Eine wichtige Eigenschaft der Plasmamembran eukaryotischer Zellen ist die stabile transversale Asymmetrie der Phospholipide. Sie wird energieabhängig durch die Aktivität einer Aminophospholipidtranslokase aufrechterhalten und gilt als wichtige Voraussetzung für die Homöostasis der Zellen. Die Plasmamembran einiger Säugerzellen weist zudem laterale Lipiddomänen auf, denen eine wesentliche Bedeutung bei der Signaltransduktion zugeschrieben wird. Während der Genese durchlaufen die Membranen der Säugerspermien intensive Veränderungen. Um die Bedeutung der Phospholipidasymmetrie für die Funktion der Spermien zu untersuchen, wurde die Lokalisation und Dynamik von Phosphatidylserin in der Zell- und Akrosommembran von Eberspermien im Verlauf von Kapazitation und Akrosomreaktion betrachtet. Unter Ausnutzung der selektiven, kalziumabhängigen Bindung von AnnexinV an endogenes Phosphatidylserin konnte dessen Lokalisation an morphologisch differenzierten Zellen verfolgt werden. Eine Markierung der Zellen mit NBD-markierten Phospholipidanaloga lieferte zudem Informationen zur Dynamik der Phospholipide in der Plasmamembran. Die Differenzierung der Zellen erfolgte entweder am Durchflusszytometer oder fluoreszenz- bzw. elektronenmikroskopisch. Die Ergebnisse der vorliegenden Arbeit weisen sowohl auf eine transversale als auch laterale Ungleichverteilung der Lipide in der Zell- und Akrosommembran während der Genese der Spermien hin. Neben der stabilen transversalen Phospholipidasymmetrie der Plasmamembran konnte erstmals eine zytoplasmatische Lokalisation von Phosphatidylserin auf der äußeren Akrosommembran nachgewiesen werden. Somit akkumulieren die beiden einander zugewandten zytoplasmatischen Monolayer von Plasmamembran und äußerer Akrosommembran Phosphatidylserin. Kapazitationsbedingt kommt es zu einer engen Wechselwirkung zwischen Plasmamembran und äußerer Akrosommembran. Die Ausbildung lateraler Membrandomänen, in denen Phosphatidylserin zytoplasmatisch akkumuliert, wird als Voraussetzung für diese enge Assoziation diskutiert. Weitere Hinweise auf eine funktionelle Bedeutung lateraler Membrandomänen lieferten die Arbeiten zur Isolation Triton-unlöslicher Lipiddomänen aus der Plasmamembran von Forellenspermien. / One of the essential qualities of cell membranes in Eucaryotae is a stable transverse phospholipid asymmetry. It is regulated and maintained by ATP-dependent action of an aminophospholipid translocase and is a major prerequisite for cell homeostasis. The plasma membranes of several mammalian cells show moreover lateral lipid domains, which are imputed to play a significant role in signal transduction. The membranes of mammalian spermatozoa undergo significant changes during genesis. The localisation and dynamics of phosphatidylserine in the cell as well as acrosome membranes of boar sperm cells was studied during capacitation and acrosome reaction to assess the relevance of lipid asymmetry for sperm function. The localisation of endogenous phosphatidylserine in morphologically differentiated cells was followed using the selective calcium depending binding of annexinV. Information on the transverse dynamics of phospholipids in the plasma membrane was obtained by labelling the cells with a NBD-phospholipid analogues. The morphological status of the cells was assessed by flow cytometry, fluorescence and electron microscopy. The results of this study indicate both a transversal and lateral inhomogenous distribution of lipids in the cell membrane as well as in the outer acrosome membrane during sperm genesis. The plasma membrane of boar sperm shows a stable transversal lipid asymmetry characterised by an accumulation of phosphatidylserine in the cytoplasmic monolayer. Moreover a cytoplasmic localisation of phosphatidylserine on the outer acrosome membrane could be detected for the first time. Therefore the two facing cytoplasmic leaflets of the outer acrosome and cell membrane contain phosphatidylserine. Applying microscopy substantiated the hypothesis that there are close interactions between the cell membrane and the outer layer of the acrosome membrane because of capacitation. The cytoplasmic accumulation of phosphatidylserine in lateral lipid domains is probably essential for the strong association of plasma and outer acrosome membrane finally leading to local fusions of both membranes. An indication for the functional meaning of lateral membrane domains in sperm cells was futher deduced from the isolation of Triton-insoluble lipid domains from membranes of trout sperm cells.

