Bandwidths of vocal tract resonances in physical models compared to transmission-line simulations

This study investigated how the bandwidths of resonances simulated by transmission-line models of the vocal tract compare to bandwidths measured from physical three-dimensional printed vowel resonators. Three types of physical resonators were examined: models with realistic vocal tract shapes based on Magnetic Resonance Imaging (MRI) data, straight axisymmetric tubes with varying cross-sectional areas, and two-tube approximations of the vocal tract with notched lips. All physical models had hard walls and closed glottis so the main loss mechanisms contributing to the bandwidths were sound radiation, viscosity, and heat conduction. These losses were accordingly included in the simulations, in two variants: A coarse approximation of the losses with frequency-independent lumped elements, and a detailed, theoretically more precise loss model. Across the examined frequency range from 0 to 5 kHz, the resonance bandwidths increased systematically from the simulations with the coarse loss model to the simulations with the detailed loss model, to the tube-shaped physical resonators, and to the MRI-based resonators. This indicates that the simulated losses, especially the commonly used approximations, underestimate the real losses in physical resonators. Hence, more realistic acoustic simulations of the vocal tract require improved models for viscous and radiation losses

Carbon Capture and Storage on its way to large-scale deployment: Social acceptance and willingness to pay in Germany

Carbon Capture and Storage (CCS) is an emerging technology to mitigate greenhouse gas emissions from fossil fuel-fired power plants. In the wake of a rapidly changing German energy system, CCS can play an important role. By means of an online survey among 130 university students in Dresden, this paper investigates the level and influencing factors of social acceptance of CCS. Furthermore, the individual willingness to pay for CCS and renewable power delivery is measured and compared through a choice model. The survey results reveal that the attitude towards CCS is neutral. Moreover, it is shown that acceptance of CCS is an important factor for the willingness to pay. The level of willingness to pay for CCS technology is much lower than for renewable energy

Investigations on Strategic Element Recovery by an Underground Membrane Pilot Plant from In-Situ Extracted Bioleaching Solutions

Focusing on the selective extraction of the critical raw materials indium and germanium from real bioleaching solutions, extended studies have been carried out using Europe’s first underground hybrid membrane pilot plant (TRL6). In order to transfer former laboratory experiments to pilot scale, NF99 (Alfa Laval) was used for the evaluation of membrane permeance and ion retention. A performance test of microfiltration (MF) and nanofiltration (NF) showed high permeances with low root-mean-square deviation under feed variation (5.2% for MF, 4.7% for NF). Depending on the feed load, a significant permeance drop of up to 57% for MF (3 bar) and 26% for NF (10 bar, 1.1 m s−1) was observed. The NF retention performance showed that, without regular chemical cleaning, the selectivity between the target elements degraded. By introducing acidic-basic cleaning steps, it was possible to keep the retention behavior at an approximately constant level (In 91.0 ± 1.3%; Ge 18.2 ± 5.5%). In relation to the specified target, the best results could be achieved at low pressure (7.5 bar) and a maximum overflow velocity of 1.1 m s−1, with a retention of 88.4% for indium and 8.8% for germanium. Moreover, the investigations proved the functionality and long-term stability of the underground membrane device

Crack Monitoring on Concrete Structures using Robust Distributed Fiber Optic Sensors

The possibility to measure strains continuously using distributed fiber optic sensors (DFOS) offers enormous potential for structural health monitoring. Cracks can be automatically detected, localized and crack widths calculated. To address the relevant questions of choosing the right DFOS and appropriate application technique for monitoring existing structures, two 4 m long reinforced concrete beams were loaded under service loads in a 4-point bending test. The DFOS were either bonded directly to the smooth concrete surface or along a groove. For comparison, another DFOS was integrated into the specimen. It is demonstrated that with the used adhesive, a good strain transfer from the specimen to the DFOS can be ensured even with subsequent installation. In order to detect all cracks with high reliability, robust DFOS with a monolithic cross section should be preferred for the practical use. The accuracy of crack width measurement was verified through Reference measurement via digital image correlation. For the monolithic DFOS, all measurement deviations were within the tolerance range of ±0.05 mm. With layered sensing cables, significant misinterpretations occurred due to strongly damped strain curves

Development of novel transient Foamy Virus (TraFo) vectors - Combining ancient viruses with bacterial CRISPR nucleases for efficient genome editing

