Trinta e oito vacas leiteiras, 19 da raça Holandês e 19 da raça Girolando (½ e ¾), foram submetidas à elevadas temperaturas por restrição à sombra durante o período entre a ordenha da manhã e da tarde. Mensurações de temperatura retal, frequências respiratória e cardíaca e escore de ofegação, além de coletas de sangue, foram realizadas nos períodos pré ordenha. Produção de leite e características físico-químicas do mesmo foram acessadas. Durante os períodos de coleta se avaliou características ambientais com vistas a calcular um índice de temperatura e umidade (ITU). Primeiramente se buscou estabelecer diferenças em parâmetros fisiológicos, de leite e sangue de vacas com diferentes porcentagens de alelos oriundos da raça Holandesa no genoma (100, 75 e 50%) em função de aumentos no ITU. Animais puros Holandês apresentaram características que condizem com menor tolerância ao calor do que animais Girolando. Em um segundo momento se analisou o efeito do estresse térmico sobre a permeabilidade das junções firmes das células epiteliais da glândula mamária de vacas Holandês. Além disso, buscou-se perceber alterações na estabilidade do leite ao teste do álcool decorrentes de um possível aumento na permeabilidade de tais estruturas celulares. O parâmetro utilizado para aferir tal efeito foi o nível sanguíneo de lactose no plasma. Alterações lácteas foram percebidas em função de aumentos de ITU e se devem principalmente à reduções em produção de leite. Aumento nos parâmetros fisiológicos não influenciou a estabilidade do leite. Leite instável apresentou maior teor de lactose. Maior número de dias em lactação pode ser o responsável pela redução na estabilidade do leite. Percebeu-se relação inversamente proporcional entre permeabilidade das junções firmes e estabilidade do leite, porém o estado de estresse térmico, ao contrário do esperado, não apresentou influência nas células da glândula mamária. / Thirty-eight dairy cows, 19 Holstein and 19 Girolando (½ e ¾), were submitted to elevated temperatures due to shade deprivation between morning and evening milkings. Rectal temperature, heart and respiratory rates and panting score measurements, besides blood samples collection, were performed before each milking. Milk production and its physical-chemical characteristics were studied. Climatic variables were measured during data collection to calculate a temperature-humidity index (THI). Firstly, the study evaluated changes in physiological, blood and milk parameters according to the percentage of alleles derived from the Holstein breed (100, 75 and 50%) due to increases in THI. Pure Holstein cows presented characteristics that indicate lower heat tolerance than Girolando cows. Secondly, heat stress effects on mammary gland cells tight junctions permeability were evaluated in Holstein cows. Alterations in milk stability due to changes in permeability were also assessed. Plasma lactose was the parameter used to detect changes in tight junctions permeability. Modifications in milk characteristics were probably due to reduction in milk production. Increase in physiological parameters did not affect milk stability. Unstable milk samples presented higher lactose levels. Higher days in milk might be the main responsible for reductions in milk stability. There was an inversely proportional relation between tight junctions permeability and milk stability to the ethanol test, although heat stress, contradicting the expected, did not influenced mammary gland cells.