Submitted by Joana Azevedo (joanad@id.uff.br) on 2017-07-04T12:35:47Z No. of bitstreams: 1 Dissert Norberto Matos.pdf: 1637531 bytes, checksum: 409b4369806baa8dbe52e4ac1c2c961d (MD5) / Rejected by Biblioteca da Escola de Engenharia (bee@ndc.uff.br), reason: Item rejeitado, pois a ficha catalográfica está fora do padrão. Atenciosamente, Catarina Ribeiro Bibliotecária BEE - Ramal 5992/5993/5994 on 2017-07-05T15:15:33Z (GMT) / Submitted by Joana Azevedo (joanad@id.uff.br) on 2017-07-25T19:25:47Z No. of bitstreams: 1 Dissert Norberto Matos.pdf: 1637756 bytes, checksum: a2a04a275ae9a2cb97450fcb5110bff6 (MD5) / Approved for entry into archive by Biblioteca da Escola de Engenharia (bee@ndc.uff.br) on 2017-07-28T15:02:38Z (GMT) No. of bitstreams: 1 Dissert Norberto Matos.pdf: 1637756 bytes, checksum: a2a04a275ae9a2cb97450fcb5110bff6 (MD5) / Made available in DSpace on 2017-07-28T15:02:38Z (GMT). No. of bitstreams: 1 Dissert Norberto Matos.pdf: 1637756 bytes, checksum: a2a04a275ae9a2cb97450fcb5110bff6 (MD5) Previous issue date: 2017-04-07 / Apesar do relativo avanço no sentido de manter a igualdade entre os cidadãos, a sociedade brasileira ainda não atende às necessidades específicas de alguns grupos, como: idosos, enfermos, crianças, gestantes e pessoas com deficiência (PcDs). Porém, no caso das PcDs, atualmente existem leis que visam incluí-las no mercado de trabalho, o que vem contribuindo para a sua contratação. Este trabalho objetivou investigar os principais desafios encontrados no processo de capacitação de PcDs pertencentes ao quadro efetivo do Centro Federal de Educação Tecnológica Celso Suckow da Fonseca (Cefet-RJ), que é uma instituição pública destinada ao ambiente educacional com formação humana, científica e tecnológica. Foi realizada uma pesquisa bibliográfica a partir dos conceitos-chave que auxiliaram na identificação de publicações da temática estudada. A metodologia utilizada foi o estudo de caso com aplicação de questionários, os quais foram respondidos por grupos de funcionários com deficiência, de seus gestores, e também do setor de capacitação, sendo todos servidores públicos pertencentes ao quadro de profissionais do Cefet-RJ. O tratamento dos dados foi realizado através da análise de conteúdo. O trabalho de campo mostrou o envolvimento do profissional PcD com a instituição. Os objetivos almejados nesta pesquisa foram atingidos e direcionados para propostas de melhorias nos processos de inserção, contratação e adaptação das PcDs em seu ambiente de trabalho. / Despite relative efforts in order to sustain citizen equality, Brazilian society still remains inattentive to some groups’ especial needs, such as the elderly, children, pregnant, sick and people with disability (PwD). However, concerning the PwDs, a current legislation aiming to insert them in the labor market actually exists and have been contributing to their hiring. This paper focused on the main challenges found during the PwDs capacitation process in the Centro Federal de Educação Tecnológica Celso Suckow da Fonseca (Cefet-RJ) full-time staff. Cefet-RJ is a public institution destined to be an education environment for human, scientific and technological formation. From key concepts that helped identify publications for this studied theme, a bibliographical research was achieved. We used the case study methodology, in addition of questionnaires, answered by groups of employees with disability, their managers and also by the resource and training center. All questioned employees are from Cefet’s full-time staff. The data handling was done by analyzing the content. The fieldwork showed PwDs’ involvement with the Institution. The targets were achieved and the results directed as improvement proposals for insertion, hiring and adaptation processes in the working environment.

