L’hyperplasie et l’hypertrophie contribuent à l'augmentation de la masse de muscle lisse bronchique observée dans le souffle. Les cellules musculaires lisses (CML) présentent deux phénotypes; prolifératif ou contractile. Le serum response factor (SRF), un facteur de transcription impliqué dans l’activation de nombreux gènes, contribuerait à cette modulation phénotypique. Notamment, lorsqu'associé au cofacteur Elk-1, un phénotype prolifératif serait observé, alors qu'en présence de la myocardine (MYOCD) il y aurait induction d'un profil contractile. Récemment, il a été démontré que SRF est surexprimé dans les voies périphériques chez les chevaux atteints du souffle suite à une exposition antigénique. Cette étude vise à caractériser l'expression protéique et génique de SRF, Elk-1 et MYOCD dans les CML des voies respiratoires centrales et périphériques chez des chevaux atteints du souffle et des chevaux contrôles. L'évaluation de l’expression protéique de SRF, Elk-1 et MYOCD s’est effectuée par immunodétection sur des tissus provenant de biopsies thoracoscopiques ou endobronchiques, et ce, avant, à 1 et 30 jours du défi antigénique. L'expression génique a été étudiée par qPCR sur du muscle lisse disséqué de la trachée, et des bronches, ainsi que sur des voies respiratoires intermédiaires et périphériques. Les expressions génique et protéique de MYOCD sont augmentées uniquement dans les voies périphériques. L’expression génique de SRF et Elk-1 varient dans les voies centrales alors que le taux de protéines demeure stable. En conclusion, SRF et MYOCD pourraient être impliquées dans l’hypertrophie des voies respiratoires périphériques dans le souffle alors que l’hyperplasie ne semble pas être activée par Elk-1. / Airway smooth muscle (ASM) cells hyperplasia and hypertrophy contribute to the increased airway smooth muscle mass present in heaves. ASM cells express either a synthetic proliferative or a contractile phenotype. Serum response factor (SRF) is a transcription factor that has been shown to regulate myocyte differentiation in vitro in vascular and intestinal smooth muscles. When SRF is associated with Elk-1, it promotes ASM proliferation while myocardin (MYOCD) promotes the expression of contractile elements. Recently, SRF was shown to be overexpressed in the peripheral airways of heaves affected horses following an antigenic challenge. The objective of this study was to characterize the protein and gene expression of SRF, Elk-1 and MYOCD in ASM cells from central and peripheral airways of heaves affected horses and controls. Protein expression of Elk-1 and MYOCD was evaluated using immunohistochemistry while immunofluorescence was used for SRF detection in pulmonary peripheral and endobronchial biopsies before and at 1 and 30 days of antigenic exposure. Gene expression was investigated in ASM cells dissected from trachea and bronchi as well as from intermediate and peripheral airways using qPCR. MYOCD gene and protein expressions are increased only in peripheral airways. SRF and Elk-1 gene expression varied in the central airways while the positive cell percentage remains stable. In conclusion, the pulmonary peripheral airways hypertrophy observed in heaves seems to implicate SRF and MYOCD while the hyperplasia doesn’t seem to be activated by Elk-1.

Le muscle squelettique adulte est un tissu possédant la capacité fondamentale d'adapter sa taille à la demande fonctionnelle : il peut s'atrophier ou s'hypertrophier en réponse à une variation de la charge mécanique qui lui est appliquée. A l'heure actuelle, les facteurs impliqués dans la plasticité musculaire demeurent méconnus. D'une part, grâce à différents modèles d'atrophie musculaire, nous démontrons que le facteur de transcription Srf joue le rôle de médiateur de la mécano-transduction par la voie actine/Mrtfs/Srf. L'arrêt de l'activité mécanique provoque une accumulation nucléaire d'actine monomérique, une délocalisation de Mrtf-A, coactivateur de Srf, et une diminution de l'activité de Srf, se traduisant notamment par une baisse de la transcription Srf-dépendante. Les gènes cibles de Srf comptant un grand nombre de protéines sarcomériques, telles que l'α-actine squelettique, la réduction de leur expression pourrait participer à l'atrophie musculaire. De plus, nos travaux suggèrent que la diminution de l'activité de Srf pourrait influencer l'organisation du réseau mitochondrial et le flux autophagique par des mécanismes qui restent à élucider. D'autre part, en tirant parti d'un modèle d'invalidation conditionnelle et inductible de Srf dans les cellules satellites, nous montrons que le phénomène d'hypertrophie compensatoire requiert l'expression de Srf par les cellules satellites. L'absence de Srf n'altère ni la prolifération ni l'entrée en différenciation des myoblastes, néanmoins elle provoque un défaut de fusion des myoblastes aux fibres au cours de l'hypertrophie induite par surcharge. Ainsi, nos travaux démontrent que Srf est un acteur majeur de la plasticité musculaire, à la fois en tant que médiateur de la mécano-transduction par la voie actine/Mrtfs/Srf et par son implication dans la fusion des cellules satellites aux fibres musculaires, nécessaire à l'hypertrophie compensatoire.

