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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

An analysis concerning three organisations work with reducing child labour : – A case study within Peru’s mining industry

Sheikholeslamzadeh, Sanaz, Bergvall Bark, Marie January 2008 (has links)
<p>This thesis aim to describe and analyze different organisations’ work with reducing child labour. In order to understand the complex matter of child labour, a case study concerning children’s situation within the mining industry in Peru has been made. The first part of the thesis has been designed to be an introduction to the matter, with a description of the mining industry in Peru and the situation of child labour. The two following parts are more analytic in character. The second and third part discusses how the Millennium Development Goals (MDGs) can be used as an instrument of reducing child labour. Further, top down and bottom up-theories will be discussed as different approaches and working methods for organisations.</p><p>In addition to find the answers to our questions, interviews have been conducted with employees of the organisations (ILO, UNICEF and Save the Children), articles and literature have been analyzed and finally Internet have contributed with information about child labour, Peru, the organisations, theories and the MDGs.</p><p>This study claims that child labour is a complex matter and one possible solution to reduce it can be through promoting education. This can only be viable if organisations work together with governments, using a combination of top down and bottom up approaches.</p>
2

Water access and provisioning in UMzumbe Local Municipality under the Ugu District Municipality, KwaZulu-Natal

Duma, Clifford Mhlanyukwa January 2015 (has links)
A dissertation submitted to the Faculty of Arts in partial fulfilment of the requirement for the degree of Master’s in the Department of Anthropology and Development Studies at the University of Zululand, 2015 / According to the Constitution, Act No.108 of 1996 of the Republic of South Africa, every person has the right to clean water. The Department of Water Affairs and Forestry’s Community Water Supply Programme was established in 1994 to achieve this objective. Access to safe drinking water is the basic need to human life and wellbeing, and is a key to public health issues. The UGu District Municipality is the water distributor to the Umzumbe Local Municipality residents. The study area for this research is the Umzumbe Local Municipality, which is the second largest municipality under the UGu District Municipality. Communities, especially in rural areas under the Umzumbe Local Municipality, are still unable to access adequate safe drinking water which meets the national standards for protecting human health. The main concern of this research was to assess water access and provisioning in the Umzumbe Local Municipality, under the UGu District Municipality. The research assessed the water supply system in the UGu District Municipality by engaging with all the stakeholders involved in water provision to verify their roles in the water supply and development of water service delivery projects. The study was predicated on three theories which were assumed to explain water provisioning. The first theory is the top-down and bottom up approach to service delivery. The second approach is the privatisation concept which is based on neo-liberalism. This explains that buying what could hinder efficient delivery, mostly in poor environments. The final theory of the study deals with project management. This theory asserts that the way a project is packaged and commissioned will have implications in the delivery of services, in this case, water delivery. The research employed mixed a methods research design. The reason for employing both qualitative and quantitative methods is because the research sought to obtain a comprehensive interpretation of the occurrences in the research area. The adoption of qualitative data alone is likely to provide limited vision which would not be sufficient for the research without the inclusion of quantitative data. The population for the research consisted of the Umzumbe Local Municipality residents, both males and females from twenty one years of age, upwards. Seventy participants were chosen for the research. Interview schedules and survey questionnaires were used as data collection instruments. vii The findings showed that the UGu District Municipality infrastructure does have some challenges, which come in the form of skills challenges, and capacity challenges. Other challenges that the study identified include inability to access municipality’s infrastructure grant (M.I.G) and bulk infrastructure grant (B.I.G). The UGu District Municipality areas that are far inland are likely to have water, and also the problems because of the low capacity of engines that are used to pump water also the problem is exacerbated by power failures. The findings further indicated that unskilled contractors could be the cause of water outage in the Umzumbe Local Municipality. It has been established that contractors who are hired to run projects have a tendency of abandoning projects that have not been completed. The findings presented that the UGu District Municipality does not consider the involvement of community members when they take decisions on project matters. The research established that water tankers do not provide community with adequate water and the utilization of water tankers is an expensive option that can cause community not to be served. The findings also indicated that some of the water infrastructure is old and it can be the cause of water shortage. Lastly, the findings showed that the Umzumbe Local Municipality has low income opportunities, causing its inhabitants to be unable to pay for water services. On the basis of the findings the study recommends that the UGu District Municipality needs to adhere to the principles of monitoring and evaluation of all activities and projects that relate to water provisioning. At a broader scale the study suggests that further research be done to determine whether the Department of Water and Sanitation can consider the construction of water reservoir in UMzimkhulu River. The reservoir should be linked with the Umgeni Water Board to provide water to communities during water outage.
3

Addressing the contradiction between discourse and practice in health promotion.

