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When azobenzene-modified photosensitive polymer films are irradiated with light interference patterns, topographic variations in the film develop that follow the electric field vector distribution resulting in the formation of surface relief grating (SRG). The exact correspondence of the electric field vector orientation in interference pattern in relation to the presence of local topographic minima or maxima of SRG is in general difficult to determine. In my thesis, we have established a systematic procedure to accomplish the correlation between different interference patterns and the topography of SRG. For this, we devise a new setup combining an atomic force microscope and a two-beam interferometer (IIAFM). With this set-up, it is possible to track the topography change in-situ, while at the same time changing polarization and phase of the impinging interference pattern. To validate our results, we have compared two photosensitive materials named in short as PAZO and trimer. This is the first time that an absolute correspondence between the local distribution of electric field vectors of interference pattern and the local topography of the relief grating could be established exhaustively. In addition, using our IIAFM we found that for a certain polarization combination of two orthogonally polarized interfering beams namely SP (↕, ↔) interference pattern, the topography forms SRG with only half the period of the interference patterns. Exploiting this phenomenon we are able to fabricate surface relief structures below diffraction limit with characteristic features measuring only 140 nm, by using far field optics with a wavelength of 491 nm. We have also probed for the stresses induced during the polymer mass transport by placing an ultra-thin gold film on top (5–30 nm). During irradiation, the metal film not only deforms along with the SRG formation, but ruptures in regular and complex manner. The morphology of the cracks differs strongly depending on the electric field distribution in the interference pattern even when the magnitude and the kinetic of the strain are kept constant. This implies a complex local distribution of the opto-mechanical stress along the topography grating. The neutron reflectivity measurements of the metal/polymer interface indicate the penetration of metal layer within the polymer resulting in the formation of bonding layer that confirms the transduction of light induced stresses in the polymer layer to a metal film.
Poly(A) Polymerase 1 (PAPS1) influences organ size and pathogen response in Arabidopsis thaliana
(2014)
Polyadenylation of pre-mRNAs is critical for efficient nuclear export, stability, and translation of the mature mRNAs, and thus for gene expression. The bulk of pre-mRNAs are processed by canonical nuclear poly(A) polymerase (PAPS). Both vertebrate and higher-plant genomes encode more than one isoform of this enzyme, and these are coexpressed in different tissues. However, in neither case is it known whether the isoforms fulfill different functions or polyadenylate distinct subsets of pre-mRNAs. This thesis shows that the three canonical nuclear PAPS isoforms in Arabidopsis are functionally specialized owing to their evolutionarily divergent C-terminal domains. A moderate loss-of-function mutant in PAPS1 leads to increase in floral organ size, whereas leaf size is reduced. A strong loss-of-function mutation causes a male gametophytic defect, whereas a weak allele leads to reduced leaf growth. By contrast, plants lacking both PAPS2 and PAPS4 function are viable with wild-type leaf growth. Polyadenylation of SMALL AUXIN UP RNA (SAUR) mRNAs depends specifically on PAPS1 function. The resulting reduction in SAUR activity in paps1 mutants contributes to their reduced leaf growth, providing a causal link between polyadenylation of specific pre-mRNAs by a particular PAPS isoform and plant growth. Additionally, opposite effects of PAPS1 on leaf and flower growth reflect the different identities of these organs. The overgrowth of paps1 mutant petals is due to increased recruitment of founder cells into early organ primordia whereas the reduced leaf size is due to an ectopic pathogen response. This constitutive immune response leads to increased resistance to the biotrophic oomycete Hyaloperonospora arabidopsidis and reflects activation of the salicylic acid-independent signalling pathway downstream of ENHANCED DISEASE SUSCEPTIBILITY1 (EDS1)/PHYTOALEXIN DEFICIENT4 (PAD4). Immune responses are accompanied by intracellular redox changes. Consistent with this, the redox-status of the chloroplast is altered in paps1-1 mutants. The molecular effects of the paps1-1 mutation were analysed using an RNA sequencing approach that distinguishes between long- and short tailed mRNA. The results shown here suggest the existence of an additional layer of regulation in plants and possibly vertebrate gene expression, whereby the relative activities of canonical nuclear PAPS isoforms control de novo synthesized poly(A) tail length and hence expression of specific subsets of mRNAs.