Il a été démontré que les chevaux atteints du souffle présentent une augmentation de la masse de muscle lisse entourant les voies respiratoires comparativement à des chevaux sains (Herszberg, Ramos-Barbon et al. 2006, Leclere, Lavoie-Lamoureux et al. 2011). L’augmentation de la masse de muscle lisse ainsi observée résulte d’une hyperplasie, et possiblement, d’une hypertrophie des myocytes. Les traitements usuels du souffle ne sont que partiellement efficaces à diminuer cette augmentation. L’objectif de cette étude était d’explorer les mécanismes moléculaires impliqués dans ces changements affectant la cellule musculaire lisse dans la pathologie du souffle chez le cheval. Pour ce faire, nous avons examiné les effets d’une exposition antigénique sur l’expression du «Serum Response Factor» (SRF) dans le muscle lisse bronchique. Le SRF est un facteur de transcription localisé dans le noyau de la cellule musculaire lisse et régulant l’expression génique de celle-ci en favorisant un phénotype prolifératif ou contractile. Les résultats démontrent qu’avant exposition antigénique, les pourcentages de cellules exprimant le SRF sont faibles. Une augmentation significative du pourcentage de myocytes exprimant le SRF survient suite à une stimulation antigénique chez les chevaux atteints de souffle alors qu’aucune augmentation n’est observée chez les chevaux contrôles. Ces résultats suggèrent que le SRF pourrait contribuer au remodelage du muscle lisse péribronchique dans la pathologie du souffle. / Previous studies have shown that heaves-affected horses, when compared to age-matched control horses, have an increased smooth muscle mass surrounding their airways (Herszberg, Ramos-Barbon et al. 2006, Leclere, Lavoie-Lamoureux et al. 2011). Also, it is shown that hyperplasia, and possibly hypertrophy, contribute to this finding. Current therapies are only partially effective at reversing this finding. The goal of this study was to explore the molecular pathways involved in airway smooth muscle remodelling during heaves. We studied the Serum Response Factor (SRF), a nuclear transcriptional factor that controls gene expression in the smooth muscle cell and favour either a proliferative or a contractile phenotype. The results show that before antigenic exposition, the percentage of cells expressing SRF is low. A significant increase in the percentage of SRF-expressing myocytes occurs after antigenic stimulation in heaves-affected horses whereas no increase is observed in control subjects. These results suggest that the transcription factor SRF may contribute to airway smooth muscle remodeling in heaves-affected horses.

The cultivation of fast growing tree species on arable land is deemed to be one of the most promising alternatives amongst the approaches to the cultivation of renewable resources currently available. As the site factors influencing the growth of poplar and willow have not yet been sufficiently identified and quantified, it is not possible to provide reliable national yield expectations as a function of the prevailing soil and climate conditions at a particular site on the basis of the data currently available. The main objective of this study, therefore, was to develop a supraregional site-based yield model for the assessment of yield expectations for poplar and willow grown on arable land over short rotations. In order to achieve this goal, a total of 38 research sites were established across as many regions of Germany as possible. The clones selected for the experiment were the poplar clones Max 1, Hybride 275 (H 275) and AF 2, and the willow clones Inger and Tordis. Each site was also characterised, on the basis of climate and soil data. From the site parameters recorded, variables were defined for modelling purposes. The climate variables comprised temperature (T), precipitation (N) and aridity index (TI) derived from total or average values from selected periods during the year and during the vegetation period. Within the variables the months were indexed as numbers (January = 1), periods were separated by comma. Parameters of the German soil appraisal such as the condition grade (ZS), the soil quality index value (BZ) and the arable land quality index value (AZ) were used as soil variables. The selected texture parameters were the proportions of sand (S[%]), silt (U[%]) and clay (T[%]). Variables of the pore space parameters were the available field moisture capacity (nFK), the air capacity (LK) and the dry bulk density (TRD). Both the texture and the pore space variables referred to a soil depth of 0 to 60 cm. To increase the precision of the results, site clusters were derived, differentiated by main soil type, and the variables were aggregated. The dGZ, measured in odt ha-1 a-1,at the end of the first three-year rotation was chosen as the dependent variable. The main results of the study are outlined in the following: Negative correlations between dGZ and temperatures in the vegetation period, especially in the months July to September, were observed on the sandy and loamy soils. Precipitation variables always had a positive effect on the growth of poplar and willow across all clones and site clusters. Taking into consideration all experimental sites, precipitation in the period from May to July was most important. The parameters of the German soil appraisal revealed a moderate correlation with growth for all sites. From the appraisal of all sites and of the individual site clusters it became apparent that U[%] represents the most important texture variable for poplar and willow growth. Considered for all sites simultaneously, the nFK had the greatest significance for growth across all of the parameters examined. The site-based yield models were all univariate and often comprised aggregated variables. Under consideration of all sites, the model predictors for Max 1, Inger and Tordis were (nFK * TI5.7) and (nFK * N5.7) for AF 2. Each model was calculated with the inverse or sigmoid approach and revealed an R²korr between 0.45 and 0.64 with a RMSE of an average of 2.0 odt ha-1 a-1. The division into site clusters improved the accuracy of the models considerably. In the sandy site cluster, the models exhibited an R²korr of 0.77 to 0.97 and an RMSE of 0.95 to 1.36 odt ha-1 a-1. These comprised (U[%]* N6.7) for Max 1 and F 2, (S[%]/ TI6.7) for H 275 and (S[%]* T7.8) for Inger and Tordis. For the silty site cluster, significant models could only be determined for the clones Max 1 and Tordis. The model predictor for Max 1 was (BZ * TI4.5) and for Tordis solely (BZ). The calculated R²korr values were 0.84 and 0.95 with a corresponding RMSE of 0.22 and 0.62 odt ha-1 a-1, respectively. For the loamy soils the models for Max 1 and Inger comprised the variable nFK, for AF 2 the variables (nFK * N5.6). R²korr varied between 0.86 and 0.98 with RMSE between 0.56 and 1.21 odt ha-1 a-1.

