Os craniofaringiomas são os tumores mais frequentes da região hipotálamohipofisária na faixa etária pediátrica. Apesar de serem histologicamente benignos, sua tendência infiltrativa e seu comportamento agressivo resultam em significante morbimortalidade. Histologicamente podem ser divididos em dois subtipos: adamantinomatosos e papilíferos. A patogênese dos craniofaringiomas é pouco compreendida. Mutações no gene CTNNB1, que codifica a proteína beta-catenina, são a única alteração molecular conhecida até o momento implicada na tumorigênese dos craniofaringiomas adamantinomatosos. Tais mutações afetam o sítio de degradação da beta-catenina, que passa a se acumular no citoplasma e no núcleo, ativando excessivamente a via de sinalização WNT, através da ligação aos fatores de transcrição da família LEF/TCF, levando a tumorigênese. Recentemente foi descoberto um novo mecanismo de determinação da linhagem celular hipofisária regulado pela beta-catenina, através do qual ela interage diretamente com o PROP1 para determinar a diferenciação celular hipofisária. De acordo com esse modelo, o complexo protéico PROP1/beta- catenina atua simultaneamente como repressor do HESX1 e ativador do PIT1, dependendo dos co-fatores associados. Pacientes com mutações germinativas inativadoras no PROP1 desenvolvem hipopituitarismo e podem apresentar aumento hipofisário com imagens de ressonância nuclear magnética (RNM) da região selar muitas vezes semelhantes àquelas dos craniofaringiomas, com hiperssinal em T1. Por outro lado, camundongos com expressão persistente do Prop1 exibem defeitos na regulação da proliferação celular hipofisária, incluindo cistos da bolsa de Rathke, hiperplasia adenomatosa e tumores, sugerindo que mutações com ganho de função no PROP1 também poderiam contribuir para a patogênese de tumores hipofisários em seres humanos. A semelhança entre as imagens de RNM dos pacientes com craniofaringiomas e daqueles com aumento hipofisário devido a mutações inativadoras no PROP1, e o fato de que camundongos transgênicos com expressão persistente do Prop1 apresentam aumento da susceptibilidade a tumores hipofisários, deram base a nossa hipótese de que uma desregulação na expressão do PROP1 em humanos poderia estar envolvida na patogênese dos craniofaringiomas adamantinomatosos. Esse trabalho teve como objetivo avaliar a presença de mutação somática no exon 3 do CTNNB1 e avaliar a expressão desse gene e do gene PROP1 em craniofaringiomas adamantinomatosos. Foram obtidas 14 amostras desse tipo de tumor por meio da ressecção terapêutica. As amostras foram submetidas à extração do RNA e posterior transcrição reversa para obtenção de cDNA. A partir do cDNA foi realizada PCR e sequenciamento do exon 3 do CTNNB1 em todas as amostras. Porém, a avaliação por PCR em tempo real foi realizada apenas em 12 amostras, devido à qualidade inadequada de 2 amostras para submissão a essa metodologia. Foram encontradas mutações missense, em heterozigose em 9 das 14 amostras, sendo 5 previamente descritas e 2 ainda não descritas em craniofaringiomas adamantinomatosos. Hiperexpressão do CTNNB1 foi encontrada em 7 amostras, sendo 5 com mutação e 2 sem mutação no CTNNB1.A hiperexpressão variou de 2,5 a 6,2 vezes maior que o pool de hipófise normal. Contudo, a expressão do PROP1 foi indetectável em todas as amostras. Concluímos que o aumento da expressão do CTNNB1 presente em 58% das amostras sugere o envolvimento também da hiperexpressão desse gene na etiopatogenia do craniofaringioma adamantinomatoso, enquanto a ausência de expressão do PROP1 afasta a participação desse gene na etiopatogenia do craniofaringioma adamantinomatoso / Craniopharyngiomas are the the commonest tumors to involve the hypothalamo-pituitary regions in childhood population. Histologically they are benign, and can be divided in two primary subtypes: the adamantinomatous and the papillary. Although histologically benign, their infiltrative tendency and aggressive behavior can result in great morbidity. The pathogenesis of craniopharyngiomas is poorly understood. To date, beta-catenin gene (CTNNB1) mutations have been identified only in the adamantinomatous subtype. These mutations affect the degradation target box of beta-catenin that accumulates in the cytoplasm and the nucleus increasing the transcriptional activity of WNT pathway through interaction with the transcription factors of LEF/TCF family, leading to tumorigenesis. Recently, an interaction between beta-catenin and PROP1 was described as a new mecanism for beta-catenindependent regulation of pituitary cell-lineage determination. According to this novel model, the PROP1/beta-catenin proteic complex would act as a binary switch to simultaneously repress the transcription factor HESX1 and to activate expression of transcription factor PIT1, depending on the associated cofactors. Patients with loss-of-function mutations in PROP1 present combined pituitary hormonal deficiency generally associated with pituitary enlargement and the magnetic resonance imaging (MRI) of the sellar region in these patients sometimes resembles that of the craniopharyngiomas, with T1 hyperintense signal. On the other hand, transgenic mice with persistent Prop1 expression exhibit defects consistent with misregulation of pituitary cell proliferation, including adenomatous hyperplasia with formation of Rathke\'s cleft cysts and tumors suggesting that misregulation of PROP1 expression in human could contribute to pathogenesis of pituitary tumors. The similarity between the MRI images of craniopharyngiomas patients and that of patients with loss-of-function mutations in PROP1, associated with the fact that transgenic mice with persistent Prop1 expression exhibit increased susceptibility to pituitary tumors gave rise to our hypothesis that a misregulation of PROP1 expression could be involved in the pathogenesis of adamantinomatous craniopharyngiomas. The aim of this study was to analyze the presence of somatic mutations in exon 3 of CTNNB1 and the expression pattern of this gene and the PROP1 gene in adamantinomatous craniopharyngiomas. Fourteen samples were obtained from therapeutic surgery and submitted to RNA extraction and reverse transcription in order to produce the cDNA. The cDNA was used as a template to CTNNB1 exon 3 PCR reaction followed by direct sequencing of all samples. However, the real-time RT-PCR analysis was realized only in 12 samples, since 2 of them had an insufficient quality for this method. Missence, heterozygous mutations were found in 9 out of 14 samples; five were previously described and 2 not yet described in adamantinomatous craniopharyngiomas. Overexpression of CTNNB1 was found in 7 samples, which them 5 with CTNNB1 mutation 2 whitout. The overexpression ranged from 2.5 to 6.2 fold more than pituitary normal pool. However, the PROP1 expression was undetectable in all the samples. We could conclude that the amount of 58% CTNNB1 overexpressed samples suggest also a role of this overexpression in the pathogenesis of adamantinomatous craniopharingiomas, while the undetectable levels of PROP1 exclude a role of this gene in the pathogenesis of adamantinomatous craniopharingiomas

