<p>T lymphocytes regulate the initiation, duration, and magnitude of adaptive immune responses and function as effector cells in cell mediated immunity. To become immunologically competent they must generate functional antigen receptors. This process takes place in the thymus and requires somatic recombination of T cell receptor (TCR) genes. It is mediated by the endonucleases recombination activating gene-1 (RAG1) and RAG2. Although the thymus regresses at puberty, T cells are present throughout life implying that other tissues must provide the proper milieu for T cell development. This thesis describes extrathymic T cell maturation in man. RAG1, RAG2, and the preTα-chain (pTα), which is exclusively utilized in developing T cells, were used as markers for TCR gene rearrangement. Two new exons (1A and 1B) encoding sequences in the 5’ untranslated region (5’UTR) of mRNA were discovered in the human RAG1 gene. The previously described 5’UTR exon (renamed 1C) was located between the new exons and exon 2, the latter containing the entire coding sequence. We found that small intestinal lymphocytes of the T cell lineage expressed the new exons in three different splice forms. RAG1 mRNA containing the 1C exon was not expressed in small intestinal lymphocytes. In contrast, splice forms containing the 1A exon were not expressed in thymocytes. RAG1 and pTα mRNA expressing lymphocytes were seen both within the epithelium and in lamina propria. Thymocyte-like CD2⁺CD7⁺CD3^-, CD4⁺CD8⁺, CD1a⁺, and IL7-R+ lymphocytes were identified in the small intestinal mucosa. CD2⁺CD7⁺CD3^- cells had the highest expression levels of mRNA for RAG1 and pTα, suggesting that the small intestinal mucosa is indeed a site for T cell maturation. Small intestinal T lymphocytes were also shown to kill via the Fas/FasL pathway in a TCR/CD3 independent manner and via the perforin/granzyme pathway in a TCR/CD3 dependent manner. The Fas/FasL-mediated cytotoxicity may reflect an ongoing selection process of extrathymically maturated T cells. </p><p>The nasopharyngeal tonsil is the major inductive site for immune reactions against inhaled antigens. Previous demonstration of RAG1 expression in tonsillar B cells was interpreted as antigen driven receptor revision. The present study confirms the expression of RAG1 in B cells. We also found that RAG1, RAG2, and pTa mRNAs were expressed in lymphocytes of the T cell lineage. A small population of cells with the immature phenotype CD2+CD7+CD3- was demonstrated. This population had the highest expression levels of mRNA for RAG1, RAG2, pTα and terminal deoxynucleotidyl transferase. All four splice-forms of RAG1 mRNA were expressed. RAG1 and pTα mRNA expressing cells were mainly located in the proximity of the surface epithelium and in the outer rim of the follicles. These results suggest that the nasopharyngeal tonsil is a site where extrathymic T cell development and antigen driven TCR revision are occurring in parallel. </p><p>Celiac disease (CD) is a small intestinal enteropathy characterized by permanent intolerance to gluten. Gluten reactive intestinal T cells are central in the pathogenesis and CD can be regarded as a failure to maintain tolerance to this food antigen. Expression of the RAG1 1A/2 splice form was significantly decreased in small intestinal T cell subsets of CD patients suggesting that impaired TCR gene rearrangement could contribute to failure of maintain tolerance in CD. </p><p>Together, these findings show that both small intestinal and nasopharyngeal tonsillar lymphocytes of T cell lineage have the molecular machinery for antigen receptor rearrangement and that thymocyte-like lymphocytes are present in both tissues. Thus these organs are likely sites of T lymphocyte ontogeny as well as for secondary T cell receptor rearrangement in man. </p>

T lymphocytes regulate the initiation, duration, and magnitude of adaptive immune responses and function as effector cells in cell mediated immunity. To become immunologically competent they must generate functional antigen receptors. This process takes place in the thymus and requires somatic recombination of T cell receptor (TCR) genes. It is mediated by the endonucleases recombination activating gene-1 (RAG1) and RAG2. Although the thymus regresses at puberty, T cells are present throughout life implying that other tissues must provide the proper milieu for T cell development. This thesis describes extrathymic T cell maturation in man. RAG1, RAG2, and the preTα-chain (pTα), which is exclusively utilized in developing T cells, were used as markers for TCR gene rearrangement. Two new exons (1A and 1B) encoding sequences in the 5’ untranslated region (5’UTR) of mRNA were discovered in the human RAG1 gene. The previously described 5’UTR exon (renamed 1C) was located between the new exons and exon 2, the latter containing the entire coding sequence. We found that small intestinal lymphocytes of the T cell lineage expressed the new exons in three different splice forms. RAG1 mRNA containing the 1C exon was not expressed in small intestinal lymphocytes. In contrast, splice forms containing the 1A exon were not expressed in thymocytes. RAG1 and pTα mRNA expressing lymphocytes were seen both within the epithelium and in lamina propria. Thymocyte-like CD2+CD7+CD3-, CD4+CD8+, CD1a+, and IL7-R+ lymphocytes were identified in the small intestinal mucosa. CD2+CD7+CD3- cells had the highest expression levels of mRNA for RAG1 and pTα, suggesting that the small intestinal mucosa is indeed a site for T cell maturation. Small intestinal T lymphocytes were also shown to kill via the Fas/FasL pathway in a TCR/CD3 independent manner and via the perforin/granzyme pathway in a TCR/CD3 dependent manner. The Fas/FasL-mediated cytotoxicity may reflect an ongoing selection process of extrathymically maturated T cells. The nasopharyngeal tonsil is the major inductive site for immune reactions against inhaled antigens. Previous demonstration of RAG1 expression in tonsillar B cells was interpreted as