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- Helmholtz Institute for RNA-based Infection Research (HIRI) (7)
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The Church is mandated by Jesus Christ to continue the mission for which he was sent into the world. The mission of Christ, which consists in “bringing the good news to the poor, proclaiming liberty to captives, restoring sight to the blind, setting the downtrodden free and the proclamation of the Lord’s year of favour” , remains the fundamental basis of the missionary and evangelising vocation of the Church. She has a message to proclaim and that message is the proclamation of making the kingdom of God present in the lives of the people. Through the ages the Church has responded to this command of the Lord to evangelise, using various methods according to different situations and times. Dialogue is a conditio sine qua non in the Church’s evangelisation. By con-voking the Second Vatican Council, Pope John gave special attention to the Church’s self-knowledge, that is the knowledge of her nature and vocation as well as the realisation of the necessity of dialogue in the Church’s pursuit of Church unity and healthy relationships with non-Christian religions and bod-ies. Besides the emphases on the importance of dialogue in the Church’s exe-cution of her mission and apostolate of building up the people of God, evi-dence from the human sciences portray the indispensable and invaluable roles of dialogue and communication in a globalised world.
Elevation of intracellular cAMP in T lymphocytes, induced by agents such as IL-1α, prostaglandins or forskolin, inhibits Th1-type cytokine production but stimulates Th2-type cytokine production. The signaling pathway engaged in cAMP-mediated induction of Th2 lymphokines remains obscure and therefore my doctoral work was focused on the elucidation of cAMP pathway in primary Th lymphocytes. While forskolin treatment of EL-4 cells led both to an activation of Th2 lymphokines and inhibition of Th1 lymphokines, ectopic expression of catalytically active PKA stimulated Th2 lymphokines but failed to inhibit Th1 lymphokine expression. Thus, the PKA activity is selectively involved in the stimulation of Th2 lymphokine expression whereas other cAMP-dependent pathway(s) appears to downregulate Th1 lymphokines. By investigating different types of primary murine Th cells, it was found that active PKA enhanced IL-5 expression only in Th0 and Th2 but not in Th1 cells. This is likely due to the different levels of GATA-3 whose expression is high in Th2, moderate in Th0 and very low in Th1 cells. Ectopic expression of GATA-3 in Th1 cells induced Th2 lymphokine expression which could be further enhanced by increased cAMP levels or PKA activity. Investigations on the role of increased cAMP levels on Th2 lymphokines in D10 cells, a Th2-type cell line, led to the conclusion that elevated cAMP concentrations do not stimulate PKA but p38 activity which, through phosphorylation of GATA-3, appeared to induce IL-5 and IL-13 expression (Chen et al., 2000). While focusing on primary Th lymphocytes, it was observed that expression of the catalytic subunit α of PKA is sufficient for optimal IL-5 expression in primary Th0 cells. In addition, downregulation of IL-5 production in primary Th2 cells by the treatment with low concentrations of H-89, a PKA specific inhibitor, as well as by the ectopic expression of a negatively acting version of regulatory PKA subunit I demonstrates that active PKA plays an important role in IL-5 gene regulation. These findings using different types of primary CD4+ T lymphocytes, including Th2 cells, the one likely to represent the native IL-5 producers in vivo, demonstrates that the adenylyl cyclase/cAMP/PKA signaling pathway plays an important role in IL-5 gene expression in primary Th2 cells. Thus the importance of cAMP/PKA signaling pathway in Th2 effector function was established during this doctoral research work.
