INTRODUÇÃO: Este trabalho avalia a capacitação dos multiplicadores do Projeto \"Nossas Crianças: Janelas de Oportunidades\", a partir da percepção dos sujeitos participantes. Tal Projeto tem como tema o desenvolvimento infantil, na perspectiva da Promoção da Saúde, por meio da atuação das equipes do PSF, junto às famílias com gestantes e crianças na faixa etária de 0 a 6 anos. O estudo considera a consonância entre a Metodologia Pedagógica empregada na capacitação e os referenciais teóricos que embasaram o Projeto, tendo em vista o empowerment dos profissionais. A estratégia escolhida para a capacitação foi a Oficina Problematizadora. OBJETIVO: Avaliar o processo de capacitação dos multiplicadores do Projeto Janelas a partir da percepção dos mesmos, por meio do grau de apreensão dos conteúdos apresentados, e da adequação da estratégia pedagógica empregada à finalidade de reproduzirem-na junto às equipes. MATERIAL E MÉTODO: A pesquisa realizada foi de natureza qualitativa na modalidade estudo de caso, com a utilização de entrevistas semi-estruturadas e posterior análise temática. A amostra foi intencional e o critério de inclusão dos participantes foi o maior grau de manutenção de suas atividades, relacionadas à implementação do Projeto em suas unidades de origem. Foram entrevistados oito participantes, número suficiente para contemplar o critério de saturação dos dados. RESULTADOS: Foram entrevistados sete mulheres e um homem, nas faixas etárias entre 25 e 50 anos, sendo cinco enfermeiras, uma técnica em enfermagem, uma assistente social e um médico, dentre eles, dois bacharéis, duas especialistas em Saúde da Família, uma sanitarista, um Mestre e uma Doutora. A grande maioria permanece de 3 a 4 anos na estratégia. A análise das falas demonstrou uma avaliação fortemente positiva pelos entrevistados quanto à sistematização e apreensão de conteúdos como: cuidados, resiliência, competências familiares, rede social e família, que os instrumentalizou para o trabalho. A Metodologia Problematizadora possibilitou um resgate de suas realidades, da experiência de grupo, da participação e da vivificação do trabalho. CONCLUSÃO: A avaliação demonstra que houve apreensão dos conteúdos pelos participantes, bem como adequação entre a metodologia e os referenciais teóricos do Projeto, possibilitando aos mesmos transmitirem seus conteúdos junto às suas equipes / INTRODUCTION: This work evaluates the qualification of the multipliers of the Project Our Children: Windows of Chances, from the perception of the participant citizens. This Project has as subject the infantile development, in the perspective of the Health Promotion, by means of the performance of the PSF teams, together to the families with pregnant women and children aged from 0 to 6 years. The study considers the accord between the employed Pedagogical Methodology in the theoretical qualification and referenciais that had based the Project, considering the empowerment of the professionals. The strategy chosen for the qualification was the Problemized Workshop. OBJECTIVE: To evaluate the process of qualification of the multipliers of the Project Windows from the perception of the same ones, by means of the degree of apprehension of the presented contents, and the adequacy of the pedagogical strategy used to the purpose to reproduce it together to the teams. MATERIAL AND METHOD: The research was of qualitative nature in the modality of case study, with the use of half-structuralized interviews and posterior thematic analysis. The sample was intentional and the criterion of inclusion of the participants was the biggest degree of maintenance of its activities, related to the implementation of the Project in its units of origin. Eight participants had been interviewed, number enough to contemplate the criterion of saturation of the data. RESULTS: Seven women had been interviewed and a man, in the age from 25 to 50 years, being five nurses, one technique in nursing, a social assistant and a doctor, amongst them, two graduated people, two specialists in Family Health, a sanitary agent, a Master and a Doctor. The great majority remains from 3 to 4 years in the strategy. The analysis of the conversations demonstrated a positive evaluation for the interviewed ones related to the systematization and apprehension of contents as: familiar cares, resilience, abilities, social net and family, who instrumentalized them for the work. The Problematized Methodology made possible a rescue of its realities, of the experience of group, the participation and the experience of the work. CONCLUSION: The evaluation demonstrates that the apprehension of the contents happened between the participants, as well as adequacy between the methodology and the theoretical referenciais of the Project, making possible the same ones to transmit its contents together to its teams