O fósforo é um dos minerais mais abundantes no corpo além de ser essencial para muitos processos biológicos. A homeostase do fósforo depende da absorção no intestino, da excreção renal, da remodelação óssea e de hormônios como o paratormônio, calcitriol e FGF-23. Nos pacientes com doença renal crônica, a excreção urinária de fósforo está comprometida levando à hiperfosfatemia, que contribui para o aumento da morbidade e mortalidade desses pacientes. A absorção intestinal de fósforo no intestino delgado, particularmente via cotransportadores NaP-IIb e PiT-1, é pouco estudada. O objetivo desse estudo foi avaliar os efeitos de dietas com diferentes concentrações de fósforo na expressão proteica e gênica dos cotransportadores NaP-IIb e PiT-1, bem como na apoptose dos enterócitos dos diferentes segmentos do intestino delgado de animais controles e urêmicos. Estudamos setenta e seis ratos Wistar machos, inicialmente divididos em dois grupos: controles (C) e urêmicos (Nx). Cada grupo foi subdividido em outros três, de acordo com a concentração de fósforo (P) na dieta: dieta Baixa (0,2%), dieta Padrão (0,54%) e dieta Alta (0,9%). Analisamos parâmetros bioquímicos (creatinina, P, Cai, PTH e FGF-23), a expressão proteica dos cotransportadores, através de Western Blotting, ELISA e imunofluorescência, a expressão gênica por PCR em tempo real e a apoptose dos enterócitos pela técnica TUNEL. Os resultados mostraram que os níveis séricos de creatinina, P, PTH e FGF-23 foram significativamente mais elevados nos animais Nx. Nos animais C com dieta baixa em P observamos aumento da expressão proteica do cotransportador NaP-IIb em todos os segmentos do intestino enquanto, no Nx Baixo, a expressão desse cotransportador foi menor somente no jejuno. Quanto ao PiT-1, sua expressão foi menor no íleo do grupo Nx Alto comparado ao seu respectivo controle. A expressão gênica do cotransportador NaP-IIb foi menor no jejuno do Nx Alto e maior no íleo desse mesmo grupo em relação aos seus respectivos controles. A expressão gênica do PiT-1 foi maior em todos os segmentos do grupo Nx Baixo em relação aos seus controles. Detectamos uma correlação direta entre a expressão gênica do NaP-IIb no jejuno com o fósforo sérico. A expressão gênica do PiT-1 no jejuno correlacionou-se com o fósforo sérico e no duodeno e jejuno, com os níveis séricos de FGF-23. No duodeno e no jejuno, a porcentagem de enterócitos apoptóticos foi maior nos animais Nx Alto comparados aos controles, particularmente no duodeno a porcentagem dessas células também foi maior no grupo Nx Baixo e controle padrão. Já no jejuno, tanto o grupo Nx baixo quanto no Nx Padrão observamos menos células apoptóticas que nos seus respectivos controles. Quando analisamos conjuntamente todos os segmentos intestinais o grupo Nx alto apresentou mais células apoptóticas que o controle e o oposto foi observado nos grupos Nx Baixo e Nx Padrão. Em conclusão, dietas com distintas concentrações de fósforo promovem modificações na expressão proteica e gênica dos cotransportadores NaP-IIb e PiT-1 no intestino delgado que não seguem um padrão uniforme. A sobrecarga de fósforo aumenta a porcentagem de enterócitos apoptóticos nos animais urêmicos / Phosphorus is one of the most abundant minerals in the body besides being essential for many biological processes. Phosphorus homeostasis depends on absorption in the small intestine, renal excretion, bone remodeling and hormones such as parathyroid hormone, calcitriol and FGF-23. In patients with chronic kidney disease phosphorus urinary excretion is compromised leading to hyperphosphatemia, which contributes to increased morbidity and mortality of these patients. Intestinal absorption of phosphorus in the small intestine, particularly of NaP-IIb and PiT-1 cotransporters is poorly studied. The aim of this study was to evaluate the effects of diets with different concentrations of phosphorus in protein expression and gene of NaP-IIb and PiT-1 cotransporters as well as in apoptosis of enterocytes of different segments of intestine in control and uremic animals. We studied seventy-six male Wistar rats initially divided into two groups: controls (C) and uremic (Nx). Each group was subdivided into three others, according to the phosphorus concentration in the diet: Low diet (0.2% P), Standard diet (0.54% P) and High diet (0.9% P). We analyzed biochemical parameters (creatinine, P, iCa, PTH and FGF-23), protein expression of cotransporters, using Western Blot, ELISA, immunofluorescence, real-time PCR and apoptosis of enterocytes using the TUNEL technique. Results showed that serum creatinine, P, PTH and FGF-23 were significantly higher in Nx animals. In C animals with a diet low in P we observed increase in protein expression of NaP-IIb cotransporter in all segments of the intestine while in low Nx expression of this cotransporter was lower only in jejunum. As for PiT-1 expression was lower in the ileum of the high Nx group compared to their respective control. Gene expression of NaP-IIb cotransporter was lower in the jejunum of high Nx and higher in the ileum of the same group as compared to their respective controls. PiT-1 gene expression was higher in all segments of the low Nx group compared to their controls. We detected a direct correlation between the gene expression of NaP-IIb in jejunum and serum phosphorus. Gene expression of PiT-1 correlated with serum phosphorus in the jejunum, and correlated with the serum levels of FGF-23 in the jejunum and duodenum. In the duodenum and jejunum the percentage of apoptotic enterocytes was higher in high Nx animals compared to controls; particularly in the duodenum the percentage of these cells was also higher in low Nx group and standard control. In the jejunum in both low Nx group and standard Nx we observed fewer apoptotic cells than in their respective controls. When we analyze all intestinal segments together the high Nx group had more apoptotic cells than the control, and the opposite was observed in the low Nx group and standard Nx group. In conclusion, diets with different phosphorus concentrations promote changes in protein expression and gene of NaP-IIb and PiT-1 cotransporters in the small intestine that do not follow a uniform pattern. Phosphorus overhead increases the percentage of apoptotic enterocytes in uremic animals