Laverack, Glenn, kimg@deakin.edu.au,jillj@deakin.edu.au,mikewood@deakin.edu.au,wildol@deakin.edu.au January 1999 (has links)
The main theme of this thesis is the contradiction between discourse and practice in health promotion. Many health promoters continue to exert power-over the community through top-down programming whilst at the same time using an emancipatory discourse. The thesis has addressed this contradiction in three parts. The first part determines how the emancipatroty discourse has evolved and eplores the role of social movements in the development of contemporary health discourses and their influence on the legitimisation of empowerment. Central to this discourse is the empowerment of communities. To understand the role of this concept the thesis provides an interpretation of the different meanings of power and community, and the different levels of analysis of empowerment in the context of health promotion programming. The second part identifies the nature of health programming and the dominance of top-down, and to a much lesser extent, bottom-up approaches. The thesis argues that these two approaches are not, and do not have to be, mutually exclusive. To address this issue the thesis presents a new methodology is situated within a framework developed for the accomodation of empowerment goals within health promotion programmes. The study also identifies the organisational areas of influence on the processs of community empowerment and it is these which are used for the assessment of this concept. Both the framework and the methodology address the contradiction in health promotion by making community empowerment operational within a programme context. The third part of the thesis supports the rationale for the design of the methodology with field work in rural Fijian communities. The findings are presented as a composite case study to highlight the experiences of implementing the methodolgy and the main themes that emerged during the field work. the final chapter of the thesis brings together the central themes of the study and draws from these and 'emergent agenda' as a way forward for health promotion research and practice.
4

An analysis concerning three organisations work with reducing child labour : – A case study within Peru’s mining industry

Sheikholeslamzadeh, Sanaz, Bergvall Bark, Marie January 2008 (has links)
This thesis aim to describe and analyze different organisations’ work with reducing child labour. In order to understand the complex matter of child labour, a case study concerning children’s situation within the mining industry in Peru has been made. The first part of the thesis has been designed to be an introduction to the matter, with a description of the mining industry in Peru and the situation of child labour. The two following parts are more analytic in character. The second and third part discusses how the Millennium Development Goals (MDGs) can be used as an instrument of reducing child labour. Further, top down and bottom up-theories will be discussed as different approaches and working methods for organisations. In addition to find the answers to our questions, interviews have been conducted with employees of the organisations (ILO, UNICEF and Save the Children), articles and literature have been analyzed and finally Internet have contributed with information about child labour, Peru, the organisations, theories and the MDGs. This study claims that child labour is a complex matter and one possible solution to reduce it can be through promoting education. This can only be viable if organisations work together with governments, using a combination of top down and bottom up approaches.
5

Top-down and bottom-up excursions beyond the standard model : the example of left-right symmetries in supersymmetry

Alloul, Adam 20 September 2013 (has links) (PDF)
The field of high-energy physics has been living a very exciting period of its history with the Large Hadron Collider (LHC) at CERN collecting data. Indeed, this enormous machine able to collide protons at a center of mass energy of 14 TeV promises to unveil the mystery around the physics at such energy scales. From the physicists side, the expectations are very strong as it isnowadays a certitude that the Standard Model of particle physics is incomplete and should, in fact, be interpreted as the effective theory of a more fundamental one. Unfortunately, the 7 and 8 TeV runs of the LHC did not provide any sign of new physics yet but there has been at least one major discovery in 2010, namely the discovery of a scalar particle with a mass of 125 GeV and whichproperties are very close to those of the Standard Model Higgs boson. Since then, many questions have come up as we now want to understand if it really is the Standard Model Higgs boson or if it exhibits any deviations. It is in this peculiar context that my research work was carried. In a first project, we, my supervisors, our collaborator and I, have wanted to explore thephenomenology associated with the neutralinos and charginos sector of the left-right symmetric supersymmetric model. Such an analysis can be motivated by several reasons such as the fact that the supersymmetric nature of these models provides a natural explanation for the infamous hierarchy problem, implies the unification of the gauge coupling constants at very high energy and provides a natural candidate for dark matter. In addition to these nice features, the left-right symmetry introduces a natural framework for explaining the smallness of neutrino masses but also helps in addressing several other unresolved issues in the Standard Model framework. Only focusing on the lightest charginos and neutralinos decaying into one or more light leptons, we have shown in our study that these models can be easily discovered in multi-leptonic final states as theylead to signatures very different from those induced by the Standard Model or its supersymmetric version.[...]
6

