Regenerative medicine and cell therapies hold great potential to treat a variety of medical conditions. Product characterisation of these therapies is particularly difficult as they pose regulatory challenges due to donor heterogeneity and the lack of standardised lot release tests that can reliably predict in vivo function. Human mesenchymal stem cells (hMSCs), also called multipotent stem cells or mesenchymal stromal cells, are a viable option in cell therapies due to their immunosuppressive and pro-angiogenic functions. Currently there are no standardised methods or potency assays to quantify these properties. To address this, five individual hMSCs lines from different donors were created and characterised based upon growth rate, differentiation capability and extracellular surface protein expression. A novel multiparameter flow cytometry method to characterise the cells based upon extracellular surface markers was developed that supports high-throughput and high-content analyses. Three candidate lines were taken forward and assessed in multiple in vitro bioassays that examined the hMSC immunosuppressive response to a defined inflammatory environment, effect on T-cell proliferation, and effect on a mixed lymphocyte population. Next, the angiogenic properties were assessed using human umbilical vein endothelial cells (HUVECs) tube formation as a model for cardiac regeneration. This involved utilising automated time lapse microscopy techniques coupled with image analysis software to quantify endothelial to tube formation. Further analysis of the hMSC secretome revealed differences in the levels of pro-angiogenic cytokines such as vascular endothelial growth factor, hepatocyte growth factor and IL-8. Significant differences in angiogenic potency were found between the hMSC lines. This thesis highlights the need to develop specific assays that reflect the intended clinical action. Taken together, these quantitative approaches provide valuable tools to measure hMSC quality and potency, and supports continued efforts to improve characterisation strategies for cellular therapies.

Made available in DSpace on 2014-06-11T19:27:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-06-22Bitstream added on 2014-06-13T18:55:49Z : No. of bitstreams: 1 souza_fjp_me_jabo.pdf: 289543 bytes, checksum: 6ec4fdfeb3984ff7356fa03db7b0aed6 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / No Brasil, a potência das vacinas veterinárias contra a raiva é avaliada pelo teste NIH (“National Institutes of Health”), no qual o desempenho de uma vacina, medido pela DE50 em camundongos, é comparado ao desempenho de uma vacina de referência. São consideradas aprovadas as partidas com potência igual ou superior a 1,0 UI. O presente estudo comparou o desafio viral com vírus fixo CVS, utilizado como padrão no teste de potência NIH, a desafios com três amostras virais isoladas de bovinos naturalmente infectados. O objetivo foi verificar se as amostras de vírus de campo apresentariam virulência maior que a do CVS, o que poderia sugerir a inadequação do teste NIH para a avaliação das vacinas contra a raiva e, portanto, inferir que as vacinas aprovadas por esse teste poderiam não proteger suficientemente o rebanho. Apesar da grande variabilidade que o teste NIH pode apresentar, as três repetições desafiadas com CVS apresentaram semelhança em um intervalo de confiança de 95%, e nos desafios realizados com três amostras de vírus de campo a vacina utilizada protegeu mais do que nos desafios com CVS. Conclui-se que a virulência das amostras de vírus de campo utilizadas não foi maior que a virulência do CVS, em camundongos, e que o rigor do desafio padrão mostrou-se adequado para a avaliação da potência pelo teste NIH e, portanto, para o controle da qualidade de vacinas contra a raiva. / The potency of veterinary rabies vaccines in Brazil is evaluated by the NIH (National Institutes of Health) potency test, in which the ability of a vaccine to induce protection in mice (ED50) is compared with a reference vaccine. The batches are approved when the potency is equal or superior to 1.0 IU. The present study compared CVS strain challenges (standard strain for the potency test) with challenges that used three wild isolates of natural occurrence in cattle. The aim was to verify if the wild strains could be more virulent than the CVS. If this occurred, the NIH potency test would prove inadequate to evaluate rabies vaccines. Therefore, one could infer that the approved vaccines might not provide sufficient herd protection. Despite the great variability of NIH test, all three repetitions that were challenged with CVS were similar at a 95% confidence-level; and when challenged with wild strains, the vaccine provided better protection than the one achieved at CVS challenge. In conclusion, wild strains virulence was not greater than CVS virulence in mice. The strictness of standard challenge proved to be adequate for potency evaluation by NIH test, and consequently, for the quality control evaluation of rabies vaccines.