Zars and co-workers were able to localize an engram of aversive olfactory memory to the mushroom bodies of Drosophila (Zars et al., 2000). In this thesis, I followed up on this finding in two ways. Inspired by Zars et al. (2000), I first focused on the whether it would also be possible to localize memory extinction.While memory extinction is well established behaviorally, little is known about the underlying circuitry and molecular mechanisms. In extension to the findings by Zars et al (2000), I show that aversive olfactory memories remain localized to a subset of mushroom body Kenyon cells for up to 3 hours. Extinction localizes to the same set of Kenyon cells. This common localization suggests a model in which unreinforced presentations of a previously learned odorant intracellularly antagonizes the signaling cascades underlying memory formation. The second part also targets memory localization, but addresses appetitive memory. I show that memories for the same olfactory cue can be established through either sugar or electric shock reinforcement. Importantly, these memories localize to the same set of neurons within the mushroom body. Thus, the question becomes apparent how the same signal can be associated with different events. It is shown that two different monoamines are specificaly necessary for formation of either of these memories, dopamine in case of electric shock and octopamine in case of sugar memory, respectively. Taking the representation of the olfactory cue within the mushroom bodies into account, the data suggest that the two memory traces are located in the same Kenyon cells, but in separate subcellular domains, one modulated by dopamine, the other by octopamine. Taken together, this study takes two further steps in the search for the engram. (1) The result that in Drosophila olfactory learning several memories are organized within the same set of Kenyon cells is in contrast to the pessimism expressed by Lashley that is might not be possible to localize an engram. (2) Beyond localization, a possibible mechanism how several engrams about the same stimulus can be localized within the same neurons might be suggested by the models of subcellular organisation, as postulated in case of appetitive and aversive memory on the one hand and acquisition and extinction of aversive memory on the other hand.
Living apart together
(2002)
Cohesiveness between members of a social unit is a defining characteristic of animal social organization. Dispersed social organizations, where members of a social unit spend the main part of their activity period apart, have only recently been distinguished from cohesive social organizations and are still poorly understood with respect to their ecological basis and reproductive consequences. The general goal of this dissertation was to study the three components of the social system of fork-marked lemurs (Phaner furcifer), a small nocturnal primate from Madagascar living in dispersed pairs. First, I characterise their social organization, focusing on behavioural mechanisms of cohesion between pair partners. Second, through application of van Schaik's ecological model, I investigate predictions about the ecological basis of female intra-sexual avoidance, male-female social relationships and the determinants of differential female reproductive success. Finally, I analyse behavioural and genetic aspects of the mating system to test a recent hypothesis that proposes high extra-pair paternity in dispersed primate pairs resulting from constraints on male mate guarding. The study was conducted in Kirindy Forest in Madagascar between September 1998 and April 2001 during three field seasons for a total of 20 months. During more than 1400 hours of focal animal protocols, I sampled year-round data on space use, feeding ecology, time budgets, and social behaviour of all adults and three subadults of 8 families, complemented by simultaneous focal follows of both pair partners, year-round information on sleeping site use, measures on food abundance in each territory, morphological measurements, and DNA-microsatellite data for seven newly discovered polymorphic loci. Across eight social units and three breeding seasons, pairs were the prevailing grouping pattern (18 of 21 family years). Most pairs were stable for more than three mating seasons and used well defined stable territories. Although both pair partners used the same territory in a fairly similar fashion, average distance between pair partners was 100m, which was far considering that many territories measure only 200m in diameter. Pair partners spent only about 20% of activity time in less than 25m distance of each other and shared a sleeping site on average only every third day. Females were found to be dominant over their partner as well as over neighbouring males in all behavioural contexts. Most important food resources were exudates of a small number of tree species. Major food resources were distributed in small, defendable patches characterized by fast depletion and rapid renewal. In accordance with the ecological model, this led to strong within-group contest and scramble competition and weak between-group contest competition over food, as indicated by a positive dominance effect and a negative group size effect on female physical condition. Female reproductive success was determined mainly by family size. Paternity likelihood and exclusion analyses revealed that four out of seven offspring were most likely sired by an extra-pair male. Behaviour during the mating season implied that females as well as males take an active part in obtaining extra-pair copulations and that males try to guard their mates. Dispersed social organization in itself, i.e. low cohesion between pair partners, cannot explain high extra-pair paternity. I propose instead that several other factors common to most primates living in dispersed pairs constrain mate guarding and lead to high EPP. The ecological settings determine the mode of food competition and have shaped the social system of fork-marked lemurs in several ways. Intense within-group competition for food may have ultimately led to female intra-sexual avoidance and range exclusivity which represents an evolutionary precursor of pair-living. Although it remains elusive why females ultimately associated with single males, patterns of within-group contest competition for food explain why pair partners avoid each other during nocturnal activity. The limited number of food resources that is used in repetitive fashion and incomplete knowledge about the pair partners position explain why pair partners meet relatively often and why most encounters involve agonistic conflict. Rigid feeding itineraries characteristic of exudate feeders are likely to pose high costs to offspring dispersing to unfamiliar areas. Feeding ecology can, therefore, explain why parents tolerate delayed natal dispersal despite a negative effect on actual female reproductive success. In conclusion, the present study successfully applied existing socio-ecological theory to a new area of research, refined a recent evolutionary model and contributed important comparative data to our understanding of dispersed pairs in particular and primate and animal societies in general.
