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No description available.

But : La perte de l’intégrité de la barrière hémo-encéphalique (BHE) est l’une des caractéristiques principales de la sclérose en plaques. Cette augmentation de la perméabilité est associée à une désorganisation des molécules de jonction serrée et à une augmentation de l’expression de molécules d’adhérence essentielles à l’extravasation des cellules immunitaires. Identifier de nouvelles molécules impliquées dans ce processus est donc crucial pour le développement de nouvelles thérapies contre la sclérose en plaques visant à promouvoir l’intégrité de la BHE et à diminuer la migration des leucocytes dans le système nerveux central (SNC) au cours du processus neuro-inflammatoire. Dans cette étude, le rôle spécifique de la molécule d’adhérence ALCAM, qui est exprimé à la surface des cellules endothéliales de la BHE (CE-BHE) et de certains sous-types de leucocytes, a été évalué. Méthodologie : À l’aide d’une analyse protéomique exhaustive, notre laboratoire a identifié ALCAM comme étant une molécule d’adhérence surexprimée par les CE-BHE mises en culture dans un milieu pro-inflammatoire. Dans le but d’étudier le rôle spécifique d’ALCAM durant la diapédèse leucocytaire, nous avons induit chez des souris de type sauvages et des souris ALCAM déficientes l’encéphalite auto-immune expérimentale (EAE), le modèle animal de la sclérose en plaques. Le rôle d’ALCAM a aussi été étudié à l’aide d’un système d’adhérence sous flux laminaire. Cet appareil, qui imite un capillaire cérébral, permet de suivre en temps réel le mouvement des leucocytes, soumis à une pression physiologique, dans un tube couvert à sa base par des CE-BHE. Résultats : En utilisant ce système d’adhérence, j’ai pu démontrer que des anticorps dirigés contre ALCAM réduisent de façon significative le roulement et l’adhérence de monocytes CD14+ humains à la surface de CE-BHE. Par ailleurs, ces anticorps préviennent de façon marquée la diminution de la vitesse moyenne des cellules au cours de l’expérience. Par le fait même, j’ai aussi observé une réduction significative de l’extravasation des monocytes traités avec de l’anti-ALCAM au travers de CE-BHE dans un modèle statique de migration. Subséquemment, j’ai démontré que ces monocytes migrent plus rapidement et en plus grand nombre au travers d’une barrière constituée de cellules endothéliales méningées à comparer à des CE-BHE. Bien que des observations similaires ont été effectuées en utilisant des lymphocytes T CD4+ humains ex vivo, j’ai été incapable de reproduire ces résultats à l’aide de cellules Th1 et Th17 réactivées in vitro. Par opposition à nos données in vitro, j’ai découvert que les souris déficientes en ALCAM développent une EAE active plus sévère que celle observée chez des souris de type sauvages. Cette EAE est par ailleurs associée à une infiltration périvasculaire de lymphocytes T pro-inflammatoires et de monocytes/macrophages de type M1 plus marqué chez les souris ALCAM déficientes. L’induction d’une EAE par transfert adoptif, dans laquelle des cellules immunitaires de type sauvage réactivées par du MOG sont injectées à des souris déficientes en ALCAM, suggère que la pathophysiologie observée durant l’EAE active serait liée à l’absence d’ALCAM au niveau de la BHE. Une caractérisation de la barrière des souris ALCAM déficientes non immunisées a par la suite révélé une réduction de l’expression de certaines molécules de jonction serrée. Une analyse plus poussée a par ailleurs démontré qu’ALCAM est lié indirectement à des molécules de jonction serrée des CE-BHE, ce qui expliquerait l’augmentation de la perméabilité de celle-ci chez les souris déficientes en ALCAM. Une analyse de la perméabilité intercellulaire de la BHE effectuée in vitro a d’autre part corrélé ces résultats. Conclusion : Collectivement, nos données prouvent qu’ALCAM joue un rôle prépondérant dans la diapédèse des monocytes, mais pas des lymphocytes