L’embranchement Hemichordata regroupe les classes Enteropneusta et Pterobranchia. Hemichordata constitue, avec l’embranchement Echinodermata, le groupe-frère des chordés. Les entéropneustes sont des organismes vermiformes solitaires qui vivent sous ou à la surface du substrat et s’alimentent généralement par déposivorie, alors que les ptérobranches sont des organismes coloniaux filtreurs habitant dans un réseau de tubes appelé coenecium. Ce mémoire présente trois études dont le point commun est l’utilisation des hémichordés actuels pour répondre à des questions concernant l’évolution des hémichordés, des chordés, et du super-embranchement qui les regroupe, Deuterostomia. Notre première étude démontre que les fentes pharyngiennes, l’organe pré-oral cilié (POCO) et le pharynx de l’entéropneuste Protoglossus graveolens sont utilisés pour l’alimentation par filtration. Le système de filtration de P. graveolens permet la capture de particules jusqu’à 1.3 um, à un débit de 4.05 mm.s-1, pour une demande énergétique de 0.009 uW. Les similarités structurales et fonctionnelles avec le système de filtration des céphalochordés suggèrent que la filtration pharyngienne est ancestrale aux deutérostomes. Lors de notre deuxième étude, nous avons exploré l’hypothèse selon laquelle le POCO des entéropneustes, une structure ciliée pré-buccale au rôle possiblement chémorécepteur, serait homologue au « wheel organ » des céphalochordés et à l’adénohypophyse des vertébrés. Pour cela, nous avons déterminé par immunohistochimie l’expression de Pit-1, un facteur de transcription spécifique à ces deux structures, chez l’entéropneuste Saccoglossus pusillus. Pit-1 est exprimé dans des cellules sensorielles du POCO, mais aussi dans des cellules épithéliales distribuées dans le proboscis, collet et tronc. Ce patron d’expression ne permet pas de confirmer ou rejeter l’homologie du POCO et de l’adénohypophyse des vertébrés. Lors de notre troisième étude, nous avons caractérisé l’ultrastructure du coenecium des ptérobranches Cephalodiscus hodgsoni, Cephalodiscus nigrescens et Cephalodiscus densus par microscopie électronique à transmisison et à balayage. Cephalodiscus est le groupe frère de Graptolithina, un groupe qui inclut les graptolithes éteints ainsi que les ptérobranches du genre Rhabdopleura. Nous avons décrit les types de fibrilles de collagène présents, leur taille et leur organisation, ainsi que l’organisation globale du coenecium. Nous avons ainsi démontré la présence chez Cephalodiscus d’une organisation similaire au paracortex, pseudocortex et eucortex des graptolithes. La présence chez Cephalodiscus de ce type d’organisation suggère que le cortex est ancestral à la classe Pterobranchia. Ces trois études illustrent plusieurs axes importants de la recherche sur les hémichordés, qui en intégrant des données morphologiques, fonctionnelles et moléculaires permet de reconstruire certains évènements clés de l’évolution des deutérostomes. / The phylum Hemichordata comprises the classes Enteropneusta and Pterobranchia. Together with echinoderms, hemichordates are the sister-group to chordates. Enteropneusts are worm-shaped solitary deposit feeders. Pterobranchs are colonial filter feeders that live in a secreted collagenous domicile called a coenecium. In this thesis, three studies are presented. These studies are based on observations of extant hemichordates, and adress a variety of issues relating to the evolution of hemichordates, chordates, and the super-phylum to which they belong: Deuterostomia. Our first study demonstrates that the gill slits, pre-oral ciliary organ (POCO), and lining of the pharynx of the enteropneust Protoglossus graveolens are used in filter feeding. The filter-feeding system of P. graveolens enables particle capture down to 1.3 um, at a rate up to 4.05 mm.s-1, with a power consumption of 0.009 uW. Structural and functional similarities with the cephalochordate filter-feeding system suggest that pharyngeal filter-feeding is ancestral to the deuterostomes. In our second study, we address the hypothesis that the enteropneust POCO, a putative chemosensory structure located anterior to the mouth, is homologous to the cephalochordate wheel organ and vertebrate adenohypophysis. We characterized the expression pattern of the adenohypophysis-specific transcription factor Pit-1 in the adult enteropneust Saccoglossus pusillus with immunohistochemistry. Pit-1 is expressed in sensory cells of the POCO and in scattered epithelial cells of the proboscis, collar and trunk. This expression pattern does not allow to confirm or reject the homology of the POCO with the vertebrate adenohypophysis. In our third study, we characterized the ultrastructure of the coenecium of the pterobranchs Cephalodiscus hodgsoni, Cephalodiscus nigrescens and Cephalodiscus densus using transmission and scanning electron microscopy. Cephalodiscus is the sister-group to the Graptolithina, which includes the extinct graptolites and the extant pterobranch genus Rhabdopleura. We described the fibril types, size and organization, as well as the general organization of the coenecium. We demonstrated that the coenecium of Cephalodiscus shows similarities with the graptolite eucortex, paracortex and pseudocortex. The cortical-like organization of the coenecium of Cephalodiscus suggests that the cortex is ancestral to the Pterobranchia. Together, these three studies illustrate different axes of hemichordate research, and show how integrating morphological, functional and molecular data allows us toinfer key events in the evolution of deuterostomes.

Made available in DSpace on 2014-12-17T14:09:16Z (GMT). No. of bitstreams: 1 JardelDC_TESE_Parcial.pdf: 1739659 bytes, checksum: 19974b9395e0d9fcb23b07c3997dab2f (MD5) Previous issue date: 2012-07-30 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The resistance of aluminum and their alloys, to the corrosion phenomenon, in aqueous solutions, is a result of the oxide layer formed. However, the corrosion process in the aluminum alloy is associated with the presence a second phase of particles or the presence of chloride ions which promote the disruption of the oxide layer located producing the corrosion process. On the other hand, the term water produced is used to describe the water after the separation of the oil and gas in API separators. The volumes of produced water arrive around 5 more times to the volume of oil produced. The greatest feature of the water is the presence of numerous pollutants. Due to the increased volume of waste around the world in the current decade, the outcome and the effect of the discharge of produced water on the environment has recently become an important issue of environmental concern where numerous treatments are aimed at reducing these contaminants before disposal. Then, this study aims to investigate the electrochemical corrosion behavior of aluminum alloy 6060 in presence of water produced and the influence of organic components as well as chloride ions, by using the electrochemical techniques of linear polarization. The modification of the passive layer and the likely breakpoints were observed by atomic force microscopy (AFM). In the pit formation potential around -0.4 to -0.8 V/EAg/AgCl was observed that the diffusion of chloride ions occurs via the layer formed with the probable formation of pits. Whereas, at temperatures above 65 ?C, it was observed that the range of potential for thepit formation was -0.4 to -0.5 V/EAg/AgCl. In all reactions, the concentration of Al(OH)3 in the form of a gel was observed / A resistencia do aluminio e suas ligas a corros?o em meio aquoso ? resultado da camada de oxido formada. Entretanto, o processo corrosivo nas ligas de alum?nio esta associado a presen?a de part?culas de segunda fase ou a presen?a de ions cloreto que promovem a ruptura da camada de ?xido produzindo o processo