Top-down and bottom-up excursions beyond the standard model : the example of left-right symmetries in supersymmetry / Excursions « top-down » et « bottom-up » au-delà du modèle standard : l'exemple des symétries gauches-droites en supersymétrie

Alloul, Adam 20 September 2013 (has links)
Une très grande effervescence secoue le monde de la physique des particules depuis le lancement du grand collisionneur de hadrons (LHC) au CERN. Cette énorme machine capable de faire se collisionner des protons à des énergies égales à 14 TeV promet de lever le voile sur la physique régissant les interactions à ces échelles d’énergies. Ces résultats sont d’autant plus attendus que l’on a acquis la certitude que le Modèle Standard de la physique des particules est incomplet et devrait, en fait, être interprété comme la théorie effective d’une théorie plus fondamentale. Toutefois, depuis le lancement des expériences au LHC avec des énergies de 7 puis de 8 TeV aucun signe de nouvelle physique n’a été découvert. Par contre, un énorme bond en avant a été franchi avec la découverte d’une particule scalaire de masse égale à 125 GeV et dont les propriétés sont relativement proches de celles du boson de Higgs telles que prédites par le Modèle Standard. C’est dans ce contexte de forte émulation internationale que mon travail de thèse s’est inscrit. Dans un premier temps, nous avons voulu explorer la phénoménologie associée au secteur des neutralinos et charginos du modèle supersymétrique symétrique gauche-droit. Cette étude peut être motivée par plusieurs raisons notamment le fait que leur caractère supersymétrique apporte une solution au problème dit de la hiérarchie mais implique aussi l’unification des constantes de jauge ainsi que l’explication de la matière noire. L’introduction de la symétrie entre les fermions gauchers et les fermions droitiers permet, quant à elle, d’expliquer naturellement, via le mécanisme dit de la balançoire, la petitesse de la masse des neutrinos mais aussi de répondre à plusieurs autres questions non solubles dans le cadre du Modèle Standard. Nous concentrant uniquement sur le secteur des charginos et neutralinos les plus légers, nous avons montré que ces modèles peuvent être facilement mis en évidence dans les évènements multi-leptoniques en ce sens que les signatures qu’ils induisent sont tr`es différentes comparées à celles du Modèle Standard et de sa version supersymétrique.[...] / The field of high-energy physics has been living a very exciting period of its history with the Large Hadron Collider (LHC) at CERN collecting data. Indeed, this enormous machine able to collide protons at a center of mass energy of 14 TeV promises to unveil the mystery around the physics at such energy scales. From the physicists side, the expectations are very strong as it isnowadays a certitude that the Standard Model of particle physics is incomplete and should, in fact, be interpreted as the effective theory of a more fundamental one. Unfortunately, the 7 and 8 TeV runs of the LHC did not provide any sign of new physics yet but there has been at least one major discovery in 2010, namely the discovery of a scalar particle with a mass of 125 GeV and whichproperties are very close to those of the Standard Model Higgs boson. Since then, many questions have come up as we now want to understand if it really is the Standard Model Higgs boson or if it exhibits any deviations. It is in this peculiar context that my research work was carried. In a first project, we, my supervisors, our collaborator and I, have wanted to explore thephenomenology associated with the neutralinos and charginos sector of the left-right symmetric supersymmetric model. Such an analysis can be motivated by several reasons such as the fact that the supersymmetric nature of these models provides a natural explanation for the infamous hierarchy problem, implies the unification of the gauge coupling constants at very high energy and provides a natural candidate for dark matter. In addition to these nice features, the left-right symmetry introduces a natural framework for explaining the smallness of neutrino masses but also helps in addressing several other unresolved issues in the Standard Model framework. Only focusing on the lightest charginos and neutralinos decaying into one or more light leptons, we have shown in our study that these models can be easily discovered in multi-leptonic final states as theylead to signatures very different from those induced by the Standard Model or its supersymmetric version.[...]
7