For 2010 the European Union has a goal to increase the share of renewable energyware to 12%. For 2020 the goal is 20%. One of visions to reach this is to raise the share of solid biofuels. Politicans all over ideologycal spectrum collectively propagate, that farming and waste management sector industry has developed ways to produce solid recovered fuels with reliable qualities, which are used successfully regarding economic and environmental aspects. One of prerequisites to reach this will be to achieve the standardization of solid biofuels. This leads to a need for flexible market, which is attainable only by common understanding concerning terminology, sampling and test methods. European energetically non-intensive industries and settlements are looking for ecologically friendly alternative fuels in order to save fossil fuels. And by doing so, enforce the development of fitting technical norms to facilitate the meeting of offer and demand for the product. Pressed biofuels like pellets are today playing the key role for the small house market as well as for large scale coal substitution in local power plants. The thesis is a part of scientific contemplation MSM 06/2/4a solved by professors S.Kužel and L.Kolář and is trying to answer the question: ``Is combustion of solid biofuels ecologically friendly?{\crqq}

The cultivation of fast growing tree species on arable land is deemed to be one of the most promising alternatives amongst the approaches to the cultivation of renewable resources currently available. As the site factors influencing the growth of poplar and willow have not yet been sufficiently identified and quantified, it is not possible to provide reliable national yield expectations as a function of the prevailing soil and climate conditions at a particular site on the basis of the data currently available. The main objective of this study, therefore, was to develop a supraregional site-based yield model for the assessment of yield expectations for poplar and willow grown on arable land over short rotations. In order to achieve this goal, a total of 38 research sites were established across as many regions of Germany as possible. The clones selected for the experiment were the poplar clones Max 1, Hybride 275 (H 275) and AF 2, and the willow clones Inger and Tordis. Each site was also characterised, on the basis of climate and soil data. From the site parameters recorded, variables were defined for modelling purposes. The climate variables comprised temperature (T), precipitation (N) and aridity index (TI) derived from total or average values from selected periods during the year and during the vegetation period. Within the variables the months were indexed as numbers (January = 1), periods were separated by comma. Parameters of the German soil appraisal such as the condition grade (ZS), the soil quality index value (BZ) and the arable land quality index value (AZ) were used as soil variables. The selected texture parameters were the proportions of sand (S[%]), silt (U[%]) and clay (T[%]). Variables of the pore space parameters were the available field moisture capacity (nFK), the air capacity (LK) and the dry bulk density (TRD). Both the texture and the pore space variables referred to a soil depth of 0 to 60 cm. To increase the precision of the results, site clusters were derived, differentiated by main soil type, and the variables were aggregated. The dGZ, measured in odt ha-1 a-1,at the end of the first three-year rotation was chosen as the dependent variable. The main results of the study are outlined in the following: Negative correlations between dGZ and temperatures in the vegetation period, especially in the months July to September, were observed on the sandy and loamy soils. Precipitation variables always had a positive effect on the growth of poplar and willow across all clones and site clusters. Taking into consideration all experimental sites, precipitation in the period from May to July was most important. The parameters of the German soil appraisal revealed a moderate correlation with growth for all sites. From the appraisal of all sites and of the individual site clusters it became apparent that U[%] represents the most important texture variable for poplar and willow growth. Considered for all sites simultaneously, the nFK had the greatest significance for growth across all of the parameters examined. The site-based yield models were all univariate and often comprised aggregated variables. Under consideration of all sites, the model predictors for Max 1, Inger and Tordis were (nFK * TI5.7) and (nFK * N5.7) for AF 2. Each model was calculated with the inverse or sigmoid approach and revealed an R²korr between 0.45 and 0.64 with a RMSE of an average of 2.0 odt ha-1 a-1. The division into site clusters improved the accuracy of the models considerably. In the sandy site cluster, the models exhibited an R²korr of 0.77 to 0.97 and an RMSE of 0.95 to 1.36 odt ha-1 a-1. These comprised (U[%]* N6.7) for Max 1 and F 2, (S[%]/ TI6.7) for H 275 and (S[%]* T7.8) for Inger and Tordis. For the silty site cluster, significant models could only be determined for the clones Max 1 and Tordis. The model predictor for Max 1 was (BZ * TI4.5) and for Tordis solely (BZ). The calculated R²korr values were 0.84 and 0.95 with a corresponding RMSE of 0.22 and 0.62 odt ha-1 a-1, respectively. For the loamy soils the models for Max 1 and Inger comprised the variable nFK, for AF 2 the variables (nFK * N5.6). R²korr varied between 0.86 and 0.98 with RMSE between 0.56 and 1.21 odt ha-1 a-1.:Inhaltsverzeichnis 1 Einleitung 1 1.1 Problemstellung 1 1.2 Zielstellung 3 2 Hintergrund 4 2.1 Energieziele Deutschlands 4 2.1.1 Flächen- und Nutzungskonkurrenz mit der Forstwirtschaft 4 2.1.2 Flächen- und Nutzungskonkurrenz mit der Landwirtschaft 5 2.2 Anbau schnellwachsender Baumarten 6 2.2.1 Anbau im Kurzumtrieb 7 2.3 Verwendete Baumarten in Kurzumtriebsplantagen 11 2.3.1 Pappel 11 2.3.2 Weide 13 2.3.3 Anmerkung Züchtung 14 2.4 Leistung von Kurzumtriebsplantagen mit Pappel und Weide in zwei- bis dreijähriger Umtriebs-zeit 15 2.5 Ertragsdynamik und leistungsbeeinflussende