez les éleveurs français et montre, face à une concurrence italienne et chinoise importante, la nécessité d’une meilleure organisation de la filière via une meilleure valorisation du LA. L’objectif de cette thèse était d’évaluer un potentiel effet santé du LA et d’apporter des preuves scientifiques robustes pour permettre une meilleure valorisation de ce dernier. Ainsi, l’effet d’une consommation orale chronique de LA a été évalué sur deux modèles précliniques murins distincts : un modèle d’iléite expérimentale induite par une administration per os d’indométacine et un modèle de stress psychologique chronique (Water Avoidance Stress, WAS). Dans ce travail, le rôle clé de l’activité du lysozyme dans les effets observés a également été évalué. Des essais de traitements thermiques ont aussi été réalisés pour optimiser un barème de pasteurisation permettant de proposer un LA répondant aux normes réglementaires microbiologiques et sanitaires tout en conservant l’activité du lysozyme contenue dans le lait. Les travaux de cette thèse montrent que le LA possède des propriétés anti-inflammatoires se traduisant par une réduction significative des lésions inflammatoires macroscopiques et microscopiques de l’iléon. Cet effet est associé à une réduction de la dysbiose intestinale et normalise les niveaux de PAMs dans les cellules de Paneth drastiquement réduits lors de l’iléite. Dans le modèle WAS, le LA et la fraction contenant l’activité en lysozyme réduisent l’hypersensibilité viscérale, l’état micro-inflammatoire induites par le stress et restaurent les niveaux de PAMs dans les cellules de Paneth réduits par le stress. Un traitement thermique de 2 min/72°C permet d’augmenter la durée de vie du LA tout en conservant l’activité du lysozyme et ses propriétés santé sur la muqueuse intestinale chez la souris. / Gut pathologies are multifactorial with a constant increasing incidence. Organic diseases include inflammatory bowel diseases such as Crohn's disease (CD) and functional pathologies include various subtypes of irritable bowel syndrome (IBS). These pathologies present common features such as transit disorders, abdominal pain, dysfunction of the intestinal barrier associated with bidirectional gut-brain axis modifications and alterations of the gut microbiota composition (dysbiosis). Dysbiosis is generally associated with a lack of production of antimicrobial peptides and proteins (AMPs) by Paneth cells. Among the alternative treatment strategies, it has been proposed to reduce gut inflammation by correcting intestinal dysbiosis. Recent studies show the efficacy of a suitable diet, probiotics or prebiotics interventions on gut microbiota and on the integrity of their intestinal barrier in CD and IBS patients. Among interesting food matrices, donkey milk (DM) is a good candidate with a nutritional composition close to human breast milk and contains high levels of AMPs like lysozyme. An economic and social observatory of the donkey industry was ordered by the Institut National Âne et Mulet for a better support of growing French activity around DM against Italian and Chinese competitions. The objective of this thesis was to evaluate the potential beneficial healthy effect of DM and to provide robust scientific evidences for a better valorization of this product. Thus, the effect of chronic oral intake of DM was evaluated on two distinct murine preclinical models: a model of experimental ileitis induced by an oral administration of indomethacin and a model of chronic psychological stress (Water Avoidance Stress, WAS). In these studies, the key role of lysozyme activity in the observed effects was highlighted. In order to develop the DM market in France, thermal treatments have been carried out to optimize a pasteurization process to obtain a DM in accordance with the regulatory microbiological and sanitary standards while preserving the activity of the lysozyme contained in milk. The work of this thesis shows that DM exerts anti-inflammatory properties resulting in a significant reduction of the macroscopic and microscopic inflammatory lesions of the ileum. This effect is associated with a reduction of gut dysbiosis while normalizing the level of drastically reduced AMPs contained Paneth cells in the ileitis model. In the WAS model, DM and the fraction containing activity of lysozyme reduce visceral hypersensitivity, stress-induced gut microinflammatory status and also restore the level of AMPs in Paneth cells . A heat treatment at 2 min/72 °C allows to increase the shelf-life of DM while preserving the activity of lysozyme and its gut beneficial healthy properties in mouse.