Submitted by Estagiário SPT BMHS (spt@fgv.br) on 2011-11-09T10:14:40Z No. of bitstreams: 1 000384143.pdf: 9419150 bytes, checksum: 93ff81a6759584a612f6608a70d4f254 (MD5) / Approved for entry into archive by Estagiário SPT BMHS (spt@fgv.br) on 2011-11-09T10:14:56Z (GMT) No. of bitstreams: 1 000384143.pdf: 9419150 bytes, checksum: 93ff81a6759584a612f6608a70d4f254 (MD5) / Approved for entry into archive by Estagiário SPT BMHS (spt@fgv.br) on 2011-11-09T10:15:05Z (GMT) No. of bitstreams: 1 000384143.pdf: 9419150 bytes, checksum: 93ff81a6759584a612f6608a70d4f254 (MD5) / Made available in DSpace on 2011-11-09T10:15:15Z (GMT). No. of bitstreams: 1 000384143.pdf: 9419150 bytes, checksum: 93ff81a6759584a612f6608a70d4f254 (MD5) Previous issue date: 2006 / As micro e pequenas empresas representam 99,2% das empresas no Brasil, empregam 57,2% da força de trabalho por isso são imprescindíveis para a economia e o desenvolvimento do país, apesar desses números representativos a grande preocupação é manter essas empresas ativas devido ao grande índice de mortalidade que representa em média 50%, tendo como um dos principais fatores condicionantes a falha gerencial na condução da empresa. Micro e pequenas empresas: A importância de apreender a empreender é o tema desta dissertação, que tem, por objetivo avaliar o conhecimento que os micro e pequenos empresários têm sobre a importância do desenvolvimento pessoal no gerenciamento de seus negócios. Por tratar-se de micro e pequenas empresas elegemos o Sebrae, instituição de apoio às micro e pequenas empresas de São Luís do Maranhão para investigar através de pesquisa de campo qual o conhecimento que os micro e pequenos empresários tem sobre está instituição seus produtos e serviços. O referencial teórico demonstrou que treinamento, capacitação, planejamento e aprendizado contínuo são ferramentas eficazes na administração organizacional de uma empresa. Na pesquisa de campo identificou-se que 76% dos empresários não buscaram nenhuma ajuda especializada antes de abrir seus negócios, apenas 45% dos empresários conhecem a marca do Sebrae e 54% não conhecem os produtos e serviços que o Sebrae São Luís oferece. / Small and middle-sized businesses represent 99,2% of the companies in Brazil. They employ 57,2% of the workforce and are indispensable for the country's economy and development, notwithstanding these representative numbers, the great concern is to keep these companies active owing to the high mortality rate, which means 50% on average as one ofthe chief conditioning factors for managerial failures in the conduct of the company. Small and middle-sized businesses. The importance to become an entrepreneur' is the theme of this present dissertation whose objectives are to assess the understanding that small and middlesized businessmen have about the importance of personal development in their business management. Because it is a question of small and middle-sized businesses, SEBRAE, an institution in support for small and middle-sized businesses in São Luis do Maranhão, has been elected to investigate through field research what knowledge the small and middle sized businesses have about SEBRAE and its products and services. The referential theory has shown that training, professional capacitation, planning and constant learning are effective tools in a company's organizational management and it has been identified by the field research that 77% of businessmen did not seek for any specialized help before going into business, that only 45% of businessmen know about SEBRAE and 54% are not familiar with the products and services that SEBRAE - São Luis offer.