O presente estudo visou investigar o efeito da refrigeração do sêmen da jaguatirica sobre o Índice de Motilidade Espermática [IME=(%M+MPx5)/2; %M = proporção de espermatozóides móveis; MP = motilidade progressiva], integridade acrossomal (IA) e capacitação espermática; assim como avaliar a eficácia da técnica FITC-PNA/IP na avaliação simultânea da viabilidade espermática (VE) e IA. Sete jaguatiricas foram eletroejaculadas, sendo utilizados apenas ejaculados (n=16) apresentando %M>=60% e MP>=3. Avaliou-se a IA por meio da Coloração Simples. Os ejaculados foram diluídos 1:1 na Variante do Diluente de PLatz e submetidos aos Protocolos de Transporte: Temperatura Ambiente e Refrigeração, - 0,23ºC/min, (Experimento 1); ou apenas Temperatura Ambiente (Experimentos 2 e 3). Após 2h, as alíquotas foram reaquecidas, reavaliando-se os parâmetros observados antes do transporte. Os espermatozóides foram lavados por centrifugação em meio F10 de Ham, ressuspensos nesse meio e processados conforme o experimento: (1) após pré-incubação (38ºC; 5%CO2) durante 0, 1, 2 e 4 horas, foram retiradas alíquotas a cada intervalo para serem incubadas (30 min) na ausência e na presença do cálcio ionóforo A23187 (Ca2+Ion) (1mM), avaliando-se IA e IME; (2) após pré-incubação por 0, 1 e 2h, foram incubadas alíquotas na ausência e presença de 1 e 2mM de Ca2+Ion, avaliado-se IA e IME; (3) pré-incubados por 9h, sendo retiradas alíquotas a cada hora, para as avaliações da IA e VE, (a) separadamente através da Coloração Simples e do IME, ou (b) simultaneamente através da técnica FITC-PNA/IP. A refrigeração causou declínio (p<0,02) da IA (71,0%) e IME (67,1), em comparação aos valores observados antes do transporte (88,5%; 85,4), enquanto a manutenção das amostras à temperatura ambiente não afetou (p>0,1) essas variáveis (84,8%; 76,4). Dentre as amostras refrigeradas, aquelas expostas ao Ca2+Ion sofreram redução (p<0,01) na IA (52,4%) frente ao controle (55,56%). Já nas amostras transportadas à temperatura ambiente, não foi observada diferença (p>0,1) entre os grupos com e sem ionóforo (64,41% vs. 63,87%). Quando analisados os tempos separadamente, o único tratamento em que houve efeito (p<0,05) do Ca2+Ion sobre a IA foi aquele refrigerado e pré-incubado por 2h. Foi verificada redução (p<0,05) nos valores de IME e IA devida à simples incubação, mesmo na ausência do Ca2+Ion. A concentração de 2µM dessa substância foi mais efetiva na indução da reação acrossômica que 1µM. Apesar dos fluorocromos FITC-PNA/IP terem se ligado aos espermatozóides, nas regiões esperadas, a proporção de células marcadas variou aleatoriamente durante pré-incubação, sem correlação (p>0,1) com IME. A IA avaliada pela Coloração Simples apresentou correlação positiva (r=0,77; p<0,0001) com IME, decrescendo (p<0,0001) durante pré-incubação. A refrigeração mostrou-se desvantajosa frente à manutenção do sêmen à temperatura ambiente, pois foi deletéria à função e às membranas dos espermatozóides. A refrigeração tornou-os capazes de responder ao estímulo do Ca2+Ion, característica observada nos espermatozóides capacitados. O ensaio de reação acrossômica induzida pelo Ca2+Ion deve ser aperfeiçoado para permitir avaliação acurada da capacitação espermática na jaguatirica. A Coloração Simples associada à avaliação do IME foi mais eficiente e menos laboriosa, frente á técnica FITC-PNA/IP, na avaliação da IA e VE. / This study aimed to investigate the effect of ocelot semen refrigeration on Sperm Motility Index [SMI=(%M+PMx5)/2; %M = proportion of motile spermatozoa ; PM = Progressive Motility], acrossomal integrity (AI) and sperm capacitation. Another objective was to evaluate the FITC-PNA/IP technique efficacy on evaluating simultaneously sperm viability (SV) and AI. Five ocelots, were electroejaculated, the semen was evaluated and only ejaculates (n=16) presenting %M>=60% and PM>=3 were used. Sperm AI was evaluated using Fast Green / Rose Bengal staining (FGRB). The ejaculates were diluted 1:1 in Platz Diluent Variant and subjected to the transportation protocols: Room Temperature and Cooling, -0.23ºC/min, (experiment 1); or only Room Temperature (experiments 2 and 3). After 2 hours, the aliquots were rewarmed and samples were taken to re-evaluate the parameters observed before the transport. The spermatozoa were washed in Ham’s F10 medium, ressuspended in fresh medium and processed differently, according the experiment: (1) after pre-incubation (38ºC; 5%CO2) during 0, 1, 2 and 4 hours, samples were taken at each time point to be incubated in the absence and presence of 1mM calcium ionophore A23187 (Ca2+Ion), SMI and AI were evaluated; (2) after pre-incubation during 0, 1 and 2h, aliquots were incubated in the absence and presence of 1 and 2 mM Ca2+Ion; SMI and AI were evaluated; (3) after pre-incubation during 9h, aliquots were taken every hour to compare the evaluation of SV and AI (a) separately by the FGRB staining and SMI or (b) simultaneously by the FITC-PNA / IP technique. Cooling caused decline (p<0.02) on AI (71.0%) and SMI (67.1), when compared to values observed before transportation (88.5%; 85.4). Maintenance at room temperature didn’t affect (p>0.1) these variables (84.8%; 76.4). Among