Made available in DSpace on 2015-03-03T15:16:23Z (GMT). No. of bitstreams: 1 DarlanAM.pdf: 1538525 bytes, checksum: af051bdcc4894d0bf236b2e6f943baa9 (MD5) Previous issue date: 2008-09-29 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Esta tese apresenta um abrangente e atualizado estudo de algumas propriedades f?sicas da mol?cula do DNA, tais como seus aspectos termodin?micos (calor espec?fico) e eletr?nicos (transmissividade eletr?nica, fator de localiza??o, entre outros). A mol?cula do DNA pode ser considerada uma seq??ncia simb?lica de quatro letras: guanina (G), adenina (A), citosina (C) e timina (T). Ela ? usualmente descrita como uma cadeia bidimensional aleat?ria com correla??o de curto-alcance, mas n?o h? impedimentos para que a cadeia seja crescida seguindo seq??ncias quasi-peri?dicas, como por exemplo, as sequ?ncias de Fibonacci e Rudin-Shapiro. Com o intuito de investigar a relev?ncia das correla??es subjacentes nas distribui??oes dos nucleot?deos, comparamos os resultados para a sequ?ncia gen?mica do DNA (Ch22) com as duas seq??ncias artificiais citadas acima, que possuem correla??ao de longo alcance. A an?lise do calor espec?fico ? feita considerando-se formalismos apropriados; o cl?ssico, utilizando a distribui??o de Maxwell-Boltzmann; a descri??oo qu?ntica, utilizando a distribui??o de Fermi-Dirac; e o formalismo da n?o-extensividade, usando a entropia de Tsallis. Os espectros de energias s?o calculados utilizando-se a equa??o de Schrodinger unidimensional na aproxima??o de liga??o forte. N?s calculamos tamb?m a transmissividade eletr?nica, o comprimento de localiza??o, bem como I (corrente) vs V (potencial), curva que caracteriza as propriedades el?tricas de uma mol?cula de DNA dupla fita. O modelo te?rico considerado faz uso de um Hamiltoniano efetivo com aproxima??o de liga??o-forte descrevendo um el?tron movendo-se em uma cadeia com um simples orbital por s?tio e intera??es entre vizinhos mais pr?ximos, juntamente com a equa??o de Schrodinger, e a muito conveniente t?cnica da matriz de transfer?ncia

Trinta e oito vacas leiteiras, 19 da raça Holandês e 19 da raça Girolando (½ e ¾), foram submetidas à elevadas temperaturas por restrição à sombra durante o período entre a ordenha da manhã e da tarde. Mensurações de temperatura retal, frequências respiratória e cardíaca e escore de ofegação, além de coletas de sangue, foram realizadas nos períodos pré ordenha. Produção de leite e características físico-químicas do mesmo foram acessadas. Durante os períodos de coleta se avaliou características ambientais com vistas a calcular um índice de temperatura e umidade (ITU). Primeiramente se buscou estabelecer diferenças em parâmetros fisiológicos, de leite e sangue de vacas com diferentes porcentagens de alelos oriundos da raça Holandesa no genoma (100, 75 e 50%) em função de aumentos no ITU. Animais puros Holandês apresentaram características que condizem com menor tolerância ao calor do que animais Girolando. Em um segundo momento se analisou o efeito do estresse térmico sobre a permeabilidade das junções firmes das células epiteliais da glândula mamária de vacas Holandês. Além disso, buscou-se perceber alterações na estabilidade do leite ao teste do álcool decorrentes de um possível aumento na permeabilidade de tais estruturas celulares. O parâmetro utilizado para aferir tal efeito foi o nível sanguíneo de lactose no plasma. Alterações lácteas foram percebidas em função de aumentos de ITU e se devem principalmente à reduções em produção de leite. Aumento nos parâmetros fisiológicos não influenciou a estabilidade do leite. Leite instável apresentou maior teor de lactose. Maior número de dias em lactação pode ser o responsável pela redução na estabilidade do leite. Percebeu-se relação inversamente proporcional entre permeabilidade das junções firmes e estabilidade do leite, porém o estado de estresse térmico, ao contrário do esperado, não apresentou influência nas células da glândula mamária. / Thirty-eight dairy cows, 19 Holstein and 19 Girolando (½ e ¾), were submitted to elevated temperatures due to shade deprivation between morning and evening milkings. Rectal temperature, heart and respiratory rates and panting score measurements, besides blood samples collection, were performed before each milking. Milk production and its physical-chemical characteristics were studied. Climatic variables were measured during data collection to calculate a temperature-humidity index (THI). Firstly, the study evaluated changes in physiological, blood and milk parameters according to the percentage of alleles derived from the Holstein breed (100, 75 and 50%) due to increases in THI. Pure Holstein cows presented characteristics that indicate lower heat tolerance than Girolando cows. Secondly, heat stress effects on mammary gland cells tight junctions permeability were evaluated in Holstein cows. Alterations in milk stability due to changes in permeability were also assessed. Plasma lactose was the parameter used to detect changes in tight junctions permeability. Modifications in milk characteristics were probably due to reduction in milk production. Increase in physiological parameters did not affect milk stability. Unstable milk samples presented higher lactose levels. Higher days in milk might be the main responsible for reductions in milk stability. There was an inversely proportional relation between tight junctions permeability and milk stability to the ethanol test, although heat stress, contradicting the expected, did not influenced mammary gland cells.