Faktoren von Kurzumtriebsplantagen 17 2.5.1 Baumart und Klon 18 2.5.2 Umtriebszeit und Bestandesdichte 19 2.5.3 Standortanforderungen 21 2.5.3.1 Reichliche Wasserversorgung 21 2.5.3.2 Lockere Böden 22 2.5.3.3 Ausreichende Mineralstoffzufuhr 23 2.5.3.4 Wachstumsangepasste Bodenreaktion 23 2.5.3.5 Wärme 23 2.5.3.6 Bodenschätzungskennwerte 24 2.5.3.7 Textur 24 2.5.3.8 Standorte der Pappel 24 2.6 Waldwachstumsmodellierung mit Schwerpunkt in Kurzumtriebsplantagen 25 2.6.1 Empirische Modelle 26 2.6.1.1 Pappelmodell nach ALI (2009) 26 2.6.1.2 Pappel- und Weidenmodell nach AYLOTT et al. (2008) 27 2.6.1.3 Leistungsbeeinflussende Standorteigenschaften für Pappel und Weide nach BERGANTE et al. (2010) 28 2.6.2 Mechanistische Modelle 29 3 Material und Methoden 33 3.1 Flächenanlage und Versuchsdesign 33 3.1.1 Flächenvorbereitung und Pflegemaßnahmen 37 3.1.2 Prüfglieder 39 3.1.2.1 Pappelklon Max 1 39 3.1.2.2 Pappelklon H 275 39 3.1.2.3 Pappelklon AF 2 40 3.1.2.4 Weidenklon Inger 41 3.1.2.5 Weidenklon Tordis 41 3.2 Klimatische Standortcharakterisierung 42 3.2.1 Temperatur 44 3.2.2 Niederschlag 46 3.3 Bodenkundliche Standortcharakterisierung 47 3.3.1 Profilansprache und allgemeine Standortinformationen 48 3.3.2 Durchführung der Probennahme 48 3.3.3 Bodenphysikalische Untersuchungen 49 3.3.4 Grundwassereinfluss 49 3.3.5 Bodenchemische Untersuchungen 50 3.3.6 Bodenschätzungskennwerte 50 3.3.7 Zusammenfassende Betrachtung der bodenkundlichen Charakteristika 51 3.4 Bestandesaufnahmen und Ernte 55 3.4.1 Anwuchs- und Überlebensrate 55 3.4.2 Anzahl an Höhentrieben 55 3.4.3 Höhe 55 3.4.4 Durchmesser 55 3.4.5 Ertragsbestimmung 55 3.4.5.1 Biomassefunktionen 56 3.4.5.2 Systematische Teilbeernteung der Kernparzelle 56 3.5 Identifizierung von leistungsbeeinflussenden Standorteigenschaften und Entwicklung standortbasierter Ertragsmodelle 57 3.5.1 Variablenwahl und -bildung 58 3.5.1.1 Klima 58 3.5.1.2 Bodenschätzung 59 3.5.1.3 Textur 59 3.5.1.4 Porenraum 59 3.5.1.5 Aggregierte Variablen 60 3.5.1.6 Weitere Einflussgrößen 60 3.5.1.7 Abhängige Variable 61 3.5.2 Standortcluster 61 3.6 Datenbasis der standortbasierten Ertragsmodellierung 61 3.7 Ertragsteigerungfaktor 63 3.8 EDV und Statistik 65 3.8.1 Varianzanalyse 65 3.8.2 Korrelationsanalyse 66 3.8.3 Regressionsanalyse 66 3.8.3.1 Nicht-lineare Regression 68 3.8.3.2 Evaluierungsgrößen 69 4 Ergebnisse 71 4.1 Waldwachstumskundliche Ergebnisse 71 4.1.1 Anwuchs- und Überlebensrate 71 4.1.2 Anzahl an Höhentrieben 72 4.1.3 Übersicht Bestandesdimensionen und Wuchsleistung 73 4.1.4 Höhe 75 4.1.5 Durchmesser 77 4.1.6 Durchschnittlicher Gesamtzuwachs 81 4.1.7 Biomassefunktionen 84 4.1.7.1 Wahl der unabhängigen Variablen 84 4.1.7.2 Einfluss der Gattung und des Klons 85 4.1.7.3 Einfluss der Bestandesmittelhöhe 85 4.1.7.4 Allgemeingültige Biomassefunktionen 87 4.2 Leistungsbeeinflussende Standorteigenschaften 88 4.2.1 Temperatur 88 4.2.2 Niederschlag 90 4.2.3 Trockenheitsindex 92 4.2.4 Bodenschätzungskennwerte 94 4.2.5 Textur 96 4.2.6 Porenraum 99 4.2.7 Korrelation zwischen unabhängigen Variablen 102 4.2.8 Aggregierte Variablen 102 4.2.9 Weitere Einflussgrößen 105 4.2.9.1 Vornutzung 105 4.2.9.2 Bodentyp 106 4.2.9.3 Grundwasser 106 4.3 Standortbasierte Ertragsmodellierung 107 4.3.1 Standortcluster ALL 107 4.3.2 Standortcluster S 110 4.3.3 Standortcluster U 113 4.3.4 Standortcluster L 115 4.4 Ertragssteigerungsfaktoren und Ertragssteigerung in Folgerotationen 118 5 Diskussion 122 5.1 Material und Methodik 122 5.1.1 Versuchsflächen 122 5.1.2 Modellierung und Variablenbildung 123 5.2 Waldwachstumskundliche Ergebnisse 124 5.2.1 Anwuchs- und Überlebensrate 124 5.2.2 Anzahl an Höhentrieben 125 5.2.3 Höhe 125 5.2.4 Durchmesser 126 5.2.5 Durchschnittlicher Gesamtzuwachs 126 5.2.6 Biomassefunktionen 128 5.3 Leistungsbeinflussende Standorteigenschaften 129 5.3.1 Temperatur 129 5.3.2 Niederschlag 130 5.3.3 Trockenheitsindex 132 5.3.4 Bodenschätzungskennwerte 132 5.3.5 Textur 133 5.3.6 Porenraum 134 5.3.7 Aggregierte Variablen 135 5.3.8 Weitere Einflussgrößen 136 5.4 Standortbasierte Ertragsmodellierung 136 5.4.1 Standortcluster ALL 137 5.4.2 Standortcluster S 138 5.4.3 Standortcluster U 138 5.4.4 Standortcluster L 138 5.4.5 Vergleich mit anderen Modellen 138 5.5 Ertragssteigerungsfaktoren und Ertragsteigerung in Folgerotationen 140 6 Schlussfolgerungen und Ausblick 141 7 Zusammenfassung 144 8 Literatur 156 9 Anhang 176

Cachexie est syndrome multifactoriel associé a une maladie chronique ou incurable et caractérisé par un importante fonte musculaire. En fait, l’exercice physique améliore la qualité de vie et la survie des patients cancéreux. Dans un modèle animal de cachexie dû au cancer, nous avons démontré que la course sur roue contre la cachexie par la libération du flux d’autophagie. Les effets de l’exercice pléitropique incluent la modification des facteurs circulants en faveur d’un environnement anti-inflammatoire et l’activation des voies de mécanotransduction dans les cellules musculaires. Notre but est d'évaluer si la mécanotransduction est suffisante à elle seule pour mimer l’exercice en présence de facteurs pro-cachétiques d'origine tumorale. Le facteur de réponse au sérum (SRF est un facteur de transcription important pour homéostasie musculaire, qui est activé avec son co-facteur MRTF par la mécanotransduction de façon dépendant à la polymérisation de l'actin. Nous utilisons une culture mixte de C2C12 myotubes et myoblastes traitée avec un milieu condition par des C26 (CM) en absence ou en présence d'étirements cycliques qui miment la stimulation mécanique. Nous avons démontré in vitro que le CM a un effet négatif sur les cultures de cellules musculaires, sur l'atrophie des myotubes, sur le recrutement et la fusion des myoblastes, et que ces effets sont contrecarrés par l'étirement mécanique. Nous avons montré que le CM inhibe l'activité transcriptionnelle de SRF-MRTF, alors que l'étirement mécanique rétablit cette activité ; en plus, des expériences de perte de fonction ont démontré que SRF est nécessaire pour médier les effets bénéfiques des stimulations mécaniques sur les cellules musculaires. Au moins une part des effets de l’exercice observés étaient