Th1 et Th17 au travers de la BHE. Par ailleurs, nos résultats suggèrent qu’ALCAM remplit une fonction biologique cruciale favorisant le maintien de l’intégrité de la BHE en agissant comme molécule adaptatrice intermédiaire entre les molécules de jonction serrées et le cytosquelette. De cette façon, l’absence d’ALCAM au niveau des CE-BHE promeut indirectement le recrutement de leucocytes pro-inflammatoires dans le SNC des souris atteintes de l’EAE en augmentant la perméabilité des vaisseaux sanguins de la BHE. / Aim: The loss of blood-brain barrier (BBB) integrity is a hallmark of multiple sclerosis. It is associated with a disorganization of junctional molecules and an upregulation of cell adhesion molecules essential for immune cell transmigration. Identifying novel key players involved in this process is thus crucial for the development of MS therapies aimed at promoting BBB integrity and decreasing leukocytes trafficking into the central nervous system (CNS) during neuroinflammation. In this study, the specific role of the adhesion molecule ALCAM, found on BBB endothelial cells (BBB-ECs) and subsets of leukocytes, was assessed. Methods: We first identified ALCAM as an important molecule upregulated during inflammation in a proteomic screen of in vitro cultured primary human BBB-ECs. In order to study the effects of ALCAM on leukocyte transmigration, both active and passive experimental autoimmune encephalomyelitis (EAE) was induced in ALCAM KO and WT animals. The specific role of ALCAM during leukocyte transmigration was also assessed using a modified adhesion assay under sheer-stress, in which leukocytes flow across a capillary-like channel lined with a monolayer of BBB-ECs under physiological pressure. Results: Using the modified adhesion assay, we demonstrated that anti-ALCAM blocking antibodies significantly reduce the rolling and the adhesion of human CD14+ monocytes interacting with primary human BBB-ECs, as well as prevent their overall decrease in velocity. Concurrently, we also observed a significant reduction in the migration of ex vivo CD14+ monocytes, across a monolayer of human BBB-ECs. These monocytes also migrated more rapidly and in higher number across meningeal endothelial cells, as compared to BBB-ECs. While similar observations were made using ex vivo CD4+ T lymphocytes, we failed to reproduce these results using in vitro activated Th1 and Th17 cells. In opposition to our in vitro data, ALCAM KO mice developed a more severe active EAE associated with a significant increase in perivascular infiltration of pro-inflammatory lymphocytes (Th1/Th17) and M1 monocytes/macrophages, as compared to WT controls. In addition, EAE transfer experiments, in which ALCAM KO mice received WT MOG-reactivated splenocytes, suggested that the pathophysiology observed in active EAE was linked to the absence of ALCAM on BBB-ECs. Phenotypic characterization of un-immunized ALCAM KO mice revealed a reduced expression of BBB junctional proteins. Further analysis showed that ALCAM is indirectly associated with tight junction molecules of the BBB-ECs, which explains the increased CNS parenchymal blood vessel in vivo permeability in ALCAM KO animals. Correlating with these data, primary culture of mouse brain BBB-ECs was shown to possess a lower TEER and an increased permeability coefficient. Conclusion: Collectively, our data provide evidence of the implication of ALCAM in monocyte transmigration, but not Th1 or Th17 lymphocyte diapedesis across CNS endothelium. Our results also point to a biologically crucial function of ALCAM in maintaining BBB integrity by acting as an adaptor molecule between tight junctions and the cytoskeleton. As such, the absence of ALCAM at the level of BBB-ECs indirectly promotes the recruitment of pro-inflammatory leukocytes in the CNS of EAE animals by increasing the BBB vessels permeability.

Thesis (M.Pharm. (Pharmacy Practice))--North-West University, Potchefstroom Campus, 2008.