Made available in DSpace on 2014-12-17T14:44:19Z (GMT). No. of bitstreams: 1 MarcosARSJ_DISSERT.pdf: 3313809 bytes, checksum: 12cc0e861a6a05e85364b57b637c08d6 (MD5) Previous issue date: 2014-02-28 / To mesure the human performance is a challenge, mainly due to the multidimensional factor movement. Instruments, mostly only assess one dimension of it. Objective: To develop a prototype capable of measuring the skills of human performance and check its validity using method comparison approach. Methods: The study was divided into two stages. The first Prototype was developed and tested simultaneously with an instrument to study the Rectilinear Uniform Motion and Uniformly Miscellaneous. In the second phase the sample consisted of Paralympic basketball athletes (n=09) and karate athletes (n=31) and all agreed to the terms of participation in the research. The evaluation of performance measures was performed with the prototype, the results obtained were compared with the data calculated by a statistical package is used as a reference. Results: All variables calculated by the prototype showed no significant differences when compared with the results calculated by the reference instrument and statistical package. Conclusion: The prototype has been developed and the results obtained in laboratory and field indicate that the prototype can be used for measuring human performance measures, with immediate results without the need for a conventional computer return, provided they fulfill the criteria described / mesurar as desempenho humano ? um desafio, principalmente devido ao fator multidimensional do movimento. Os instrumentos, em sua maioria, avaliam apenas uma dimens?o do mesmo. Objetivo: desenvolver um prot?tipo capaz de mensurar as habilidades do desempenho humano e verificar sua validade utilizando abordagem de compara??o de m?todos. M?todos: O estudo foi dividido em duas etapas. A primeira o Prot?tipo foi desenvolvido e testado simultaneamente com um instrumento utilizado para estudo do movimento Retil?neo Uniforme e Uniformemente Variado. Na segunda fase a amostra constituiu-se de atletas paral?mpicos de basquete (n=09) e atletas de Karate (n=31) e todos concordaram com o termos da participa??o da pesquisa. A avalia??o das medidas do desempenho foi realizada com o Prot?tipo, os resultados obtidos foram comparados com os dados calculados por um Pacote Estat?stico sendo utilizado como refer?ncia. Resultados: todas as vari?veis calculadas pelo Prot?tipo n?o apresentaram diferen?as significativas quando comparadas com os resultados calculados pelo instrumento de refer?ncia e pacote estat?stico. Conclus?o: O prot?tipo foi desenvolvido e os resultados obtidos em laborat?rio e campo indicam que o prot?tipo pode ser usado para mensurar as medidas do desempenho humano, com retorno imediato dos resultados sem necessidade de um computador convencional, desde que observados os crit?rios descritos

Orientadora: Adolorata Aparecida Bianco Carvalho / Banca: Maria da Gloria Buzinaro / Banca: Luzia Helena Queiroz / Resumo: No Brasil, a potência das vacinas veterinárias contra a raiva é avaliada pelo teste NIH ("National Institutes of Health"), no qual o desempenho de uma vacina, medido pela DE50 em camundongos, é comparado ao desempenho de uma vacina de referência. São consideradas aprovadas as partidas com potência igual ou superior a 1,0 UI. O presente estudo comparou o desafio viral com vírus fixo CVS, utilizado como padrão no teste de potência NIH, a desafios com três amostras virais isoladas de bovinos naturalmente infectados. O objetivo foi verificar se as amostras de vírus de campo apresentariam virulência maior que a do CVS, o que poderia sugerir a inadequação do teste NIH para a avaliação das vacinas contra a raiva e, portanto, inferir que as vacinas aprovadas por esse teste poderiam não proteger suficientemente o rebanho. Apesar da grande variabilidade que o teste NIH pode apresentar, as três repetições desafiadas com CVS apresentaram semelhança em um intervalo de confiança de 95%, e nos desafios realizados com três amostras de vírus de campo a vacina utilizada protegeu mais do que nos desafios com CVS. Conclui-se que a