The contractile vacuole (CV) is an osmoregulatory organelle found exclusively in algae and protists. In addition to expelling excessive water out of the cell, it also expels ions and other metabolites and thereby contributes to the cell's metabolic homeostasis. The interest in the CV reaches beyond its immediate cellular roles. The CV's function is tightly related to basic cellular processes such as membrane dynamics and vesicle budding and fusion; several physiological processes in animals, such as synaptic neurotransmission and blood filtration in the kidney, are related to the CV's function; and several pathogens, such as the causative agents of sleeping sickness, possess CVs, which may serve as pharmacological targets. The green alga Chlamydomonas reinhardtii has two CVs. They are the smallest known CVs in nature, and they remain relatively untouched in the CV-related literature. Many genes that have been shown to be related to the CV in other organisms have close homologues in C. reinhardtii. We attempted to silence some of these genes and observe the effect on the CV. One of our genes, VMP1, caused striking, severe phenotypes when silenced. Cells exhibited defective cytokinesis and aberrant morphologies. The CV, incidentally, remained unscathed. In addition, mutant cells showed some evidence of disrupted autophagy. Several important regulators of the cell cycle as well as autophagy were found to be underexpressed in the mutant. Lipidomic analysis revealed many meaningful changes between wild-type and mutant cells, reinforcing the compromised-autophagy observation. VMP1 is a singular protein, with homologues in numerous eukaryotic organisms (aside from fungi), but usually with no relatives in each particular genome. Since its first characterization in 2002 it has been associated with several cellular processes and functions, namely autophagy, programmed cell-death, secretion, cell adhesion, and organelle biogenesis. It has been implicated in several human diseases: pancreatitis, diabetes, and several types of cancer. Our results reiterate some of the observations in VMP1's six reported homologues, but, importantly, show for the first time an involvement of this protein in cell division. The mechanisms underlying this involvement in Chlamydomonas, as well as other key aspects, such as VMP1's subcellular localization and interaction partners, still await elucidation.
Mars is one of the best candidates among planetary bodies for supporting life. The presence of water in the form of ice and atmospheric vapour together with the availability of biogenic elements and energy are indicators of the possibility of hosting life as we know it. The occurrence of permanently frozen ground – permafrost, is a common phenomenon on Mars and it shows multiple morphological analogies with terrestrial permafrost. Despite the extreme inhospitable conditions, highly diverse microbial communities inhabit terrestrial permafrost in large numbers. Among these are methanogenic archaea, which are anaerobic chemotrophic microorganisms that meet many of the metabolic and physiological requirements for survival on the martian subsurface. Moreover, methanogens from Siberian permafrost are extremely resistant against different types of physiological stresses as well as simulated martian thermo-physical and subsurface conditions, making them promising model organisms for potential life on Mars. The main aims of this investigation are to assess the survival of methanogenic archaea under Mars conditions, focusing on methanogens from Siberian permafrost, and to characterize their biosignatures by means of Raman spectroscopy, a powerful technology for microbial identification that will be used in the ExoMars mission. For this purpose, methanogens from Siberian permafrost and non-permafrost habitats were subjected to simulated martian desiccation by exposure to an ultra-low subfreezing temperature (-80ºC) and to Mars regolith (S-MRS and P-MRS) and atmospheric analogues. They were also exposed to different concentrations of perchlorate, a strong oxidant found in martian soils. Moreover, the biosignatures of methanogens were characterized at the single-cell level using confocal Raman microspectroscopy (CRM). The results showed survival and methane production in all methanogenic strains under simulated martian desiccation. After exposure to subfreezing temperatures, Siberian permafrost strains had a faster metabolic recovery, whereas the membranes of non-permafrost methanogens remained intact to a greater extent. The strain Methanosarcina soligelidi SMA-21 from Siberian permafrost showed significantly higher methane production rates than all other strains after the exposure to martian soil and atmospheric analogues, and all strains survived the presence of perchlorate at the concentration on Mars. Furthermore, CRM analyses revealed remarkable differences in the overall chemical composition of permafrost and non-permafrost strains of methanogens, regardless of their phylogenetic relationship. The convergence of the chemical composition in non-sister permafrost strains may be the consequence of adaptations to the environment, and could explain their greater resistance compared to the non-permafrost strains. As part of this study, Raman spectroscopy was evaluated as an analytical technique for remote detection of methanogens embedded in a mineral matrix. This thesis contributes to the understanding of the survival limits of methanogenic archaea under simulated martian conditions to further assess the hypothetical existence of life similar to methanogens on the martian subsurface. In addition, the overall chemical composition of methanogens was characterized for the first time by means of confocal Raman microspectroscopy, with potential implications for astrobiological research.