技術標準化與相關智慧財產權保護，一直以來為智慧財產權法與競爭法的交集與爭議的話題。其中又以標準必要專利侵權糾紛為主。基於標準必要專利權人與前在被授權人雙方的立場，其中目前最具爭議的問題應該涉及禁制令救濟的適用性或以F/RAND授權原則為基礎的抗辯來排除侵權。 標準制訂組織（Standard Setting Organization, SSO）訂定F/RAND授權原則承諾（Fair, Reasonable And Non-Discriminatory）於其智慧財產權政策，要求標準專利權人應以公平、合理且無歧視的授權條件，向所有標準實施者提供授權。F/RAND授權原則承諾之發展，目前趨向於強調專利權人的契約義務，以第三方受益人的立場來平衡授權當事人的談判地位；如何「符合F/RAND授權原則之授權」，目前各國尚未有明文法律解釋，對於F/RAND授權原則承諾之清楚定義與規範，目前僅有法院及競爭法主管機關之見解。 在標準必要專利訴訟中，台灣廠商處於被告之身分的狀況居多。面對禁制令的威脅，如何更清楚地了解目前各管轄法院的看法以決定訴訟或談判策略更是重要。本文整理美國、歐洲及亞洲國家之管轄法院案例，加上對競爭法架構下的標準專利授權規範的分析，最後整理如何讓F/RAND授權原則承諾成為對抗禁制令有效抗辯。希望本文能為涉及標準專利訴訟之台灣廠商提供有價值的參考意見。 / Technology standardization and intellectual property protection has been an overlapping and controversial issue between Intellectual Property laws and Competition Law, particularly when it comes to infringement on F/RAND encumbered Standard Essential Patent, SEP. From both standard essential patent owner and potential licensee’ perspectives, the most questionable issue is whether injunctive relief should be available to the holder of F/RAND encumbered SEP who committed to license on fair, reasonable and non-discriminatory (F/RAND) terms, in order to prevent a third-party implementer from practicing a standard reading on that SEP, when such implementer is willing to take a license but the parties disagree on the terms of the license. Furthermore, the definition of F/RAND has never been clearly defined by statutes or interpreted by any judiciary; interested parties could only refer to decisions or guidelines made by the judiciaries or competition authorities in different countries. It is rather common for Taiwanese companies to face F/RAND encumbered SEP law suits as the defendants. Given the even severer threat of injunctive relief, it becomes more important to understand the position each judiciary takes on this issue to have appropriate strategies on law suits and negotiation. This thesis is accordingly written on the following perspectives: firstly, starting with discussion about F/RAND-encumbered SEP law suits in the United States, Europe and Asia; secondly, bringing in SEP encumbered disputes or investigations into framework of Competition Law from competition authorities among different countries and lastly trying to present possibilities that F/RAND commitment as a cause of action under Contract Law can be applied as defense to overcome injunctive relief sought by F/RAND-encumbered SEP licensors. Meanwhile, this thesis is expected to provide Taiwanese companies valuable strategies to law suits or disputes involving F/RAND-encumbered SEPs.

Acid-related disorders are common, chronic conditions that have considerable impact on a patient's quality of life. In a study conducted by Majumdar et al. (2003:2411) the prevalence of chronic acid-related disorders was 2.3%. Acid-related disorders represent a major financial consideration with respect to the costs of drug prescribing (Whitaker, 1998:6). Health care cost increases each year. This leads to an increased interest in economic evaluation of health care and medical technologies (Anell & Svarvar, 2000:175). Health care providers no longer make treatment decisions independent of the consideration of the resultant cost. The treatment provided must not only provide value but the value must be documented to justify spending money. Economic evaluation research has emerged to offer guidance to policy makers, practitioners, health plans and institutions facing difficult treatment and coverage decisions (Ellis era/., 2002:271). The main objectives of this study were to investigate the prescribing patterns and cost of acid reducing medicine with special reference to proton pump inhibitors and histamine-2 receptor antagonists in a section of the private health care sector of South Africa from 2001 to 2006. A longitudinal retrospective drug utilisation study was done on acid reducing medicine items claimed through a national medicine claims database. The five study years were 2001, 2002, 2004, 2005 and 2006. All the study years stretched from 1 January to 31 December. It was determined that acid reducing medicine items prescribed decreased from 2.74% during 2001 to 2.50% during 2006 of all medicine items claimed. The same decreasing trend was observed regarding the cost of acid reducing medicine items. The cost percentage decreased from 4.89% (2001) to 3.72% (2006). However, the average cost per medicine item for the acid reducers increased by 5.35% from 2001 (R230.04 ± 176.29) to 2002 (R243.72 ± 184.18) and then decreased by 15.23% from 2002 to 2004. It again decreased with 15.05% from 2004 (R206.19 ± 