antigen driven receptor revision. The present study confirms the expression of RAG1 in B cells. We also found that RAG1, RAG2, and pTa mRNAs were expressed in lymphocytes of the T cell lineage. A small population of cells with the immature phenotype CD2+CD7+CD3- was demonstrated. This population had the highest expression levels of mRNA for RAG1, RAG2, pTα and terminal deoxynucleotidyl transferase. All four splice-forms of RAG1 mRNA were expressed. RAG1 and pTα mRNA expressing cells were mainly located in the proximity of the surface epithelium and in the outer rim of the follicles. These results suggest that the nasopharyngeal tonsil is a site where extrathymic T cell development and antigen driven TCR revision are occurring in parallel. Celiac disease (CD) is a small intestinal enteropathy characterized by permanent intolerance to gluten. Gluten reactive intestinal T cells are central in the pathogenesis and CD can be regarded as a failure to maintain tolerance to this food antigen. Expression of the RAG1 1A/2 splice form was significantly decreased in small intestinal T cell subsets of CD patients suggesting that impaired TCR gene rearrangement could contribute to failure of maintain tolerance in CD. Together, these findings show that both small intestinal and nasopharyngeal tonsillar lymphocytes of T cell lineage have the molecular machinery for antigen receptor rearrangement and that thymocyte-like lymphocytes are present in both tissues. Thus these organs are likely sites of T lymphocyte ontogeny as well as for secondary T cell receptor rearrangement in man.

The genus Quercus L. is represented with more than 400 species in the world and 18 of these species are found naturally in Turkey. Although its taxonomical, phytogeographical and dendrological importance, the genus Quercus is still taxonomically one of the most problematical woody genus in Turkish flora. In this study, multivariate morphometric approach was used to analyze oak specimens collected from an isolated forest (Beynam Forest, Ankara) where Quercus pubescens Willd., Q. infectoria Olivier subsp. boissieri (Reuter) O. Schwarz and Q. macranthera Fisch. &amp / C. A. Mey. ex Hohen. subsp. syspirensis (C.Koch) Menitsky taxa are belonging to section Quercus sensu stricto (s.s.) are found. Additional oak specimens were included in the analysis for comparison. Morphometric study was based on 52 leaf characters such as, distance, angle, and area as well as counted, descriptive and calculated variables. Morphometric variables were calculated automatically by use of landmark and outline data. Random forest classification method was used to select discriminating variables and predict unidentified specimens by use of pre-identified training group. The results of the random forest variable selection procedure and the principal component analysis (PCA) showed that the morphometric variables could distinguish the specimens of Q. pubescens and Q. macranthera subsp. syspirensis mostly based on the overall leaf size and number of intercalary veins while the specimens of Q. infectoria subsp. boissieri were separated from others based on lobe and lamina base shape. Finally, micromorphological observations of abaxial lamina surface have been performed by scanning electron microscope (SEM) on selected specimens which were found useful to differentiate, particularly the specimens of Q. macranthera subsp. syspirensis and its putative hybrids from other taxa.

Existe uma necessidade urgente em monitorar as respostas da biodiversidade e dos processos ecológicos a distúrbios e mudanças no uso da terra. Diversos estudos tem indicado que o uso da abordagem funcional da biodiversidade, baseada em atributos funcionais, facilita a compreensão sobre a organização dos organismos nas comunidades, da funcionalidade dos ecossistemas, e desta forma, auxilia na identificação das suas relações com fatores ambientais. No primeiro capítulo desta tese eu apresento o arcabouço teórico da ecologia funcional aplicada ao estudo dos invertebrados terrestres, e reviso o que são atributos funcionais, como usá-los na descrição e monitoramento das comunidades e o porquê, ressaltando a ligação dos invertebrados com processos ecológicos. Nos próximos capítulos eu apresento estudos experimentais avaliando as respostas funcionais de dois grupos de invertebrados - aranhas (Araneae) e tripes (Thysanoptera) - e de processos ecológicos influenciados por invertebrados (detritivoria e decomposição de serapilheira) ao distúrbio por fogo nos Campos Sulinos. As queimadas representam uma estratégia de manejo frequente nas pastagens naturais do RS, e carecem em estudos avaliando seus efeitos no ambiente, biodiversidade e processos. A investigação foi realizada com base em um experimento de queima controlada na Estação Agronômica da UFRGS, em Eldorado do Sul, RS. O design foi em blocos casualizados, com 14 parcelas experimentais de 10 x 10 m pareadas; sendo uma parcela de cada bloco queimada no final da primavera de 2009, e a outra, considerada como controle. Informações sobre invertebrados, processos ecológicos e propriedades abióticas do hábitat foram obtidas em todas parcelas antes do fogo (para a caracterização inicial), e 1 mês, 6-7 meses e 12 meses após o fogo. Para o estudo de caso com aranhas, eu classifiquei as comunidades em famílias, atributos de estratégias de caça e medi todos os indivíduos (1755) em atributos funcionais morfológicos, sendo incorporada toda a variabilidade intra e interespecífica, e fenológica das comunidades. Por outro lado, no estudo de caso com tripes, descrevi atributos funcionais (hábitos alimentares, requerimentos de hábitat e atributos morfológicos) por espécie de tripes (48 spp.), assumindo que a variação nos atributos funcionais é sempre maior entre espécies do que dentro da mesma espécie. Nos dois estudos de caso, eu encontrei padrões indicativos de filtragem ambiental, ou seja, o fogo e/ou as mudanças no hábitat induzidas pelo fogo, parecem ter atuado como um filtro, selecionando organismos com