Le dioxyde de carbone, principal gaz à effet de serre lié aux activités humaines, est considéré comme l’un des gaz les plus problématiques pour notre environnement ces dernières années, principalement à cause du réchauffement climatique qu’il engendre. C’est pour cette raison que l’augmentation de sa teneur dans l’atmosphère nous concerne tous quant aux conséquences futures pour notre planète. Afin de limiter l’émission de CO2, sa conversion en composés à valeur ajoutée présente un grand intérêt et est possible notamment via des procédés plasmas. Plus particulièrement, les décharges à barrière diélectrique (DBD) sont utilisées depuis quelques années pour générer des plasmas froids opérant à pression atmosphérique, principalement pour des applications en traitement de surface, mais également pour le traitement d’effluents gazeux.Lors de cette thèse, nous nous sommes focalisés sur le processus de dissociation du CO2 en CO et O2 via un réacteur DBD à flux continu et avons analysé sa conversion et son efficacité énergétique via différentes études. Celles-ci ont été réalisées grâce à plusieurs méthodes de diagnostic, comme par exemple la spectrométrie de masse utilisée pour déterminer la conversion et l’efficacité du processus, la spectroscopie d’émission optique, l’oscilloscope pour une caractérisation électrique, etc. afin d’avoir une meilleure compréhension du comportement des décharges CO2.Dans un premier temps, nous avons réalisé une étude détaillée d’un plasma CO2 pur où nous avons fait varier différents paramètres, tels que le temps de résidence, la fréquence, la puissance, la pulsation de la haute tension et l’épaisseur et la nature du diélectrique. Le CO2 donne lieu généralement à une décharge filamentaire, consistant en de nombreuses microdécharges réparties au niveau de la zone du plasma. Celles-ci constituent la principale source de réactivité dans une DBD. Un aperçu détaillé de l’aspect physique de ces microdécharges a été réalisé grâce à la caractérisation électrique, permettant de mieux comprendre les propriétés électriques de la décharge et des microdécharges. En effet, nous avons pu déterminer l’importance de la tension présente au niveau du plasma, de l’intensité du courant plasma, du nombre de microdécharges et de leur temps de vie sur l’efficacité du processus de dissociation de CO2.Ensuite, nous avons conclu ce travail avec des études combinant le CO2 en phase plasma avec de l’eau ou du méthane afin de produire des molécules à valeur ajoutée telles que les syngas (CO et H2), mais aussi des hydrocarbures (C2H6, C2H4, C2H2 et CH2O) dans le cas de l’ajout du méthane. A travers ces études, nous avons obtenu une meilleure connaissance de la chimie et de la physique qui ont lieu dans ce type de plasma. / Carbon dioxide appears as one of the most problematic gases for the environment, mostly because it is responsible for global warming. This is why its increasing concentration into the atmosphere, mainly due to anthropogenic activities, is a real concern for planet Earth. In order to prevent the release of large amounts of CO2, its conversion into value-added products is of great interest. In this context, plasma-based treatments using dielectric barrier discharges (DBDs) are nowadays more and more used for the conversion of this gas. In this thesis, we investigated the CO2 splitting process into CO and O2 via a flowing cylindrical DBD and we studied its conversion and energy efficiency by means of several diagnostic methods, such as mass spectrometry to determine the conversion and energy efficiency of the process, optical emission spectroscopy for gas temperature measurements, and an oscilloscope for electrical characterization, in order to obtain a better understanding of the CO2 discharge itself.First, we focused on an extensive experimental study of a pure CO2 plasma where different parameters were varied, such as the gas residence time, the operating frequency, the applied power, the pulsation of the AC signal, the thickness and the nature of the dielectric. CO2 discharges typically exhibit a filamentary behavior, consisting of many microdischarges, which act as the main source of reactivity in a DBD. A detailed insight in the physical aspects was achieved by means of an in-depth electrical characterization, allowing more insight in the electrical properties of the discharge and more specifically in the microdischarges, which are spread out throughout the active zone of the plasma. It was found throughout this work that the plasma voltage, which reflects the electric field and thus determines how the charged particles are accelerated, the plasma current, which reflects the electron density, but also the number of microdischarges and their average lifetime, play an important role in the efficiency of the CO2 dissociation process. It was revealed that the microdischarge number is important as it represents the repartition of the locations of reactivity. Indeed, as the microfilaments are more spread out in the same discharge volume, the probability for the CO2 molecules to pass through the reactor and interact with at least one microdischarge filament becomes more important at a larger number of microfilaments.The second part of the thesis was dedicated to discharges combining CO2 and H2O or CH4, both being hydrogen source molecules. The combined CO2/H2O or CO2/CH4 conversion allows forming value-added products like syngas (CO and H2), but also hydrocarbons (C2H6, C2H4, C2H2 and CH2O), at least in the presence of methane. Throughout this study, we tried to obtain a better knowledge of the chemistry and physic behind these conversion processes. / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