Knowledge on the human genome and specific sequences associated with human diseases is continuously growing. The ability to connect human genetics to cellular mechanisms and physiology raises the need for medicine to get to gene specific therapeutics. In order to achieve gene-specific modification, tools are required to enable sequence-specific DNA cleavage. Not long ago, the RNA-guided endonuclease Cas9 was shown to effectively facilitate gene editing in humans. Cas9 endonuclease, which is naturally part of an adaptable bacterial immune system, can be easily adjusted to recognize and cleave specific DNA sequences in a 20 nt RNA-DNA complementary manner. The easy adjustability and high efficiency of Cas9 gave rise to hopes that this genome engineering tool could pave the way to ‘gene surgery’ in humans. However, to achieve DNA cleavage, the endonuclease and its guiding RNA need to be sufficiently accessible in the nucleus of target cells. Viruses, which evolution has made well adapted to transfer their own genetic information into cells can be exploited for transfer of foreign genetic material. Replication deficient retroviruses therefore represent interesting vehicles for gene delivery. Retroviruses preferentially incorporate their own genetic information in the form of RNA into viral particles. Typically, viral RNA of retroviruses is reverse transcribed into DNA during viral infection and integrated into host cell chromosomes. In this respect, integration-competent or integration-deficient lentiviral (HIV-derived) vectors (ICLV/IDLV) were reported to be efficient ‘gene shuttles’ for Cas9 delivery. In contrast, up to now Foamy viruses (FV), which represent a distinct subfamily in the family of retroviruses have not previously been tested for their efficiency to transduce CRISPR/Cas9 components. FV show several unique characteristics some of which make them interesting candidates for gene therapy, such as high transduction efficiency on a wide variety of human cell lines or a special capability to efficiently transfer and provide non-viral RNA in target cells. In this thesis the unique characteristic of FVs, which allow for the efficient transduction of non-viral RNAs, was exploited for transient FV mediated (TraFo) Cas9 expression. It is shown in this thesis that gene knock-out (KO) achieved with TraFo Cas9 particles appears to have several advantages over ICLV or IDLV mediated Cas9 transduction. In this work, it could be demonstrated that a single application of TraFo Cas9 supernatant results in high efficiency of GFP KO in osteosarcoma cells (U2OS). The efficiency of gene KO with TraFo Cas9 particles exceeded gene KO frequencies achieved with similar volumes of ICLV or IDLV supernatant for Cas9 transduction. In addition, transient Cas9 delivery achieved with TraFo particle supernatant resulted in remarkably reduced Cas9 off-target cleavage compared to corresponding infections with ICLV or IDLV particles. The results show, that TraFo Cas9 represents an interesting addition to the currently utilized methods for transient Cas9 delivery. One particular feature of TraFo particle transduction is especially noteworthy – TraFo mediated transduction does not depend on any particular adjustment on the encapsidated non-viral RNA sequence (such RNA only needs to be present in sufficient amounts during virus assembly) nor does it depend on any modification of viral proteins. The easy adaptability of TraFo mediated non-viral RNA transfer is an especially remarkable feature, since science continues to both developing new variants of Cas9 and continues to find new and interesting members of the pool of CRISPR enzymes. In this regard TraFo particles represent interesting vehicles to transiently provide mRNA transcripts of such new protein candidates in cells. The ability of TraFo particles to provide the RNA sequence needed to guide Cas9 (termed sgRNA) to its target DNA sequence in cells was additionally investigated. It was assumed that typically engaged RNA polymerase (RNAP) III transcription of sgRNAs hampers transduction with TraFo particles, since RNAP III-derived transcripts are not actively exported into the cytoplasm and show low stability. An additional CRISPR enzyme Csy4 was used, which is able to specifically cleave RNA. This enabled TraFo mediated transfer of RNAP II transcripts (with active nuclear export and higher stability than RNAP III transcripts) with embedded sgRNA sequences. It was demonstrated that a simultaneous infection of cells with TraFo particles providing bicistronic transcripts of Cas9 and Csy4 on the one side and RNAP II-derived transcripts with embedded sgRNA sequences on the other, enabled reasonable GFP gene inactivation in U2OS cells. Gene KO with RNAP II transcripts as a result significantly