médiés par la balance des facteurs pro- et anti-myogeniques de la superfamille TGF-b. Nous proposons que les effets positifs de l’exercice sur les patients cancéreux et les souris pourraient être spécifiquement dûs a la réponse mécanique des fibres musculaires affectant la sécrétion des myokines. / Cachexia is a multifactorial syndrome associated to chronic or acute disease (cancer, HIV,…) and characterized by severe muscle wasting. In fact, exercise training improves quality of life and survival of cancer patients. In an animal model of cancer cachexia we demonstrated that wheel running counteracts cachexia by releasing the autophagic flux. Exercise pleitropic effects include the alteration of circulating factors in favour of an anti-inflammatory environment and the activation of mechanotransduction pathways in muscle cells. Our goal is to assess whether mechanostransduciton per se is sufficient to elicit exercise effects in the presence of pro-cachectic factors of tumor origin. Serum response factor (SRF) is a transcription factor of pivotal importance for muscle homeostasis, which is activated with its co-factor MRTF by mechanostranduction in a way dependent on actin polymerisation. We use mixed cultures of C2C12 myotubes and myoblasts treated with C26 conditioned medium (CM) in the absence or presence of cyclic stretch to mimic the mechanical stimulation occurring upon exercise. In vitro we showed that CM had a negative effect on muscle cell cultures, both in terms of myotube atrophy and of myoblast recruitment and fusion, and that these effects were counteracted by cyclic stretch. We showed that CM repressed SRF-MRTF transcriptional activity, while mechanical stretch rescued their transcriptional activity; in addition, loss of function experiments demonstrated that SRF was necessary to mediate the beneficial effects of mechanical stimulation on muscle cells. At least part of the observed effects was mediated by the balance of pro- and anti-myogenic factor of the TGF-b superfamily. We propose that the positive effects of exercise on cancer patients and mice may be specifically due to a mechanical response of muscle fibers affecting the secretion of myokines. / Cachexia è una sindrome di multifactorial associata a malattia cronica o acuta (cancro, HIV.) e caratterizzò da spreco di muscolo severo. Infatti, l'esercizio addestrando migliora qualità della vita e sopravvivenza di pazienti di cancro. In un modello animale di cachexia di cancro noi dimostrammo quello ruota correndo contrattacca cachexia rilasciando il flusso di autophagic. Gli effetti di pleitropic di esercizio includono la modifica di fattori circolanti nel favore di un ambiente antinfiammatorio e l'attivazione di sentieri di mechanotransduction in celle di muscolo. La nostra meta è stimare se mechanostransduciton per se è sufficiente per suscitare effetti di esercizio nella presenza di pro-cachectic fattori di origine di tumore. Il fattore (SRF) di risposta di siero è un fattore di trascrizione dell'importanza importantissima per omeostasi di muscolo che è attivato col suo co-coefficiente MRTF da mechanostranduction in un modo dipendente su polymerisation di actin. Noi usiamo le culture mescolate del myotubes di C2C12 e myoblasts trattate con C26 condizionarono mezzo (Cm) nell'assenza o presenza di stiramento ciclico a mimico la stimolazione meccanica che accade su esercizio. In vitro noi mostrammo che il Cm aveva un effetto negativo su culture di cella di muscolo, ambo nelle condizioni di atrofia di myotube e di assunzione di myoblast e fusione, e che questi effetti furono contrattaccati da stiramento ciclico. Noi mostrammo che il Cm represse l'attività di transcriptional di SRF-MRTF, mentre lo stiramento meccanico liberò la loro attività di transcriptional; in somma, perdita di esperimenti di funzione dimostrò, che SRF era necessario per interporre gli effetti benefici di stimolazione meccanica su celle di muscolo. Almeno parte degli effetti osservati fu interposta dall'equilibrio di pro - ed anti-myogenic fattore del TGF. il superfamily. Noi proponiamo che gli effetti positivi di esercizio su pazienti di cancro e topi specificamente possono essere a causa di una risposta meccanica di fibre di muscolo che colpiscono l'occultamento di myokines.

Transkriptionelle Netzwerke (Transkriptionsfaktoren, epigenetische Modulatoren und spezifische Zielgene) stellen die unterste Ebene der zellinternen Signalübertragung dar. Eingebettet in verschiedene stimulusabhängige Signalwege bedienen sich ihre Komponenten genetischer und epigenetischer Mechanismen, um Zielgene transkrip-tionell zu regulieren und Veränderungen der Chromatinstruktur hervorzurufen. In der vorliegenden Arbeit wurde die hierarchische Organisation und Zusam-mensetzung des MEK-ERK-abhängig gesteuerten transkriptionellen Netzwerks und seine Veränderung im Zuge der HRAS-vermittelten onkogenen Transformation von HA1-Zellen untersucht. Viele Arbeiten haben sich bereits eingehend mit der Charak-terisierung einzelner Komponenten und Zielgene beschäftigt (Wagner et al. 2005, Reddy et al. 2002, Sun et al. 2006, Kapitel 1). Im Unterschied zu den zitierten Studien wurde in der vorliegenden Arbeit ein umfassendes Protokoll zur genomweiten De-chiffrierung transkriptioneller Netzwerke unter Kombination von experimentellen und bioinformatischen Methoden entwickelt und durchgeführt. Die Analyse ge-nomweiter Expressionsprofile un- und U0126-behandelter immortaler und HRASV12-transformierter humaner Nierenepithelzellen (HA1EB, HA1ER) erlaubte die Identifi-zierung von 138 auf- und 103 abregulierten genspezifischen IDs der RAS-ERK-abhängig gesteuerten Signalkaskade. Regulierte Transkriptionsfaktoren wurden i-dentifiziert und im Westernblot, sowie zum Teil mittels Durchflusszytometrie und RT-PCR validiert und nachfolgend transienten siRNA-Experimenten unterzogen. Für die Transkriptionsfaktoren ELK3, SRF und den hierarisch darunter liegenden Faktor FRA1 wurden Expressionsprofile der spezifischen siRNA-vermittelten Hemmung in beiden Zelllinien erstell und mit bioinformatischen Methoden (TRAP, GSEA-GO) a-nalysiert um direkte und indirekte sowie gemeinsame Zielgene