cooled samples, spermatozoa exposed to Ca2+Ion showed smaller (P<0.01) AI value (52.4%) compared to the group incubated without that substance (55.56%). For samples transported at room temperature, it wasn’t observed difference (P>0.05) between the groups with and without ionophore (64.41% vs. 63.87%). When time intervals were analysed separately, the only treatment in which there was effect (p<0,05) of Ca2+Ion on AI was the group refrigerated and pre-incubated for 2h. There was a reduction (p<0,05) on SMI and AI due simply to incubation, even in the absence of Ca2+Ion. The 2µM concentration of this substance was more effective to induce acrosome reaction than 1µM. FITC-PNA and IP fluorocromes bound spermatozoa at the expected sites. However, proportion of marked cells varied randomly during pre-incubation, and didn’t correlate (p>0,1) with SMI. IA evaluated by FGRB staining showed positive correlation (r=0,77; p<0,0001) with SMI, decreasing (p<0,0001) during incubation. Cooling was disadvantageous compared to maintaining semen at room temperature, since it was deleterious to spermatozoa membranes and function, and made those cells capable to answer the Ca2+Ion challenge, a characteristic observed in capacitated spermatozoa. Ca2+Ion induced acrosome reaction assay must be improved to allow accurate evaluation of sperm capacitation on ocelots. FGRB staining associated to SMI evaluation was more efficient and easier to perform, than FITC-PNA/IP technique, for AI and SV investigation.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-06-23T18:37:14Z No. of bitstreams: 1 danielleteodorosantoscamarota.pdf: 1062198 bytes, checksum: 5004c4eb4251e7f7721ed54547e0b364 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-07-13T15:41:19Z (GMT) No. of bitstreams: 1 danielleteodorosantoscamarota.pdf: 1062198 bytes, checksum: 5004c4eb4251e7f7721ed54547e0b364 (MD5) / Made available in DSpace on 2016-07-13T15:41:19Z (GMT). No. of bitstreams: 1 danielleteodorosantoscamarota.pdf: 1062198 bytes, checksum: 5004c4eb4251e7f7721ed54547e0b364 (MD5) Previous issue date: 2012-10-31 / Com o intuito de reafirmar a importância da formação no exercício da gestão escolar, este Plano de Ação Educacional (PAE) tem como objetivo a proposição de um programa que visa capacitar os professores, da rede municipal de ensino de Juiz de Fora, MG, que anseiam concorrer ao cargo de gestor escolar, para o exercício da função. Resta claro que, atualmente, com uma sociedade cada vez mais complexa e com o arcabouço de informações que fazem parte do cotidiano escolar, os diretores estão sendo cada vez mais exigidos. Sendo assim, é possível observar que muitos diretores, ainda que eleitos por processos democráticos, encontrem dificuldades para desempenhar as inúmeras demandas advindas do cargo. Tal fato se evidencia, principalmente, no que tange à compreensão das políticas públicas implementadas pelas esferas federais e estaduais, como, por exemplo, as políticas de avaliação. Isso pode ser vislumbrado na forma como os gestores lidam com os indicadores educacionais disponibilizados para suas unidades. Também é possível identificar outras dificuldades, como a falta de habilidade no uso de ferramentas tecnológicas, bem como a dificuldade para lidar com a questão orçamentária e de gestão de pessoas. Isso posto, a proposta é criar um programa de formação e capacitação de gestores escolares para a rede municipal de ensino de Juiz de Fora, tendo o Centro de Formação do Professor como espaço propício para estímulo e aperfeiçoamento das habilidades inerentes ao cargo de gestor educacional. / Aiming to reaffirm the importance of the in-service training of school managers, this Educational Action Plan (PAE, in Portuguese) has as its goal the proposal of a program designed to capacitate teachers from the municipal education network of Juiz de Fora, who aspire to run for school managers, exercising such function. It is clear that nowadays with a society ever growing in complexity, and with the framework of information pertaining to the school daily life, principals are being under increasing demand. As such, it is possible to observe that many principals, even though elected through democratic processes, find difficulty in playing the roles that come with the position. Such fact is made evident, in the scope of understanding public policies implemented by the federal and state spheres, as for example, evaluation policies. This may be observed in the way that managers deal with educational indicators made available to their units. It is also possible to identify other difficulties, such as the lack of skills in utilizing technological tools, as well as difficulty in dealing with the budgetary issue of the people management one. Therefore, the proposal is to create a training and capacitating program to school managers in the municipal educational network of Juiz de For a, having the Center for Teacher Formation as the favourable setting to stimulate and perfect the skills inherent to the position of school manager.