Originally renowned for their spectacular epigaeic raids, army ants have captured scientific attention for almost two centuries. They now belong to one of the best studied group of ants. However, most of our knowledge about army ants was derived from the study of the minority of specialized, epigaeicly active species. These species evolved probably rather recently from hypogaeic ancestors. The majority of army ant species still leads a hypogaeic life and is almost completely unknown in its entire sociobiology. It thus remained speculative, whether the assumed 'general' characteristics of army ants represent an adaptation to epigaeic activity or apply also to the majority of hypogaeic species. Based on the recent observation that the hypogaeic Asian army ant Dorylus (Dichthadia) laevigatus recruits predictably to palm oil baits, I developed and tested an oil-baiting method for the study of hypogaeic (army)ants. Prior to my study, nothing was known about the sociobiology of the assumed rare D. laevigatus. Throughout my work, I showed D. laevigatus to be very common and abundant in a wide range of habitats in West-Malaysia and on Borneo. Investigating its foraging behavior, I revealed D. laevigatus to differ from epigaeicly active species in several ways. Never demonstrated for any of the epigaeic species, D. laevigatus established stable trunk trail systems. Such a trail system contradicted the perception of army ant foraging, which was believed to be characterized by raids with constantly alternating trail directions. The trunk trail system further enabled a near omnipresence of D. laevigatus within its foraging area, which was also believed to be atypical for an army ant. Raids differed in structure and composition of participating workers from those of epigaeic species. Also, bulky food sources could be exploited over long periods of time. The foraging system of D. laevigatus resembled in several ways that of e.g. leaf-cutter and harvester ants. Likewise contrary to the assumptions, D. laevigatus had a wide food spectrum and showed only little effect on local arthropod communities, even falling itself prey to other ants. Strong aggressive behavior was observed only towards ant species with similar lifestyles, enabling me to provide the first detailed documentation of interspecific fights between two sympatric Dorylus species. Similar to foraging habits or ecological impact, nothing was known about colony size and composition, nesting habits, or worker polymorphism for D. laevigatus or any other hypogaeic Dorylus species prior to my work. By observing and eventually excavating a colony, I showed D. laevigatus to have a much smaller colony size and to lack the large sized workers of epigaeic Dorylus species. Similar to epigaeic Dorylinae, I showed D. laevigatus to have a non-phasic brood production, to emigrate rarely, and to alter its nest form along with habitat conditions. Detailed morphological and geographical descriptions give an impression of the Asian Dorylus species and are expected to aid other researchers in the difficult species identification. The genetic analysis of a male collected at a light trap demonstrated its relation to D. laevigatus. Confirming the male and queen associations, D. laevigatus is now one of five Dorylus species (out of a total of 61), for which all castes are known. In cooperation with D. Kistner, I provide a morphological and taxonomical description of nine Coleopteran beetles associated with D. laevigatus. Behavioral observations indicated the degree of their integration into the colony. The taxonomic position of the beetles further indicated that D. laevigatus emigrated from Africa to Asia, and was accompanied by the majority of associated beetles. The diversity of D. laevigatus guests, which included a number of unidentified mites, was rather low compared to that of epigaeic species. Overall, I demonstrated the developed baiting containers to effectively enable the study of hypogaeic ants. I showed several other hypogaeic ant species to be undersampled by other methods. Furthermore, the method enabled me to documented a second hypogaeic Dorylus species on Borneo. A detailed description of this species' morphology, ecology, and interactions with D. laevigatus is provided. My study indicated D. laevigatus to be an ecologically important species, able to influence soil structure and organisms of tropical regions in many ways. Relating the observed traits of D. laevigatus to epigaeicly active species, I conclude that our assumption of 'general' army ant behavior is erroneous in several aspects and needs to be changed. The oil-baiting method finally provides a tool enabling the location and study of hypogaeic (army)ant species. This opens a broad field for future studies on this cryptic but nonetheless important group of ants.