179.42) to 2006 (R175.70 ± 172.55). The changes in the average cost of acid reducers were of no practical significance. Proton pump inhibitors represented about half of the acid reducing medicine items prescribed and more than 70% of the total cost of acid reducing medicine items during the study years. The average cost of PPIs revealed a practical significant decrease (d > 0.8) from 2002 (R372.42 ± 156.62) to 2006 (R241.56 ± 177.21). H2RAs contributed between 15.00% and 18.26% of all acid reducing medicine items while contributing to between 9.68% and 16.85% of the total cost of all acid reducers. The active ingredient most often prescribed was lansoprazole during 2001 and 2002, esomeprazole during 2004 and omeprazole during 2005 and 2006. Lanzor® 30mg was the acid reducer with the highest cost from 2001 to 2005, while Pariet® 20mg took the lead in 2006. Zantac® 150mg effervescent tablets were the H2RA, with the highest cost, during the five study years. The percentage innovator items decreased by 4.50% from 2001 to 2002, increased by 1.01% from 2002 to 2004 and decreased again by 31.06% from 2004 to 2006. The slight increase in the percentage innovator medicine items claimed from 2002 to 2004 may be explained by the introduction of Nexiam® (esomeprazole) into the market in 2002. The total number of generic medicine items claimed contributed between 9.62% (n = R1 788 242.25) in 2001 and 30.75% (n = R3 196 163.34) in 2006 of the total cost of acid reducing medicine items. The average cost per day of innovator medicine items was higher than the average cost per day of generic medicine items. This might be explained by a lower average cost for generic medicine items. It was also determined that the prevalence of the two-drug regimens was the highest during the five study years. The Helicobacter pylori (H.pylori) eradication treatments, which included different antibiotics, increased from 2.72% in 2001 to 5.05% in 2006. The PDD for most of the active ingredients of H2RAs and PPIs remained stable during the study years. However, it appears that the PDDs, of the PPIs, active ingredients were more constant than the PDDs, or the H2RAs, active ingredients. The median of the different PPI active ingredients was reasonably more constant than the median of the different H2RA active ingredients. Thus the changes between the PPIs' and H2RAs' active ingredients might be explained by the variation in the median (the number of days the relevant medicine item was claimed for). It is then also recommended that the aspects of generic substitution as well as the usage of H2RAs as prescribed vs. self medication should be further investigated to increase possible cost savings. / Thesis (M.Pharm. (Pharmacy Practice))--North-West University, Potchefstroom Campus, 2008.

Acid-related disorders are common, chronic conditions that have considerable impact on a patient's quality of life. In a study conducted by Majumdar et al. (2003:2411) the prevalence of chronic acid-related disorders was 2.3%. Acid-related disorders represent a major financial consideration with respect to the costs of drug prescribing (Whitaker, 1998:6). Health care cost increases each year. This leads to an increased interest in economic evaluation of health care and medical technologies (Anell & Svarvar, 2000:175). Health care providers no longer make treatment decisions independent of the consideration of the resultant cost. The treatment provided must not only provide value but the value must be documented to justify spending money. Economic evaluation research has emerged to offer guidance to policy makers, practitioners, health plans and institutions facing difficult treatment and coverage decisions (Ellis era/., 2002:271). The main objectives of this study were to investigate the prescribing patterns and cost of acid reducing medicine with special reference to proton pump inhibitors and histamine-2 receptor antagonists in a section of the private health care sector of South Africa from 2001 to 2006. A longitudinal retrospective drug utilisation study was done on acid reducing medicine items claimed through a national medicine claims database. The five study years were 2001, 2002, 2004, 2005 and 2006. All the study years stretched from 1 January to 31 December. It was determined that acid reducing medicine items prescribed decreased from 2.74% during 2001 to 2.50% during 2006 of all medicine items claimed. The same decreasing trend was observed regarding the cost of acid reducing medicine items. The cost percentage decreased from 4.89% (2001) to 3.72% (2006). However, the