zu ermitteln. Zusätz-lich wurde der Effekt auf phänotypischer Ebene (Softagar, MTT) überprüft. In der vorliegenden Arbeit ließ sich keine direkte Hierarchie der drei Transkrip-tionsfaktoren SRF, FRA1 und ELK3 bestätigen. Allerdings konnte zum ersten Mal eine gemeinsam regulierte Gruppe von Genen identifiziert werden, die darauf schließen lässt, dass die drei Transkriptionsfaktoren sowohl in HA1EB, als auch in HA1ER Teile eines gemeinsam regulierenden Netzwerks sind. Aus den Proliferationsexperimenten wurde zudem bestätigt, dass jeder Transkriptionsfaktor individuell eine essentielle Rolle bei der Promotion maligner Eigenschaften spielt. Für alle drei Transkriptionsfak-toren konnte eine RAS-abhängige starke Verschiebung der spezifisch angesteuerten Gene nachgewiesen werden. Diese Verschiebung wurde mittels TRAP und GSEA auch für alternative Regulatoren der spezifischen Zielgene festgestellt. Die nähere Analyse der FRA1-abhängigen Zielgene führte zu neuen Erkenntnis-sen zur Umordnung des Transkriptoms im Zuge der onkogenen Transformation. Die FRA1-spezifischen Zielgene in HA1EB und HA1ER weisen unterschiedliche Funktio-nalitäten auf. So wurden in HA1EB viele Gene identifiziert, die im Rahmen der Im-munantwort eine Rolle spielen und in HA1ER nicht reguliert werden. In den RAS-transformierten HA1ER konnten dagegen Gene identifiziert werden, die in der Tu-morprogression eine Rolle spielen (FRA1, STAT3, MTA1, TCFL5). Die Verifizierungen mittels qPCR und ChIP bestätigten 5 der 38 möglichen FRA1-Zielgene. Von diesen, FRA1, AEBP1, FRA1, TCFL5, NPAS2 und YWHAZ ist lediglich FRA1 bereits als FRA1-Zielgen beschrieben. Die Funktionen der neu identifizierten RAS-abhängigen FRA1-Zielgene untermauerten bereits bekannte Funktionen der FRA1-vermittelten Transkription (Differenzierung, Proliferation, zirkadiane Rhythmen, Apoptose) und erweitern sie um verschiedene Aspekte wie Metabolismus und Rückkopplungen in die Signaltransduktion, die noch nicht für die RAS-abhängige FRA1-vermittelte Transktiption beschrieben worden sind. Dazu gehören unter anderem Interaktionen mit TGFbeta, WNT, JAK/STAT und JNK. Daneben sind in den HA1ER eine Vielzahl von Regulatoren des RHO-Signalwegs identifiziert worden, was für FRA1 auf bisher unbekannte Interaktionen mit RAC/RHO-Signalwegen schließen lässt. / Transcriptional networks represent the final level of internal signal transmission. They are embedded in different signalling pathways and use genetic as well as epi-genetic mechanisms to regulate their according target genes. During oncogenic trans-formation they are undergoing massive rearrangements in composition, regulation and interaction. This leads to radical changes in the transcriptome and drives the on-cogenic phenotype of the according cells. My thesis employs the composition of the MEK-ERK-dependent transcriptional net-work and its alteration during the HRAS-oncogene-mediated transformation in HA1-cells. By commencing from already known components: SRF, Ternary Complex Fac-tors (TCF: SAP1, SAP2/ELK3, ELK1) and members of the AP1-complex (JUN, FOS-proteins) I analyzed the alteration in expression of secondary targets and their inter-action as well as their relation to the superior factors. Therefore I compared genome wide expression profiles (Affymetrix, HG-U133A) of immortal HA1EB and HRASV12-oncogene-transformed HA1ER-cells with and without U0126-induced MEK/ERK-inhibition and extracted several MEK/ERK-dependent transcription factors. Among them where FRA1 and ELK3, two transcription factors already known to be involved in oncogenesis and proliferation associated processes. ELK3 needs SRF as crucial binding partner to function. Therefor I also included SRF into the subsequent analysis. The three transcription factors function in different time-dependent hierarchy states so we supposed a putative hierachical network be-tween them. I established transient knockdown cells deriving from HA1EB and HA1ER for all three transcription factors and generated further expression profiles from them. Additionally I verified the importance of these transcription factors on survival and proliferation via MTT and Softagar experiments. Using different statis-tically and bioinformatical methods (GSEA, TRAP) in collaboration with the Max-Planck-Institute for molecular Genetics Berlin, several direct and indirect targets of these transcription factors were predicted. These were partially overlapping in all transcription factors. Also, in comparison of the immortal and the transformed cell line, a shift of functionalities and composition of the different target gene populations and collaborating factors could be detected for all three transcription factors. It was found that in HA1EB FRA1 seems more likely to regulate immunresponsive genes as well as genes associated with the cytoskeleton and nucleus organisation whereas in HA1ER FRA1 regulates a large group of transcription- and signalling-associated genes. Additionally it could be shown that in both cell lines FRA1 regulates genes in-volved in epigenetic processes as well as circadian rhythms which are known to be important aspects in oncogenic transformation. I verified 37 different putative target genes of FRA1 using qRT-PCR (Taqman) and partially also ChIP-analysis. Of these 37genes, 5 were fully validated as directly regu-lated targets of FRA1: FRA1, AEBP1, YWHAZ, NPAS2 and TCFL5. They imply functionalities connected to proliferation and differentiation (AEBP1, FRA1, TCFL5) as well as apoptosis (YWHAZ) cell cycle control and circadian rhythm (NPAS2, AEBP1), feedbacks into the signalling (YWHAZ, AEBP1) and metabolism (NPAS2, AEBP1). Summarised the work of this thesis contributes to the decipherment of the direct and indirect targets of the according transcription factors and strengthens the argument of a general and massive shift of the transcriptional network during oncogenic trans-formation of cells. The importance of all three transcription factors on the survival of genes could be proved via proliferation assays. Additionally the functionality of their according targets could be integrated into processes connected to oncogenic trans-formation.