TRAF4 est un gène fréquemment surexprimé dans les carcinomes suggérant qu’il y joue un rôle. Tandis que la protéine TRAF4 est majoritairement localisée dans les jonctions serrées (JS) des cellules épithéliales mammaires (CEM) normales, elle s’accumule dans le cytoplasme des CEM malignes. Dans cette étude, nous montrons que TRAF4 possède un nouveau domaine liant les phosphoinositides (PIP) et que ce dernier est requis pour son recrutement aux JS. Des analyses moléculaires et structurales ont montré que le domaine TRAF de TRAF4 forme un trimère pouvant lier jusqu’à trois molécules de lipides grâce à des résidus basiques présents à la surface. Des études cellulaires indiquent que TRAF4 régule négativement les JS et augmente la migration cellulaire. Ces deux fonctions sont dépendantes de sa capacité à lier les PIPs. Notre travail suggère que la surexpression de TRAF4 pourrait contribuer à la progression des cancers du sein en déstabilisant les JS et en favorisant la migration cellulaire. / TRAF4 (tumor necrosis factor (TNF) receptor-associated factor 4) is frequently overexpressed in carcinomas suggesting a specific role in cancer. While TRAF4 protein is predominantly found at tight junctions (TJ) in normal mammary epithelial cells (MEC), it accumulates in the cytoplasm of malignant MEC. How TRAF4 is recruited and functions at TJ is unclear. Here we show that TRAF4 possesses a novel phosphoinositide (PIP)- binding domain crucial for its recruitment to TJ. Molecular and structural analyses revealed that the TRAF domain of TRAF4 exists as a trimer which binds up to 3 lipids using basic residues exposed at its surface. Cellular studies indicated that TRAF4 acts a negative regulator of TJ and increases cell migration. These functions are dependent from its ability to interact with PIPs. Our results suggest that TRAF4 overexpression might contribute to breast cancer progression by destabilizing TJ and favoring cell migration.

Trinta e oito vacas leiteiras, 19 da raça Holandês e 19 da raça Girolando (½ e ¾), foram submetidas à elevadas temperaturas por restrição à sombra durante o período entre a ordenha da manhã e da tarde. Mensurações de temperatura retal, frequências respiratória e cardíaca e escore de ofegação, além de coletas de sangue, foram realizadas nos períodos pré ordenha. Produção de leite e características físico-químicas do mesmo foram acessadas. Durante os períodos de coleta se avaliou características ambientais com vistas a calcular um índice de temperatura e umidade (ITU). Primeiramente se buscou estabelecer diferenças em parâmetros fisiológicos, de leite e sangue de vacas com diferentes porcentagens de alelos oriundos da raça Holandesa no genoma (100, 75 e 50%) em função de aumentos no ITU. Animais puros Holandês apresentaram características que condizem com menor tolerância ao calor do que animais Girolando. Em um segundo momento se analisou o efeito do estresse térmico sobre a permeabilidade das junções firmes das células epiteliais da glândula mamária de vacas Holandês. Além disso, buscou-se perceber alterações na estabilidade do leite ao teste do álcool decorrentes de um possível aumento na permeabilidade de tais estruturas celulares. O parâmetro utilizado para aferir tal efeito foi o nível sanguíneo de lactose no plasma. Alterações lácteas foram percebidas em função de aumentos de ITU e se devem principalmente à reduções em produção de leite. Aumento nos parâmetros fisiológicos não influenciou a estabilidade do leite. Leite instável apresentou maior teor de lactose. Maior número de dias em lactação pode ser o responsável pela redução na estabilidade do leite. Percebeu-se relação inversamente