determinado conjunto de atributos em detrimento de outros. Esses padrões foram dependentes do período após a queima considerado, por exemplo: um mês após o fogo aranhas construtoras de teias irregulares e tripes fungívoros foram prejudicados, enquanto que aranhas caçadoras da vegetação, com grandes quelíceras, tripes fitófagos e associados com gramíneas aumentaram suas proporções. Para aranhas ainda verifiquei convergência de atributos ao gradiente de biomassa induzido pelo fogo (filtragem ambiental), e divergência de atributos ao gradiente de heterogeneidade da vegetação, indicativo de limitação de similaridade (i.e. aumento da coexistência de organismos funcionalmente diferentes e diminuição de competição). Finalmente com relação aos processos ecológicos, encontrei correlações interessantes acerca de seus agentes modeladores, e.g. entre a atividade de detritivoria com a decomposição de serapilheira, e desta com a equidade da vegetação em campo não queimado. Também mostro que os processos são sensíveis, mas resilientes, ao distúrbio por fogo, seguindo a recuperação da estrutura de hábitat. Adicionalmente, eu incorporei à tese um capítulo sobre uma técnica para avaliar o processo de detritivoria, denominado bait-lamina test, o qual avalia diretamente as taxas de alimentação de invertebrados detritívoros do solo. Neste estudo pioneiro nos campos do RS, esclareço alguns aspectos metodológicos que possam constituir uma base para futuras pesquisas. Contudo, com esta tese espero estar contribuindo com o conhecimento sobre as respostas das comunidades de invertebrados e dos processos ecológicos ao distúrbio por fogo nos Campos Sulinos. Em geral, posso concluir que a abordagem funcional trouxe muitos benefícios ao monitoramento da biodiversidade, e que há elevada resiliência das assembléias de aranhas e tripes, e dos processos de detritivoria e decomposição às queimadas de pequena escala nos campos. / There is a huge need of monitoring the responses of biodiversity and ecological processes to disturbances and land use changes. Several studies have indicated that the functional approach of biodiversity, based on functional traits, facilitates the understanding of community assembly and ecosystem functionality, and thus assists on the identification of their relationships with environmental factors. In the first chapter of this Thesis I present the functional ecology theory applied to studies on terrestrial invertebrates, and I review what functional traits are, how to use them in community description and monitoring, and why, highlighting the connections between invertebrates and ecological processes. In the following chapters I present experimental studies evaluating the functional responses of two invertebrate groups- spiders (Araneae) and thrips (Thysanoptera) – and ecological processes influenced by invertebrates (detritivory and litter decay) to fire disturbance in Campos grasslands. Burning represents a common strategy for pasture management in Rio Grande do Sul State, and there is a lack of studies regarding their effects on the environment, biodiversity and processes. Our investigation was performed based in a burning experiment at Estação Agronômica of UFRGS, Eldorado do Sul, RS. The design was blocked, with 14 experimental paired plots of 10 x 10 m. One plot from each block was burned at the end of spring 2009, and the other is considered a control plot. Information concerning invertebrates, ecological processes and abiotic habitat proprieties were obtained from all plots before fire (initial characterization) and 1 month, 6-7 months and 12 months after fire. For the spider study case, I classified the organisms in families, feeding strategy traits, and measured all spider individuals (1755) thus incorporating all intra and interspecific variability of the communities. On the other hand, in the thrips study case I described functional traits (feeding habits, habitat requirements, and morphological traits) per species (48 spp.), assuming that the variation on functional traits should be higher between species than within species. In both cases, I found patterns indicating environmental filtering, which means that fire and/or the fire-induced habitat changes acted as a filter, sorting organisms with a determined group of traits instead of others. These patterns were dependent on the period after fire considered. For example, one month after fire irregular web spider builders and fungivorous thrips were more uncommon than expected, while vegetation hunter spiders, with large chelicerae, and leaf feeder thrips and thrips associated to grasses increased their proportions. For spiders we also verified a trait-convergence pattern along the plant biomass gradient induced by fire (environmental filtering), and trait-divergence pattern along the vegetation heterogeneity gradient indicating limiting similarity (i.e. increase in coexistence of functionally different organisms, and reduction of competition). Finally, regarding ecological processes, I found interesting correlations on their drivers, e.g. between detritivory and litter decay, and between these with plant evenness in unburned grasslands. Also, I show that ecological processes are sensitive but resilient to fire, following habitat structure recovery. Additionally, I incorporate to the Thesis a methodology chapter about the bait-lamina test, which evaluates the feeding rates of soil detritivore invertebrates directly. In this pioneer study, I clarify some methodological aspects of measuring invertebrate detritivory that could help future research. Nevertheless, with this Thesis I hope to contribute to the understanding of invertebrate community and ecological processes responses to fire disturbance in South Brazilian grasslands. Overall, I can conclude that a functional approach brings many benefits to biodiversity monitoring, and also that spiders and thrips communities, and detritivory and decomposition processes are very resilient to the change brought by small scale grassland burning.