corrosivo localizado. Por outro lado, o termo ?gua produzida ? usado para descrever a ?gua ap?s a sua separa??o do ?leo e g?s nos separadores API. Os volumes de ?gua produzida chegam em m?dia de 5 vezes o volume do ?leo produzido. A maior caracter?stica dessa ?gua ? a presen?a de in?meros contaminantes. Devido ao aumento do volume dos res?duos em todo o mundo na d?cada atual, o desfecho e do efeito da descarga de ?gua produzida sobre o meio ambiente tem recentemente tornar-se uma quest?o importante de preocupa??o ambiental onde in?meros tratamentos s?o destinados a redu??o destes contaminantes antes do descarte. Ent?o, o presente estudo visa investigar o comportamento eletroquimico de corros?o da liga de aluminio 6060 em presen?a de ?gua produzida e a influencia dos componentes organicos assim como dos ions cloreto, atrav?s do uso das tecnicas eletroqu?micas de polariza??o linear. A modifica??o da camada passiva e os prov?veis pontos de ruptura foram observados por microscopia de for?a atomica (AFM). No potencial de forma??o de pit de -0,4 a -0,8 V/EAg/AgCl observa-se que ocorre a difus?o dos ?ons cloreto pela camada formada com prov?vel forma??o de pites. Entretanto, em temperaturas superiores a 65?C observou-se que a faixa de potencial de forma??o de pit foi de -0,4 a -0,5V/ EAg/AgCl. Em todas as rea??es observou-se o aumento na concentra??o de Al(OH)3 sob a forma de um gel / 2020-01-01

Os craniofaringiomas são os tumores mais frequentes da região hipotálamohipofisária na faixa etária pediátrica. Apesar de serem histologicamente benignos, sua tendência infiltrativa e seu comportamento agressivo resultam em significante morbimortalidade. Histologicamente podem ser divididos em dois subtipos: adamantinomatosos e papilíferos. A patogênese dos craniofaringiomas é pouco compreendida. Mutações no gene CTNNB1, que codifica a proteína beta-catenina, são a única alteração molecular conhecida até o momento implicada na tumorigênese dos craniofaringiomas adamantinomatosos. Tais mutações afetam o sítio de degradação da beta-catenina, que passa a se acumular no citoplasma e no núcleo, ativando excessivamente a via de sinalização WNT, através da ligação aos fatores de transcrição da família LEF/TCF, levando a tumorigênese. Recentemente foi descoberto um novo mecanismo de determinação da linhagem celular hipofisária regulado pela beta-catenina, através do qual ela interage diretamente com o PROP1 para determinar a diferenciação celular hipofisária. De acordo com esse modelo, o complexo protéico PROP1/beta- catenina atua simultaneamente como repressor do HESX1 e ativador do PIT1, dependendo dos co-fatores associados. Pacientes com mutações germinativas inativadoras no PROP1 desenvolvem hipopituitarismo e podem apresentar aumento hipofisário com imagens de ressonância nuclear magnética (RNM) da região selar muitas vezes semelhantes àquelas dos craniofaringiomas, com hiperssinal em T1. Por outro lado, camundongos com expressão persistente do Prop1 exibem defeitos na regulação da proliferação celular hipofisária, incluindo cistos da bolsa de Rathke, hiperplasia adenomatosa e tumores, sugerindo que mutações com ganho de função no PROP1 também poderiam contribuir para a patogênese de tumores hipofisários em seres humanos. A semelhança entre as imagens de RNM dos pacientes com craniofaringiomas e daqueles com aumento hipofisário devido a mutações inativadoras no PROP1, e o fato de que camundongos transgênicos com expressão persistente do Prop1 apresentam aumento da susceptibilidade a tumores hipofisários, deram base a nossa hipótese de que uma desregulação na expressão do PROP1 em humanos poderia estar envolvida na patogênese dos craniofaringiomas adamantinomatosos. Esse trabalho teve como objetivo avaliar a presença de mutação somática no exon 3 do CTNNB1 e avaliar a expressão desse gene e do gene PROP1 em craniofaringiomas adamantinomatosos. Foram obtidas 14 amostras desse tipo de tumor por meio da ressecção terapêutica. As amostras foram submetidas à extração do RNA e posterior transcrição reversa para obtenção de cDNA. A partir do cDNA foi realizada PCR e sequenciamento do exon 3 do CTNNB1 em todas as amostras. Porém, a avaliação por PCR em tempo real foi realizada apenas em 12 amostras, devido à qualidade inadequada de 2 amostras para submissão a essa metodologia. Foram encontradas mutações missense, em heterozigose em 9 das 14 amostras, sendo 5 previamente descritas e 2 ainda não descritas em craniofaringiomas adamantinomatosos. Hiperexpressão do CTNNB1 foi encontrada em 7 amostras, sendo 5 com mutação e 2 sem mutação no CTNNB1.A hiperexpressão variou de 2,5 a 6,2 vezes maior que o pool de hipófise normal. Contudo, a expressão do PROP1 foi indetectável em todas as amostras. Concluímos que o aumento da expressão do CTNNB1 presente em 58% das amostras sugere o envolvimento também da hiperexpressão desse gene na etiopatogenia do craniofaringioma adamantinomatoso, enquanto a ausência de expressão do PROP1 afasta a participação desse gene na etiopatogenia do craniofaringioma adamantinomatoso / Craniopharyngiomas are the the commonest tumors to involve the hypothalamo-pituitary regions in childhood population. Histologically they are benign, and can be divided in two primary subtypes: the adamantinomatous and the papillary. Although histologically benign, their infiltrative tendency and aggressive behavior can result in great morbidity. The pathogenesis of craniopharyngiomas is poorly understood. To date, beta-catenin gene (CTNNB1) mutations have been identified only in the adamantinomatous subtype. These mutations affect the degradation target box of beta-catenin that accumulates in the cytoplasm and the nucleus increasing the transcriptional activity of WNT pathway through interaction with the transcription factors of LEF/TCF family, leading to tumorigenesis. Recently, an interaction between beta-catenin and PROP1 was described as a new mecanism for beta-catenindependent regulation of pituitary cell-lineage determination. According to this novel model, the PROP1/beta-catenin proteic complex would act as a binary switch to simultaneously repress the transcription factor HESX1 and to activate expression of transcription factor PIT1, depending on the associated cofactors. Patients with loss-of-function mutations in PROP1 present combined pituitary hormonal deficiency generally associated with pituitary enlargement and the magnetic resonance imaging (MRI) of the sellar region in these patients sometimes resembles that of the craniopharyngiomas, with T1 hyperintense signal. On the other hand, transgenic mice with persistent Prop1 expression exhibit defects consistent with misregulation of pituitary cell proliferation, including adenomatous hyperplasia with formation of Rathke\'s cleft cysts and tumors suggesting that misregulation of PROP1 expression in human could contribute to pathogenesis of pituitary tumors. The similarity between the MRI images of craniopharyngiomas patients and that of patients with loss-of-function mutations in PROP1, associated with the fact that transgenic mice with persistent Prop1 expression exhibit increased