>Magister Scientiae - MSc / Cissampelos capensis, is commonly known by the Afrikaans name ‟dawidjies” or ‟dawidjieswortel”. C. capensis is the most important and best known medicinal plant of the family Menispermaceae used by the Khoisan and other rural people in the western regions of South Africa. Among numerous other ailments, it is traditionally taken to treat male fertility problems. Yet, no studies have investigated the effects of this plant or its extracts on human spermatozoa. The aim of study was to investigate the effects of C. capensis rhizome extracts on sperm function.

Made available in DSpace on 2016-05-02T13:55:22Z (GMT). No. of bitstreams: 1 Laila Pereira da Silva Dissertacao.pdf: 2576409 bytes, checksum: d8ddb33c748f481b6487512d8f098802 (MD5) Previous issue date: 2015-02-27 / Fundacao de Amparo a Pesquisa do Estado de Minas Gerais / The in vitro production (IVP) of bovine embryos despite already used on a commercial scale, presents high variability in the results, that can often be attributed to the fertilization step. Therefore, it is important to research factors related to semen, such as sperm capacitation process. The aim of the study was to evaluate theophylline or its combination with heparin as possible replacement capacitation agent in the development of in vitro produced embryos and in acrosome reaction of sperm cells. The experiment was carried out with 4 bulls and 3 treatments, with 12 experimental groups. Each bull was evaluated in all treatments as described below: Treatment 1 (T1): Heparin - 10mg / ml; Treatment 2 (T2): Theophylline - 5 mM; Treatment 3 (T3): Heparin (10 mg / ml) + theophylline (5mM). The bulls semen was thawed and exposed to incubation in three treatments for 0, 6, 12 and 18h, subsequently stained with Trypan blue / Giemsa and analyzed by electron microscopy for assessment of the acrosome reaction. For IVP of embryos capacitation agents added to fertilization medium were evaluated utilizing the same bulls. In sperm analysis, real acrosome reaction rate was higher (p<0,05) at 0h time while the higher dead spermatozoa rate (p<0,05) was observed at 12 h (84,46 ± 5,82 ) and 18h (86,75 ± 4,19). The IVP embryos rate (37,97 ± 13) and the hatching rate (33,50 ± 14) were higher (p<0,05) in heparin treatment. There was no influence of bull or treatments (p> 0.05) in the acrosome reaction analysis. The use of theophylline as a capacitation agent decreased embryo production rates in IVF, however was as efficient as heparin in acrosome reaction induction. / A produção in vitro de embriões (PIVE) bovinos, apesar de já utilizada em escala comercial, apresenta elevada variabilidade em seus resultados, que pode frequentemente ser atribuída à etapa de fertilização. Sendo assim, torna-se importante o estudo dos fatores relacionados ao sêmen, como o processo de capacitação espermática. O objetivo do estudo foi avaliar a teofilina como agente de capacitação substituto ou associado à heparina no desenvolvimento de embriões produzidos in vitro e sobre a reação acrossômica das células espermáticas. O experimento foi realizado com 4 touros e 3 tratamentos, totalizando 12 grupos experimentais. Cada touro foi avaliado em cada tratamento descrito a seguir: Tratamento 1 (T1): Heparina - 10µg/mL; Tratamento 2 (T2): Teofilina 5 mM; Tratamento 3 (T3): Heparina (10µg/mL) + Teofilina (5mM). O sêmen dos touros foi descongelado e submetido à incubação com os três tratamentos por 0, 6, 12 e 18h, posteriormente corados com Trypan blue/ Giemsa e analisados em microscopia eletrônica para avaliação da reação acrossômica. Para a PIVE os agentes de capacitação adicionados aos meios de fertilização foram testados utilizando os mesmos touros. Na análise espermática a taxa de reação acrossômica verdadeira foi maior (p<0,05) no tempo 0h enquanto para os espermatozoides mortos foi observada maior taxa (p<0,05) nos tempos de 12h (84,46 ± 5,82) e 18h (86,75 ± 4,19). A taxa de embriões produzidos na PIVE (37,97 ± 13) e a taxa de eclosão (33,50 ± 14) foram maiores (p<0,05) para o tratamento heparina. Não houve influência de touro ou de tratamentos (p>0,05) na análise de reação acrossômica. A utilização da teofilina como agente de capacitação reduziu as taxas de produção embrionária na fertilização in vitro, no entanto foi tão eficiente quanto à heparina na indução da reação do acrossoma.