No description available.

Existe uma necessidade urgente em monitorar as respostas da biodiversidade e dos processos ecológicos a distúrbios e mudanças no uso da terra. Diversos estudos tem indicado que o uso da abordagem funcional da biodiversidade, baseada em atributos funcionais, facilita a compreensão sobre a organização dos organismos nas comunidades, da funcionalidade dos ecossistemas, e desta forma, auxilia na identificação das suas relações com fatores ambientais. No primeiro capítulo desta tese eu apresento o arcabouço teórico da ecologia funcional aplicada ao estudo dos invertebrados terrestres, e reviso o que são atributos funcionais, como usá-los na descrição e monitoramento das comunidades e o porquê, ressaltando a ligação dos invertebrados com processos ecológicos. Nos próximos capítulos eu apresento estudos experimentais avaliando as respostas funcionais de dois grupos de invertebrados - aranhas (Araneae) e tripes (Thysanoptera) - e de processos ecológicos influenciados por invertebrados (detritivoria e decomposição de serapilheira) ao distúrbio por fogo nos Campos Sulinos. As queimadas representam uma estratégia de manejo frequente nas pastagens naturais do RS, e carecem em estudos avaliando seus efeitos no ambiente, biodiversidade e processos. A investigação foi realizada com base em um experimento de queima controlada na Estação Agronômica da UFRGS, em Eldorado do Sul, RS. O design foi em blocos casualizados, com 14 parcelas experimentais de 10 x 10 m pareadas; sendo uma parcela de cada bloco queimada no final da primavera de 2009, e a outra, considerada como controle. Informações sobre invertebrados, processos ecológicos e propriedades abióticas do hábitat foram obtidas em todas parcelas antes do fogo (para a caracterização inicial), e 1 mês, 6-7 meses e 12 meses após o fogo. Para o estudo de caso com aranhas, eu classifiquei as comunidades em famílias, atributos de estratégias de caça e medi todos os indivíduos (1755) em atributos funcionais morfológicos, sendo incorporada toda a variabilidade intra e interespecífica, e fenológica das comunidades. Por outro lado, no estudo de caso com tripes, descrevi atributos funcionais (hábitos alimentares, requerimentos de hábitat e atributos morfológicos) por espécie de tripes (48 spp.), assumindo que a variação nos atributos funcionais é sempre maior entre espécies do que dentro da mesma espécie. Nos dois estudos de caso, eu encontrei padrões indicativos de filtragem ambiental, ou seja, o fogo e/ou as mudanças no hábitat induzidas pelo fogo, parecem ter atuado como um filtro, selecionando organismos com determinado conjunto de atributos em detrimento de outros. Esses padrões foram dependentes do período após a queima considerado, por exemplo: um mês após o fogo aranhas construtoras de teias irregulares e tripes fungívoros foram prejudicados, enquanto que aranhas caçadoras da vegetação, com grandes quelíceras, tripes fitófagos e associados com gramíneas aumentaram suas proporções. Para aranhas ainda verifiquei convergência de atributos ao gradiente de biomassa induzido pelo fogo (filtragem ambiental), e divergência de atributos ao gradiente de heterogeneidade da vegetação, indicativo de limitação de similaridade (i.e. aumento da coexistência de organismos funcionalmente diferentes e diminuição de competição). Finalmente com relação aos processos ecológicos, encontrei correlações interessantes acerca de seus agentes modeladores, e.g. entre a atividade de detritivoria com a decomposição de serapilheira, e desta com a equidade da vegetação em campo não queimado. Também mostro que os processos são sensíveis, mas resilientes, ao distúrbio por fogo, seguindo a recuperação da estrutura de hábitat. Adicionalmente, eu incorporei à tese um capítulo sobre uma técnica para avaliar o processo de detritivoria, denominado bait-lamina test, o qual avalia diretamente as taxas de alimentação de invertebrados detritívoros do solo. Neste estudo pioneiro nos campos do RS, esclareço alguns aspectos metodológicos que possam constituir uma base para futuras pesquisas. Contudo, com esta tese espero estar contribuindo com o conhecimento sobre as respostas das comunidades de invertebrados e dos processos ecológicos ao distúrbio por fogo nos Campos Sulinos. Em geral, posso concluir que a abordagem funcional trouxe muitos benefícios ao monitoramento da biodiversidade, e que há elevada resiliência das assembléias de aranhas e tripes, e dos processos de detritivoria e decomposição às queimadas de pequena escala nos campos. / There is a huge need of monitoring the responses of biodiversity and ecological processes to disturbances and land use changes. Several studies have indicated that the functional approach of biodiversity, based on functional traits, facilitates the understanding of community assembly and ecosystem functionality, and thus assists on the identification of their relationships with environmental factors. In the first chapter of this Thesis I present the functional ecology theory applied to studies on terrestrial invertebrates, and I review what functional traits are, how to use them in community description and monitoring, and why, highlighting the connections between invertebrates and ecological processes. In the following chapters I present experimental studies evaluating the functional responses of two invertebrate groups- spiders (Araneae) and thrips (Thysanoptera) – and ecological processes influenced by invertebrates (detritivory and litter decay) to fire disturbance in Campos grasslands. Burning represents a common strategy for pasture management in Rio Grande do Sul State, and there is a lack of studies regarding their effects on the environment, biodiversity and processes. Our investigation was performed