susceptibility to pituitary tumors gave rise to our hypothesis that a misregulation of PROP1 expression could be involved in the pathogenesis of adamantinomatous craniopharyngiomas. The aim of this study was to analyze the presence of somatic mutations in exon 3 of CTNNB1 and the expression pattern of this gene and the PROP1 gene in adamantinomatous craniopharyngiomas. Fourteen samples were obtained from therapeutic surgery and submitted to RNA extraction and reverse transcription in order to produce the cDNA. The cDNA was used as a template to CTNNB1 exon 3 PCR reaction followed by direct sequencing of all samples. However, the real-time RT-PCR analysis was realized only in 12 samples, since 2 of them had an insufficient quality for this method. Missence, heterozygous mutations were found in 9 out of 14 samples; five were previously described and 2 not yet described in adamantinomatous craniopharyngiomas. Overexpression of CTNNB1 was found in 7 samples, which them 5 with CTNNB1 mutation 2 whitout. The overexpression ranged from 2.5 to 6.2 fold more than pituitary normal pool. However, the PROP1 expression was undetectable in all the samples. We could conclude that the amount of 58% CTNNB1 overexpressed samples suggest also a role of this overexpression in the pathogenesis of adamantinomatous craniopharingiomas, while the undetectable levels of PROP1 exclude a role of this gene in the pathogenesis of adamantinomatous craniopharingiomas

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2017-02-20T13:51:05Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Dissertação CD Biblioetca Denise Moraes 2016 FINAL.pdf: 2561125 bytes, checksum: 30370d4b25d98afe00850c78927ba185 (MD5) / Made available in DSpace on 2017-02-20T13:51:05Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Dissertação CD Biblioetca Denise Moraes 2016 FINAL.pdf: 2561125 bytes, checksum: 30370d4b25d98afe00850c78927ba185 (MD5) Previous issue date: 2016-08-19 / FACEPE / O conhecimento dos parâmetros de crescimento e mortalidade em braquiúros é fundamental para a compreensão da sua dinâmica e para o manejo de suas populações. Neste sentido, foram estimados os parâmetros de crescimento e mortalidade para Cardisoma guanhumi Latreille, 1825 (guaiamum), uma espécie com elevada importância socioeconômica no nordeste brasileiro e atualmente considerada pelo Ministério do Meio Ambiente como Criticamente em Perigo de extinção. As amostragens foram efetuadas durante um ano, entre abril de 2015 a março de 2016, na margem superior do mangue do CMA/ICMBio/CEPENE na ilha de Itamaracá, Pernambuco, Brasil. Foram capturados, medidos e pesados 1078 indivíduos (572 machos e 506 fêmeas). Destes, 291 indivíduos foram marcados com microchips PIT ("Passive Integrated Transponder"), para determinação dos parâmetros de crescimento através dos incrementos de peso e tamanho e do crescimento individual. Para a análise dos dados, foram utilizados o método ELEFAN I (baseado na distribuição de frequência de largura de carapaça de 1078 indivíduos), inserido no pacote computacional Fisat II e através da função GrowthTraject (baseada em incrementos individuais de 291 indivíduos marcados com PITs, utilizando o pacote fishmethods (Ambiente de programação “R”). Os indivíduos de C.guanhumi apresentaram largura da carapaça entre 20,9 e 70,0 mm (média: 43,45 mm, desvio padrão: 8,53 mm, mediana: 44,05 mm) o peso total entre 4 e 162 g (média: 45,85 g, desvio padrão: 25,34 g, mediana: 44,0 g). Não houve diferenças significativas, em tamanho médio e peso médio, entre machos e fêmeas. Os parâmetros de crescimento estimados com a função GrowthTraject para 130 incrementos (machos e fêmeas) foram: Linf (tamanho assintótico) = 108,03 mm (largura da carapaça); K (coeficiente de crescimento) = 0,145 ano-1. A Mortalidade total (Z) da população, estimada pelo método Length – converted Catch Curve (pacote FISAT II) usando os parâmetros de crescimento do GrowthTraject foi de = 2,39 ano-1. Não ocorrem capturas comerciais nesta área fechada, portanto, este valor equivale à mortalidade total e natural (Z=M). A idade dos indivíduos capturados variou de 1,49 anos (20,9mm) a 7,02 anos (70,0mm). O recrutamento, calculado no pacote FISAT II e estimado através da presença de juvenis, mostrou-se contínuo durante todo o ano. Os métodos de análises de frequência e comprimento inseridos no pacote FISAT (Bhattacharya, ELEFAN I e Shepherd’s) não foram capazes de determinar os parâmetros de crescimento, provavelmente devido ao crescimento lento e recrutamento contínuo durante todo ano. O tamanho total da população no manguezal do CMA foi estimado em 1262 indivíduos (+- 401ind.), baseado nos dados de marcação e recaptura. / Knowledge on the growth parameters and mortality in brachyuran is fundamental to the understanding of the dynamics and management of their populations. Growth and mortality parameters were estimated for Cardisoma guanhumi Latreille, 1825 (locally known as”guaiamum”), a species with high socio-economic importance in northeastern Brazil and currently considered by the Ministry of the Environment as Critically Endangered of Extinction. The samples were taken over one year, from April 2015 to March 2016, at the upper margin of a small, isolated mangrove patch at CMA-ICMBio-CEPENE on Itamaracá Island, Pernambuco, Brazil. A total of 1078 individuals (572 males and 506 females) were captured, measured and weighed. Of these, 291 individuals were marked with PIT tags (Passive Integrated Transponder tags), as to determine the growth parameters through the analysis of the increments in weight and size and individual growth. For the data analysis, we used the ELEFAN I method (based on the frequency distribution of carapace width 1078 individuals), inserted into the computer package FISAT II and through the GrowthTraject function (based on individual increments of 291 individuals marked with PIT tags, using the fishmethods package ("R” Environment). C. guanhumi showed carapace widths between 20.9 and 70.0 mm (mean: 43,45 mm, standard deviation: 8,53 mm, median: 44,05 mm) and total weight between 4 and 162 g (mean: 45,85 g, standard deviation: 25,34 g, median: 44,0 g). There were no significant differences in average size and average weight between males and females. Growth parameters estimated with the fishmethods package, based on 130 increments (males and females), were: L∞ (asymptotic carapace width) = 108.03 mm; K (coefficient growth) = 0.145 y-1. Total mortality (Z) of the population, estimated through the Length-converted catch Curve method (FISAT II package), using the fishmethods growth parameters, was 2.39 y-1. Since this is a restricted area without regular commercial catches, so this value is roughly equivalent to the total and natural mortality (Z = M). The age of the individuals captured ranged from 1.49 years (20,9 mm) to 7.02 years (70,0 mm). Recruitment, calculated in FISAT II package and estimated by the presence of juveniles, showed to be continuous throughout the year. The methods of length-frequency analysis inserted in the FISAT package (Bhattachary of protective measures for this species.