proporcional entre permeabilidade das junções firmes e estabilidade do leite, porém o estado de estresse térmico, ao contrário do esperado, não apresentou influência nas células da glândula mamária. / Thirty-eight dairy cows, 19 Holstein and 19 Girolando (½ e ¾), were submitted to elevated temperatures due to shade deprivation between morning and evening milkings. Rectal temperature, heart and respiratory rates and panting score measurements, besides blood samples collection, were performed before each milking. Milk production and its physical-chemical characteristics were studied. Climatic variables were measured during data collection to calculate a temperature-humidity index (THI). Firstly, the study evaluated changes in physiological, blood and milk parameters according to the percentage of alleles derived from the Holstein breed (100, 75 and 50%) due to increases in THI. Pure Holstein cows presented characteristics that indicate lower heat tolerance than Girolando cows. Secondly, heat stress effects on mammary gland cells tight junctions permeability were evaluated in Holstein cows. Alterations in milk stability due to changes in permeability were also assessed. Plasma lactose was the parameter used to detect changes in tight junctions permeability. Modifications in milk characteristics were probably due to reduction in milk production. Increase in physiological parameters did not affect milk stability. Unstable milk samples presented higher lactose levels. Higher days in milk might be the main responsible for reductions in milk stability. There was an inversely proportional relation between tight junctions permeability and milk stability to the ethanol test, although heat stress, contradicting the expected, did not influenced mammary gland cells.

Abstract It is a generally accepted dogma that sepsis disturbs skin function and wound healing, but surprisingly there is only remote pathophysiological evidence available behind that presumption. As the skin is the largest defensive barrier, the skin dysfunction in severe sepsis deserves more attention. In this study, the suction blister model was used to create experimental wounds. The study population included 44 patients with severe sepsis and 15 controls. The blister fluid was collected to analyse cytokine profile of the skin. The transepidermal water loss and blood flow from the wound were measured. A 4mm biopsy was taken under local anaesthesia on the first and the eighth day of the study from the healthy looking skin. Then 15 healing suction blisters were excised. Serum samples were also collected on the first day of the study. The barrier restoration was diminished, and the inflammation in the wound was more intense in severe sepsis than in the controls. The expression of the basement membrane components Laminin-332 and type IV collagen decreased during the septic disease, but increased over the next 3 months without achieving the level oft he controls. The expression of tight junction proteins remained nearly intact in the healing wound in severe sepsis compared to the controls. The expression of occludin on the leading edge of the migrating keratinocytes was more restricted and late in severe sepsis compared to the controls. The levels of the tumour necrosis factor (TNF), interleukin-10 (IL-10) and IL-6 in skin blister fluid were higher in the sepsis compared to controls. The blister fluid and serum cytokine response in the sepsis differed since the levels of epidermal growth factor, vascular endothelial growth factor, TNF and basic fibroblastic growth factor (bFGF) in the blister fluid did not correlate with the levels of serum. The septic patients