based in a burning experiment at Estação Agronômica of UFRGS, Eldorado do Sul, RS. The design was blocked, with 14 experimental paired plots of 10 x 10 m. One plot from each block was burned at the end of spring 2009, and the other is considered a control plot. Information concerning invertebrates, ecological processes and abiotic habitat proprieties were obtained from all plots before fire (initial characterization) and 1 month, 6-7 months and 12 months after fire. For the spider study case, I classified the organisms in families, feeding strategy traits, and measured all spider individuals (1755) thus incorporating all intra and interspecific variability of the communities. On the other hand, in the thrips study case I described functional traits (feeding habits, habitat requirements, and morphological traits) per species (48 spp.), assuming that the variation on functional traits should be higher between species than within species. In both cases, I found patterns indicating environmental filtering, which means that fire and/or the fire-induced habitat changes acted as a filter, sorting organisms with a determined group of traits instead of others. These patterns were dependent on the period after fire considered. For example, one month after fire irregular web spider builders and fungivorous thrips were more uncommon than expected, while vegetation hunter spiders, with large chelicerae, and leaf feeder thrips and thrips associated to grasses increased their proportions. For spiders we also verified a trait-convergence pattern along the plant biomass gradient induced by fire (environmental filtering), and trait-divergence pattern along the vegetation heterogeneity gradient indicating limiting similarity (i.e. increase in coexistence of functionally different organisms, and reduction of competition). Finally, regarding ecological processes, I found interesting correlations on their drivers, e.g. between detritivory and litter decay, and between these with plant evenness in unburned grasslands. Also, I show that ecological processes are sensitive but resilient to fire, following habitat structure recovery. Additionally, I incorporate to the Thesis a methodology chapter about the bait-lamina test, which evaluates the feeding rates of soil detritivore invertebrates directly. In this pioneer study, I clarify some methodological aspects of measuring invertebrate detritivory that could help future research. Nevertheless, with this Thesis I hope to contribute to the understanding of invertebrate community and ecological processes responses to fire disturbance in South Brazilian grasslands. Overall, I can conclude that a functional approach brings many benefits to biodiversity monitoring, and also that spiders and thrips communities, and detritivory and decomposition processes are very resilient to the change brought by small scale grassland burning.

Existe uma necessidade urgente em monitorar as respostas da biodiversidade e dos processos ecológicos a distúrbios e mudanças no uso da terra. Diversos estudos tem indicado que o uso da abordagem funcional da biodiversidade, baseada em atributos funcionais, facilita a compreensão sobre a organização dos organismos nas comunidades, da funcionalidade dos ecossistemas, e desta forma, auxilia na identificação das suas relações com fatores ambientais. No primeiro capítulo desta tese eu apresento o arcabouço teórico da ecologia funcional aplicada ao estudo dos invertebrados terrestres, e reviso o que são atributos funcionais, como usá-los na descrição e monitoramento das comunidades e o porquê, ressaltando a ligação dos invertebrados com processos ecológicos. Nos próximos capítulos eu apresento estudos experimentais avaliando as respostas funcionais de dois grupos de invertebrados - aranhas (Araneae) e tripes (Thysanoptera) - e de processos ecológicos influenciados por invertebrados (detritivoria e decomposição de serapilheira) ao distúrbio por fogo nos Campos Sulinos. As queimadas representam uma estratégia de manejo frequente nas pastagens naturais do RS, e carecem em estudos avaliando seus efeitos no ambiente, biodiversidade e processos. A investigação foi realizada com base em um experimento de queima controlada na Estação Agronômica da UFRGS, em Eldorado do Sul, RS. O design foi em blocos casualizados, com 14 parcelas experimentais de 10 x 10 m pareadas; sendo uma parcela de cada bloco queimada no final da primavera de 2009, e a outra, considerada como controle. Informações sobre invertebrados, processos ecológicos e propriedades abióticas do hábitat foram obtidas em todas parcelas antes do fogo (para a caracterização inicial), e 1 mês, 6-7 meses e 12 meses após o fogo. Para o estudo de caso com aranhas, eu classifiquei as comunidades em famílias, atributos de estratégias de caça e medi todos os indivíduos (1755) em atributos funcionais morfológicos, sendo incorporada toda a variabilidade intra e interespecífica, e fenológica das comunidades. Por outro lado, no estudo de caso com tripes, descrevi atributos funcionais (hábitos alimentares, requerimentos de hábitat e atributos morfológicos) por espécie de tripes (48 spp.), assumindo que a variação nos atributos funcionais é sempre maior entre espécies do que dentro da mesma espécie. Nos dois estudos de caso, eu encontrei padrões indicativos de filtragem ambiental, ou seja, o fogo e/ou as mudanças no hábitat induzidas pelo fogo, parecem ter atuado como um filtro, selecionando organismos com determinado conjunto de atributos em detrimento de outros. Esses padrões foram dependentes do período após a queima considerado, por exemplo: um mês após o fogo aranhas construtoras de teias irregulares e tripes fungívoros foram prejudicados, enquanto que aranhas caçadoras da vegetação, com