Abstract The living circumstances of the elderly are closely related to the social changes in society. The aim of this research is to firstly, identify the impact of social changes on the spatial organization of dwellings and housing from the perspective of the elderly within the framework of social ageing, and secondly to examine the reasons for relocation of the elderly to sheltered housing. Using this data, some recommendations are made about a suitable barrier-free housing model for the elderly based on traditional Finnish rural housing. In addition to the need for barrier-free design criteria in the design process of future dwellings, understanding the social and psychological aspects of the traditional housing of the northern outreach can provide a new perspective for developing and modifying current living environments of the network society. Dwelling architecture already contains a versatile spatial order and a productive identity to constitute different levels of social order and integration. Applied in the network society, the traditional Finnish housing feature known as "tupa" has the capability of exceeding the physical boundaries of home to integrate the service possibilities of the cyberworld. This "tupa" model, both on the dwelling unit level and on the housing level, is not only suitable for the lifestyle of the network society, but also for the elderly who are gradually growing fragile. The "tupa" model functions as a suitable space for the elderly; firstly, as a place to grow old as productive members of the society after retirement and, secondly, as a place to function as active members fostered by the housing community and by the facilities of the network society. "Theme housing" model which is a reinterpretation of the principles of Finnish rural housing, provides a model which can both facilitate social interaction, and the productivity of different generations, while avoiding isolation and social exclusion in the network society.

Three wastewater treatment systems (WWTS) situated on Cayes in the Belize Barrier Reef System were assessed in terms of the unique public health and environmental circumstances of being a tourist destination surrounded by fragile coral reef. Laughing Bird Caye, Silk Caye, and Little Water Caye are three small cayes that are the staging points for local diving, fishing, and other recreational tourism. All three systems are based upon pour-flush toilets, semi-anaerobic biodigesters and drainage fields. Limitations in cost, available resources, useable area, high infiltration rates of the sand, and salinity of the water have played a major factor in the construction and performance of the WWTS on the Cayes. This thesis aims to form an understanding of treatment efficiency of the WWTS, investigate the effectiveness of decentralized saltwater-based WWTS in comparison to freshwater-based WWTS, and provide recommendations to improve the performance and resource recovery in a manner appropriate for the context in which the systems are deployed. A mathematical model was developed to predict the performance of the WWTS based on available operational and water-quality input data. The model is based on the mass balances of six species: inert solids, fecal solids, bacterial biomass, soluble substrate (i.e. dissolved organic carbon), ammonium and nitrate. Effects of salinity were estimated for the two saltwater-based WWTS. The model predicted the effluent concentrations of fecal solids, soluble biological oxygen demand (BOD), ammonium, and nitrate. A sensitivity analysis was also performed on the predicted effluent treatment efficiency based upon influent load, oxygen concentration and system salinity. Results from Silk Caye and Laughing Bird Caye indicate that varying the number of visitors from seasonal lows to highs has a moderate impact on the effluent fecal solids and soluble BOD in the effluent. Due to the relatively large volume of the WWTS at Little Water Caye, and thus high HRT, varying the number of visitors did not have a significant effect. The model predicted a reduction of nitrogen from the effluent due to settled solids and the assimilation of the nitrogen into bacteria. However the model consistently projected an effluent nitrate concentration (as mg/L as N) between 60 and 63 across the three WWTS. The oxygen concentration within the WWTS had the greatest effect on effluent BOD of the three parameters tested in the sensitivity analysis. Results from the sensitivity analysis indicate that a minimum concentration of 0.95 mg/L of oxygen is required before the model can accurately predict the effluent BOD concentration. The concentration of effluent fecal solids did not significantly change with changes in oxygen concentration. Salinity had a significant effect on the predicted fecal solids and soluble BOD in the effluent. Predicted fecal solids in the effluent wastewater increased approximately 60 percent from freshwater conditions to 4 percent salinity. Similarly, effluent BOD concentration increased strongly with increasing salinity. The increase in concentration is due to the major reduction of substrate-consuming bacteria by cell-die-off. The model predicts that a significant increase in cell die-off begins to occur at 2.4 percent salinity. The predicted effluent of the freshwater-based WWTS on Little Water Caye was compared to 166 wastewater treatment plants operating in Brazil. Comparison between the WWTS on the Caye and the decentralized WWTS in Brazil indicate that the predicted removal efficiencies of total suspended solids and soluble BOD are higher than the measured efficiencies of the WWTS. However, the total nitrogen removal efficiency for the WWTS on the Caye was the least effective; most-likely because the model does not account for denitrification within the biodigester. The comparison between the WWTS illustrates that the predicted removal efficiency of BOD and TSS solids is most likely less in the actual measurement than predicted value from the model. The WWTS on the Cayes were constructed to mitigate the impacts of the wastewater produced by visitors on the general health of the pubic and the environment. Considering the reports of the eutrophication affecting the coral reefs surrounding the Cayes, the WWTS have largely failed in at least one aspect of their purpose. The effluent water quality predicted by the model also suggests that significant concentrations of nitrogen are entering the surrounding ocean habitat as ammonia and nitrate. Recommendations to improve the effluent wastewater quality were separated into three categories based upon the required level of input to realize the recommendation. The input includes the capital cost and labor of the change, the level of buy-in from the users of the system, and the resulting maintenance requirements. The implementation of a urine separation toilet system was proposed as a method to reduce effluent nitrogen entering the environment and to create a resource recovery system (RR) from the already constructed WWTS.