In Chapter 1 of the dissertation, the role of social networks is analyzed as an important determinant in the search behavior of the unemployed. Based on the hypothesis that the unemployed generate information on vacancies through their social network, search theory predicts that individuals with large social networks should experience an increased productivity of informal search, and reduce their search in formal channels. Due to the higher productivity of search, unemployed with a larger network are also expected to have a higher reservation wage than unemployed with a small network. The model-theoretic predictions are tested and confirmed empirically. It is found that the search behavior of unemployed is significantly affected by the presence of social contacts, with larger networks implying a stronger substitution away from formal search channels towards informal channels. The substitution is particularly pronounced for passive formal search methods, i.e., search methods that generate rather non-specific types of job offer information at low relative cost. We also find small but significant positive effects of an increase of the network size on the reservation wage. These results have important implications on the analysis of the job search monitoring or counseling measures that are usually targeted at formal search only. Chapter 2 of the dissertation addresses the labor market effects of vacancy information during the early stages of unemployment. The outcomes considered are the speed of exit from unemployment, the effects on the quality of employment and the short-and medium-term effects on active labor market program (ALMP) participation. It is found that vacancy information significantly increases the speed of entry into employment; at the same time the probability to participate in ALMP is significantly reduced. Whereas the long-term reduction in the ALMP arises in consequence of the earlier exit from unemployment, we also observe a short-run decrease for some labor market groups which suggest that caseworker use high and low intensity activation measures interchangeably which is clearly questionable from an efficiency point of view. For unemployed who find a job through vacancy information we observe a small negative effect on the weekly number of hours worked. In Chapter 3, the long-term effects of participation in ALMP are assessed for unemployed youth under 25 years of age. Complementary to the analysis in Chapter 2, the effects of participation in time- and cost-intensive measures of active labor market policies are examined. In particular we study the effects of job creation schemes, wage subsidies, short-and long-term training measures and measures to promote the participation in vocational training. The outcome variables of interest are the probability to be in regular employment, and participation in further education during the 60 months following program entry. The analysis shows that all programs, except job creation schemes have positive and long-term effects on the employment probability of youth. In the short-run only short-term training measures generate positive effects, as long-term training programs and wage subsidies exhibit significant locking-in'' effects. Measures to promote vocational training are found to increase the probability of attending education and training significantly, whereas all other programs have either no or a negative effect on training participation. Effect heterogeneity with respect to the pre-treatment level education shows that young people with higher pre-treatment educational levels benefit more from participation most programs. However, for longer-term wage subsidies we also find strong positive effects for young people with low initial education levels. The relative benefit of training measures is higher in West than in East Germany. In the evaluation studies of Chapters 2 and 3 semi-parametric balancing methods of Propensity Score Matching (PSM) and Inverse Probability Weighting (IPW) are used to eliminate the effects of counfounding factors that influence both the treatment participation as well as the outcome variable of interest, and to establish a causal relation between program participation and outcome differences. While PSM and IPW are intuitive and methodologically attractive as they do not require parametric assumptions, the practical implementation may become quite challenging due to their sensitivity to various data features. Given the importance of these methods in the evaluation literature, and the vast number of recent methodological contributions in this field, Chapter 4 aims to reduce the knowledge gap between the methodological and applied literature by summarizing new findings of the empirical and statistical literature and practical guidelines for future applied research. In contrast to previous publications this study does not only focus on the estimation of causal effects, but stresses that the balancing challenge can and should be discussed independent of question of causal identification of treatment effects on most empirical applications. Following a brief outline of the practical implementation steps required for PSM and IPW, these steps are presented in detail chronologically, outlining practical advice for each step. Subsequently, the topics of effect estimation, inference, sensitivity analysis and the combination with parametric estimation methods are discussed. Finally, new extensions of the methodology and avenues for future research are presented.