virulência das amostras de vírus de campo utilizadas não foi maior que a virulência do CVS, em camundongos, e que o rigor do desafio padrão mostrou-se adequado para a avaliação da potência pelo teste NIH e, portanto, para o controle da qualidade de vacinas contra a raiva. / Abstract: The potency of veterinary rabies vaccines in Brazil is evaluated by the NIH (National Institutes of Health) potency test, in which the ability of a vaccine to induce protection in mice (ED50) is compared with a reference vaccine. The batches are approved when the potency is equal or superior to 1.0 IU. The present study compared CVS strain challenges (standard strain for the potency test) with challenges that used three wild isolates of natural occurrence in cattle. The aim was to verify if the wild strains could be more virulent than the CVS. If this occurred, the NIH potency test would prove inadequate to evaluate rabies vaccines. Therefore, one could infer that the approved vaccines might not provide sufficient herd protection. Despite the great variability of NIH test, all three repetitions that were challenged with CVS were similar at a 95% confidence-level; and when challenged with wild strains, the vaccine provided better protection than the one achieved at CVS challenge. In conclusion, wild strains virulence was not greater than CVS virulence in mice. The strictness of standard challenge proved to be adequate for potency evaluation by NIH test, and consequently, for the quality control evaluation of rabies vaccines. / Mestre

O uso de Objetos Digitais Aprendizagem – ODA– em ambientes educacionais é uma necessidade inerente à atual época tecnológica que vivemos, uma vez que o acesso a recursos computacionais é ampliado a cada momento, tanto nas escolas como no cotidiano dos cidadãos. Portanto, criar ferramentas, ou seja, ODA, ganha a dimen-são de possibilitar que a educação ultrapasse os muros da escola em qualquer ins-tante ou lugar. Nesse contexto sociohistórico, o presente trabalho apresenta uma proposta de abordar os conceitos de Energia Elétrica e Potência Elétrica bem como as suas relações com Tensão, Corrente e Resistência Elétricas, através de uma bre-ve explanação sobre essas relações e com uma posterior proposta de resolução de um Teste Conceitual envolvendo esses conceitos através de recursos Hipermídia. O diferencial e, por consequência, a contribuição que o presente estudo agrega à co-munidade acadêmica está no fato de que, durante a interação dos aprendizes com um Objeto Digital de Aprendizagem, o erro adquire status de ponto essencial na evo-lução da Estrutura Cognitiva do aprendiz amparado pela Teoria da Aprendizagem de David Ausubel e pela Epistemologia de Gaston Bachelard. Os resultados aqui obti-dos revelam que os ODA quer sejam utilizados isoladamente, quer estejam agrega-dos a outras atividades, em Ambientes Virtuais de Aprendizagem – AVA – podem ampliar as possibilidades do professor no processo de Ensino Aprendizagem. / O uso de Objetos Digitais Aprendizagem – ODA– em ambientes educacionais é uma necessidade inerente à atual época tecnológica que vivemos, uma vez que o acesso a recursos computacionais é ampliado a cada momento, tanto nas escolas como no cotidiano dos cidadãos. Portanto, criar ferramentas, ou seja, ODA, ganha a dimen-são de possibilitar que a educação ultrapasse os muros da escola em qualquer ins-tante ou lugar. Nesse contexto sociohistórico, o presente trabalho apresenta uma proposta de abordar os conceitos de Energia Elétrica e Potência Elétrica bem como as suas relações com Tensão, Corrente e Resistência Elétricas, através de uma bre-ve explanação sobre essas relações e com uma posterior proposta de resolução de um Teste Conceitual envolvendo esses conceitos através de recursos Hipermídia. O diferencial e, por consequência, a contribuição que o presente estudo agrega à co-munidade acadêmica está no fato de que, durante a interação dos aprendizes com um Objeto Digital de Aprendizagem, o erro adquire status de ponto essencial na evo-lução da Estrutura Cognitiva do aprendiz amparado pela Teoria da Aprendizagem de David Ausubel e pela Epistemologia de Gaston Bachelard. Os resultados aqui obti-dos revelam que os ODA quer sejam utilizados isoladamente, quer estejam agrega-dos a outras atividades, em Ambientes Virtuais de Aprendizagem – AVA – podem ampliar as possibilidades do professor no processo de Ensino Aprendizagem