蛋白激酶 CK2 是一種具有多種功能的絲胺酸/蘇胺酸蛋白質激酶，CK2 作用的受質眾多且廣泛表現在哺乳類動物細胞中，對於細胞週期的發展、轉錄作用以及抗細胞凋亡等機制扮演非常重要的角色。在神經系統中，CK2 已知可以保護神經細胞以抵抗外來的傷害，但是其分子層面的機制目前尚未釐清。本篇論文的研究重點在於探討 CK2 保護作用可能參與的細胞分子機制。血清反應因子 SRF 是一種哺乳類動物細胞的轉錄因子，調控基因的轉錄作用來促進細胞的存活。Mcl-1 是抗細胞凋亡家族 Bcl-xL 家族蛋白成員之一，可以促進細胞的存活能力。先前研究指出，SRF 會受到 CK2 的磷酸化作用而增加本身的 DNA 結合能力。在其他研究也指出，Mcl-1 會受到 SRF 的調控。在本篇論文的第一部份，著重於 Mcl-1 的表現是否會受到 CK2 調控 SRF 的路徑所影響，實驗結果顯示，轉染野生型 CK2α 質體 DNA 可以增加海馬迴 CA1 腦區的 SRF 磷酸化，而轉染不活化的突變型 CK2αΑ156 質體 DNA 則會減少 SRF 的磷酸化。更進一步，轉染野生型 CK2α 會增加 Mcl-1 的 mRNA 及蛋白質表現，轉染突變型 CK2αΑ156 則減少 Mcl-1 的表現。此外，轉染突變型 SRF99A 也會減少 Mcl-1 的 mRNA 及蛋白質表現；而且在共同轉染實驗中，SRF99A 會拮抗野生型 CK2α 對促進的 Mcl-1 蛋白質表現的作用。 另一方面，DARPP-32 是一個在新紋狀體神經細胞中具有調控多巴胺訊息效力的訊息傳遞分子。先前研究指出，DARPP-32 具有抗細胞凋亡的功能，且發現在 DARPP-32 Ser102 氨基酸會受 CK2 的磷酸化作用。因此，本篇論文的第二部份主要是探討 CK2 的抗細胞凋亡能力是否是透過磷酸化 DARPP-32 來調控。實驗結果顯示，轉染野生型 CK2α 可以增加紋狀體 DARPP-32 的磷酸化，而轉染不活化的突變型 CK2αΑ156 則會減少 DARPP-32 的磷酸化。此外，轉染 CK2α 的小干擾 RNA (siRNA) 可以抑制內生性的 CK2 表現，同時也會減少 DARPP-32 的磷酸化以及抗細胞凋亡蛋白, Bcl-xL 的表現。綜合這些實驗結果，CK2α可以分別透過 SRF 或 DARPP-32 調控的訊息傳遞來促進 Mcl-1 或 Bcl-xL 的表現進而調控神經系統的抗細胞凋亡機制。 / Protein kinase CK2 is a multifunctional serine/threonine protein kinase with many protein substrares and is ubiquitously expressed in mammalian cells to play an important role in cell cycle progression, transcription, and anti-apoptosis. In the nervous system, CK2 is shown to protect neurons against injury, but the cellular mechanisms are not well studies. In the present studies, we investigate which cellular mechanism might involve in the CK2 protection effects. The serum response factor (SRF) is a mammalian transcription factor which mediates some gene transcriptions relevent to promote the cell survival. The Myeloid cell leukemin 1 (Mcl-1) is one of the anti-apoptotic Bcl-2 family members and is involved in promoting cell viability. Previous studied have revealed that the SRF phosphorylation by CK2 can enhance its DNA-binding activity. The regulation of Mcl-1 by SRF has also been reported in other studies. In the first part of the present studies, we investigate whether the Mcl-1 expression is regulated by CK2 through SRF mediated pathway. The results from wildtype CK2α plasmid DNA transfection revealed that the phosphorylated SRF were increased in hippocampus CA1 region, whereas transfection of the catalytically inactive CK2αA156 mutant plasmid DNA decreased phosphorylated SRF. Further, wildtype CK2α increased, whereas CK2αA156 mutant decreased the mRNA and protein levels of Mcl-1. Moreover, transfection of the mutant SRF99A also decreased the mRNA and protein levels of Mcl-1. Furthermore, the mutant SRF99A antagonized the upregulatory effects of wildtype CK2α on Mcl-1 protein level in the co-transfection experiments. In the other side, DARPP-32 (dopamine- and cAMP-regulated phosphoprotein of Mr 32 kDa) is a signal transduction molecule that regulates the efficacy of dopamine signaling in neostriatal neurons. Previous studies have revealed that DARPP-32 might involve in the anti-apoptosis and its Ser102 residue is phosphorylated by CK2. Therefore, in the second part of this study, we investigate whether one of the anti-apoptotic effects of CK2 is through DARPP-32 phosphorylation by CK2 in the present study. The results revealed that the phosphorylated DARPP-32 is increased in stratum by wildtype CK2α transfection and decreased by catalytically inactive CK2αA156 mutant transfection. Further, transfection of CK2α siRNA can inhibit endogenous CK2 expression and also decrease phosphorylation of DARPP-32 as well as the anti-apoptotic protein, Bcl-xL. These results together suggest that CK2α-mediated anti-apoptotic effects are partially through SRF mediated or DARPP-32 mediated signaling to regulate Mcl-1 or Bcl-xL expression, respectively.