In the 21st century, there is a growing realisation that providing principals with the necessary opportunities of training in leadership and management skills has become increasingly important as a way to increase school effectiveness and achieve quality performance. Central to this argument is the fact that principals are merely accorded a status and role without the necessary training. The purpose of this study was therefore to explore the effectiveness of state-funded professional development programmes of school principals with specific reference to Soshanguve secondary schools and also to ascertain the extent to which principals’ training meets the schools’ and principals’ needs given the changed conditions that exist in the country. The study employed a quantitative research methods approach to collect data from 100 educators and 20 Head of Departments (HoDs). This was done through administration of questionnaires. Data was analysed using the Statistical Package for the Social Sciences (SPSS), which was used to generate the frequencies and descriptive statistics that were used to answer research questions. While it is expected that principals should effectively and rigorously be developed to enable them to improve teaching and learning at their schools, the study, however, found that training of school principals in Soshanguve was elusive and ineffective, and as result, the majority of these principals lack basic management training prior to and after their entry into principalship. The study has shown that apart from being faced with various challenges and being promoted to principalship without the necessary leadership experience, principals rely on trial-and-error experience and common sense in leading and managing schools. The study also showed that a majority of principals are in great need of being professionally developed in some specific areas. Furthermore, the study discovered that the Department of Basic Education does not seem to be making efforts in ensuring that principals are professionally developed. Consequently, principals develop on their own through informal professional development strategies. In conclusion, the study recommended, amongst others, that principals should be encouraged to take personal responsibility and initiative in preparing and developing school leadership through self-study, reading literature, attending seminars and workshops out of their own personal volition. That Department of Basic Education (DoBE) needs to look into ways of subsidising the training of school principals. DoBE should also find ways of formally incorporating more of the private sector and non-governmental organisations to help in the preparation and development of principals for school leadership by allowing them to offer in-service courses for potential principals and serving principals. / Educational Leadership and Management / D. Ed. (Education Management)

Made available in DSpace on 2014-06-11T19:35:12Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-11-24Bitstream added on 2014-06-13T20:06:44Z : No. of bitstreams: 1 azevedo_hc_dr_botfmvz.pdf: 1090855 bytes, checksum: bd7f7a04aa00c86d1200aecb917ca794 (MD5) / Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Objetivando testar a ação do colesterol, desmosterol, ácido oléico-linoléico ou Q-Iactoalbumina sobre os espermatozóides ovinos, alíquotas de sêmen de 25 carneiros foram submetidas à criopreservação após tratamento com essas substâncias. O sêmen foi avaliado antes do processamento e após a refrigeração, descongelação e incubação quanto à, cinética pela análise subjetiva e computadorizada (CASA), morfologia, avaliação simultânea da integridade da membrana plasmática (IMP) e do acrossomo (IAC) e do potencial de membrana mitocondrial (PMM) pela associação da aglutinina de Pisum sativum conjugada ao isotiocianato de fluoresceína (FITC-PSA), iodeto de propídio (PI) e JC-1 e, status de capacitação pela c10rtetraciclina (CTC). A influência dos aditivos em poucos parâmetros não foi suficiente para sugerir a superioridade de qualquer um dos tratamentos. A criopreservação provocou prejuízos à cinética, IMP, IAC, PMM, além de induzir a capacitação e reação do acrossomo. Foram observadas diferenças entre grupos de carneiros quanto ao status de capacitação no sêmen in natura, assim como na sensibilidade desse sêmen à crioinjúria e criocapacitação. Após a descongelação, as mudanças semelhantes à capacitação foram maiores nos animais de menor IMP. A segregação dos carneiros de acordo com a crioresistência da membrana plasmática norteou o comportamento de vários outros parâmetros. Conclui-se, que o colesterol, desmosterol, ácido oléico-linoléico e alactoalbumina não protegem o sêmen ovino contra as crioinjúrias, que a congelação-descongelação é mais danosa que a refrigeração e, que existem diferenças entre grupos de indivíduos relacionadas à crioresistência e criocapacitação espermática. / In order to evaluate the proteeting aetion of some substanceson ram spermatozoa, semen samples of 25 rams were colleeted and submitted to cryopreservation after treatment with cholesterol, desmosterol, oleic-linoleic acid or Q-Iactalbumin. Before the processing and after the procedures of cooling, freezing-thawing and incubation, semen was evaluated as to kínetcs by subjective and computerized analyses (CASA), morphology, simultaneously to plasma membrane integrity (PMI), acrosome integrity (ACI) and mitochondrial membrane potential (MMP) by isothiocyanate-conjugated Pisum sativum (FITCPSA), propidium iodide (PI) and JC-1 combination and as to sperm capacitation and acrosome reaction determined by chlortetracycline (CTC) assay. The influence resulted from the additives in a few parameters was not enough to suggest the superiority of any treatments. The cryopreservation process, especially the freezing-thawing, promotes damages to kinetics, PMI, ACI, MMP and induees accelerated eapacitation and acrosome reaction in spermatozoa. Differenees among groups of rams were observed regarding capacitation status in fresh semen as well as the sensibility of their semen to cryoinjury and cryocapacitation. Higher and more accelerated capacitation-like ehanges were observed in groups of rams presenting lower plasmatic membrane cryoresistance after thawing. The segregation of rams in different levels of PMI influenced the behavior of several other parameters. It was concluded that cholesterol, desmosterol, oleic-Iinoleic acid and Q-Iactalbumin do not protect the ram semen against the cryoinjuries as well as freezing-thawing is more harmful than cooling. Important differenees among groups of individuais were noticed as for the spermatozoa resistance to the damages provoked by cryopreservation such as the eryocapacitation susceptibility.