工作犬計畫執行成效及關鍵因素分析─以A市為例 / Effectiveness and Crucial Affecting Factors Analysis for Working Dogs Program Execution in City A

陳筠凡, Chen, Yun Fan Unknown Date (has links)
2017年,零撲殺政策在未考量國內現況下貿然實施,造成收容所過度擁擠、收容品質驟降。為舒緩收容壓力及兼顧動物權益,中央政府積極提倡多元認養政策作為地方施政重點。然而認養行為涉及民眾觀念、意願及資源,難以在短期內提升成效,加上家戶飼養能量似瀕臨飽和,未來發展成疑。 A市以「工作犬計畫」為推動流浪犬認養之策略,且短期內認養率升幅最高,率先達成零撲殺目標,故本文以此為個案,藉「由下而上」政策執行分析途徑,透過文獻分析、深度訪談、參與觀察、問卷調查及焦點團體訪談等研究方法,了解工作犬計畫之內容及執行過程、工作犬佔個人認養率之比例與後續飼養品質等情形,並剖析影響前述成效之關鍵,以供其他縣市參考。最後總結上述發現,對相關政策行動者提出建議,並就整體流浪犬政策議題提出未來研究思考方向。 / 英文文獻 Ajzen, I. (1985). From Intention to Action: The Theory of Planned Behavior. In J. Kuhl, & J. Beckmann (Ed.), Action Control From Cognition to Behavior (pp. 11-39), New York: Springer Verlag. Barrett, S., & C. Fudge (1981). Examining the Policy-Action Relationship. Fudge & Barrett, Policy and Action: Essays on the Implementatino of Public Policy (pp.3-32). London: Methuen. DeLeon, P. (1999). The Missing Link Revisited: Contemporary Implementation Research. Policy Studies Review, Vol.16, 311-338. Downey, H. & S. Ellis (2008). Tails of animal attraction: incorporating the feline into the family. J. Journal of Business Research, Vol. 61, 434–441. 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(3rd ed.) Berkeley: University of California Press. Marston, L. C., P. C. Bennett, & G. J. Coleman (2005). Adopting shelter dogs: Owner experiences of the first month post-adoption. Anthrozoos, 18, 358-378. Mazmanian, D., & P. Sabatier. (1983). Implementation and Public Policy. Glendale, IL: Scott, Foresman. McLaughlin, M. W., (1976). Implementation as Matual Adaptation: Change in Classroom Organization. Teacher College Record,Vol. 77(3) : 339-351. Mjern, B., & C. Hull. (1982). Implementation Research as Empirical Constitutionalism. European Journal of Political Research, Vol. 10(2), 105-115. Raupp, C. (1999). Treasuring, trashing, or terrorizing: Adult outcomes of childhood socialization about companion animals. Society & Animals, Vol. 7(2), 141–149. Smith, T.B., (1973). The Policy Implementation Process. Policy Sciences, Vol. 4(2), 203. Soto, F. R., F. Ferreira, S. R. Pinheiro, F. Nogari, M. R. Risseto, O. de Souza, & Amaku M. (2005). 