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2015-01-27T13:50:07Z No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Dissertação - Diego Eterno de Oliveira Mendonça - 2014.pdf: 4971681 bytes, checksum: 7063bb73e6f0a7553c28f116269050f0 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2015-01-28T11:43:39Z (GMT) No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Dissertação - Diego Eterno de Oliveira Mendonça - 2014.pdf: 4971681 bytes, checksum: 7063bb73e6f0a7553c28f116269050f0 (MD5) / Made available in DSpace on 2015-01-28T11:43:39Z (GMT). No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Dissertação - Diego Eterno de Oliveira Mendonça - 2014.pdf: 4971681 bytes, checksum: 7063bb73e6f0a7553c28f116269050f0 (MD5) Previous issue date: 2014-07-25 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The use of low level lasers in the burning wounds healing has been described in the literature with a great variety of treatment parameters such as multiplicity of wave lenghts, energetic dosages and time of use. Objective: the aim of this study was to compare the effect of low potency AsGaAl laser in the third degree burning wounds healing in Wistar rats with and without diabets. Methodology: this project was approved by the ethics committee in Animals use from UFG, protocol number 007/2012.100. The animals were distributed into 4 groups: a) control group non diabetic; b) treated group non diabetic; c) diabetic control group; d) diabetic treated group. The treated groups received an energetic dosage of 3 J/cm2 in the first 7 days and 6 J/cm2 after the seventh experimental day. The animals received changes of the occlusive bandages embedded with silver sulphadiazine and all of them were accompanied throughout 3, 7, 14, 21 and 30 days for the macroscopic, morphometric and microscopic aspects evaluation. The parametric "t test" was used for the statistical analysis. Results: the comparison of the wounds from the diabetics and non diabetics animals showed that the treatment with low laser induced a significant reduction (p<0.05) of the contraction area; macroscopically there were no differences between groups. Regarding the microscopic findings there was a significant increase in angiogenesis (p<0.05) and increase in the fibrin presence (p<0.05) when compared the treated groups from diabetic and non diabetic animals in 3rd and 14th days of experiment. In the treated diabetic animals the angiogenesis in the 14th day and the number of fibroblasts in the 14th and 21st days of experiment presented a significant difference (p<0.05) when compared to the non treated group. Also there was greater collagen disposition throughout the 14th, 21st and 30th days of experiment in all treated groups when compared to the control groups. In conclusion the low laser treatment reduced the injury in long term, stimulating the angiogenesis, fibrin deposition, fibroblasts and collagen deposition. / A utilização do laser de baixa potência na cicatrização de feridas por queimaduras tem sido descrita na literatura com grande variedade de parâmetros de tratamento, tais como, a multiplicidade dos comprimentos de onda, dosagens energéticas e tempo de utilização. Objetivo: O objetivo desse trabalho foi comparar o efeito do laser em baixa potência AsGaAl na cicatrização de queimaduras de 3° grau em ratos Wistar com e sem diabetes. Metodologia: Este projeto foi aprovado pela Comissão de Ética no uso de animais da UFG, protocolo número 007/2012.100. Os animais foram distribuídos em 4 grupos, isto é: grupo controle não diabético ; b) grupo tratados não diabéticos ; c) Grupo controle diabético, e d) Grupo tratado diabético . Os grupos tratados receberam a dosagem energética de 3 J/cm² nos primeiros 3 dias e 6 J/cm² após o sétimo dia experimental. Os animais receberam trocas com curativos oclusivos embebidos com Sulfadiazina de Prata e todos foram acompanhados ao longo de 3, 7, 14, 21 e 30 dias para avaliação