In the first part of this doctoral thesis, a new analytical HPLC-MS/MS method for monitoring of concentration changes of 17β-estradiol (βE2) during in vitro mouse sperm capacitation was developed. Capacitation was performed for three initial concentrations of βE2 (200, 20 and 2 μg/L). For all the concentrations a similar trend for the total unbound βE2 was observed. In general, the βE2 concentration decreased to reach its minimum and then increased again. The position of the minimum differed for the individual tested βE2 concentrations. Experimentally obtained results were subjected to the kinetic analysis. The curves fitted through the experimentally determined points displayed an autocatalytic character. For the agreement between the curves obtained by fitting through the experimental points and the theoretical calculated curves, it is necessary to assume that the first step is adsorption of βE2 onto the surface of the sperm controlled by Langmuir isotherm. The kinetic study was also used to study the effects of fluorides and aluminium fluoride complexes on the capacitation of mouse sperm. The experimental points were in very good agreement with the shape of the theoretical curves and this fact verifies the mechanism of the mouse sperm capacitation kinetics. In the second part of this work, two...

No description available.

Jamais nos modes de vie et les produits qui nous entourent n'ont évolué aussi rapidement. Cette évolution est un facteur nouveau à prendre en compte dans le développement de produits pour s'assurer de leur appropriation par les utilisateurs. C'est ainsi que les approches de conception s'orientent petit à petit vers l'intégration des usages. L'approche Living Lab est axée autour de quatre principes : le réalisme de la situation d'usage étudiée ; la continuité de la collaboration avec les utilisateurs ; la collecte de données d'usage spontanées ; l'augmentation de la capacité des utilisateurs à influencer le développement du produit. Ces principes facilitent le passage de la conception centrée-utilisateurs vers la conception centrée-usages, mais sont difficiles à réaliser concrètement. Pour accompagner et inspirer les concepteurs, nous proposons une méthode de pilotage de projet Living Lab composée de dix-huit opérations visant chacune l'atteinte d'un de ces principes. La méthode proposée est représentée sous forme de diagrammes NIAM-ORM, facilement compréhensibles car proche du langage naturel binaire. Deux projets urbains et deux projets industriels ont permis de tester notre méthode. La pertinence des opérations sur l'atteinte des principes est évaluée par le biais des réseaux bayésiens : selon les résultats obtenus, les opérations sont validées, ajustées ou reformulées puis ré-implémentées dans le modèle NIAM-ORM / Our way of life and the products that surround us have never evolved so quickly. This evolution is a new factor to take into account in the product development to ensure the product appropriation by the users. For that reason the design approaches try to integrate usages. The Living Lab approach is organised around four principles: the realism of the studied situation; the continuity in the collaboration with the users; the collection of spontaneous usage data; the empowerment of users in the design process. These principles facilitate the path from "user-centred design" to "usage-centred design" but are difficult to realise practically. To guide and inspire the designers we propose a Living Lab project method made of eighteen operations; each operation is supposed to reach one of the four principles. The proposed method is represented with NIAM-ORM diagrams which are easily understandable because of its similarity with natural language. We tested our method on two urban projects and two industrial projects. The relevance of the tested operations on the principle realisation is evaluated according to bayesian networks: depending on the results the operations are validated, adjusted or rephrased and implemented again in the NIAM-ORM model