O fósforo é um dos minerais mais abundantes no corpo além de ser essencial para muitos processos biológicos. A homeostase do fósforo depende da absorção no intestino, da excreção renal, da remodelação óssea e de hormônios como o paratormônio, calcitriol e FGF-23. Nos pacientes com doença renal crônica, a excreção urinária de fósforo está comprometida levando à hiperfosfatemia, que contribui para o aumento da morbidade e mortalidade desses pacientes. A absorção intestinal de fósforo no intestino delgado, particularmente via cotransportadores NaP-IIb e PiT-1, é pouco estudada. O objetivo desse estudo foi avaliar os efeitos de dietas com diferentes concentrações de fósforo na expressão proteica e gênica dos cotransportadores NaP-IIb e PiT-1, bem como na apoptose dos enterócitos dos diferentes segmentos do intestino delgado de animais controles e urêmicos. Estudamos setenta e seis ratos Wistar machos, inicialmente divididos em dois grupos: controles (C) e urêmicos (Nx). Cada grupo foi subdividido em outros três, de acordo com a concentração de fósforo (P) na dieta: dieta Baixa (0,2%), dieta Padrão (0,54%) e dieta Alta (0,9%). Analisamos parâmetros bioquímicos (creatinina, P, Cai, PTH e FGF-23), a expressão proteica dos cotransportadores, através de Western Blotting, ELISA e imunofluorescência, a expressão gênica por PCR em tempo real e a apoptose dos enterócitos pela técnica TUNEL. Os resultados mostraram que os níveis séricos de creatinina, P, PTH e FGF-23 foram significativamente mais elevados nos animais Nx. Nos animais C com dieta baixa em P observamos aumento da expressão proteica do cotransportador NaP-IIb em todos os segmentos do intestino enquanto, no Nx Baixo, a expressão desse cotransportador foi menor somente no jejuno. Quanto ao PiT-1, sua expressão foi menor no íleo do grupo Nx Alto comparado ao seu respectivo controle. A expressão gênica do cotransportador NaP-IIb foi menor no jejuno do Nx Alto e maior no íleo desse mesmo grupo em relação aos seus respectivos controles. A expressão gênica do PiT-1 foi maior em todos os segmentos do grupo Nx Baixo em relação aos seus controles. Detectamos uma correlação direta entre a expressão gênica do NaP-IIb no jejuno com o fósforo sérico. A expressão gênica do PiT-1 no jejuno correlacionou-se com o fósforo sérico e no duodeno e jejuno, com os níveis séricos de FGF-23. No duodeno e no jejuno, a porcentagem de enterócitos apoptóticos foi maior nos animais Nx Alto comparados aos controles, particularmente no duodeno a porcentagem dessas células também foi maior no grupo Nx Baixo e controle padrão. Já no jejuno, tanto o grupo Nx baixo quanto no Nx Padrão observamos menos células apoptóticas que nos seus respectivos controles. Quando analisamos conjuntamente todos os segmentos intestinais o grupo Nx alto apresentou mais células apoptóticas que o controle e o oposto foi observado nos grupos Nx Baixo e Nx Padrão. Em conclusão, dietas com distintas concentrações de fósforo promovem modificações na expressão proteica e gênica dos cotransportadores NaP-IIb e PiT-1 no intestino delgado que não seguem um padrão uniforme. A sobrecarga de fósforo aumenta a porcentagem de enterócitos apoptóticos nos animais urêmicos / Phosphorus is one of the most abundant minerals in the body besides being essential for many biological processes. Phosphorus homeostasis depends on absorption in the small intestine, renal excretion, bone remodeling and hormones such as parathyroid hormone, calcitriol and FGF-23. In patients with chronic kidney disease phosphorus urinary excretion is compromised leading to hyperphosphatemia, which contributes to increased morbidity and mortality of these patients. Intestinal absorption of phosphorus in the small intestine, particularly of NaP-IIb and PiT-1 cotransporters is poorly studied. The aim of this study was to evaluate the effects of diets with different concentrations of phosphorus in protein expression and gene of NaP-IIb and PiT-1 cotransporters as well as in apoptosis of enterocytes of different segments of intestine in control and uremic animals. We studied seventy-six male Wistar rats initially divided into two groups: controls (C) and uremic (Nx). Each group was subdivided into three others, according to the phosphorus concentration in the diet: Low diet (0.2% P), Standard diet (0.54% P) and High diet (0.9% P). We analyzed biochemical parameters (creatinine, P, iCa, PTH and FGF-23), protein expression of cotransporters, using Western Blot, ELISA, immunofluorescence, real-time PCR and apoptosis of enterocytes using the TUNEL technique. Results showed that serum creatinine, P, PTH and FGF-23 were significantly higher in Nx animals. In C animals with a diet low in P we observed increase in protein expression of NaP-IIb cotransporter in all segments of the intestine while in low Nx expression of this cotransporter was lower only in jejunum. As for PiT-1 expression was lower in the ileum of the high Nx group compared to their respective control. Gene expression of NaP-IIb cotransporter was lower in the jejunum of high Nx and higher in the ileum of the same group as compared to their respective controls. PiT-1 gene expression was higher in all segments of the low Nx group compared to their controls. We detected a direct correlation between the gene expression of NaP-IIb in jejunum and serum phosphorus. Gene expression of PiT-1 correlated with serum phosphorus in the jejunum, and correlated with the serum levels of FGF-23 in the jejunum and duodenum. In the duodenum and jejunum the percentage of apoptotic enterocytes was higher in high Nx animals compared to controls; particularly in the duodenum the percentage of these cells was also higher in low Nx group and standard control. In the jejunum in both low Nx group and standard Nx we observed fewer apoptotic cells than in their respective controls. When we analyze all intestinal segments together the high Nx group had more apoptotic cells than the control, and the opposite was observed in the low Nx group and standard Nx group. In conclusion, diets with different phosphorus concentrations promote changes in protein expression and gene of NaP-IIb and PiT-1 cotransporters in the small intestine that do not follow a uniform pattern. Phosphorus overhead increases the percentage of apoptotic enterocytes in uremic animals

L’embranchement Hemichordata regroupe les classes Enteropneusta et Pterobranchia. Hemichordata constitue, avec l’embranchement Echinodermata, le groupe-frère des chordés. Les entéropneustes sont des organismes vermiformes solitaires qui vivent sous ou à la surface du substrat et s’alimentent généralement par déposivorie, alors que les ptérobranches sont des organismes coloniaux filtreurs habitant dans un réseau de tubes appelé coenecium. Ce mémoire présente trois études dont le point commun est l’utilisation des hémichordés actuels pour répondre à des questions concernant l’évolution des hémichordés, des chordés, et du super-embranchement qui les regroupe, Deuterostomia. Notre première étude démontre que les fentes pharyngiennes, l’organe pré-oral cilié (POCO) et le pharynx de l’entéropneuste Protoglossus graveolens sont utilisés pour l’alimentation par filtration. Le système de filtration de P. graveolens permet la capture de particules jusqu’à 1.3 um, à un débit de 4.05 mm.s-1, pour une demande énergétique de 0.009 uW. Les similarités structurales et fonctionnelles avec le système de filtration des céphalochordés suggèrent que la filtration pharyngienne est ancestrale aux deutérostomes. Lors de notre deuxième étude, nous avons exploré l’hypothèse selon laquelle le POCO des entéropneustes, une structure ciliée pré-buccale au rôle possiblement chémorécepteur, serait homologue au « wheel organ » des céphalochordés et à l’adénohypophyse des vertébrés. Pour cela, nous avons déterminé