with multiple organ failure had higher levels of several cytokines than patients without organ failure. Survivors had lower levels of IL-10 and bFGF in blister fluid than the non-survivors. This study offers novel findings for skin and wound healing in sepsis. Together, all the findings suggest that skin dysfunction in severe sepsis exists even when the most profound structures remain intact. Understanding these mechanisms of impaired wound healing can improve future treatments, such as the timing of surgery. / Tiivistelmä Sepsiksen ajatellaan heikentävän haavanparanemista, mutta tieteellistä näyttöä on niukasti. Iholla on keskeinen osa elimistön puolustuksessa ja tasapainon ylläpidossa, joten sen toiminnan häiriintyminen systeemisessä tulehduksessa ansaitsee suuremman huomion. Imurakkulahaavat tehtiin 44 septiselle potilaalle ja 15 kontrollille. Haavoista mitattiin veden haihtumista ja veren virtausta sekä otettiin imurakkulaneste näytteeksi sytokiinimäärityksiä varten. Tutkimuksen ensimmäisenä ja kahdeksantena päivänä otettiin 4mm biopsiat terveeltä iholta ja 15 potilaalta poistettiin näytteeksi paraneva imurakkulahaava. Seeruminäytteet otettiin tutkimuksen ensimmäisenä päivänä. Veden haihtuminen haavalta oli voimakkaampaa eli ihon barrierin palautuminen oli hidastunut septisillä potilailla verrattuna kontrolleihin. Haavassa havaittu tulehdus oli sepsiksessä voimakkaampi. Tyvikalvon komponenttien Laminiini-332:n ja tyypin IV kollageenin ilmentyminen oli vähäisempää sepsiksen aikana ja lisääntyi 3kk kohdalla, mutta ei kuitenkaan saavuttanut kontrollien tasoa. Tiivisliitosproteiinien ilmentyminen oli lähes muuttumatonta sepsiksessä kontrolleihin verrattuna. Okludiinin ilmentyminen sen sijaan paranevassa haavassa vaeltavien keratinosyyttien etureunassa oli rajoittuneempaa ja myöhäisempää sepsiksessä kuin kontrolleilla. Sytokiineistä tuumorinekroositekijä (TNF), interleukiini-10 (IL-10) ja IL-6 olivat koholla imurakkulanesteessä verrattuna kontrolleihin. Epidermaalinen kasvutekijä, verisuonten endoteelikasvutekijä, TNF ja perusfibroplastinen kasvutekijä (bFGF) pitoisuudet rakkulanesteessä erosivat seerumin pitoisuuksista eli ihon sytokiiniprofiili erosi systeemisestä sytokiiniprofiilista. Potilailla, joilla oli monielinvaurio, todettiin korkeampia sytokiinipitoisuuksia. Potilailla, jotka menehtyivät 30 vrk kuluessa, oli korkeammat pitoisuudet IL-10 ja bFGF rakkulanesteessä. Tämä tutkimus tarjoaa uutta tietoa ihosta ja haavanparanemisesta sepiksessä. Tulosten perusteella voidaan todeta, että ihon toimintahäiriö on sepsiksessä todellinen, vaikka kaikkein perustavimmat rakenteet säilyvät muuttumattomina. Toimintahäiriön mekanismien ymmärtäminen voisi auttaa septisen potilaan hoidossa, kuten kirurgisten toimenpiteiden ajoittamisessa paranemisen kannalta mahdollisimman otolliseen aikaan.

Abstract Lung cancers and chronic obstructive pulmonary disease (COPD) are the most common smoking-related lung diseases and both have high mortality rate. Tight junctions (TJ) are apical junctions between epithelial cells that regulate the permeability of epithelium and form the tight junction along with occludin. Dysfunction of the TJ and dysregulation of TJ proteins leads to a loss of cell-cell adhesion and a loss of cohesion as well as epitheliomesenchymal transition. These increase invasion of lung carcinomas and possibly predispose to the exacerbations in COPD. Therefore, the aim of this thesis was to study expression and regulation of claudins in different lung carcinomas and COPD. Carcinomas expressed claudins 1, 2, 3, 4, 5 and 7 in different variations. Claudin 5 expression was weak in all carcinoma types. Strong