L’infertilité affecte jusqu’à 15-20% des couples en âge de se reproduire. C’est pourquoi, mieux comprendre les mécanismes à la base de la fécondation est essentiel pour l’identification de nouvelles causes d’infertilité et l’optimisation des techniques de reproduction assistée. La capacitation est une étape de la maturation des spermatozoïdes qui se déroule dans le tractus génital femelle. Elle est requise pour la fécondation d’un ovocyte. Notre laboratoire a démontré que des protéines du plasma séminal bovin, appelées protéines Binder of SPerm (BSP), se lient aux phospholipides portant des groupements choline à la surface de la membrane des spermatozoïdes lors de l’éjaculation et promeuvent la capacitation. Ces protéines exprimées par les vésicules séminales sont ubiquitaires chez les mammifères et ont été étudiées chez plusieurs espèces dont l’étalon, le porc, le bouc et le bélier. Récemment, l’expression de gènes homologues aux BSP a été découverte dans les épididymes d’humains (BSPH1) et de souris (Bsph1 et Bsph2). Notre hypothèse est que les BSP chez ces deux espèces sont ajoutées aux spermatozoïdes lors de la maturation épididymaire et ont des rôles dans les fonctions spermatiques, similaires à ceux des protéines BSP bovines. Les protéines BSP humaines et murines représentent une faible fraction des protéines totales du plasma séminal. Pour cette raison, afin d’étudier leurs caractéristiques biochimiques et fonctionnelles, des protéines recombinantes ont été produites. Les protéines recombinantes ont été exprimées dans des cellules Escherichia coli origami B(DE3)pLysS en utilisant un vecteur d’expression pET32a. Suivant la lyse cellulaire, les protéines ont été dénaturées avec de l’urée et purifiées par chromatographie d’affinité sur ions métalliques immobilisés. Une fois liées à la colonne, les protéines ont été repliées à l’aide d’un gradient d’urée décroissant avant d’être éluées. Cette méthode a mené à la production de trois protéines recombinantes (rec-BSPH1 humaine, rec-BSPH1 murine et rec-BSPH2 murine) pures et fonctionnelles. Des expériences de chromatographie d’affinité et de co-sédimentation nous ont permis de démontrer que les trois protéines peuvent se lier à des ligands connus des protéines BSP comme la gélatine et l’héparine en plus de pouvoir se lier aux spermatozoïdes. Nos études ont également révélées que les deux protéines rec-BSPH1 peuvent se lier aux liposomes de phosphatidylcholine (PC) et sont capable de promouvoir la capacitation des spermatozoïdes. À l’opposé, rec-BSPH2 ne peut ni se lier aux liposomes de PC, ni stimuler la capacitation. Finalement, les protéines recombinantes n’ont aucun effet sur la réaction acrosomique ou sur la motilité des spermatozoïdes. Chez les bovins, les protéines BSP induisent la capacitation grâce des interactions avec les lipoprotéines de haute densité (HDL) et les glycosaminoglycanes. Puisque le HDL est également un joueur important de la capacitation chez la souris, le rôle de la protéine native BSPH1 murine au niveau de la capacitation induite par le HDL a été étudié. Les résultats obtenus suggèrent que, in vivo, la protéine BSPH1 de souris serait impliquée dans la capacitation via une interaction directe avec le HDL. Comme les protéines BSPH1 humaines et murines sont orthologues, ces résultats pourraient aussi s’appliquer à la fertilité humaine. Les résultats présentés dans cette thèse pourraient mener à une meilleure compréhension de la fertilité masculine et aider à améliorer les techniques de reproduction assistée. Ils pourraient également mener au développement de nouveaux tests diagnostiques ou de contraceptifs masculins. / Infertility can affect as much as 15-20% of couples of reproductive age. Therefore, elucidating mechanisms occurring during fertilization is needed to resolve cases of infertility and optimize assisted reproductive technology procedures. Sperm capacitation is a maturation step that takes place in the female genital tract and is deemed to be essential for sperm to fertilize an oocyte. Our laboratory has demonstrated that proteins from bovine seminal plasma called Binder of SPerm (BSP) proteins bind to choline phospholipids on the sperm membrane upon ejaculation and promote capacitation. These proteins expressed in seminal vesicles are ubiquitous amongst mammals and have been studied in many species including stallion, boar, ram and goat. More recently, the expression of BSP-homologous genes has been discovered in the epididymis of humans (BSPH1) and in mice (Bsph1 and Bsph2). We hypothesized that the BSP homologs in these two species are added to sperm during epididymal maturation and play similar roles in sperm functions as bovine BSP proteins. BSP proteins in humans and mice constitute only a minute percentage of the seminal plasma proteins. Thus, to study their biochemical and functional characteristics recombinant proteins were produced. Recombinant proteins were expressed in Escherichia coli origami B(DE3)pLysS cells using a pET32a expression vector. Following cell lysis, proteins were denatured using urea and purified by immobilized metal ion affinity chromatography. Once bound to the resin, proteins were refolded using a decreasing urea gradient after which they were eluted. This method led to the production of three pure, functional recombinant proteins (human rec-BSPH1, mouse rec-BSPH1 and mouse rec-BSPH2). Using affinity chromatography