exceeds TraFo transduction of RNAP III-derived sgRNA. Interestingly, with regard to gene KO, it was found that de novo transcription of sgRNAs from viral DNA (by integration-competent or integration-deficient retroviral vector [ICRV/IDRV] transduction) when combined with TraFo Cas9 transduction was superior to a TraFo transduction of sgRNA transcripts. IDRV mediated transduction was optimized in order to minimize the risk of unfavorable genome modification of cells by viral DNA integration. By adding the coding sequence of a fluorescent marker to the viral vector, it was demonstrated that a smaller number of viral particles helps to significantly lower the frequency of viral DNA integration. In addition, the expression of a fluorescent marker opened up the opportunity to further reduce the cell fraction with continuous marker gene expression by flow cytometric cell sorting. The IDRV/ICRV sgRNA and TraFo Cas9 delivery system was then challenged for use on immortalized and primary T cells. Primary T cells represent interesting targets for genetic engineering since modified T cells can be utilized as ‘living drugs’ (by expression of chimeric antigen receptors – CARs) against cancer cells. Efficient gene inactivation was observed on the immortalized T cell line – Jurkat. Transduction of primary T cells pointed to certain restrictions of the split two-virus delivery system for sgRNA and Cas9 transduction. However, despite certain limitations, it was possible to demonstrate that this FV-derived Cas9 delivery system is also feasible on primary tissue, and further optimization could make it an interesting alternative delivery method for CAR therapy. The ability of IDRV vector genomes to provide repair template donor DNA to induce homologous recombination (HR) was additionally investigated. DNA double-strand breaks in eukaryotic cells are typically repaired by the error prone non-homologous end joining pathway (often leading to frame-shift mutations by small insertions or deletions) or HR. Delivery of a homologous DNA sequence during DNA cleavage enables site-specific integration of exogenous DNA sequences. The work of this thesis showed that IDRV vector genomes providing repair template donor DNA allow for HR in a homology length dependent manner. Besides the length of homology, it was also observed, that the length of sequence which should be integrated (KI) remarkably influences the frequency of HR. HR is therefore engaged significantly more frequently if single nucleotides, rather than a whole gene, are provided as sequences within a repair template. In addition, viral vectors were augmented with additional fluorescent marker sequences. It could subsequently be demonstrated that the majority of cells showed accurate sequence-specific DNA integration. Furthermore, several indications were found, which lead to the assumption that the ratio of KI to homologous sequence markedly influences the accuracy of HR. Using the previously obtained knowledge it was further possible to tag an essential human protein by FV vector mediated transient Cas9 and repair template transduction. It was found that the large packaging capacity of FV vectors can be exploited to enable selection and flow cytometric sorting of cells with correct site-specific DNA integration. In summary, the results of this thesis demonstrate for the first time that FV mediated non-viral mRNA Cas9 transduction in combination with retroviral delivery of sgRNA (and repair template sequence) are a promising basis for several different interesting applications with relevance for not only basic research, but also for gene therapy.:1. Introduction 1 1.1 Gene therapy 1 1.2 Viral vectors for gene therapy 2 1.3 History of retroviral research 2 1.4 Taxonomy of Retroviruses 3 1.5 Foamy Viruses 4 1.5.1 Morphology of Foamy Virus 6 1.5.2 Foamy Virus replication 7 1.5.3 Foamy virus proteins, as part of a viral vector system 10 1.6 Genetic engineering 14 1.6.1 ‘DNA scissors’ – Zinc-finger and Transcription-activator like effector nucleases 15 1.6.2 History of CRISPR/Cas9 as a tool for genetic engineering 16 1.7 CRISPR/Cas immunity in prokaryotes 18 1.8 CRISPR/Cas9 functioning 21 1.9 Double-strand break repair in eukaryotic cells 21 1.9.1 Classical NHEJ 23 1.9.2 Homologous recombination 24 1.9.3 DSB repair in vertebrates 26 1.10 DSBs in context of CRISPR/Cas9 cleavage 27 1.11 Thesis Aim: CRISPR/Cas9 transduction with FV particles 28 2. Materials and Methods 30 2.1 Materials 30 2.1.1 Chemicals 30 2.1.2 Buffers and Solutions 30 2.1.3 Bacterial Growth Media 33 2.1.4 Cell Culture Media 34 2.1.5 Antibodies 34 2.1.6 Enzymes 35 2.1.7 Commercial Kits and additional reagents 36 2.1.8 Size Markers 36 2.1.9 Antibiotics 36 2.1.10 Bacterial strains 37 2.1.11 Cell lines 37 2.1.12 