average cost per medicine item for the acid reducers increased by 5.35% from 2001 (R230.04 ± 176.29) to 2002 (R243.72 ± 184.18) and then decreased by 15.23% from 2002 to 2004. It again decreased with 15.05% from 2004 (R206.19 ± 179.42) to 2006 (R175.70 ± 172.55). The changes in the average cost of acid reducers were of no practical significance. Proton pump inhibitors represented about half of the acid reducing medicine items prescribed and more than 70% of the total cost of acid reducing medicine items during the study years. The average cost of PPIs revealed a practical significant decrease (d > 0.8) from 2002 (R372.42 ± 156.62) to 2006 (R241.56 ± 177.21). H2RAs contributed between 15.00% and 18.26% of all acid reducing medicine items while contributing to between 9.68% and 16.85% of the total cost of all acid reducers. The active ingredient most often prescribed was lansoprazole during 2001 and 2002, esomeprazole during 2004 and omeprazole during 2005 and 2006. Lanzor® 30mg was the acid reducer with the highest cost from 2001 to 2005, while Pariet® 20mg took the lead in 2006. Zantac® 150mg effervescent tablets were the H2RA, with the highest cost, during the five study years. The percentage innovator items decreased by 4.50% from 2001 to 2002, increased by 1.01% from 2002 to 2004 and decreased again by 31.06% from 2004 to 2006. The slight increase in the percentage innovator medicine items claimed from 2002 to 2004 may be explained by the introduction of Nexiam® (esomeprazole) into the market in 2002. The total number of generic medicine items claimed contributed between 9.62% (n = R1 788 242.25) in 2001 and 30.75% (n = R3 196 163.34) in 2006 of the total cost of acid reducing medicine items. The average cost per day of innovator medicine items was higher than the average cost per day of generic medicine items. This might be explained by a lower average cost for generic medicine items. It was also determined that the prevalence of the two-drug regimens was the highest during the five study years. The Helicobacter pylori (H.pylori) eradication treatments, which included different antibiotics, increased from 2.72% in 2001 to 5.05% in 2006. The PDD for most of the active ingredients of H2RAs and PPIs remained stable during the study years. However, it appears that the PDDs, of the PPIs, active ingredients were more constant than the PDDs, or the H2RAs, active ingredients. The median of the different PPI active ingredients was reasonably more constant than the median of the different H2RA active ingredients. Thus the changes between the PPIs' and H2RAs' active ingredients might be explained by the variation in the median (the number of days the relevant medicine item was claimed for). It is then also recommended that the aspects of generic substitution as well as the usage of H2RAs as prescribed vs. self medication should be further investigated to increase possible cost savings. / Thesis (M.Pharm. (Pharmacy Practice))--North-West University, Potchefstroom Campus, 2008.

Mon poste de psychologue clinicienne au sein d’une maison de retraite m’a sensibilisée à la souffrance psychique des aides-soignantes. A partir de l’écoute des aides-soignantes dans le cadre de ma mission de support technique à l’équipe, émerge la plainte récurrente d’un déficit égotiste de désinvestissement émotionnel et affectif et la non reconnaissance de la pénibilité de leur tâche, qui me conduit à poser l’hypothèse d’un burn out. La recherche porte donc sur le Syndrome d’Épuisement Professionnel. Elle étudie les relations complexes entre le personnel médico-social aides-soignantes et la cadre de santé, et entre la personne âgée et sa fille. Un questionnaire clinique a fait ressortir l’ampleur du ressenti subjectif d’être épuisé. L’échelle standardisée MSP de Louise Lemyre, celle du ressenti subjectif d’être stressé. Le stress étant convoqué dans la position conceptuelle théorique retenue du Syndrome d’Épuisement Professionnel.L’analyse de ces deux ressentis a révélé les rapports complexes entre soignante–soignée et la cadre et entre fille–mère et grand-mère. C’est ainsi que l’enjeu narcissique du personnel fait ressurgir les origines archaïques de la sexualité infantile dans le lien intime au corps. En même temps, à cause d’une féminisation généralisée de la profession, la spécificité de la fonction du Féminin Psychique s’impose.Cette recherche souhaite apporter, grâce à ce Féminin Psychique, un éclairage autre sur la position intermédiaire de la Cadre. Le statut de bonne ou mauvaise mère que les aides-soignantes lui reconnaissent, aggrave ou