claudin 1, 4 and 7 expression was associated with better survival in squamous cell carcinoma and adenocarcinoma. Claudin 3 expression was associated with COPD in large airways. Claudins 3 and 4 was found to be stronger in small airways of smokers and COPD patients than in non-smokers. Transcription factor snail had prognostic value in lung carcinomas. Negative snail expression was associated with longer life expectancy in lung carcinoma patients. Negative snail expression was associated with up-regulated claudin 5 and 7 expression, while strong expression was associated with low claudin 1 and 3 expression. Transcription factors slug and twist were inversely associated with claudins 3 and 4 in small and large airways. Slug expression was higher in non-smokers than in COPD patients and smokers. Transcription factor knockdown increased claudin expression in normal bronchial cell line. Except for claudin 2 and 7, which were decreased. Adenocarcinoma-like cell line was not affected by snail knockdown and in squamous cell carcinoma-like cell line claudin 3, 4 and 7 expression was increased. Transcription factor snail knockdown inhibited invasion of cell lines. Twist knockdown increased transepithelial resistance in normal bronchial cell line indicating higher barrier function in cell layer. / Tiivistelmä Keuhkosyöpä ja keuhkoahtaumatauti ovat yleisiä tupakoinnin aiheuttamia keuhkosairauksia, joissa on korkea kuolleisuus. Tupakointi aiheuttaa muutoksia keuhkojen epiteelisoluissa ja solujen välisissä liitoksissa. Tiivisliitokset solujen välillä säätelevät epiteelin rakennetta ja läpäisevyyttä. Klaudiinit ovat proteiineja, jotka muodostavat tiivisliitoksen yhdessä okkludiinin kanssa. Tiivisliitos proteiinien toimintahäiriöt voivat johtaa solujen välisten liitosten katoamiseen ja epiteelin hajoamiseen sekä epiteelisolujen muuntumiseen mesenkymaalisten solujan kaltaisiksi. Nämä seikat lisäävät invaasiota keuhkosyövissä ja saattavat altistaa pahenemisvaiheisiin keuhkoahtaumataudissa. Väitöskirjassa tutkittiin klaudiinien ilmentymistä ja säätelyä keuhkosyövässä ja keuhkoahtaumataudissa. Klaudiinien1, 2, 3, 4, 5 ja 7 esiintyminen keuhkosyövän histologisissa alatyypeissä vaihteli. Klaudiinien 1, 4 ja 7 voimakas ilmentyminen voitiin yhdistää pidempään elinikään potilailla, joilla oli levyepiteeli- tai adenokarsinooma. Klaudiini 3:n ilmentyminen liittyi keuhkoahtaumatautiin suurissa hengitysteissä. Klaudiinien 3 ja 4 voimakas ilmeneminen pienissä ilmateissä oli yleisempää keuhkoahtaumatautipotilailla ja tupakoitsijoilla kuin tupakoimattomilla henkilöillä. Transkriptiotekijä snailin puuttuminen keuhkosyövässä liittyi potilaiden pidempään elinaikaan. Klaudiinien 5 ja 7 ilmeneminen oli voimakkaampaa, kun snailin määrä oli vähäinen. Klaudiinien 1 ja 3 ilmeneminen väheni snail:in ollessa voimakas keuhkosyövässä. Traskriptiotekijöiden (slug ja twist) ilmeneminen liittyi käänteisesti klaudiinien ilmentymiseen pienissä ja suurissa ilmateissä. Slugin ilmeneminen oli voimakkaampaa tupakoimattomilla henkilöillä kuin tupakoivilla tai keuhkoahtaumatautia sairastavilla. Transkriptiotekijöiden snail, slug ja twist toiminnan estäminen lisäsi klaudiinien määrää normaaleissa keuhkon epiteelisoluissa. Poikkeuksen muodostivat klaudiinit 2 ja 7, joiden määrä väheni kun snail:in toiminta oli estetty. Adenokarsinooma-soluissa snailin estolla ei ollut vaikutusta, ja levyepiteelisyövän soluissa klaudiinien 3, 4 ja 7 määrä kasvoi. Snail myös vähensi solujen invaasiota. Transkriptiotekijä twistin toiminnan esto normaaleissa keuhkoepiteelisoluissa nosti solumaton läpi kulkevan sähkön resistenssiä, mikä on osoitus tiiviistä solujen välisistä liitoksista.