and co-sedimentation experiments, we were able to demonstrate that all three recombinant proteins bind known ligands of BSP proteins including gelatin, heparin and have the ability to bind to sperm. Studies also revealed that both rec-BSPH1 proteins bind to phosphatidylcholine (PC) liposomes and promote sperm capacitation. However, rec-BSPH2 neither binds to PC liposomes nor stimulates capacitation. Recombinant proteins had no effect on acrosome reaction or sperm motility. In bovine, BSP proteins promote sperm capacitation through interactions with high-density lipoproteins (HDL) and glycosaminoglycans. Since in mice HDL is also a major factor implicated in capacitation, the role of the native murine BSPH1 protein in HDL-induced capacitation was investigated. Results obtained suggest that, in vivo, murine BSPH1 protein could act in capacitation via a direct interaction with HDL. As human and murine BSPH1 are orthologs, these results could possibly also apply to human fertility. The results presented in this thesis could lead to a better understanding of male fertility and help improve assisted reproduction technology procedures. They could also lead to the development of diagnostic tests as well male contraceptives.

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.

Made available in DSpace on 2016-04-27T14:32:27Z (GMT). No. of bitstreams: 1 Maria Angelica Batista.pdf: 1600735 bytes, checksum: c46ff59830bbc4f1e6bbd6610b582358 (MD5) Previous issue date: 2005-06-13 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This work aims at analyzing and critically understanding the capacitation model applied by the Programa Alfabetização Solidária (PAS) in São Paulo´s Alto Tietê region; it presents an approach and interpretation of what is theorized about the program, what is actually done and what is said about the praxis . This research intends to contribute to the evolution of public policies in the area of youth and adult education in Brazil as well as to the area of teacher formation in general. It focuses on the tutor, his/her formation, and the teaching he/she provides, contrasted to the program´s official guidelines which represent the discourse of power in this context. The methodology includes different instruments for collecting data such as: visits to the literacy centers, conversations during capacitation sessions, documentary analysis of the program, analysis of texts produced by the tutors in sessions, and the technique of group discussion. The data have been dealt with on the basis of the theoretical frame which supports the research, resulting in three major categories for analysis: Capacitation - constructing necessary knowledge for the tutor , which discusses the understanding tutors have about their role in the program, their pedagogical knowledge, and expected changes in their lives after the capacitation; Literacy tutoring practice in large urban centers , which deals with the pedagogical practice of the tutor, how they plan and organize their teaching, how they evaluate the process, and their routine actions; The tutor s view of the world and his/her social and cultural intervention in the community , which discusses the implications of the tutor s work beyond the classroom, in the community life, during and after the literacy term. The analysis has been able to produce a new view of the capacitation program which may be used to its improvement. Unequal distribution of knowledge and power in society creates a tension line which must be surpassed though varied strategies. In education, although some laws have brought some improvement, overall, government policies have not been effective in solving the problem of illiteracy which has been bandaged, deferred or simply dealt with as statistic figures. This studiy reveals that tutors have a degree of knowledge which, due to its limitations, is not enough to encompass the difficult task of adequately bringing literacy to the unattended masses of youths and adults in Brazilian society. Another model in the formation of educators for adults must be devised in order to answer to that challenge / Este trabalho busca analisar e compreender criticamente o modelo de capacitação do Programa Alfabetização Solidária nos grandes centros urbanos da Região do Alto Tietê em São Paulo, propondo modos de aproximação e de interpretação entre o teorizado, o praticado e o dito sobre o praticado. A pesquisa pretende contribuir com as políticas públicas de Educação de Jovens e Adultos no país e a formação docente de um modo geral, concentrando-se na figura do alfabetizador, na ação formadora que incide sobre ele e na formação ele que realiza em seus alunos. Como contraponto, é analisado o discurso oficial do programa que se constrói como lugar discursivo de poder. A metodologia utilizada fez uso de diferentes instrumentos de coleta de dados tais como: visitas realizadas nos núcleos de alfabetização, conversas informais durante a capacitação, análise de documentos do programa, análise de produções escritas dos alfabetizadores nas atividades de capacitação e aplicação da técnica de grupo focal. Foi feita a discussão dos dados conforme o quadro teórico de apoio à pesquisa, a partir do quê elegeram-se três grandes categorias de análise, a saber: Capacitação: período de construção de saberes necessários ao alfabetizador , na qual buscou-se discutir a compreensão que os alfabetizadores