grandes quelíceras, tripes fitófagos e associados com gramíneas aumentaram suas proporções. Para aranhas ainda verifiquei convergência de atributos ao gradiente de biomassa induzido pelo fogo (filtragem ambiental), e divergência de atributos ao gradiente de heterogeneidade da vegetação, indicativo de limitação de similaridade (i.e. aumento da coexistência de organismos funcionalmente diferentes e diminuição de competição). Finalmente com relação aos processos ecológicos, encontrei correlações interessantes acerca de seus agentes modeladores, e.g. entre a atividade de detritivoria com a decomposição de serapilheira, e desta com a equidade da vegetação em campo não queimado. Também mostro que os processos são sensíveis, mas resilientes, ao distúrbio por fogo, seguindo a recuperação da estrutura de hábitat. Adicionalmente, eu incorporei à tese um capítulo sobre uma técnica para avaliar o processo de detritivoria, denominado bait-lamina test, o qual avalia diretamente as taxas de alimentação de invertebrados detritívoros do solo. Neste estudo pioneiro nos campos do RS, esclareço alguns aspectos metodológicos que possam constituir uma base para futuras pesquisas. Contudo, com esta tese espero estar contribuindo com o conhecimento sobre as respostas das comunidades de invertebrados e dos processos ecológicos ao distúrbio por fogo nos Campos Sulinos. Em geral, posso concluir que a abordagem funcional trouxe muitos benefícios ao monitoramento da biodiversidade, e que há elevada resiliência das assembléias de aranhas e tripes, e dos processos de detritivoria e decomposição às queimadas de pequena escala nos campos. / There is a huge need of monitoring the responses of biodiversity and ecological processes to disturbances and land use changes. Several studies have indicated that the functional approach of biodiversity, based on functional traits, facilitates the understanding of community assembly and ecosystem functionality, and thus assists on the identification of their relationships with environmental factors. In the first chapter of this Thesis I present the functional ecology theory applied to studies on terrestrial invertebrates, and I review what functional traits are, how to use them in community description and monitoring, and why, highlighting the connections between invertebrates and ecological processes. In the following chapters I present experimental studies evaluating the functional responses of two invertebrate groups- spiders (Araneae) and thrips (Thysanoptera) – and ecological processes influenced by invertebrates (detritivory and litter decay) to fire disturbance in Campos grasslands. Burning represents a common strategy for pasture management in Rio Grande do Sul State, and there is a lack of studies regarding their effects on the environment, biodiversity and processes. Our investigation was performed based in a burning experiment at Estação Agronômica of UFRGS, Eldorado do Sul, RS. The design was blocked, with 14 experimental paired plots of 10 x 10 m. One plot from each block was burned at the end of spring 2009, and the other is considered a control plot. Information concerning invertebrates, ecological processes and abiotic habitat proprieties were obtained from all plots before fire (initial characterization) and 1 month, 6-7 months and 12 months after fire. For the spider study case, I classified the organisms in families, feeding strategy traits, and measured all spider individuals (1755) thus incorporating all intra and interspecific variability of the communities. On the other hand, in the thrips study case I described functional traits (feeding habits, habitat requirements, and morphological traits) per species (48 spp.), assuming that the variation on functional traits should be higher between species than within species. In both cases, I found patterns indicating environmental filtering, which means that fire and/or the fire-induced habitat changes acted as a filter, sorting organisms with a determined group of traits instead of others. These patterns were dependent on the period after fire considered. For example, one month after fire irregular web spider builders and fungivorous thrips were more uncommon than expected, while vegetation hunter spiders, with large chelicerae, and leaf feeder thrips and thrips associated to grasses increased their proportions. For spiders we also verified a trait-convergence pattern along the plant biomass gradient induced by fire (environmental filtering), and trait-divergence pattern along the vegetation heterogeneity gradient indicating limiting similarity (i.e. increase in coexistence of functionally different organisms, and reduction of competition). Finally, regarding ecological processes, I found interesting correlations on their drivers, e.g. between detritivory and litter decay, and between these with plant evenness in unburned grasslands. Also, I show that ecological processes are sensitive but resilient to fire, following habitat structure recovery. Additionally, I incorporate to the Thesis a methodology chapter about the bait-lamina test, which evaluates the feeding rates of soil detritivore invertebrates directly. In this pioneer study, I clarify some methodological aspects of measuring invertebrate detritivory that could help future research. Nevertheless, with this Thesis I hope to contribute to the understanding of invertebrate community and ecological processes responses to fire disturbance in South Brazilian grasslands. Overall, I can conclude that a functional approach brings many benefits to biodiversity monitoring, and also that spiders and thrips communities, and detritivory and decomposition processes are very resilient to the change brought by small scale grassland burning.