dos aspectos macroscópicos, morfométricos e microscópicos. Para a análise estatística utilizou-se o ―teste t’’ paramétrico. Resultados: Ao se comparar as feridas dos animais sem diabetes e com diabetes, observouse que no grupo tratado houve redução significativa (p<0,05) na área de contração; macroscopicamente não foram observadas diferenças entre os grupos; em relação aos achados microscópicos houve aumento significativo (p<0,05) na neoângiogenese (p<0,05) e na presença de fibrina (p<0,05) no grupo tratado comparado ao controle nos animais não diabéticos no 3° dia e 14° dia respectivamente; e nos animais com diabetes a neoângiogenes (p<0,05) ao 14° dia e o número de fibroblastos (p<0,05) no 14° e 21° dia foram estatisticamente significante no grupo tratado em relação ao controle, houve maior deposição de colágenos ao longo do 14°, 21° e 30° dia em todos os grupos irradiados comparados ao controle. Conclui-se que o tratamento reduziu a ferida ao longo do tempo, estimulando a neoangiogênese, fibrina, aumento de fibroblastos e maior deposição de colágenos.

Magister Psychologiae - MPsych / South Africans are exposed to many traumatic events and exposure to such events is associated with negative emotional and behavioural outcomes. However, there are those who are still able to remain psychologically well amidst their difficulties. Resilience is the motivation to be strong in the face of unwarranted demands and this study proposes that it could serve as a buffer against the harmful effects of traumatic events. It is assumed that traumatic events present with demanding circumstances and further that resilience plays a role in the relationship between trauma, the challenges these events present and one’s ability to function in these circumstances. Furthermore, ‘race’ may have a particular influence on this relationship as the level and severity of trauma among different ‘racial’ groups may vary. The link between resilience and trauma has been investigated, but little focus has been given to how ‘race’ may influence this relationship among students in the South African context. The aim of the current study was therefore to investigate the interaction between these three variables and add to existing knowledge related to resilience. Constructs related to resilience include sense of coherence, potency, hardiness, learned resourcefulness and fortitude. The two ‘racial’ groups included in the sample include ‘African’ and ‘Coloured’ students (categories created by the past apartheid government). Resilience was measured by the Resilience Scale for Adults (RSA), trauma was measured by the Harvard Trauma Scale (HTS) and violence exposure was measured by the Childhood Exposure to Community Violence Scale (CECV). Participants included 249 students at the University of the Western Cape. Results indicate that ‘Coloured’ students report similar levels of violence exposure and trauma symptoms as ‘African’ students do. ‘Coloured’ students report higher scores on overall resilience and resilience sub-scales (except the structured style sub-scale) than ‘African’ students as measured on the RSA. With regard to the trauma versus no trauma groups, the results indicate that ‘African’ students who form part of the no trauma group score higher on resilience than those who form part of the trauma group and within the trauma group ‘Coloured’ students score higher on resilience than ‘African’ students. In analyses amongst the high and the low trauma groups, the results indicate that those who form part of the low trauma group score higher on resilience than those who are among the high trauma group; ‘African’ students who form part of the low trauma group score higher on resilience than those who form part of the high trauma group; and ‘Coloured’ students who form part of the low trauma group score higher on resilience than those who form part of the high trauma group. The results yielded in the current study are both similar to and differ from findings presented in previous studies and highlight the complexity of the construct of resilience. Limitations of the study are outlined and recommendations for future research are also provided.