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賴亦德(2017b)。談2017年TNVR入法(2):談2017年TNVR入法(2):要學保加利亞動保法就學徹底一點可以嘛?。2018年1月21日,取自:http://yitelai.blogspot.tw/2017/09/2017tnvr2.html。 賴亦德(2017c)。談2017年TNVR入法(3):最後三點終於出現比較合理的修法建議了。2018年1月21日,取自:http://yitelai.blogspot.tw/2017/09/2017tnvr3.html。 賴亦德(2017d)。談2017年TNVR入法(4):法條新瓶裝舊酒,三年前的問題現在也還在。2018年1月21日,取自:http://yitelai.blogspot.tw/2017/09/2017tnvr.html。 賴亦德(2017e)。談2017年TNVR入法(5):我不反TNVR但反對這樣的入法,更討厭不誠實的倡議。2018年1月21日,取自:http://yitelai.blogspot.tw/2017/09/2017tnvrtnvr.html。 鄧仲華(2003a)。流浪動物收容及政策檢討─認領養問題。2016年7月29日,取自關懷生命協會官方網站http://www.lca.org.tw/book/670。 鄧仲華(2003b)。流浪動物收容及政策檢討─收容所管理問題,2016年7月29日,取自關懷生命協會官方網站http://www.lca.org.tw/book/672。 關懷生命協會(1997)。犬殤專題之一:步履「犬殤」。2016年8月6日,取自關懷生命協會官方網站http://www.lca.org.tw/column/node/934。 關懷生命協會(2000)。步履犬殤─八十七年臺灣公立收所現況調查報告及具體改善建議。2016年8月6日,取自關懷生命協會官方網站http://www.lca.org.tw/column/node/937。 關懷生命協會(2001)。公立流浪犬收容所體檢報告。2016年8月6日,取自關懷生命協會官方網站http://www.lca.org.tw/column/node/942。 關懷生命協會(2015)。動保網站評比:《為牠找個家─臺灣各縣市動物收容業務網站分析報告》,2016年8月5日,取自關懷生命協會官方網站http://www.lca.org.tw/news/node/5781。 關懷生命協會(2016)。公民觀察收容所 開拓動保心思路。2016年8月6日,取自關懷生命協會官方網站http://www.lca.org.tw/avot/6000。 盧思岑(2016)。誰會是下一位自殺的前線獸醫師?,2016年7月25日,取自:https://www.twreporter.org/a/opinion-veterinarian-suicide。 陳佳利、紀岳君(2012)。【我們的島】街頭的救贖,2016年6月5日,取自:公視新聞議題中心http://pnn.pts.org.tw/main/2012/01/16/%E3%80%90%E6%88%91%E5%80%91%E7%9A%84%E5%B3%B6%E3%80%91%E8%A1%97%E9%A0%AD%E7%9A%84%E6%95%91%E8%B4%96/。 國際伴侶動物管理聯盟(2007)。人道犬隻族群管理指南,2016年7月17日,取自:http://www.icam-coalition.org/downloads/ICAM_Co_Humane_Dog_Population_Management_Guidance_Chinese.pdf。 蕭士塔(2015)。TN(V)R促成零撲殺?動保處不認可,2016年6月2日,取自:臺灣動物新聞網http://www.tanews.org.tw/info/9152。 A市政府動物防疫保護處:http://www.tainan.gov.tw/ahipo/default.asp。 政府資料 監察院(2011)。流浪狗調查報告。100年2月17日院台財字第10022300770號調查報告,未出版。 農委會(2014)。104至107年度加強動物保護行政效能計畫。未出版。 期刊雜誌 林洧楨(2016)。模範城市─動保最成功在臺南,連童子賢都按讚《賴清德數字管理學 流浪犬三年少一半》。商業周刊,1496,50-52。
8