diminue le Syndrome d’Épuisement Professionnel et le stress. Enfin, cette recherche insiste une nouvelle fois sur la nécessité impérieuse de la formation permanente institutionnelle et du travail d’élaboration psychique de ces personnels et ce, à périodicités constantes. / As a psychologist in an EHPAD (a regulated home for dependent seniors), I became very much aware of the auxiliary nurses’ psychological sufferings. Listening to them during my team-supporting mission, I heard a recurring complaint emerge, that of an egotistical deficit of emotional and affective disinvestment and of a lack of recognition of the painfulness of their task. This has led me to hypothesize professional exhaustion. The research in this thesis therefore focuses on the burn out syndrome. It studies the complex relations between the auxiliary nurses as medico-social staff and the health manager in charge as well as between the elderly person and his/her daughter.A clinical questionnaire highlighted the depth of the subjective feeling of exhaustion while Louise Lemyre’s standardized scale highlighted the depth of the subjective feeling of stress, an operational notion in the theoretical concept of burn out.The analysis of both feelings revealed the complex relations between patient-auxiliary nurse and manager as well as between daughter-mother and grandmother. The narcissism at stake with the staff reactivates the archaic origins if child sexuality in the nursing place. A t the same time, because of the general feminization of the profession, the specificity of the feminine psyche is of foremost importance.Thanks to this notion, the research paper aims to shed a different light on the intermediary position of the manager. The status of good or bad mother figure that auxiliary nurses grant her worsens or lessens the burn out and stress.Last, this research paper further insists on the absolute necessity of permanent institutional training and of psychic elaborative work for the staff at regular intervals.

La barrière hémo-encéphalique (BHE) est formée des cellules endothéliales microvasculaires cérébrales reliées entre elles par des jonctions serrées. Grâce à sa perméabilité restreinte et sélective, la BHE entrave le passage des molécules et cellules du sang vers le système nerveux central (SNC). Chez les patients atteints de sclérose en plaques (SEP), une maladie inflammatoire du SNC, la rupture de la BHE permet aux cellules immunes actives d'infiltrer le tissu cérébral. Il s'ensuit une réaction inflammatoire excessive au cours de laquelle d'autres leucocytes sont recrutés dans le cerveau et qui culmine par la formation des plaques de démyélinisation caractéristiques de la SEP. On dénote au niveau de ces lésions une présence importante de lymphocytes T CD4⁺ activés et de cytokines pro-inflammatoires propres à une réponse de type TH1, tels l’IFN-γ et l’IL-1. Curieusement cependant, l’inhibition de la voie TH1 n’empêche pas l’apparition de la maladie dans le modèle murin de la SEP et en aggrave même les symptômes. On attribue maintenant aux lymphocytes TH17, nommées en raison de leur capacité à produire de l’IL-17, un rôle clé dans le développement de la maladie. L’objectif de ce travail de thèse visait à caractériser les lymphocytes TH17 chez l’humain et définir leur contribution exacte dans la fragilisation de la BHE, une étape décisive dans la formation des lésions de SEP. Pour ce faire, nous avons mis au point une méthode expérimentale permettant l’expansion in vitro de populations de lymphocytes TH17 à partir de cellules mononuclées du sang de donneurs sains. Nos travaux démontrent que l’IL-23 induit la production d’IL-17, d’IL-22 et de granzyme B par les lymphocytes T CD4⁺CD45RO⁺ mémoires humains et qu’une proportion des cellules exprime de manière concomitante de l’IL-17 et de l’IFN-γ. La fréquence des lymphocytes T CD4⁺ IL17⁺, IL-22⁺ et des doubles positifs IL-17⁺IFN-γ⁺ est significativement plus élevée dans les lignées de lymphocytes TH17 provenant de patientes en poussée que dans celles de contrôles. Nos analyses démontrent que les cellules endothéliales de la BHE expriment de faibles niveaux des récepteurs de l’IL-17 et de l’IL-22 à l’état basal mais que leur présence est accrue dans le cerveau de patients atteints de SEP. L’activation du récepteur de l’IL-17 entraîne une augmentation de la perméabilité de la BHE et une perturbation de l’organisation des protéines de jonction occludine et ZO-1. Finalement, nous démontrons que la migration des lymphocytes TH17 à travers la BHE est régie en grande partie par la molécule d’adhérence ICAM-1 et