par immunohistochimie l’expression de Pit-1, un facteur de transcription spécifique à ces deux structures, chez l’entéropneuste Saccoglossus pusillus. Pit-1 est exprimé dans des cellules sensorielles du POCO, mais aussi dans des cellules épithéliales distribuées dans le proboscis, collet et tronc. Ce patron d’expression ne permet pas de confirmer ou rejeter l’homologie du POCO et de l’adénohypophyse des vertébrés. Lors de notre troisième étude, nous avons caractérisé l’ultrastructure du coenecium des ptérobranches Cephalodiscus hodgsoni, Cephalodiscus nigrescens et Cephalodiscus densus par microscopie électronique à transmisison et à balayage. Cephalodiscus est le groupe frère de Graptolithina, un groupe qui inclut les graptolithes éteints ainsi que les ptérobranches du genre Rhabdopleura. Nous avons décrit les types de fibrilles de collagène présents, leur taille et leur organisation, ainsi que l’organisation globale du coenecium. Nous avons ainsi démontré la présence chez Cephalodiscus d’une organisation similaire au paracortex, pseudocortex et eucortex des graptolithes. La présence chez Cephalodiscus de ce type d’organisation suggère que le cortex est ancestral à la classe Pterobranchia. Ces trois études illustrent plusieurs axes importants de la recherche sur les hémichordés, qui en intégrant des données morphologiques, fonctionnelles et moléculaires permet de reconstruire certains évènements clés de l’évolution des deutérostomes. / The phylum Hemichordata comprises the classes Enteropneusta and Pterobranchia. Together with echinoderms, hemichordates are the sister-group to chordates. Enteropneusts are worm-shaped solitary deposit feeders. Pterobranchs are colonial filter feeders that live in a secreted collagenous domicile called a coenecium. In this thesis, three studies are presented. These studies are based on observations of extant hemichordates, and adress a variety of issues relating to the evolution of hemichordates, chordates, and the super-phylum to which they belong: Deuterostomia. Our first study demonstrates that the gill slits, pre-oral ciliary organ (POCO), and lining of the pharynx of the enteropneust Protoglossus graveolens are used in filter feeding. The filter-feeding system of P. graveolens enables particle capture down to 1.3 um, at a rate up to 4.05 mm.s-1, with a power consumption of 0.009 uW. Structural and functional similarities with the cephalochordate filter-feeding system suggest that pharyngeal filter-feeding is ancestral to the deuterostomes. In our second study, we address the hypothesis that the enteropneust POCO, a putative chemosensory structure located anterior to the mouth, is homologous to the cephalochordate wheel organ and vertebrate adenohypophysis. We characterized the expression pattern of the adenohypophysis-specific transcription factor Pit-1 in the adult enteropneust Saccoglossus pusillus with immunohistochemistry. Pit-1 is expressed in sensory cells of the POCO and in scattered epithelial cells of the proboscis, collar and trunk. This expression pattern does not allow to confirm or reject the homology of the POCO with the vertebrate adenohypophysis. In our third study, we characterized the ultrastructure of the coenecium of the pterobranchs Cephalodiscus hodgsoni, Cephalodiscus nigrescens and Cephalodiscus densus using transmission and scanning electron microscopy. Cephalodiscus is the sister-group to the Graptolithina, which includes the extinct graptolites and the extant pterobranch genus Rhabdopleura. We described the fibril types, size and organization, as well as the general organization of the coenecium. We demonstrated that the coenecium of Cephalodiscus shows similarities with the graptolite eucortex, paracortex and pseudocortex. The cortical-like organization of the coenecium of Cephalodiscus suggests that the cortex is ancestral to the Pterobranchia. Together, these three studies illustrate different axes of hemichordate research, and show how integrating morphological, functional and molecular data allows us toinfer key events in the evolution of deuterostomes.

The production of raw materials through mining projects is nowadays very challenging, mainly due to the rapid progress in the industrial and technological fields. On the one hand, they have to fulfill industries' requirements in their demand for materials while making a profit based on the current technologies. On the other hand, they should consider all other limitations, primarily environmental and social challenges that are confronting. The transportation system in any mining project is one of the most significant parts, especially in the technical and economic issues. It must transfer the planned volume of ore/waste that the whole stream of the mining process would not be interrupted and, it can cover the technical challenges and the costs imposed on the project. Additionally, it should be designed and selected to have the lowest environmental impact and the highest safety during the operation. Accordingly, a transportation system selection process that considers all these factors is one of the challenging issues in any mining project. Although the Truck-Shovel system is known as the conventional transportation in open-pit mines, which is preferable because of the low capital cost and high flexibility, it still imposes a high rate of operating costs, safety issues as well as environmental footprints. In-Pit Crushing and Conveying (IPCC) systems are the alternative transportation systems for the Truck-Shovel systems, in which the material is crushed inside the mine’s pit limit and transferred into the outside through conveyor belts. Although these systems are not new, they are mostly neglected as a transportation option basically due to the high capital cost and low flexibility. On the contrary, they can offer more environmentally friendly and safer working areas and a lower operating cost. According to these facts, each transportation system is preferable in a couple of technical, economic, environmental, safety, and social issues. Accordingly, in each circumstance, one or more of these systems can be used in the mining project. However, there is not yet a way or tool that investigates the transportation system selection along with the mine life that takes into account all of these factors. To fill this gap, this project aims to define a model to introduce all these elements while it is interactively connected throughout the mine life. For this and as the first step, the system dynamics modeling is defined and used to build the model for all the technical, economic, environmental, safety, and social factors. As an output of this step, software entitled “TEcESaS Indexes” is designed and produced through Venapp that makes working with the model comfortable. As the second step, a selection method based on the Analytical Hierarchy Process (AHP) is performed that the transportation system selection regarding all the mentioned factors can be made. As the output in this step, the “Sustainability Index” software programmed in the Java language is developed. Considering a hypothetical copper open-pit mine as the case study and implementing the designed software, the results show although the Truck-Shovel system should be used in the first two years of the project (2016 and 2017) in the single expert and deterministic mode, the Fully Mobile In-Pit Crushing and Conveying (FMIPCC) system shows the highest sustainability index among other transportation systems from 2018 until the end of the mine life. While in the group decision making and deterministic simulation, the Truck-Shovel system should be utilized from 2016 to 2020. Additionally, in the group decision making and stochastic mode, the FMIPCC is the selected transportation system with the highest sustainability index probability.