Contextualizing discretion : micro-dynamics of Canada’s refugee determination system

Bayrak, Sule 03 1900 (has links)
À une époque où l'immigration internationale est de plus en plus difficile et sélective, le statut de réfugié constitue un bien public précieux qui permet à certains non-citoyens l'accès et l'appartenance au pays hôte. Reposant sur le jugement discrétionnaire du décideur, le statut de réfugié n’est accordé qu’aux demandeurs qui établissent une crainte bien fondée de persécution en cas de retour dans leur pays d'origine. Au Canada, le plus important tribunal administratif indépendant, la Commission de l'immigration et du statut de réfugié du Canada (CISR), est chargé d’entendre les demandeurs d'asile et de rendre des décisions de statut de réfugié. Cette thèse cherche à comprendre les disparités dans le taux d’octroi du statut de réfugié entre les décideurs de la CISR qui sont politiquement nommés. Au regard du manque de recherches empiriques sur la manière avec laquelle le Canada alloue les possibilités d’entrée et le statut juridique pour les non-citoyens, il était nécessaire de lever le voile sur le fonctionnement de l’administration sur cette question. En explorant la prise de décision relative aux réfugiés à partir d'une perspective de Street Level Bureaucracy Theory (SLBT) et une méthodologie ethnographique qui combine l'observation directe, les entretiens semi-structurés et l'analyse de documents, l'étude a d'abord cherché à comprendre si la variation dans le taux d’octroi du statut était le résultat de différences dans les pratiques et le raisonnement discrétionnaires du décideur et ensuite à retracer les facteurs organisationnels qui alimentent les différences. Dans la lignée des travaux de SLBT qui documentent la façon dont la situation de travail structure la discrétion et l’importance des perceptions individuelles dans la prise de décision, cette étude met en exergue les différences de fond parmi les décideurs concernant les routines de travail, la conception des demandeurs d’asile, et la meilleure façon de mener leur travail. L’analyse montre comment les décideurs appliquent différentes approches lors des audiences, allant de l’interrogatoire rigide à l’entrevue plus flexible. En dépit des contraintes organisationnelles qui pèsent sur les décideurs pour accroître la cohérence et l’efficacité, l’importance de l’évaluation de la crédibilité ainsi que l’invisibilité de l’espace de décision laissent suffisamment de marge pour l’exercice d’un pouvoir discrétionnaire. Même dans les environnements comme les tribunaux administratifs où la surabondance des règles limite fortement la discrétion, la prise de décision est loin d’être synonyme d’adhésion aux principes de neutralité et hiérarchie. La discrétion est plutôt imbriquée dans le contexte de routines d'interaction, de la situation de travail, de l’adhésion aux règles et du droit. Même dans les organisations qui institutionnalisent et uniformisent la formation et communiquent de façon claire leurs demandes aux décideurs, le caractère discrétionnaire de la décision est par la nature difficile, voire impossible, à contrôler et discipliner. Lorsqu'ils sont confrontés à l'ambiguïté des objectifs et aux exigences qui s’opposent à leur pouvoir discrétionnaire, les décideurs réinterprètent la définition de leur travail et banalisent leurs pratiques. Ils formulent une routine de rencontre qui est acceptable sur le plan organisationnel pour évaluer les demandeurs face à eux. Cette thèse montre comment les demandeurs, leurs témoignages et leurs preuves sont traités d’une manière inégale et comment ces traitements se répercutent sur la décision des réfugiés. / In an era where international immigration is increasingly difficult and selective, refugee status constitutes a valuable public good that enables some non-citizens access and membership to the host country. Based on the discretionary judgment of the decision-maker, refugee status is only granted to claimants who establish well-founded fear of persecution if returned to their home country. Canada’s largest independent administrative tribunal, Immigration and Refugee Board of Canada (IRB), is charged to hear refugee claimants and make refugee status determinations. This dissertation investigates why significant disparities exist among IRB’s politically appointed decision-makers’ refugee status grant rates. As little was known about the concrete ways Canada allocates opportunities for entry and legal status for non-citizens, lifting the blanket of administration was necessary. By exploring refugee decision-making from a Street Level Bureaucracy Theory (SLBT) perspective, and an ethnographic methodology that combined direct observation, semi-structured interviews and document analysis, the study sought first to understand whether the variation in grant rates were a result of differences in decision-makers’ discretionary practices and reasoning and second to trace the organizational factors that foster variation. In line with previous scholarship on SLBT that document how the work situation structure discretion and how individual views play in decision-making; this study demonstrates substantive differences among decision-makers in terms of their work routines, conceptions of refugee claimants and the best way to conduct their work. The analysis illustrates how decision-makers apply not a singular but a variety of approaches to the refugee hearing, ranging from rigid interrogation to the more resilient interview style. Despite clear organizational constraints on decision-makers that target to increase consistency and efficiency of refugee determinations, the significance of credibility-assessment and the invisibility of the decision-making space leave ample room for discretionary behavior. Even in rule-saturated environments like administrative tribunals which extensively regulate discretion; decision-making hardly means neutral and hierarchical rule adherence. Instead discretion is nested within the context of interaction routines, work situation, rule adherence and law. It is inherently difficult if not improbable to control and discipline discretionary decision-making even in organizations that institutionalize and standardize training and communicate their demands clearly to decision-makers. When faced with goal ambiguity and with demands that they consider run against their discretionary authority, decision-makers reinterpret their job definition and routinize their practices. They formulate an encounter routine that is organizationally acceptable to assess the people in front of them. This dissertation illustrates how unevenly the claimants, their testimony and evidence are treated and how these treatments are reflected on the refugee decision.
9

Novel surface-tethered estrogen polymeric platforms in cardiovascular regenerative medicine