Les nanocomposites à base de polymères sont utilisés depuis plusieurs années dans différents domaines industriels. Ils permettent une amélioration des propriétés des polymères et/ou l'accès à certaines propriétés presque inexistantes sur le polymère de base. Cependant, les polymères utilisés sont souvent d’origine pétrochimique. Au cours des dernières années, l'accumulation de déchets toxiques a, entre autres, conduit à un remplacement progressif de ces matières par des polymères issus des agro-ressources et/ou biodégradables. Notre étude porte sur deux polymères biodégradables utilisés comme matrice dans des nanocomposites, le poly (acide lactique) (PLA) et le poly (butylène succinate) (PBS). Elle a pour but, d'une part, d’améliorer les propriétés barrière aux gaz et à la vapeur d'eau, et d'autre part, de les rendre conducteur de chaleur. Pour ce faire, deux différents types de charges ont été utilisés. La première, la montmorillonite organiquement modifiée (OMMT) a permis d’obtenir une amélioration significative des propriétés mécaniques et barrière du polymère. La seconde, le graphite expansé (EG) a prouvé sa capacité à augmenter les propriétés de transport de chaleur dans les polymères. Une étude bibliographique a révélé que les conditions de mise en œuvre et les affinités entre les deux composés influencent considérablement les propriétés finales du nanocomposite. Ainsi les conditions de mise en œuvre ont été optimisées en faisant varier le temps et les températures de mélange de même que la méthode de mise en œuvre en fonction des charges utilisées. Pour améliorer les interactions entre la matrice et les charges, ces dernières ont été fonctionnalisées par un alkoxysilane, le 3-AminoPropylTrimethoxy-Silane. Les effets de cette fonctionnalisation sur les propriétés mécaniques, barrière, thermiques et thermophysiques des nanocomposites sont présentés. Pour les nanocomposites chargés en EG, l’effet des tailles et de la distribution des tailles des particules sur les conductivités thermiques des nanocomposites a été discuté en se basant sur un modèle de type Maxwell-Garnett basé sur la théorie des champs moyen. / Polymer based nanocomposites have been used for several years in various industrial fields. They allow an improvement of the polymer properties and/or an access to some almost non-existent properties of the neat material. However, the used polymers originated mostly from petrochemical industry. In recent years, the accumulation of toxic waste, among others, led to a gradual replacement of these materials by polymers from agro-resources and/or biodegradables. Our study focuses on two biodegradable polymers used as matrix in nanocomposites, poly (lactic acid) (PLA) and poly (butylene succinate) (PBS). It aims, on one hand, to improve their barrier properties to gases and water vapor, and on the other, to make them heat conductors. To achieve this, two different kinds of fillers were used. The first one, organically modified montmorillonite (OMMT), has achieved a significant improvement in mechanical and barrier properties of the polymer. The second one, expanded graphite (EG), has proven its ability to increase heat transport properties of polymers. A literature review has revealed that the conditions of nanocomposite preparation and the affinities between the two materials greatly influence the final properties of the nanocomposite. Thus the preparation parameters were optimized by varying the melting time and temperature as well as the preparation method according to the fillers used. To improve the interaction between the matrix and the fillers, they were functionalized with an alkoxysilane, 3-aminopropyltrimethoxy-Silane. The effects of this functionalization on the mechanical, barrier, thermal and thermophysical properties of nanocomposites are presented. For EG based nanocomposites, the EG size effect and size distribution on the thermal conductivity of nanocomposites were discussed based on a theoretical model using the Maxwell-Garnett mean-field theory.