Organizations try to gain competitive advantages, and to increase customer satisfaction. To ensure the quality and efficiency of their business processes, they perform business process management. An important part of process management that happens on the daily operational level is process controlling. A prerequisite of controlling is process monitoring, i.e., keeping track of the performed activities in running process instances. Only by process monitoring can business analysts detect delays and react to deviations from the expected or guaranteed performance of a process instance. To enable monitoring, process events need to be collected from the process environment. When a business process is orchestrated by a process execution engine, monitoring is available for all orchestrated process activities. Many business processes, however, do not lend themselves to automatic orchestration, e.g., because of required freedom of action. This situation is often encountered in hospitals, where most business processes are manually enacted. Hence, in practice it is often inefficient or infeasible to document and monitor every process activity. Additionally, manual process execution and documentation is prone to errors, e.g., documentation of activities can be forgotten. Thus, organizations face the challenge of process events that occur, but are not observed by the monitoring environment. These unobserved process events can serve as basis for operational process decisions, even without exact knowledge of when they happened or when they will happen. An exemplary decision is whether to invest more resources to manage timely completion of a case, anticipating that the process end event will occur too late. This thesis offers means to reason about unobserved process events in a probabilistic way. We address decisive questions of process managers (e.g., "when will the case be finished?", or "when did we perform the activity that we forgot to document?") in this thesis. As main contribution, we introduce an advanced probabilistic model to business process management that is based on a stochastic variant of Petri nets. We present a holistic approach to use the model effectively along the business process lifecycle. Therefore, we provide techniques to discover such models from historical observations, to predict the termination time of processes, and to ensure quality by missing data management. We propose mechanisms to optimize configuration for monitoring and prediction, i.e., to offer guidance in selecting important activities to monitor. An implementation is provided as a proof of concept. For evaluation, we compare the accuracy of the approach with that of state-of-the-art approaches using real process data of a hospital. Additionally, we show its more general applicability in other domains by applying the approach on process data from logistics and finance.
The Adana Basin of southern Turkey, situated at the SE margin of the Central Anatolian Plateau is ideally located to record Neogene topographic and tectonic changes in the easternmost Mediterranean realm. Using industry seismic reflection data we correlate 34 seismic profiles with corresponding exposed units in the Adana Basin. The time-depth conversion of the interpreted seismic profiles allows us to reconstruct the subsidence curve of the Adana Basin and to outline the occurrence of a major increase in both subsidence and sedimentation rates at 5.45 – 5.33 Ma, leading to the deposition of almost 1500 km3 of conglomerates and marls. Our provenance analysis of the conglomerates reveals that most of the sediment is derived from and north of the SE margin of the Central Anatolian Plateau. A comparison of these results with the composition of recent conglomerates and the present drainage basins indicates major changes between late Messinian and present-day source areas. We suggest that these changes in source areas result of uplift and ensuing erosion of the SE margin of the plateau. This hypothesis is supported by the comparison of the Adana Basin subsidence curve with the subsidence curve of the Mut Basin, a mainly Neogene basin located on top of the Central Anatolian Plateau southern margin, showing that the Adana Basin subsidence event is coeval with an uplift episode of the plateau southern margin. The collection of several fault measurements in the Adana region show different deformation styles for the NW and SE margins of the Adana Basin. The weakly seismic NW portion of the basin is characterized by extensional and transtensional structures cutting Neogene deposits, likely accomodating the differential uplift occurring between the basin and the SE margin of the plateau. We interpret the tectonic evolution of the southern flank of the Central Anatolian Plateau and the coeval subsidence and sedimentation in the Adana Basin to be related to deep lithospheric processes, particularly lithospheric delamination and slab break-off.
Transcription factors (TFs) are ubiquitous gene expression regulators and play essential roles in almost all biological processes. This Ph.D. project is primarily focused on the functional characterisation of MYB112 - a member of the R2R3-MYB TF family from the model plant Arabidopsis thaliana. This gene was selected due to its increased expression during senescence based on previous qRT-PCR expression profiling experiments of 1880 TFs in Arabidopsis leaves at three developmental stages (15 mm leaf, 30 mm leaf and 20% yellowing leaf). MYB112 promoter GUS fusion lines were generated to further investigate the expression pattern of MYB112. Employing transgenic approaches in combination with metabolomics and transcriptomics we demonstrate that MYB112 exerts a major role in regulation of plant flavonoid metabolism. We report enhanced and impaired anthocyanin accumulation in MYB112 overexpressors and MYB112-deficient mutants, respectively. Expression profiling reveals that MYB112 acts as a positive regulator of the transcription factor PAP1 leading to increased anthocyanin biosynthesis, and as a negative regulator of MYB12 and MYB111, which both control flavonol biosynthesis. We also identify MYB112 early responsive genes using a combination of several approaches. These include gene expression profiling (Affymetrix ATH1 micro-arrays and qRT-PCR) and transactivation assays in leaf mesophyll cell protoplasts. We show that MYB112 binds to an 8-bp DNA fragment containing the core sequence (A/T/G)(A/C)CC(A/T)(A/G/T)(A/C)(T/C). By electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation coupled to qPCR (ChIP-qPCR) we demonstrate that MYB112 binds in vitro and in vivo to MYB7 and MYB32 promoters revealing them as direct downstream target genes. MYB TFs were previously reported to play an important role in controlling flavonoid biosynthesis in plants. Many factors acting upstream of the anthocyanin biosynthesis pathway show enhanced expression levels during nitrogen limitation, or elevated sucrose content. In addition to the mentioned conditions, other environmental parameters including salinity or high light stress may trigger anthocyanin accumulation. In contrast to several other MYB TFs affecting anthocyanin biosynthesis pathway genes, MYB112 expression is not controlled by nitrogen limitation, or carbon excess, but rather is stimulated by salinity and high light stress. Thus, MYB112 constitutes a previously uncharacterised regulatory factor that modifies anthocyanin accumulation under conditions of abiotic stress.