Introdução: As Distrofias Musculares Congênitas (DMCs) são um grupo clínica e geneticamente heterogêneo de doenças musculares que se manifestam ao nascimento ou no primeiro ano de vida, sendo caracterizadas por hipotonia, fraqueza muscular, retardo do desenvolvimento motor e retrações fibrotendíneas. O músculo esquelético apresenta-se distrófico, mas sem alterações estruturais específicas. Em quase metade dos casos a doença é causada pela deficiência da laminina alfa;-2 (merosina). Outras deficiências proteicas descritas incluem: colágeno VI, selenoproteína N1, várias glicosiltransferases responsáveis pela glicosilação da alfa- distroglicana e lamina A/C. Vários genes já foram identificados. Objetivo: o objetivo deste estudo foi a caracterização clínica, histológica e molecular das crianças com DMC por deficiência de lamina A/C. Método: Foram incluídos 13 pacientes com diagnóstico clínico e histológico de DMC, com expressão muscular normal para distrofina, sarcoglicanas, merosina, colágeno 6 e disferlina. Os pacientes foram reavaliados segundo protocolo clínico e neurológico. As biópsias musculares realizadas previamente foram revisadas e o estudo das mutações no gene da lamina A/C foi realizado através de sequenciamento de toda região codificadora do gene. Resultados: Identificamos mutações em 30,7% dos pacientes (quatro casos) com fenótipo clínico de DMC por deficiência de lamina A/C. Todas as mutações encontradas (p.E358K, p.R249W, e p.N39S) ocorreram em heterozigose e de novo e já haviam sido descritas na literatura em pacientes com distrofias musculares. Em geral, estes pacientes apresentavam um grave comprometimento motor com o característico aspecto de cabeça caída, com início dos sintomas nos primeiros dois anos de vida. A CPK estava elevada entre 2 a 6 vezes o padrão superior da normalidade. O padrão histológico variou desde um músculo levemente até gravemente distrófico. Curiosamente, no estudo histológico do músculo, um dos pacientes apresentou agregados intracitoplasmáticos. Um outro paciente apresentava associadamente alterações neurogênicas ao estudo eletroneuromiográfico. Em todos os casos observamos complicações respiratórias, cardíacas e distúrbios de deglutição. Houve um caso de morte súbita, provavelmente em decorrência de arritmia cardíaca. Conclusões: A correlação genótipo-fenótipo permanece difícil, mas todos os casos apresentaram sinal da cabeça caída, comprometimento respiratório, cardíaco e biópsia muscular distrófica. A ampliação do conhecimento clínico e histológico pode orientar o diagnóstico e direcionar para o estudo molecular adequado, além de permitir o diagnóstico precoce das complicações, tão frequentes na DMC por deficiência de lamina A/C. Os exons 1, 4 e 6 são os mais frequentemente mutados e devem ser pesquisados inicialmente. Esta série de casos contribui também por demonstrar a distribuição universal da doença / Background: The Congenital Muscular Dystrophies (CMD) are a clinically and genetically heterogeneous group of myopathies characterized by muscle hypotonia, delayed motor development and early onset of progressive muscle weakness with dystrophic pattern on muscle biopsy. The clinical course is broadly variable and can comprise the involvement of the brain and eyes. Almost half of the cases is caused by deficiency of laminin-alfa 2 (merosin). Other protein deficiencies described include: collagen VI, selenoprotein N1, several glycosyltransferases responsible for glycosylation of alfa-dystroglycan and lamin A/C. Several genes have been identified and the increased knowledge of new clinical and histological forms of CMD can guide diagnosis and direct appropriate molecular studies. LMNA-related CMD is often characterized by muscle weakness and a dropped head developed in the early years of life. Regarding lamin A/C deficiency, the immunohistochemical findings can be normal, probably because the protein change is functional only; this makes diagnosis using muscle samples more difficult. Objectives: The aim of this study was to characterize the clinical, histological and molecular aspects in patients with CMD related to deficiency of lamin A/C. Methods: thirteen children with clinical and histological diagnosis of CMD with normal muscle expression for dystrophin, merosin, collagen 6, sarcoglycans and dysferlin were included in this study. The LMNA gene was sequenced after amplification of all coding exons. In addition, the muscle biopsies were revised. Results: In 30.7% (four cases) of our patients with typical clinical phenotype of lamin A/C deficiency were detected mutations on LMNA gene and all of them presented dropped-head syndrome, restrictive ventilator insufficiency, cardiac changes, increased serum CPK level and myopathic/dystrophic aspect on muscle biopsy. Two of the patients had normal motor development milestones in the first months of life and subsequently developed cervical and limb weakness. The other two patients presented a more severe motor involvement and failure to walk. One patient showed associated peripheral neuropathy. Curiously one case had myofibrillar aggregates on muscle biopsy. All mutations (p.E358K, p.R249W and p.N39S) were heterozygous and de novo and had been previously described in patients with muscular dystrophy. Conclusion: Genotype/phenotype correlation in CMD remains difficult. However patients with LMNA mutation and CMD seems to have a more homogeneous phenotype characterized by dropped head, severe motor disability, and cardiac and pulmonary involvement. Mutations on exons 1, 4 and 6 should be tested first. This case series also contributes for showing the universal distribution of the disease

This work explores the use of carbon-infiltrated carbon nanotube (CI-CNT) forests as a material for fabricating compliant MEMS devices. The impacts of iron catalyst layer thickness and carbon infiltration time are examined. An iron layer of 7nm or 10nm with an infiltration time of 30 minutes produces CI-CNT best suited for compliant applications. Average maximum strains of 2% and 2.48% were observed for these parameters. The corresponding elastic moduli were 5.4 GPa and 4.1 GPa, respectively. A direct comparison of similar geometry suggested CI-CNT is 80% more flexible than single-crystal silicon. A torsional testing procedure provided an initial shear modulus of about 5 GPa for the 7-nm, 30-min CI-CNT. The strain and elastic modulus values were used to design numerous functional devices which were then fabricated in CI-CNT. A series of compliant cell restraint mechanisms were developed, assessed, and revised. A passive restraint with no moving parts was found to be both the most effective design and the easiest design to produce economically. A refined version of the passive restraint has been released commercially. Another series of designed devices successfully demonstrates the implementation of CI-CNT LEM designs.