Qi, Baowen 07 1900 (has links)
L’estradiol (E2) est une hormone femelle qui joue un rôle essentiel, à la fois dans la régulation et dans la détermination de certaines conditions physiologiques in vivo, telle que la différenciation et la prolifération cellulaire. Lorsque l’E2 est donné en supplément, par exemple dans le cas de thérapie hormonale, deux effets sont observés, un effet génomique et un effet non-génomique, de par son interaction avec les récepteurs à œstrogène du noyau ou de la membrane cellulaire, respectivement. L’effet non-génomique est plus difficile à étudier biologiquement parce que l’effet se produit sur une échelle de temps extrêmement courte et à cause de la nature hydrophobe de l’E2 qui réduit sa biodisponibilité et donc son accessibilité aux cellules cibles. C’est pourquoi il est nécessaire de développer des systèmes d’administration de l’E2 qui permettent de n’étudier que l’effet non-génomique de l’œstrogène. Une des stratégies employée consiste à greffer l’E2 à des macromolécules hydrophiles, comme de l’albumine de sérum bovin (BSA) ou des dendrimères de type poly(amido)amine, permettant de maintenir l’interaction de l’E2 avec les récepteurs d’œstrogène de la membrane cellulaire et d’éviter la pénétration de l’E2 dans le noyau des cellules. Toutefois, ces systèmes macromolécules-E2 sont critiquables car ils sont peu stables et l’E2 peut se retrouver sous forme libre, ce qui affecte sa localisation cellulaire. L’objectif de cette thèse est donc de développer de nouvelles plateformes fonctionnalisées avec de l’E2 en utilisant les approches de synthèses ascendantes et descendantes. Le but de ces plateformes est de permettre d’étudier le mécanisme de l’effet non-génomique de l’E2, ainsi que d’explorer des applications potentielles dans le domaine biomédical. L’approche ascendante est basée sur un ligand d’E2 activé, l’acide 17,α-éthinylestradiol-benzoïque, attaché de façon covalente à un polymère de chitosan avec des substitutions de phosphorylcholine (CH-PC-E2). L’estradiol est sous forme de pro-drogue attachée au polymère qui s’auto-assembler pour former un film. L’effet biologique de la composition chimique du film de chitosan-phosphorylcholine a été étudié sur des cellules endothéliales. Les films de compositions chimiques différentes ont préalablement été caractérisés de façon physicochimique. La topographie de la surface, la charge de surface, ainsi que la rhéologie des différents films contenant 15, 25, ou 40% molaires de phosphorylcholine, ont été étudiés par microscopie à force atomique (AFM), potentiel zêta, résonance plasmonique de surface et par microbalance à cristal de quartz avec dissipation (QCM-D). Les résultats de QCM-D ont montré que plus la part molaire en phosphorylcholine est grande moins il y a de fibrinogène qui s’adsorbe sur le film de CH-PC. Des cellules humaines de veine ombilicale (HUVECs) cultivées sur des films de CH-PC25 et de CH-PC40 forment des amas cellulaire appelés sphéroïdes au bout de 4 jours, alors que ce n’est pas le cas lorsque ces cellules sont cultivées sur des films de CH-PC15. L’attachement de l’estradiol au polymère a été caractérisé par plusieurs techniques, telles que la résonance magnétique nucléaire de proton (1H NMR), la spectroscopie infrarouge avec transformée de Fourier à réfraction totale atténuée (FTIR-ATR) et la spectroscopie UV-visible. La nature hydrogel des films (sa capacité à retenir l’eau) ainsi que l’interaction des films avec des récepteurs à E2, ont été étudiés par la QCM-D. Des études d’imagerie cellulaires utilisant du diacétate de diaminofluoresceine-FM ont révélé que les films hydrogels de CH-PC-E2 stimulent la production d’oxyde nitrique par les cellules endothéliales, qui joue un rôle protecteur pour le système cardiovasculaire. L’ensemble de ces études met en valeur les rôles différents et les applications potentielles qu’ont les films de type CH-PC-E2 et CH-PC dans le cadre de la médecine cardiovasculaire régénérative. L’approche descendante est basée sur l’attachement de façon covalente d’E2 sur des ilots d’or de 2 μm disposés en rangées et espacés par 12 μm sur un substrat en verre. Les ilots ont été préparés par photolithographie. La surface du verre a quant à elle été modifiée à l’aide d’un tripeptide cyclique, le cRGD, favorisant l’adhésion cellulaire. L’attachement d’E2 sur les surfaces d’or a été suivi et confirmé par les techniques de SPR et de QCM-D. Des études d’ELISA ont montré une augmentation significative du niveau de phosphorylation de la kinase ERK (marqueur important de l’effet non-génomique) après 1 heure d’exposition des cellules endothéliales aux motifs alternant l’E2 et le cRGD. Par contre lorsque des cellules cancéreuses sont déposées