The number of males in animal groups is an essential determinant of male and female reproductive strategies. Females may benefit from living with several males, whereas males generally strive to monopolize a group of females. Due to male intrasexual competition, the sex ratio of groups of anthropoid primates is generally female-biased. Gregarious Malagasy lemurs deviate from theoretical expectations derived from sexual selection theory and from patterns found among anthropoids because they live in relatively small groups with an even or male-biased adult sex ratio and lack sexual dimorphism. The aim of this thesis was to investigate sex-specific reproductive strategies relating to the unusual group composition of redfronted lemurs (Eulemur fulvus rufus) by combining behavioral, demographic and endocrinological data. In the first of a set of four studies I investigated the applicability of non-invasive endocrine measurements for monitoring ovarian function in wild redfronted lemur females in order to evaluate the degree of estrus synchrony. Further, I tested the prediction that males living in multi-male groups rely on indirect mechanisms of intrasexual competition, such as physiological suppression of testicular function. Several possible benefits gained from living with many males have been proposed and the hypothesis that additional males improve social thermoregulation was tested in the third study. Finally, I examined the proximate determinants of the unusual sex ratio within groups, the variation in the adult sex ratio as well as possible social benefits of the high number of males for both sexes. The study was conducted in Kirindy Forest, Madagascar, between April 1999 and July 2000. I recorded >3000 hours of focal animal data on social and sexual behavior of all adult members of five groups. Additionally, >2200 fecal samples of males and females were collected for subsequent hormone analysis using enzymeimmunoassay (EIA). Further, I analyzed demographic data from seven Eulemur fulvus rufus groups collected between 1996 and 2002. The analyses of fecal estrogen and progestogen excretion in wild and captive females revealed that monitoring ovarian function is principally possible in redfronted lemurs, as demonstrated by the analysis of samples from captive females. Characterization of ovarian cycles in wild females, however, was not possible, because of a high day-to-day variability in excreted hormones. Nevertheless, the study provided reliable information on gestation and cycle length as well as endocrine changes associated with gestation. Additionally, I established a method for prenatal sex determination using maternal fecal samples collected during late gestation. The excretion pattern of androgens in samples of males revealed no differences between dominant and subordinate males, indicating that dominant males did not suppress the endocrine function of subordinate rivals. High frequencies of matings in combination with large testes size suggest that male reproductive competition relies at least partly on sperm competition. Females did not benefit from the high number of males in their groups in terms of improved thermoregulation because surplus males did not participate frequently in huddling groups with females. Analysis of the demographic data revealed that birth and mortality rates were not sex-biased and that males migrated considerably more frequently than females, providing no proximate explanation for the unusual sex ratio. Females in this study may proximately regulate group composition by synchronizing their fertile periods, which were inferred indirectly from the temporal distribution of births within groups. Both males and females benefit from the high number of co-resident males because reduced male group size seemed to be the main predictor of take-over rate, and thus, infanticide risk. The results of these studies suggest that certain life history traits (fast maturation, short inter-birth intervals) may ultimately determine the high number of males and the lack of single-male groups seen in redfronted lemurs. An accelerated male life history may facilitate joint group transfers and take-overs of male coalitions without a transitional time outside bisexual groups. Because males and females both benefit from a high number of males the conflict of interests between the sexes is considerably defused.