Tagebaumaßnahmen führen durch Entfernung des Oberbodens, Vermischung von Bodenschichten bei der Wiederverfüllung von Restlöchern und Maschineneinsatz dazu, dass auf Tagebaufolgeflächen meist extrem schwierige Standortbedingungen für Pflanzenwachstum herrschen. Dazu zählen v.a. Bodenverdichtung und Bodenversauerung durch die Verwitterung von Eisensulfiden, was Ernährungsstörungen und Belastungen mit Schwermetallen und anderen Schadstoffen zur Folge hat. Die Bodenbedingungen sind auf kleinem Raum sehr heterogen. Das ungünstige Mikroklima der vegetationsfreien Flächen verstärkt die durch Bodenverdichtung schwierige Wasserversorgung. Mykorrhiza-Pilze fehlen oft zunächst. Ziel dieser Arbeit war die Entwicklung eines möglichst wenig aufwendigen Konzeptes, um mit Hilfe geeigneter Baumarten und Bodenhilfsstoffe zumindest auf kleiner Fläche eine Begrünung von Kippen-Rohböden zu erreichen, um so Startpunkte einer Sukzession zu etablieren. Zu diesem Zweck wurden verschiedene Baumarten (Quercus robur, Acer platanoides, Sorbus aucuparia, Alnus glutinosa, Pseudostuga menziesii und Pinus sylvestris) und Bodenhilfsstoffe zur Verbesserung der physikalischen (Perlit, Wasserspeichersubstanzen: sowohl herkömmliches, petrochemisches Hydrogel als auch neuartiger Superabsorber aus pflanzlicher Stärke), chemischen (Kalk), physikalischen und chemischen (Kompost) sowie biologischen (Mykorrhiza-Impfung) Bodeneigenschaften getestet. Die Bodenhilfsstoffe wurden einzeln und miteinander kombiniert eingesetzt. Auf auch Jahre nach der Ablagerung des Haldensubstrats noch vegetationsfreien Flächen im ehemaligen Ronneburger Uranerz-Abbaugebiet wurden Freilandversuche mit knapp 1.000 Bäumen angelegt; Topfversuche im Gewächshaus dienten der Begleitung und Vertiefung. Die physikalischen und chemischen Bodeneigenschaften der Versuchsflächen wurden analysiert, die Effekte der Bodenhilfsstoffe darauf im Topfversuch überprüft. Die Entwicklung der Versuchspflanzen wurde über zwei Jahre hinweg durch regelmäßige Vitalitätsschätzungen auf einer 5-stufigen Skala, Messungen verschiedener Wuchsparameter und Analysen der Elementgehalte in den Assimilationsorganen untersucht. Die Böden wiesen eine sehr geringe Wasserspeicherkapazität und eine stark gestörte Infiltrations- und Wasserleitfähigkeit auf. Das Nährstoffpotential war mit Ausnahme sehr geringer Stickstoff-Gehalte gut. Eine sehr starke Versauerung sowie stark belastende Kupfer- und Schwefel-Gehalte waren die Haupt-Probleme. Auffällig waren hohe Magnesium-Gehalte und -Sättigungen. Die Versuchspflanzen waren entsprechend sehr stark mit Schwefel und Kupfer sowie mit Eisen, Aluminium und teilweise Mangan belastet und zeigten auffallend hohe Magnesium-Gehalte. Sie wiesen Ernährungsdefizite auf, die auf die geringe Stickstoffversorgung, Nährstoffantagonismen/Ionenkonkurrenz und Toxizität zurückgeführt werden können. Im Verlauf des Versuchs entwickelte sich auf einigen Versuchsflächen in Teilbereichen Spontanvegetation (Moose, Calamagrostis epigejos). Bodenuntersuchungen ergaben leicht höhere pH-Werte und eine entsprechend günstigere Ernährungssituation in diesen Bereichen, vermutlich aufgrund einer unregelmäßigen Flächenkalkung einige Jahre vor Versuchsbeginn. Die Entwicklung der Versuchspflanzen wurde stark von den Standortunterschieden beeinflusst. Kalk und Kompost hatten deutlich positive Effekte auf die chemischen Bodenbedingungen und die Pflanzenentwicklung. Mit Kompost war der Einfluss vor allem bei den anspruchsloseren Baumarten Eberesche, Schwarz-Erle und Wald-Kiefer oft größer. Daneben zeigten Mykorrhiza-Impfungen eine starke Wirkung, vor allem in Kombination mit Kalk und/oder Wasserspeichersubstanzen. Die positiven Wirkungssynergismen von Mykorrhiza-Impfungen und Wasserspeichersubstanzen auch ohne Kalk-Gaben waren angesichts der sehr stark sauren Standortverhältnisse bemerkenswert. Darüber hinaus hatten Wasserspeichersubstanzen wie auch Perlit keine, teilweise sogar negative Effekte durch Herabsetzung der Kalk-Wirkung in kombinierten Varianten. Zudem wiesen die mit Perlit oder Wasserspeichersubstanzen gepflanzten Bäume teils hohe Schadstoff-Belastungen auf. Die Bodeneigenschaften wurden nicht beeinflusst; die feinbodenreichen Lehmböden mit ihrer geringen Infiltrationsfähigkeit durch Zerscherung der Grobporen bei Verdichtung schienen für den Einsatz von Wasserspeichersubstanzen weniger geeignet zu sein. Das Vorhandensein von Pilz-Partnern zur Mykorrhiza-Bildung ist auf diesen schwierigen Standorten offenbar von besonderer Bedeutung, da diese Symbiose die Wasserversorgung von Pflanzen verbessern kann, außerdem ihre Ernährung fördert und die Schadstoff-Aufnahme hemmt, wie im Rahmen der vorliegenden Arbeit bestätigt werden konnte. So konnten durch kombinierte Mykorrhiza-Impfungen teilweise selbst die extremen Schwefel-Gehalte auf ein normales Maß gesenkt werden, was mit Kalkung allein nicht der Fall war. Die obligat Mykorrhiza bildenden Rot-Eichen und Wald-Kiefern, zwei eigentlich zur Tagebaurekultivierung sehr geeignete und viel verwendete Baumarten, zeigten insgesamt hohe Ausfallraten. Bei den Eichen spielte dabei die schlechte Qualität des Pflanzenmaterials, bei den Kiefern wahrscheinlich starke Trockenheit zum Zeitpunkt der Versuchsanlage im Frühjahr 2005 eine Rolle. Bei den ebenfalls obligat Mykorrhiza bildenden Douglasien war der Standort in günstigeren Bereichen, in denen sich auch Spontanvegetation entwickeln konnte und in die nachweislich Mykorrhiza-Pilze einwanderten, von großer Bedeutung. Wie die als einzige Baumart nicht mit Kupfer belastete Eberesche und die durch ihre Symbiose mit Stickstoff-fixierenden Bakterien sehr gut geeignete Erle, die von allen Baumarten die besten Ergebnisse zeigte, konnte sich auch die Douglasie auf diesen etwas besseren Standorten selbst ohne Bodenhilfsstoffe zufrieden stellend entwickeln. Angesichts des extremen Sommers 2006 kam vermutlich auch ihre Trockenheitstoleranz zum Tragen. Die Spitz-Ahorne zeigten hohe Schadstoff-Belastungen, kaum Wachstum und eine stark gestörte Wurzel-Ausformung. Die Baumart wurde zudem stark verbissen und erwies sich insgesamt als wenig geeignet. Auf Grundlage der Ergebnisse der auf den Ronneburger Tagebaufolgeflächen durchgeführten Untersuchungen wird für die Rekultivierung dieser und vergleichbarer Standorte eine Mischung aus Erlen, Ebereschen und Douglasien empfohlen, wobei die Erle auch aufgrund ihrer Standort verbessernden Eigenschaften den größten Anteil einnehmen sollte. In stärker versauerten (pH KCl < 4), frei von Bodenvegetation bleibenden Bereichen sollte einige Zeit im Vorfeld der Pflanzungen eine Kalkung mit aufgrund der Bodengehalte magnesiumfreiem Kalk erfolgen, um die Voraussetzungen für Bodenvegetation und Mykorrhiza-Pilze zu verbessern. Bei den Ebereschen und Erlen ist eine Kompost-Gabe ins Pflanzloch als Starthilfe sinnvoll. Zumindest bei den Douglasien sollten Mykorrhiza-Impfungen durchgeführt werden, die generell auf diesen Standorten empfehlenswert sind und somit auch für die Ebereschen und Erlen sehr vorteilhaft sein können. Die Dauer der Wirkung der Bodenhilfsstoffe und die über die beobachtete Zeit hinaus gehende Entwicklung der Bäume sind unter den gegebenen, sehr schwierigen Standortbedingungen mit fortschreitender Pyrit-Verwitterung und damit Bodenversauerung offen.