que les lymphocytes qui co-expriment l’IL-17 et l’IFN-γ sont plus aptes à franchir la BHE que ceux qui produisent uniquement l’une ou l’autre de ces cytokines. Nous retrouvons d’ailleurs des cellules qui expriment simultanément les facteurs de transcription T-bet et RORC, associés respectivement aux lymphocytes TH1 et aux TH17, au sein des infiltrats péri-vasculaires des lésions actives de SEP. Les travaux présentés dans cette thèse auront permis d’affiner nos connaissances sur les mécanismes d’entrée des lymphocytes TH17 dans le SNC et les propriétés délétères des cytokines qu’ils sécrètent, notamment dans l’activation et la déstabilisation de l’endothélium cérébral. / The blood-brain barrier (BBB) plays a crucial role in protecting the central nervous system (CNS) by restricting entry of cells and molecules into the brain. In the CNS disorder multiple sclerosis (MS), breakdown of the BBB allows activated leukocytes to infiltrate the brain parenchyma, leading to the formation of the characteristic demyelinated lesions. For decades, MS was viewed as a TH1-mediated disease, a notion that was largely supported by studies in its animal model and by the abundance of prototypical TH1-associated cytokines within active MS lesions. However, over the years, accumulating evidence has highlighted the involvement of another subset of CD4⁺ T cells that express IL-17, therefore named TH17 lymphocytes, in the pathology of the disease. The goal of the work presented herein was to characterize the human TH17 lymphocyte population and define their contribution to the disruption of the BBB and leukocyte infiltration into the CNS, both important early events in the formation of MS lesions. To do so, we developed and optimized a method to successfully generate human TH17 lines in vitro from peripheral blood mononuclear cells of healthy donors. We demonstrate that in response to IL-23, human memory CD4⁺CD45RO⁺ but not naïve CD4⁺CD45RA⁺ T lymphocytes produce IL-17, IL-22, and granzyme B, with a subset of cells simultaneously expressing IL-17 and IFN-γ. Interestingly, we measure a significant increase in the percentage of T CD4⁺ IL17⁺, of IL-22⁺, and of IL-17⁺IFN-γ⁺ dual producers in TH17 cell lines expanded from the peripheral blood of acutely relapsing MS women as compared to those generated from healthy controls and remitting MS patients. We show that both IL-17 and IL-22 receptors are upregulated on BBB endothelial cells in situ during inflammation and that IL-17 enhances BBB permeability by disrupting the integrity of tight junction proteins occludin and ZO-1. Finally, we provide evidence that TH17 lymphocytes transmigrate efficiently across human brain endothelial cells via the adhesion molecule ICAM-1 and show that IL-17⁺IFN-γ⁺ double producers have an increased propensity to do so. Accordingly, we detect lymphocytes that display immunoreactivity against both the TH1- and TH17-associated transcription factors T-bet and RORC within perivascular infiltrates of active MS lesions. The work presented in this thesis has refined our understanding of the mechanisms that drive TH17 lymphocyte recruitment into the CNS and shed light on the deleterious effect of TH17-secreted cytokines, specifically in the activation and breakdown of the BBB.

The legal nature of the curriculum documents of the Czech Republic's educational system ABSTRACT This thesis treats the selected set of curriculum documents within the Czech legal framework laid down by the Act No. 561/2004 Collection of Law, on Pre-school, Basic, Secondary, Tertiary Professional and Other Education (the Education Act), as amended. The focus aims towards the Framework Educational Programmes (FEP) and School Educational Programmes (SEP), for which the relevant FEPs were issued. The National Educational Programme according to the Education Act as amended effective until 30. September 2020 is also taken into consideration on a theoretical level. In the times of expecting considerable state-level curriculum documents contents changes, and consequent changes on the school level, this thesis aims to analyse substantial legal aspects of each type of curriculum documents and their mutual effects and to assess, whether the current legal framework reflects their presumed use and meaning appropriately. With that goal, the key notions are defined first though and a brief insight into the curriculum historical development is provided. The thesis identifies an insufficiency within the valid legislation as the legal forms the curriculum documents should take are not laid down and consequently examines...