têm do seu papel no programa, seus saberes pedagógicos, e as possíveis mudanças em suas vidas após a capacitação; A prática alfabetizadora nos núcleos de alfabetização dos grandes centros urbanos , em que se discutiu aspectos da prática pedagógica desses alfabetizadores nos núcleos de alfabetização, como planejam e organizam o ensino, como realizam avaliações diagnósticas, as rotinas e modos de trabalhar no cotidiano das salas de aula; Leitura de mundo do alfabetizador e sua intervenção sócio-cultural na comunidade , que discutiu a atuação do alfabetizador na vida comunitária onde se insere o núcleo durante e após a duração do módulo de alfabetização. A análise permitiu construir um novo olhar sobre a capacitação que considero ser valioso para uma análise crítica do próprio programa, tendo em vista seu aperfeiçoamento. Saberes e poderes rigorosamente mal distribuídos entre as classes sociais são linhas de tensão a serem superadas em diferentes frentes. Na educação, embora algumas leis tenham avançado, políticas públicas não concretizaram avanços e, conseqüentemente, o problema do analfabetismo tem sido remediado, postergado ou tratado em termos de resultados numéricos apenas. O presente estudo constata que os alfabetizadores têm saberes, mas esses saberes que possuem são muito limitados e não dão conta de realizar a difícil tarefa de alfabetizar adequadamente as massas populares adultas da sociedade brasileira. Um outro modelo de formação de educador de adultos precisa ser pensado para responder a isto

Les actes juridiques unilatéraux des États, extrêmement variés dans leurs formes comme leurs contenus, contribuent à la formation et à l’évolution des normes internationales. Cependant, la présente thèse soutient que ces actes unilatéraux ne constituent pas, pris isolément, un mode de production du droit international ; tout au plus en sont-ils des éléments de formation. Parmi tous les actes adoptés par les États, certains seulement véhiculent des «prétentions normatives stricto sensu». Ces prétentions manifestent une certaine tension entre l’exercice par les États d’un pouvoir d’application du droit existant, en vertu de normes d’habilitation établies ou revendiquées, et d’un pouvoir de l’interpréter ou de le transformer. Dans ces conditions, ces prétentions bénéficient d’une existence objective et d’une opposabilité présumée tout en étant potentiellement contestables durant un certain laps de temps. L’attribution d’une valeur au silence des tiers intéressés apporte la preuve d’un tel présupposé. Ainsi, la représentation proposée met en lumière l’exercice par les États d’un véritable pouvoir de sommation : il découle des prétentions identifiées un effet autonome consistant à requérir les réactions des tiers susceptibles d’en remettre en cause le bien-fondé. Les protestations, reconnaissances ou acquiescements silencieux des tiers constituent alors la condition nécessaire et suffisante aux fins de détermination de l’(in)opposabilité du contenu normatif des prétentions. Du processus décrit, il peut être déduit que l’existence et la signification des rapports de droit produits par la confrontation des prétentions et réactions des États reposent certes sur des accords informels. Toutefois, le fonctionnement du processus révèle avant tout l’influence considérable de l’unilatéralisme sur la définition et l’évolution du droit international. Les États se voient en effet soumis à une obligation accrue de vigilance à l’égard des prétentions susceptibles d’emporter des effets sur leurs droits et obligations. / Unilateral acts of States, which are extremely varied in both form and content, contribute to the formation process and evolution of international standards. However, this thesis suggests that unilateral acts, taken in isolation, are not a means of producing international law; at most they are elements in its formation process. Among the acts adopted by States, some merely express “normative pretentions in the strict sense of the term”. These pretentions express a certain tension in a State’s exercising of a power of application of existing law, in virtue of established or claimed capacitation, and the power to interpret or transform it. Under these conditions, these pretentions benefit from objective existence and supposition of opposability, while remaining potentially contestable during a given period of time. The attribution of a value to the silence of concerned third parties is proof of such a presupposition. This means the proposed representation shows the exercising by States of an actual power of summons: the pretentions identified give rise to an independent effect requiring the action of third parties likely to question validity. Protests, recognition or acquiescence of third parties thus constitute the condition deemed necessary and sufficient to the determination of the (un)opposability of the normative content of the pretentions. From the process described, it is possible to deduce that the existence and significance of law produced by the confrontation of pretentions and reactions of States is based on informal agreements. However, the functioning of the process reveals above all the considerable influence of unilateralism on the definition of the evolution of international law. States are subject to an increased obligation for vigilance with regard to pretentions likely to impact their rights and obligations.