In my Ph.D. thesis "On the geometry and parametrization of almost invariant subspaces and observer theory" I consider the set of almost conditioned invariant subspaces of fixed dimension for a given fixed linear finite-dimensional time-invariant observable control system in state space form. Almost conditioned invariant subspaces were introduced by Willems. They generalize the concept of a conditioned invariant subspace requiring the invariance condition to hold only up to an arbitrarily small deviation in the metric of the state space. One of the goals of the theory of almost conditioned invariant subspaces was to identify the subspaces appearing as limits of sequences of conditioned invariant subspaces. An example due to {\"O}zveren, Verghese and Willsky, however, shows that the set of almost conditioned invariant subspaces is not big enough. I address this question in a joint paper with Helmke and Fuhrmann (Towards a compactification of the set of conditioned invariant subspaces, Systems and Control Letters, 48(2):101-111, 2003). Antoulas derived a description of conditioned invariant subspaces as kernels of permuted and truncated reachability matrices of controllable pairs of the appropriate size. This description was used by Helmke and Fuhrmann to construct a diffeomorphism from the set of similarity classes of certain controllable pairs onto the set of tight conditioned invariant subspaces. In my thesis I generalize this result to almost conditioned invariant subspaces describing them in terms of restricted system equivalence classes of controllable triples. Furthermore, I identify the controllable pairs appearing in the kernel representations of conditioned invariant subspaces as being induced by corestrictions of the original system to the subspace. Conditioned invariant subspaces are known to be closely related to partial observers. In fact, a tracking observer for a linear function of the state of the observed system exists if and only if the kernel of that function is conditioned invariant. In my thesis I show that the system matrices of the observers are in fact the corestrictions of the observed system to the kernels of the observed functions. They in turn are closely related to partial realizations. Exploring this connection further, I prove that the set of tracking observer parameters of fixed size, i.e. tracking observers of fixed order together with the functions they are tracking, is a smooth manifold. Furthermore, I construct a vector bundle structure for the set of conditioned invariant subspaces of fixed dimension together with their friends, i.e. the output injections making the subspaces invariant, over that manifold. Willems and Trentelman generalized the concept of a tracking observer by including derivatives of the output of the observed system in the observer equations (PID-observers). They showed that a PID-observer for a linear function of the state of the observed system exists if and only if the kernel of that function is almost conditioned invariant. In my thesis I replace PID-observers by singular systems, which has the advantage that the system matrices of the observers coincide with the matrices appearing in the kernel representations of the subspaces. In a second approach to the parametrization of conditioned invariant subspaces Hinrichsen, M{\"u}nzner and Pr{\"a}tzel-Wolters, Fuhrmann and Helmke and Ferrer, F. Puerta, X. Puerta and Zaballa derived a description of conditioned invariant subspaces in terms of images of block Toeplitz type matrices. They used this description to construct a stratification of the set of conditioned invariant subspaces of fixed dimension into smooth manifolds. These so called Brunovsky strata consist of all the subspaces with fixed restriction indices. They constructed a cell decomposition of the Brunovsky strata into so called Kronecker cells. In my thesis I show that in the tight case this cell decomposition is induced by a Bruhat decomposition of a generalized flag manifold. I identify the adherence order of the cell decomposition as being induced by the reverse Bruhat order.