La question cruciale dans l'exploration du stockage ultime à haute densité est l'anisotropie magnéto-cristalline (MCA) qui provient du couplage spin-orbite. Utilisant à la fois la méthode des liaisons fortes et les calculs « premiers principes », nous calculons la MCA de nanocristaux de fer et de cobalt qui peuvent être obtenus par croissance épitaxiale sur un substrat SrTiO3 avec un contrôle remarquable de leur taille, forme et structure. Afin de définir une décomposition locale appropriée de la MCA, nous avons implémenté le « Théorème de Force » à l'aide d'une formulation grand-canonique dans le code QUANTUM ESPRESSO ainsi que dans notre modèle de liaisons fortes. Il est intéressant de noter que pour les deux éléments, la MCA totale de nanocristaux isolés est largement dominée par les facettes (001) dont il résulte un comportement opposé: une anisotropie « hors-plan » pour les nanocristaux (contenant plusieurs centaines d'atomes) de fer et « dans le plan » pour ceux de cobalt. Nous avons également mis en évidence un fort renforcement de la MCA pour les petits clusters (contenant quelques atomes seulement) déposés sur un substrat SrTiO3. En conséquence, nous prévoyons que les nanocristaux de fer (même de très petite taille) devraient être magnétiquement plus stables et sont donc de bons candidats potentiels pour le stockage magnétique. Enfin, notre analyse MCA résolu en orbitales s'applique également à d'autres système et permet, par exemple, de prédire le comportement de la MCA de films minces magnétiques après déposition de matériaux organiques comme le graphène ou de molécules tel C60. / The crucial issue in exploring ultimate density data storage is magneto-crystalline anisotropy (MCA) which originates from spin-orbit coupling. Using both tight-binding and first-principles methods, we report the MCA of Fe and Co nanocrystals that can be grown epitaxially on SrTiO3 with a remarkable control of their size, shape and structure. In order to define the proper local decomposition of MCA, we implemented the “Force Theorem” within the grand-canonical formulation in QUANTUM ESPRESSO as well as in our tight-binding model. Interestingly, for both elements, the total MCA of free nanocrystals is largely dominated by (001) facets resulting in the opposite behavior: out-of-plane and in-plane magnetization direction is favored in Fe and Co nanocrystals (containing up to several hundred atoms), respectively. We also find a strong enhancement of MCA for small clusters (containing only several atoms) upon their deposition on a SrTiO3 substrate. As a consequence, we predict that the Fe nanocrystals (even rather small) should be magnetically stable and are thus good potential candidates for magnetic storage devices. Finally, our rather general orbital-resolved analysis of MCA applies also to other systems and allows, for example, predicting the MCA behavior of magnetic thin films upon covering by various organic materials such as graphene or C60 molecule.

La barrière épidermique du stratum corneum (SC) est doublée par une barrière secondaire des jonctions serrées (JS) qui influent sur la formation de barrière principale. Dans mes travaux, je me suis concentré sur l'étude de la présence et l'évolution des éléments jonctionnelles composants ces deux barrières ; les cornéodesmosomes au niveau du SC et les JSs au niveau de la granuleuse. En plus, je me suis intéressé à l'implication des glycanes dans la fonction barrière épidermique. Ces travaux ont été réalisés soit dans un contexte physiologique soit par la modulation de la barrière épidermique par des facteurs intrinsèques et extrinsèques. Nos résultats confirment que les JSs jouent un rôle subalterne par rapport à la barrière du SC et montrent que les glycanes persistent à la surface des cornéocytes humains. La composition et la répartition utlrastructurale des glycanes évoluent à travers les assises du SC jusqu'à la desquamation d'une manière concordante avec la répartition des cornéodesmosomes. Certaines modifications intrinsèques naturelles lors du vieillissement ou pathologiques notamment l'état pelliculaire et la dermatite atopique, ont permis d'appréhender le rôle de ces composants dans la cohésion du SC et la prestance d'une barrière fonctionnelle. Les modifications extrinsèques de la barrière par l'application de solvants, d'excipients ou de formulations perméabilisantes montrent l'importance de l'organisation utlrastructurale des composants jonctionnelles et non jonctionnelles du SC dans le maintien d'une barrière efficace / The stratum corneum (SC) barrier is doubled by the secondary barrier of tight junctions which influences the formation of