Im Rahmen der Routinediagnostik von Endometriumbioptaten der Stute fielen regel-mäßig periglanduläre mononukleäre Entzündungszellinfiltrate (PAME) auf. Ziel der vorliegenden Studie war die histomorphologische und immunhistologische Charakterisierung der PAME im klinisch-gynäkologischen, jahreszeitlichen und endometrialen Kontext. Insbesondere sollte geklärt werden, ob das Phänomen der periglandulären Entzündungszellakkumulation Ausdruck eines physiologischen endometrialen Befundes, Frühstadium (Trigger) der Endometrose oder Begleitsymptom einer Endometritis ist. Zu diesem Zweck wurden alle im Jahr 2009 mittels einer Übersichtsfärbung (H.-E.-Färbung) im Institut für Veterinär-Pathologie der Universität Leipzig untersuchten Endometriumbioptate von Stuten (n = 754) hinsichtlich des Vorkommens einer PAME überprüft. Es konnten 133 Bioptate von 131 Stuten identifiziert werden, die ausführlich histopathologisch ausgewertet wurden. 72 Bioptate wurden zusätzlich anhand einer Methylgrün-Pyronin-Färbung (MGP) und immunhistologischer Verfahren beurteilt: Neben der Differenzierung der an der PAME beteiligten Zellpopulationen (CD3, CD79 A, MAC 387, MGP), wurde das Vorkommen der Intermediärfilamente Vimentin und Desmin sowie von α-GMA in den beteiligten Epithel- und Stromazellen untersucht. Die glanduläre Basallamina wurde anhand der Basallaminakomponente Laminin dargestellt und charakterisiert. PAME kamen in 18 % aller im Jahr 2009 untersuchten Bioptate vor, bevorzugt (96 %) in entzündlich und/oder fibrotisch alterierten Endometrien. Der Entzündungszellcharakter der begleitenden Endometritis stimmte in 29 % der Fälle nicht mit dem Entzündungszellbild der PAME (mononukleär) überein. Während im übrigen Endometrium häufig (95 %) eine gering- bis mittelgradige Endometrose nachweisbar war, wiesen die PAME-Drüsen mehrheitlich (71 %) keine periglanduläre Fibrose auf. Bei der Endometrose um Drüsen mit einer PAME handelt es sich überwiegend (67 %) um die destruierende und zu 50 % um die aktive/gemischte Form. In 48 % der PAME-Lokalisationen wurde eine Infiltration der periglandulären Entzündungszellen zwischen die Drüsenepithelzellen beobachtet. Die PAME bestanden hauptsächlich aus T-Lymphozyten (CD3-positiv). Daneben fanden sich, in geringerer und variierender Anzahl, Plasmazellen (MGP), B-Zellen (CD79A-positiv) und Makrophagen (MAC 387-positiv). Anhand der Expression der Basallaminakomponente Laminin wurden in allen untersuchten PAME-Lokalisationen Alterationen der Basallamina nachgewiesen. Die Läsionen der Basallamina waren bei mittel- und hochgradigen PAME graduell stärker ausgeprägt und standen häufig (43 %) mit einer intraepithelialen Infiltration der Entzündungszellen im Zusammenhang. Vimentin wurde vereinzelt (4 %) in intraläsionalen Drüsenepithelzellen nachgewiesen. Periläsionale Stromazellen exprimieren zu 60 % Vimentin, zu 17 % Desmin und zu 28 % α-GMA. Bei der PAME handelt es sich sehr wahrscheinlich nicht ausschließlich um ein „histopathologisches Symptom“ einer chronischen nicht-eitrigen Endometritis. Die Interpretation der PAME als physiologischer Bestandteil eines Schleimhaut-assoziierten lymphatischen Gewebes (MALT) im equinen Endometrium hingegen ist denkbar und bedarf in Folgeuntersuchungen der Klärung. Hinsichtlich der Strukturfilamente wurden in den involvierten Epithel- und Stromazellen immunhistologische Expressionsmuster nachgewiesen, die mit denen in frühen Stadien der equinen Endometrose vergleichbar sind. Darüber hinaus wiesen die periläsionalen Stromazellen Differenzierungsmerkmale von Myofibroblasten auf. PAME waren eng mit dem Vorliegen von Basallaminaalterationen verknüpft, die im Entstehungsprozess der equinen Endometrose eine zentrale Bedeutung besitzen. Es ist vorstellbar, dass es sich bei der PAME möglicherweise um einen auslösenden Faktor der Endometrose handelt. Außerdem ist eine intrinsische Aufrechterhaltung im Rahmen der zytokinvermittelten Interaktion (IL-4, IL-13, MCP-1), zwischen den periglandulären Entzündungszellen und den an einer Fibrose beteiligten Stromazellen, mit daraus resultierender progredienter Fibrose denkbar. Zukünftig sollte in weiteren Untersuchungen eine diesbezügliche Analyse der potentiellen Regelkreise erfolgen.