This work presents the analysis, 3D modeling and interpretation of gravity and aeromagnetic data of Jordan and Middle East. The potential field data delineate the location of the major faults, basins, swells, anticlines, synclines and domes in Jordan. The surface geology of Jordan and the immediate area east of the Rift is dominated by two large basins, the Al-Jafr basin in the south and the Al-Azraq-Wadi as Sirhan basin to the northeast. These two basins strike southeast-northwest and are separated by an anticlinal axis, the Kilwah-Bayir swell. The Karak Wadi El Fayha fault system occurs along the western flank of the swell. The Swaqa fault occurs on the southwest hinge of Al-Azraq basin and the Fuluq fault occurs on its northeast hinge. In the south west of Jordan, Wadi Utm-Quwaira and Disi-Mudawara fault zones are shown clearly in the aeromagnetic and gravity maps. The previous major faults are well correlated with the structural map of Jordan published by Bender (1968). 3D modeling of gravity data in the Dead Sea basin (DSB) was used together with existing geological and geophysical information to give a complete structural picture of the basin. The 3D models of the DSB show that the internal structure of the Dead Sea basin (DSB) is controlled by longitudinal faults and the basin is developed as a full graben bounded by sub-vertical faults along its long sides. In the northern planes of the 3D model, the accumulation of Quaternary (salt and marl) and Mesozoic (pre-rift) sediments are thinner than in the central and southern planes of the model. In the northern planes, the thickness of the Quaternary sediments is about 4 km, 5 km in the southern planes and it exceeds 8 km in the central planes of the DSR. The thickness of the pre-rift sediments reaches 10-12 km in the northern and southern planes and exceeds 15 km in the central planes of the DSR. The planes of the 3D models show that the depth to the crystalline basement under the eastern shoulders of the DSR is shallower than those beneath the western shoulders. It is about 3-5 km beneath the eastern shoulders and 7-9 km under the western shoulder of the DSR. The gravity anomaly maps of residual and first derivative gravity delineate the subsurface basins of widely varying size, shape, and depth along the Rift Valley. The basins are created by the combination of the lateral motion along a right-tending step over and normal faulting along the opposite sides. Al Bakura basin occupies the upper Jordanian River valley and extends into the southern Tiberias Lake. Bet Shean basin to the south of Al Bakura basin plunges asymmetrically toward the east. The Damia basin, comprising the central Jordan Valley and Jericho areas to the north of the Dead Sea is shallow basin (~600-800m deep). The Lisan basin is the deepest basin in the Rift. The 3D gravity models indicate a maximum of ~12 km of basin fill. Three basins are found in Wadi Araba area, Gharandal, Timna (Qa'-Taba) and Aqaba (Elat) basin. The three basins become successively wider and deeper to the south. The three regional gravity long E-W profiles (225 km) from the Mediterranean Sea crossing the Rift Valley to the east to the Saudi Arabia borders, show the positive correlation between topography and free air anomaly and strong negative Bouguer anomaly under the central part of the Dead Sea Basin (DSB) and normal regional Bouguer anomaly outside of the DSB in the transform valley. Depth to the top of the bedrock in the under ground of Jordan was calculated from potential field data. The basement crops out in the south west of Jordan and becomes deeper to northwards and eastwards to be about ~ 8 km below ground surface in the Risha area.