the main barrier. In my work, I focused on the study of the junctional elements composing those two barriers; corneodesmosomes in the SC and the tight junction at the granular layer level. In addition, I got interested in the involvement of glycans in the epidermal barrier function. This work was carried out either in skin physiological conditions or by the modulation of the epidermal barrier by intrinsic or extrinsic factors. Our results confirm that tight junctions play a subordinate role compared to the SC barrier and that glycans remain present at the surface of human corneocytes. The composition and the ultrastructure distribution of glycans evolve from the SC compactum to the SC disjunctum, towards desquamation in a comparable manner to the repartition of corneodesmosomes. Natural intrinsic changes during aging and pathological changes, including dandruff and atopic dermatitis, helped us to understand the role of those components in the cohesion of the SC and the conservation of functional barrier. Extrinsic modulation of the barrier by the application of solvents, excipients or topical formulations shows the importance of the ultrastructural organization of junctional and non-junctional SC components in maintaining an effective barrier

Inflammatory bowel disease (IBD), which includes the subtypes Crohn's disease (CD) and ulcerative colitis (UC), is a common disease particularly in the Western world. IBD is characterised by inflammation of the small intestine and/or colon. The two subtypes affect different gut locations but both show an increased intestinal permeability or the 'leaky gut syndrome'. This led to the hypothesis that tight junction (TJ) proteins expressed in the epithelium may affect the intestinal permeability as a cause or effect of IBD.Initially, variants in the CARD15, IL23R and ATG16L1 genes, previously associated with an increased risk of IBD, were genotyped in a cohort of 500 IBD (295 CD and 205 UC) patients and 877 matched controls. These variants were significantly associated in our cohort. A random effects meta-analysis was undertaken on all previously reported CD associations with the variant rs2241880 from ATG16L1 (n=25, p=0.0017, OR: 1.36 95% CI 1.12-1.66) and with rs11209026 from IL23R (n=26, p=0.0006, OR: 0.37 95% CI 0.21-0.67), showing pooled odds ratios consistent with those reported in our cohort. Individuals carrying >1 CARD15 mutant variant were found to have a 2.5 fold increased risk of CD (p=0.0001). Candidate TJ proteins were chosen on the basis of previous reported associations and through the investigation of the claudin proteins which are abundant at TJs. Twenty one candidate genes were selected and 79 variants successfully genotyped in up to 1063 IBD (502 CD and 478 UC) and 870 control patients. Significant associations were detected with variants in the CLDN1, CLDN5 and CDH1 genes with CD; CLDN5, CLDN8 and CDH1 variants were associated to IBD; and the rs7791132 variant (between CLDN4 and ELN) and a CDH1 variant were associated to UC. The CLDN1 rs6809685 variant trended towards association in a Toronto ascertained IBD replication cohort (genotypic p=0.04, allelic p=0.06) suggesting this may be a novel IBD susceptibility variant. Small intestinal biopsies from CD patients with known rs6809685 genotypes showed a dose dependent reduced immunohistochemical staining of claudin 1 with carriage of the mutant G allele. Claudin 1 helps seal TJs and reduced levels may increase risk of CD.Peroxisome proliferator activator receptors (PPARs) can directly affect TJ proteins and could therefore affect intestinal permeability. Twelve PPARγ variants were genotyped in up to 1050 IBD (502 CD and 467 UC) and 725 control patients. Significant genotypic associations were found with the rs2067819 variant in CD (p=0.05) and IBD (p=0.02), and also the rs13099634 variant in UC (P=0.02). There was a strong gender difference particularly for rs2067819 and rs4135247, where allelic associations were highly significant and increased risk of IBD in men (p=0.01 and p=0.007 respectively). However no significant associations were found in the female cohort. Troglitazone a PPARα agonist increased Caco2 cell transepithelial electrical resistance (TEER), a marker of TJ integrity, and increased expression of claudins -3 and -4. In contrast, the PPARα antagonist GW6471 reduced the TEER without causing cell death and PPARγ ligands did not affect TEER measurements. In summary, using a robust cohort of cases and controls the data indicates that variants in genes encoding TJ proteins may affect susceptibility to IBD and that PPARs can regulate these proteins altering intestinal permeability.