Made available in DSpace on 2016-04-04T18:36:43Z (GMT). No. of bitstreams: 1 Rosemary Passos.pdf: 846677 bytes, checksum: af03c3a21f5caf359c4a396de5582270 (MD5) Previous issue date: 2003-02-21 / The main hypothesis presented here, in this researcher, is that the knowledge , the use of tools and support for the recovery of information, allied with the normalization of the technical structure used in scientific works, if applied since from the very beginning of any course (college graduation, post graduation or specialization), will be essential to the professor s formation. The main objectives of this work were to present analyse with an emphasis in the importance of the use of technics in information recovery and bibliographical normalization, in the process of professor s formation with the cooperation of the librarian. With the help of user s capacitacion seminar, idealized by Libraries System of UNICAMP - SBU, but created specifically for the educational area; to make a general evaluation about the user s capacitacion seminar and its structure; to investigate the quality of the attendance in the reference service of the Faculty of Education the UNICAMP Library. The research method utilized here was the case study, which provided an elaboration of a diagnosis with approaches of the characteristics of a group of students, about their difficulties in utilizing the resources for the information recovery and the use of the library services. The study showed during its development, the importance to make the professor more capable in what concerns to informational resources, the necessity of qualification and capacitation of librarians to work as educators to the college students, the necessity of the creation of disciplines with a special focus in this aspects inside for the classroom and principally the interactivity between librarians and professors, in order to develop a effective partnership in the educational environment. / A principal hip?tese, apresentada nesta pesquisa, ? a de que o conhecimento e o uso de ferramentas e suportes para recupera??o da informa??o, e a normaliza??o da estrutura t?cnica de trabalhos cient?ficos, se aplicados desde o in?cio de qualquer curso (p?s-gradua??o, gradua??o, especializa??o), ser?o fundamentais para a forma??o de professores. Os objetivos desse trabalho foram apresentar uma an?lise que enfatizasse a import?ncia do uso de t?cnicas de recupera??o de informa??o e normaliza??o bibliogr?fica no processo de forma??o de professores, com a colabora??o do bibliotec?rio, utilizando-se de um Curso de Capacita??o de Usu?rios, estruturado pelo Sistema de Bibliotecas da UNICAMP SBU, mas direcionado especificamente para a ?rea educacional; fazer uma avalia??o geral do Curso de Capacita??o em sua estrutura; e investigar a qualidade de atendimento no servi?o de refer?ncia da Biblioteca da Faculdade de Educa??o da UNICAMP. O m?todo de pesquisa utilizado foi o de Estudo de Caso, que possibilitou a elabora??o de um diagn?stico com abordagens das caracter?sticas do grupo de alunos que faziam parte da amostra, analisando suas dificuldades em utilizar os recursos para a recupera??o da informa??o e utiliza??o dos servi?os da biblioteca. O estudo demonstrou durante seu desenvolvimento, a import?ncia da capacita??o de professores no que se refere aos recursos informacionais, a necessidade de qualifica??o e capacita??o dos bibliotec?rios para atuarem como educadores perante os alunos dos cursos, a necessidade do estabelecimento de disciplinas que contemplem esses aspectos dentro da sala de aula, e principalmente a intera??o entre bibliotec?rios e professores, para que estes dois profissionais possam desenvolver um trabalho em parceria no contexto educacional.