Als Kapazitation bezeichnet man den im weiblichen Reproduktionstrakt stattfindenden Reifungsschritt, der Spermien das volle Fertilisierungspotential verleiht. Die molekularbiologischen Grundlagen dieses für eine erfolgreiche natürliche oder auch artifizielle Befruchtung essenziellen Prozesses sind bis heute nur unvollständig verstanden. Im Rahmen der vorliegenden Dissertation wurden die mit der Kapazitation einhergehenden funktionellen und strukturellen spermalen Veränderungen untersucht. Die kapazitative Stimulation führte zu einer gesteigerten Motilität bis hin zur Hyperaktivierung, zu einer vermehrt induzierten Akrosomenreaktion und zu einer deutlich reduzierten Apoptoseaktivität. Anhand von Inhibitionsexperimenten wurde die Rolle der potentiellen Signaltransduktoren Caspase-1, Calpain und Calmodulin analysiert. Dabei wies die Calmodulinantagonisierung auf eine ausgeprägte Calciumabhängigkeit aller untersuchten kapazitationsassoziierten Prozesse hin. Die Hemmung von Caspase-1 und Calpain führte zu einer Beeinträchtigung der Motilität und der Akrosomenreaktion ohne das Ausmaß der Apoptoseinduktion zu beeinflussen. Die vorstehend genannten Erkenntnisse wurden zur Evaluation verschiedener Ejakulataufbereitungsprotokolle genutzt. Dabei konnte gezeigt werden, dass die Kombination des modernen Verfahrens der immunomagnetische Zellseparation mit der etablierten Methode der Dichtegradientenzentrifugation dem einfachen Standard in Bezug auf die Anreicherung hochmotiler Spermien mit minimaler Apoptoseaktivität aus frischen wie auch aus kryokonservierten Ejakulaten deutlich überlegen war. Bedeutsam im Hinblick auf eine mögliche pratische Anwendung der immunomagnetischen Zellseparation erscheint der Befund, dass die durch das kombinierte Anreicherungsverfahren erhaltene Spermatozoensubpopulation im Hamsteroozytenpenetrationstest ein signifikant höheres Fertilisierungspotential zeigte.

Als Kapazitation bezeichnet man den im weiblichen Reproduktionstrakt stattfindenden Reifungsschritt, der Spermien das volle Fertilisierungspotential verleiht. Die molekularbiologischen Grundlagen dieses für eine erfolgreiche natürliche oder auch artifizielle Befruchtung essenziellen Prozesses sind bis heute nur unvollständig verstanden. Im Rahmen der vorliegenden Dissertation wurden die mit der Kapazitation einhergehenden funktionellen und strukturellen spermalen Veränderungen untersucht. Die kapazitative Stimulation führte zu einer gesteigerten Motilität bis hin zur Hyperaktivierung, zu einer vermehrt induzierten Akrosomenreaktion und zu einer deutlich reduzierten Apoptoseaktivität. Anhand von Inhibitionsexperimenten wurde die Rolle der potentiellen Signaltransduktoren Caspase-1, Calpain und Calmodulin analysiert. Dabei wies die Calmodulinantagonisierung auf eine ausgeprägte Calciumabhängigkeit aller untersuchten kapazitationsassoziierten Prozesse hin. Die Hemmung von Caspase-1 und Calpain führte zu einer Beeinträchtigung der Motilität und der Akrosomenreaktion ohne das Ausmaß der Apoptoseinduktion zu beeinflussen. Die vorstehend genannten Erkenntnisse wurden zur Evaluation verschiedener Ejakulataufbereitungsprotokolle genutzt. Dabei konnte gezeigt werden, dass die Kombination des modernen Verfahrens der immunomagnetische Zellseparation mit der etablierten Methode der Dichtegradientenzentrifugation dem einfachen Standard in Bezug auf die Anreicherung hochmotiler Spermien mit minimaler Apoptoseaktivität aus frischen wie auch aus kryokonservierten Ejakulaten deutlich überlegen war. Bedeutsam im Hinblick auf eine mögliche pratische Anwendung der immunomagnetischen Zellseparation erscheint der Befund, dass die durch das kombinierte Anreicherungsverfahren erhaltene Spermatozoensubpopulation im Hamsteroozytenpenetrationstest ein signifikant höheres Fertilisierungspotential zeigte.