Dans les systèmes biologiques, les barrières tissulaires permettent le transport sélectif de molécules du sang au tissu approprié. Un exemple de barrière tissulaire est la barrière hémato-encéphalique (BHE). La BHE protège le cerveau du sang et maintient l'homéostasie du microenvironnement du cerveau, ce qui est essentiel à l'activité et à la fonction neuronale. La caractérisation de cette BHE est importante, car un dysfonctionnement de cette barrière est souvent révélateur de toxicité ou de maladie. Bien que le nombre d'articles publiés dans le domaine du développement et de la caractérisation de la BHE ait été multiplié ces dernières années, la validité des modèles utilisés est encore un sujet de débat. L'avènement de l'électronique organique a créé une occasion unique pour coupler les mondes de l'électronique et de la biologie, à l'aide de dispositifs tels que le transistor électrochimique organique (OECT). OECT constitue un outil très sensible et économique pour diagnostiquer l’intégrité d’une barrière tissulaire. Dans cette étude, nous avons tout d’abord développé trois différents modèles de BHE. Nous avons optimisé l’adhésion des cellules endothéliales cérébrales sur la matière active du transistor. Nous avons ainsi pu établir l'intégration des OECTs avec des cellules immortalisées humaines micro vasculaires cérébrales endothéliales (h CMEC/D3) en tant que modèle in vitro de BHE. Nous avons démontré que la fonction de tissu de la BHE peut être détectée en utilisant cette nouvelle technique. En outre, par cette technique, une perturbation de la barrière (par exemple, provoquée par un composé toxique) pourra être détectée électriquement au moyen d'une mesure de courant. / In biological systems many tissue types have evolved a barrier function to selectively allow the transport of matter from the lumen to the tissue beneath; one example is the Blood Brain Barrier (BBB). The BBB protects the brain from the blood and maintains homeostasis of the brain microenvironment, which is crucial for neuronal activity and function. Characterization of the BBB is very important as its disruption or malfunction is often indicative of toxicity/disease. Though the number of published papers in the field of in vitro BBB has multiplied in recent years, the validity of the models used is still a subject of debate.The advent of organic electronics has created a unique opportunity to interface the worlds of electronics and biology, using devices such as the Organic ElectroChemical Transistor (OECT), which provide a very sensitive way to detect minute ionic currents in an electrolyte as the transistor amplifies the gate current.In this study, we test three different type of BBB in order to develop a stable BBB model. We optimize the adhesion of brain endothelial cell on OECT conducting polymer. We show the integration of OECTs with immortalized human cerebral microvascular endothelial cells as a model of human blood brain barrier, and demonstrate that the barrier tissue function can be detected. Moreover, by this technique, a disruption in the barrier (e.g. caused by a toxic compound) is assessed electrically through a measurement of the drain current. Results show the successful development and validation of an in vitro BBB model. Dynamic monitoring of the barrier properties of the BBB barrier tissue was possible using the OECT.