The interest in renewables for energy has increased in the last 2-3 decades because of the negative environmental impact caused by the burning of fossil fuels, the raising prices of traditional fuels, the dependence on foreign oil, and the decrease in fossil fuels resources. Biomass energy represents one of the most promising alternatives. Many studies worldwide were devoted to investigate growth and yield of short rotation forestry plantations for energetic use and several empirical and process-based models were developed to predict the potential production of biomass. The current work was concentrated on modelling site productivity (potential of biomass production) of specific poplar clones planted on arable Saxon land under different stocking densities. Empirical data collected from several experimental areas were used. Site productivity has been predicted depending on stand age and site variables using a two-step model. In step one age and site variables were used to model stand dominant height and in step two the constructed dominant height was involved with stocking density to predict stand oven dried biomass. Depending on data availability the model was parameterized for four different groups of poplar clones: Androscoggin (clone Androscoggin), Matrix (Matrix and hybrid 275), Max (Max 1 …Max 5) and Münden (clone Münden). Both stand dominant height and stand dry biomass were modelled for ages 2 – 9 years for clone groups: Matrix and Max and for ages 2 – 7 years for clone groups: Androscoggin and Münden. The model has been tested and validated using several statistical and graphical methods. The relative bias (ē %) in the dominant height estimates ranged between 0.5 % > ē % > - 0.5 % in all clone groups and had a maximum bias of 10.41 % in stand biomass estimates. Model accuracy (mx %) in the dominant height estimates ranged between 12.25 and 17.56 % and between 8.05 and 27.32 % in stand biomass estimates. Two different scenarios were presented to show the potential of biomass that can be produced from poplar plantations on arable and former fallow Saxon lands at different stocking densities. ArcGIS has been used to visualize model application results. In order to produce a mean annual increment ≥ 8 [dry t/ha/a] from poplar plantations (Max group) for more than 50 % of arable or former fallow lands in the first rotation at least 9 years are required under stocking density of 4000 stems/ha and 7 years for both stocking densities 8333 and 10,000 stems/ha. / Die Nachfrage nach Holz für energetische Zwecke nimmt in Deutschland und ganz Europa zu. Um diesen Bedarf künftig besser befriedigen zu können, müssen verstärkt Ressourcen aus verschiedenen Quellen wie z. B. Holz aus Niederwäldern oder Durchforstungsreserven im Hochwald mobilisiert und ergänzend Holz in Kurzumtriebsflächen produziert werden (Guericke, M. 2006). Ziel dieser Arbeit war es, das Ertragspotential von Pappelklonen in Kurzumtriebsplantagen unterschiedlicher Baumdichte auf sächsischen Ackerflächen zu untersuchen. Hierzu wurden die potentiellen Erträge anhand empirischer, auf verschiedenen Versuchsflächen erhobener Daten modelliert. Zur Schätzung des Ertragspotentials wurde ein zweistufiges Modell entwickelt: Im ersten Schritt erfolgte die Modellierung der Oberhöhe eines Bestandes (ho, m) in Abhängigkeit von Bestandesalter und Standortfaktoren unter Verwendung einer multiplen linearen Regressionsanalyse, dabei wurden Bestimmtheitsmaße (R²) von 0,975 bis 0,989 erreicht. In einem zweiten Schritt lässt sich dann der Biomassevorrat [tatro/ha/a] mittels nichtlinearer Regressionsanalyse durch die Bestandesoberhöhe schätzen. Das Bestimmtheitsmaß von R² ≥ 0,933 weist auch hier auf eine hohe Anpassungsgüte hin. Die Modellparametrisierung erfolgte für folgende vier Gruppen von Pappelklonen: • Max-Gruppe: Klone Max 1, Max 2, Max 3, Max 4 und Max 5, Altersbereich 2 – 9 Jahre, Baumdichten von 1150 – 13000 Stämmen/ha; • Matrix-Gruppe: Klon Matrix und Hybride 275, Altersbereich 2 – 9 Jahre, Baumdichte 1550 Stämme/ha; • Androscoggin-Gruppe: Klon Androscoggin, Altersbereich 2 – 7 Jahre, Baumdichte 1550 Stämme/ha; und • Münden-Gruppe: Klon Münden, Altersbereich 2 – 7 Jahre, Baumdichte 1550 Stämme/ha. Die Güte des Modells wurde mit Hilfe verschiedener statistischer Verfahren überprüft. Bei der Validierung anhand des Datensatzes, welcher für die Modellkonstruktion Verwendung fand, zeigte das Modell eine Verzerrung bzw. einen Bias von 0,5 % > ē % > - 0,5 % bei der Bestandesoberhöhenschätzung und einen maximalen Bias von 10,41 % bei der Schätzung der Bestandesbiomasse. Die Treffgenauigkeit (mx %) des Modells hingegen variierte zwischen 12,25 % und 17,56 % bzw. 8,05 und 27,32 % (bei Schätzung der Bestandesoberhöhe bzw. der Bestandesbiomasse). Zudem wies das Modell keinen systematischen Fehler zwischen den geschätzten und den realen Werten auf. Bei der Validierung mit einem unabhängigen Datensatz betrug die Treffgenauigkeit (mx %) für die Schätzung der Bestandsoberhöhe und des Biomassevorrates 15,72 bzw. 26,68 %. Um das Ertragspotenzial von Pappelplantagen für die gesamte sächsische Ackerfläche bzw. die gesamte ehemalige Stilllegungsfläche zu bestimmen, wurden die zu Schätzung erforderlichen Standortvariablen auf Gemeindebasis kalkuliert, mittels ArcGIS dargestellt sowie Simulationsrechungen für verschiedene Bestandsdichten vorgenommen und ebenfalls visualisiert. Den Ergebnissen der Simulationsrechnungen zufolge wäre bei einer Stammzahl von 4000 N/ha eine Rotationslänge von 9 Jahren, bei 8333 bis 10.000 N/ha von 7 Jahren erforderlich, um einen durchschnittlichen Gesamtzuwachs (dGz) von ≥ 8 [tatro/ha/a] auf mehr als 50 % der sächsischen Ackerflächen bzw. ehemaligen Stilllegungsflächen in erster Rotation zu erreichen. Würde die gesamte ehemalige sächsische Stilllegungsfläche mit einer Baumdichte von 10.000 Stämmen/ha bepflanzt werden, könnten Pappelplantagen im Alter 9 einen dGz von 520.000 [tatro/a] (entsprechend 250.000 Kubikmeter Diesel) erreichen. Bei Bestockung aller sächsischen Ackerflächen würde sich der Ertrag auf bis zu 9.087.000 [tatro/a] (entsprechend 4.367.000 Kubikmeter Diesel) erhöhen.