PACAP and VIP have prominent effects on cardiac function in several species, but little is known about their influence on the murine heart. Accordingly, we evaluated the expression of PACAP/VIP receptors in mouse heart and the response of isolated atria to peptide agonists. Quantitative PCR demonstrated that PAC1, VPAC1, and VPAC2 receptor mRNAs are present throughout the mouse heart. Expression of all three receptor transcripts was low, PAC1 being the lowest. No regional differences in expression were detected for individual receptor mRNAs after normalization to L32. Pharmacological effects of PACAP-27, VIP, and the selective PAC1 agonist maxadilan were evaluated in isolated, spontaneously beating atria from C57BL/6 mice of either sex. Incremental additions of PACAP-27 at 1 min intervals caused a concentration-dependent tachycardia with a log EC50=-9.08±0.15 M (n=7) and a maximum of 96.3±5.9% above baseline heart rate. VIP and maxadilan also caused tachycardia but their potencies were about two orders of magnitude less. Increasing the dosing interval to 5 min caused a leftward shift of the concentration-response curve to maxadilan but no changes in the curves for PACAP-27 or VIP. Under this condition, neither the potency nor the efficacy of maxadilan differed from those of PACAP-27. Neither PACAP-27 nor maxadilan caused tachyphylaxis, and maximal responses to maxadilan were maintained for at least 2 h. We conclude that all three VIP/PACAP family receptors are expressed by mouse cardiac tissue, but only PAC1 receptors mediate positive chronotropic responses to PACAP-27 and VIP.

PACAP and VIP have prominent effects on cardiac function in several species, but little is known about their influence on the murine heart. Accordingly, we evaluated the expression of PACAP/VIP receptors in mouse heart and the response of isolated atria to peptide agonists. Quantitative PCR demonstrated that PAC1, VPAC1, and VPAC2 receptor mRNAs are present throughout the mouse heart. Expression of all three receptor transcripts was low, PAC1 being the lowest. No regional differences in expression were detected for individual receptor mRNAs after normalization to L32. Pharmacological effects of PACAP-27, VIP, and the selective PAC1 agonist maxadilan were evaluated in isolated, spontaneously beating atria from C57BL/6 mice of either sex. Incremental additions of PACAP-27 at 1 min intervals caused a concentration-dependent tachycardia with a log EC50=-9.08±0.15 M (n=7) and a maximum of 96.3±5.9% above baseline heart rate. VIP and maxadilan also caused tachycardia but their potencies were about two orders of magnitude less. Increasing the dosing interval to 5 min caused a leftward shift of the concentration-response curve to maxadilan but no changes in the curves for PACAP-27 or VIP. Under this condition, neither the potency nor the efficacy of maxadilan differed from those of PACAP-27. Neither PACAP-27 nor maxadilan caused tachyphylaxis, and maximal responses to maxadilan were maintained for at least 2 h. We conclude that all three VIP/PACAP family receptors are expressed by mouse cardiac tissue, but only PAC1 receptors mediate positive chronotropic responses to PACAP-27 and VIP.

[pt] A presente dissertação tem como objetivo investigar e estabelecer um elo entre três perspectivas teóricas a partir de Bartleby. A fórmula preferiria não abre caminho para muitas leituras: das mais revolucionárias até as mais niilistas. Em meio a tudo isso, verificou-se a necessidade de construir uma linha de investigação crítica altamente política. Para tanto, elencou-se três vertentes de investigação denominadas de fórmulas. A primeira das três é a fórmula da potência de Giorgio Agamben. A segunda é a fórmula da parresía de Michel Foucault. E a terceira é a fórmula do inexprimível de Gilles Deleuze. Partindo do presente para construir uma constelação de pensamento em torno de Bartleby, esta pesquisa visa ainda produzir ferramentas teóricas para pensar desafios estéticos, éticos e políticos do mundo contemporâneo. / [en] The present dissertation aims to investigate and establish a link between three theoretical perspectives from Bartleby. The formula I would rather not opens the way for many readings: from the most revolutionary to the most nihilistic. In the midst of all this, there was a need to build a highly political line of critical investigation. To this end, three lines of investigation called formulas were listed. The first of the three is Giorgio Agamben s Formula of Potency. The second is Michel Foucault s formula of parrhesia. And the third is Gilles Deleuze s formula of the inexpressible. Starting from the present to build a constellation of thought around Bartleby, this research also aims to produce theoretical tools to think about aesthetic, ethical and political challenges of the contemporary world.