Cerebral ischaemia by mechanism of thrombosis is one of the leading causes of disability and/or death worldwide, the outcome thereof increasing in severity with advancing age. Cerebral ischaemia triggers a cascade of events including inflammation, blood-brain barrier disruption and apoptosis. It is well known that oestrogen is neuroprotective through various mechanisms including the interruption of inflammation, regulation of thrombosis and delay of apoptosis. This creates a strong factorial interconnection in predicting the consequences of cerebral ischaemia. Since platelets have a central role in thrombosis and inflammation, their ultrastructure being altered in conditions of inflammatory and thrombotic derivation, the question arises whether chemical analysis of coagulation factors and ultrastructural analyses of platelet morphology may provide further insight into the role of oestrogen during ischaemic insult associated with stroke. Accordingly, an exclusively hyperglycaemic modification of the two-vessel occlusion model for inducing experimental cerebral ischaemia was established, since pre-ischaemic hyperglycaemia is known to intensify the outcome of cerebral ischaemic injury. Consequent neural tissue injury levels were correlated for three experimental groups (males, cyclic and acyclic females) of Sprague Dawley rats at vital times, to the presence of oestrogen as well as changes in coagulation factors and ultrastructure. This design allowed for an association to be formed between cerebral ischaemia, inflammation and thrombotic potential. Collectively the results strongly suggest that oestrogen is indeed neuroprotective through various actions including roles in the regulation of thrombosis and inflammation, targeting neural cells through the inhibition of apoptosis and exerting anti-inflammatory and antioxidant effects. It is evident that under the influence of oestrogen in cyclic females, there is reduced neural tissue injury as well as a lesser degree of inflammation evident in coagulation factor analysis and platelet activation morphology when compared to males and acyclic females. Oestrogen therefore exerts positive effects on the outcome of cerebral ischaemia through mechanisms which regulate inflammation, thrombosis and apoptosis. Furthermore it is unmistakeable that neural injury is closely shadowed, if not preceded, by inflammatory changes in the coagulation system, particularly manifested in platelet ultrastructure. It is therefore suggested that platelets may be used successfully to follow the progression of events of cerebral ischaemia and possibly assist in the assessment of treatment strategies and their effects on haemostasis. This research advances the understanding that inflammation is evident soon after ischaemic insult and if such inflammation is not curbed, necrosis of platelets and more severe injury to neural tissue may follow. Therefore, the development of agents which not only target thrombosis, but also which control inflammation must be explored to advance treatment strategies. It is proposed that even before it is determined whether a stroke has been caused by thromboembolism or haemorrhage; it will be beneficial to immediately target inflammation in order to prevent most severe consequences in human patients. / Thesis (PhD)--University of Pretoria, 2013. / gm2013 / Anatomy / unrestricted

Polymorphic light eruption (PLE) is the most common photodermatosis, affecting ~17% of the population. PLE is a delayed-type hypersensitivity response to an antigen induced by solar ultra-violet radiation (UVR). Its effects vary between patients, but the main symptom is a non-scarring, red papular rash in areas exposed to UVR. An effective therapy is low dose ultra-violet B (NBUVB) phototherapy. It is thought that NBUVB phototherapy desensitises the skin to further UVR exposure, but the mechanism by which this happens is unknown. Current immune based studies have been unable to clarify a mechanism as to how PLE arises. However, research in other skin diseases, such as psoriasis and atopic dermatitis, has shown that the barrier function of the skin is compromised by these disorders. Furthermore, research in lesional PLE skin showed an increase in barrier permeability of the skin. Recent research has specifically linked claudin proteins of tight junctions to the barrier dysfunction. Therefore, this study used quantitative immunofluorescent staining to measure tight junction (TJ) proteins and other barrier proteins of interest. Barrier function was also measured by transepidermal water loss (TEWL); a tracer dye penetration assay was used to measure TJ barrier function specifically. All measurements were made in non-lesional PLE skin, as compared to skin from healthy human volunteers. In photoprotected PLE skin the TJ protein claudin-1 was significantly reduced compared to healthy skin. The use of a tracer dye highlighted there was a reduction in TJ barrier function in PLE skin compared to healthy individuals. PLE and healthy skin were then exposed to ultra-violet B (UVB) and 24h later TJ proteins and TJ barrier function were measured. There was no change to claudin-1 after UVB exposure in PLE skin, but claudin-7 was reduced and claudin-12 increased. In contrast, in UVB-irradiated skin in normal controls after UVB exposure claudin-7 and claudin-12 were both increased, whilst claudin-1 was reduced. In PLE patients there was no further change to TJ barrier function, however, in normal controls, skin TJ barrier function was reduced post UVB. Both in healthy and PLE skin TEWL was unchanged before and after UVB exposure. Lastly TJ proteins were investigated after NBUVB in PLE patients. There was a further reduction in claudin-1 in PLE patients as well as a reduction in the TJ protein occludin, however the stratum corneum was significantly thickened. It could be suggested that this is a compensatory measure for the reduction seen in TJ barrier proteins, however further studies are needed to understand this. These data show significant differences in the TJ skin barrier in patients with PLE as compared to healthy human volunteers before and after UVB exposure. Furthermore, in PLE skin there is a significant change to the epidermis after NBUVB phototherapy. These data demonstrate that TJ protein expression and function is altered in PLE skin and may contribute to aetiology of the disorder, however the role of TJ barrier in aetiology is yet to be firmly established.