The term Linked Data refers to connected information sources comprising structured data about a wide range of topics and for a multitude of applications. In recent years, the conceptional and technical foundations of Linked Data have been formalized and refined. To this end, well-known technologies have been established, such as the Resource Description Framework (RDF) as a Linked Data model or the SPARQL Protocol and RDF Query Language (SPARQL) for retrieving this information. Whereas most research has been conducted in the area of generating and publishing Linked Data, this thesis presents novel approaches for improved management. In particular, we illustrate new methods for analyzing and processing SPARQL queries. Here, we present two algorithms suitable for identifying structural relationships between these queries. Both algorithms are applied to a large number of real-world requests to evaluate the performance of the approaches and the quality of their results. Based on this, we introduce different strategies enabling optimized access of Linked Data sources. We demonstrate how the presented approach facilitates effective utilization of SPARQL endpoints by prefetching results relevant for multiple subsequent requests. Furthermore, we contribute a set of metrics for determining technical characteristics of such knowledge bases. To this end, we devise practical heuristics and validate them through thorough analysis of real-world data sources. We discuss the findings and evaluate their impact on utilizing the endpoints. Moreover, we detail the adoption of a scalable infrastructure for improving Linked Data discovery and consumption. As we outline in an exemplary use case, this platform is eligible both for processing and provisioning the corresponding information.
Reading is a complex cognitive task based on the analyses of visual stimuli. Due to the physiology of the eye, only a small number of letters around the fixation position can be extracted with high visual acuity, while the visibility of words and letters outside this so-called foveal region quickly drops with increasing eccentricity. As a consequence, saccadic eye movements are needed to repeatedly shift the fovea to new words for visual word identification during reading. Moreover, even within a foveated word fixation positions near the word center are superior to other fixation positions for efficient word recognition (O’Regan, 1981; Brysbaert, Vitu, and Schroyens, 1996). Thus, most reading theories assume that readers aim specifically at word centers during reading (for a review see Reichle, Rayner, & Pollatsek, 2003). However, saccades’ landing positions within words during reading are in fact systematically modulated by the distance of the launch site from the word center (McConkie, Kerr, Reddix, & Zola, 1988). In general, it is largely unknown how readers identify the center of upcoming target words and there is no computational model of the sensorimotor translation of the decision for a target word into spatial word center coordinates. Here we present a series of three studies which aim at advancing the current knowledge about the computation of saccade target coordinates during saccade planning in reading. Based on a large corpus analyses, we firstly identified word skipping as a further factor beyond the launch-site distance with a likewise systematic and surprisingly large effect on within-word landing positions. Most importantly, we found that the end points of saccades after skipped word are shifted two and more letters to the left as compared to one-step saccades (i.e., from word N to word N+1) with equal launch-site distances. Then we present evidence from a single saccade experiment suggesting that the word-skipping effect results from highly automatic low-level perceptual processes, which are essentially based on the localization of blank spaces between words. Finally, in the third part, we present a Bayesian model of the computation of the word center from primary sensory measurements of inter-word spaces. We demonstrate that the model simultaneously accounts for launch-site and saccade-type contingent modulations of within-word landing positions in reading. Our results show that the spatial saccade target during reading is the result of complex estimations of the word center based on incomplete sensory information, which also leads to specific systematic deviations of saccades’ landing positions from the word center. Our results have important implications for current reading models and experimental reading research.