sur les surfaces présentant des motifs d’E2, ces cellules ne croissent pas, ce qui suggère que l’E2 n’exerce pas d’effet génomique. Les résultats de l’approche descendante montrent le potentiel des surfaces présentant des motifs d’E2 pour l’étude des effets non-génomiques de l’E2 dans un modèle in vitro. / Estradiol (E2) is an essential female hormone in the regulation and determination of various physiological conditions in vivo, such as cell proliferation and differentiation. When supplementing exogenous E2 as a clinical strategy for hormone therapy, it generates genomic and non-genomic effect simultaneously via binding to the estrogen receptors in the cell nucleus or membrane site. Compared to the genomic effect, it is quite difficult to monitor the E2-induced non-genomic biological behavior because this effect occurs in extremely transient time scale and the bioavailability and accessibility of E2 to target cells is very low due to the hydrophobic nature of E2. As a result, it is indispensable to develop E2 delivery systems to specifically understand estrogenic non-genomic nature. One of strategies is to graft E2 to the hydrophilic macromolecules, e.g. bovine serum albumin (BSA) or poly(amido)amine dendrimer, to maintain E2 interacting with membrane estrogen receptors instead of penetrating into the cell nucleus. However, the instability of those E2-macromolecules systems, either containing free E2 leaching or discrepancies of cellular localizations, led to controversies. Herein, the objective of present thesis is to develop novel E2-functionlized platforms by the principle of bottom-up and top-down approaches for understanding the mechanism of estrogenic non-genomic effect, and further, to explore their potential applications in the biomedicine. As a bottom-up approach, an activated E2 ligand, 17α-ethinylestradiol-benzoic acid was covalently conjugated onto a phosphorylcholine substituted chitosan polymer (CH-PC-E2) as a prodrug strategy for the fabrication of self-assembled films. Through a series of combined physicochemical and cellular investigations, the relationship between various chemical compositions of chitosan-phosphorylcholine (CH-PC) films and cellular responses was also evaluated. Based on atomic force microscopy (AFM) examination, zeta-potential measurements, surface plasmon resonance (SPR), and quartz crystal microbalance with dissipation (QCM-D) measurements, surface topography, charge, and rheology of CH-PC films with 15, 25, and 40 mol% PC contents were characterized. Moreover, QCM-D measurements indicated that the amount of fibrinogen adsorbed on CH-PC films decreased significantly with increasing PC content. Finally, it was also showed that human umbilical vein endothelial cells (HUVECs) form spheroids on CH-PC25 and CH-PC40 films, but not on CH-PC15 films cultured over 4 days. In addition, the CH-PC-E2 polymer conjugates were prepared and characterized by several techniques, such as 1H nuclear magnetic resonance (1H NMR), Fourier transformed infrared-attenuated total refraction (FTIR-ATR) and UV/Vis spectra measurements. The hydrogel nature of CH-PC-E2 film as well as its interactions to estrogen receptors was further extensively investigated by QCM-D study. In the cellular study, CH-PC-E2 hydrogel films can significantly stimulate the production of nitric oxide, a protective molecule in the cardiovascular system, in the endothelial cells by a diaminofluorescein-FM diacetate imaging study. The studies above demonstrated the different roles and potential applications of CH-PC-E2 and CH-PC surfaces in the cardiovascular regenerative medicine. As a top-down approach, micropatterned substrates were used for E2 functionalization, which were prepared by photolithography via aligning ~ 2 μm in diameter gold arrays onto a glass substrate. After that, a cell adhesive peptide, cyclic RGD was introduced to the glass surface in order to induce the attachment of cells. Meanwhile, estradiol was covalently immobilized on the gold surface and the process was monitored and validated by combining SPR and QCM-D studies. In the micropatterned substrate-coupled cell ELISA study, a phosphorylation level of extracellular signal-regulated kinase (ERK), which is an important non-genomic marker, was significantly elevated by this E2-functionalized micropatterned surface after 1 hour incubation. Furthermore, E2-functionalized micropatterned substrate didn’t proliferate cancer cells indicating the absence of genomic effect stimulation. Based on these results, our E2-functionalized micropatterned substrates can function as an in vitro model for the elucidation of estrogenic non-genomic behaviors.

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