The first goal of this study was to develop cell lines with a stable expression of bio-fluorescent topo II and topo I. This was successfully achieved using a bicistronic vector system. Control experiments showed that proteins of expected size were expressed, and that GFP-tagged topos I, IIa, and IIb were active in the cells and fully integrated in the endogenous pools of the enzymes. These cell-lines provided a novel tool for investigating the cell biology of human DNA topoisomerases. Our most important finding was, that both types of mammalian topoisomerases are entirely mobile proteins that are in continuous and rapid flux between all compartments of the nucleus and between the cytososl and the chromosomes of mitotic cells. This was particularly surprising with regard to topo II, which is considered to be a structural component of the nuclear matrix and the chromosome scaffold. We must conclude that if this was the case, then these architectural structures appear to be much more dynamic than believed until now. In this context it should also be mentioned, that the alignment of topo II with the central axes of the chromosome arms, which has until now been considered a hall-mark of the enzyme’s association with the chromosomal scaffold, is not seen in vivo and can be demonstrated to be to some extent an artefact of immunohistochemistry. Furthermore, we show that the two isoforms of topo II (a and b) have a different localisation during mitotic cell division, supporting the general concept that topo II functions at mitosis are exclusively assigned to the a-form, whereas at interphase the two isoenzymes work in concert. Despite unrestricted mobility within the entire nuclear space, topoisomerases I and II impose as mostly nucleolar proteins. We show that this is due to the fact that in the nucleoli they are moving slower than in the nucleoplasm. The decreased nucleolar mobility cannot be due to DNA-interactions, because compounds that fix topoisomerases to the DNA deplete them from the nucleoli. Interestingly, the subnucleolar distribution of topoisomerases I and II was complementary. The type II enzyme filled the entire nucleolar space, but excluded the fibrial centers, whereas topo I accumulated at the fibrial centers, an allocation directed by the enzyme’s N-terminus. During mitosis, it also mediates association with the nucleolar organising regions of the acrocentric chromosomes. Thus, topo I stays associated with the rDNA during the entire cell-cycle and consistently colocalizes there with RNA-polymerase I. Finally, we show that certain cancer drugs believed to act by stabilising covalent catalytic DNA-intermediates of topoisomerases, do indeed immobilize the enzymes in living cells. Interestingly, these drugs do not target topoisomerases in the nucleoli but only in the nucleoplasm.
Transforming-Growth-Factor-beta1 (TGF-b1) is a multifunctional cytokine that regulates cell growth and differentiation in many types of cells. TGF-b1 is especially known to exert a variety of regulatory functions in the immune system, such as T cell differentiation and T cell function. Signal transduction of TGF-b1 is mediated by phosphorylation of receptorassociated Smad proteins (R-Smads). R-Smads are phosphorylated by the activated type I receptor, which is itself phosphorylated by the high affinity type II receptor upon ligand binding. The phosphorylated R-Smads then associate with Co-Smads. Heterooligomers of R- and Co-Smads translocate into the nucleus where they regulate transcription of target genes in concert with other transcription factors such as CBP/p300 or AP-1. Recent findings suggest that the pleiotropic effects of TGF-b1 are conferred by crosstalks to other signal transduction pathways such as the MAP-kinases or the STAT-pathway. Here we describe the effect of long-term exposure to TGF-b1 on the effector function of differentially stimulated primary murine splenocytes and purified primary murine CD8+ cytotoxic T cells. Long-term exposure to TGF-b1 results in non-responsiveness to TGF-b1- induced Smad2 phosphorylation. This is seen either by no phosphorylation or sustained phosphorylation of Smad2. Furthermore, we observed a strong correlation between sustained Smad2 phosphorylation and resistance to TGF-b1 mediated growth inhibition. In contrast, splenocyte cultures strongly growth inhibited by TGF-b1 showed no Smad2 phosphorylation. Lytic activity of these cultures, however, was found to be suppressed regardless of proliferation properties and Smad2 phosphorylation pattern. We also describe that a functional MEK-1 pathway is a prerequisite for rendering murine splenocytes unresponsive to TGF-b1 mediated growth inhibition, and that inhibition of the MEK-1 cascade alters the Smad2 phosphorylation pattern. In addition, we show that resistance to TGF-b1 mediated growth inhibition correlates with the activation of the JNK pathway. However, the resistant phenotype was found unable to be reverted upon administration of exogeneous IFNg and/or aCD28 antibody. In human or mouse T cell lines, however, the described correlation between the type of stimulation and TGF-b growth resistance or growth sensitivity is not present. Thus, this correlation is specific for primary T cells. We also cloned a chimeric dominantnegative TGF-b receptor which is coupled to a suicide gene, in order to render T cells resistant to TGF-b mediated effects.These findings shed light on how TGF-b1 mediates its immunosuppressive role, and may help to gain knowledge of averting these TGF-b1 effects in the course of tumor therapy.