Devices and Software 37 2.1.13 Oligonucleotides 38 2.1.14 Plasmids 46 2.1.15 sgRNA sequences 56 2.1.16 Consumable material 57 2.2 Molecular Biology Methods 58 2.2.1 Restriction of DNA 58 2.2.2 Polymerase chain reaction 59 2.2.3 Gibson assembly 60 2.2.4 Agarose gel electrophoresis 60 2.2.5 Ligation 61 2.2.6 Cultivation of bacteria 62 2.2.7 Transformation 62 2.2.8 Plasmid Preparation 63 2.2.9 Sequencing 65 2.3 Cell culture methods 66 2.3.1 Passaging of cells 66 2.3.2 Cell counting 66 2.3.3 Freezing and thawing of cells 66 2.3.4 Seeding and fixation of cells for microscopy 67 2.4 Virological Methods 67 2.4.1 Polyethyleneimine transfection 67 2.4.2 Integration-competent, integration-deficient and ‘transient’ retroviral vectors 68 2.4.3 Infection of adherent cells 70 2.4.4 Infection of suspension cells 71 2.4.5 Flow cytometry 72 2.4.6 Multiplicity of infection (MOI) 72 2.4.7 Particle preparation 73 2.5 Nucleic acid composition in viral particles and culture cells 73 2.5.1 Isolation of total RNA from viral particles 73 2.5.2 RNA isolation from culture cells 73 2.5.3 Reverse transcription of viral or cellular RNA 73 2.5.4 DNA isolation from culture cells 74 2.5.5 Quantitative PCR (qPCR) analysis 74 2.5.6 T7 endonuclease assay 75 2.6 Protein biochemistry methods 76 2.6.1 Cell lysates 76 2.6.2 Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 76 2.6.3 Semi-dry Western Blot 77 2.6.4 Immunodetection 78 2.6.5 Stripping of Western blot membranes 78 2.6.6 Immunostaining of cells for FACS analysis 78 2.7 Microscopy methods 79 2.7.1 Fluorescence microscopy 79 2.7.2 Confocal Laser scanning Microscopy (CLSM) 79 2.7.3 Live-cell imaging 79 3. Results 80 3.1 Transient foamy virus transduction of non-viral mRNA transcripts 80 3.2 Transient foamy virus transduction of Cas9-encoding mRNA transcripts 81 3.3 Cas9-encoding nucleic acids and their ‘effects’ in cells after retroviral transduction 84 3.4 Off-target analysis after TraFo Cas9 delivery 87 3.5 Transient fomy virus transduction of Cas9 and sgRNAs 89 3.6 Retroviral vectors providing sgRNAs and a fluorescent protein 92 3.6.1 Gene knock-out with retroviral vectors under saturated conditions 92 3.6.2 MOI adjusted ID sgRNA vector supernatants for comparison of residual vector integration 94 3.6.3 Gene knock-out in murine embryonic fibroblasts 95 3.7 Influence of Cas9 expression on IDRV vector genome integration 96 3.8 TraFo Cas9 mediated T cell receptor knock-out in immortalized and primary human T cells 97 3.9 Homology-directed repair after FV CRISPR/Cas9 mediated double-strand breaks 99 3.9.1 Length of homologous donor DNA and its influence on HDR 100 3.9.2 Effect of freezing viral supernatants on the frequency of HDR 102 3.9.3 Effect of donor DNA mismatches on the frequency of HDR 104 3.10 Investigation on donor DNA integration with additional fluorescent markers 105 3.11 Lentiviral and foamyviral transduction of HDR donor DNA 107 3.12 HDR mediated single nucleotide substitutions after TraFo CRISPR/Cas9 delivery 109 3.13 Tagging of an endogenous protein after TraFo CRISPR/Cas9 delivery 111 3.13.1 Specific CRISPR/Cas9 mediated cleavage of endogenous hPLK1 gene 111 3.13.2 Homology-directed repair of the hPLK1 gene for endogenous gene tagging 113 3.13.3 Confocal fluorescence microscopy analysis of GFP-Plk1 HeLa cell populations 118 4. Discussion 120 4.1 Genetic engineering – potential and risks 120 Chapter I Transient FV vectors – mRNA delivery vehicles for CRISPR/Cas9 mediated gene editing 122 4.2 Non-viral Cas9-encoding mRNA transfer in foamy virus particles 122 4.2.1 Fate of Cas9-encoding nucleic acids in cells after TraFo Cas9 transduction 124 4.2.2 Potential adjustments to further improve TraFo Cas9 transduction 125 4.2.3 Lentiviral in contrast to TraFo transduction of Cas9-encoding nucleic acids 126 4.3 Efficiency of Cas9-mediated gene knock-out with different retroviral vectors 127 4.4 Type of retroviral Cas transduction and its influence on the specificity of Cas9 cleavage 127 4.5 Alternative approaches to deliver Cas9-encoding mRNA in human cells 129 4.6 Transient sgRNA transduction with TraFo particles 131 Chapter II Delivery of foreign DNA with FV-derived vectors – enabling gene knock-out and homology-directed repair 133 4.7 Gene inactivation by TraFo Cas9 transduction and sgRNA expression from retroviral vector genomes 133 4.7.1 Gene editing in immortalized and primary T cells 135 4.8 Homology-directed repair with IDRV genomes 137 4.8.1 The influence of the length of sequence homology on HR 138 4.8.2 The influence of freezing viral supernatants on HR 139 4.8.3 Widening the applicability of TraFo vector particles for improved HR 140 4.8.4 The influence of mismatching nucleotides on HR 140 4.8.5 Visualization of inaccurate HR or additional dsDNA integration 141 4.8.6 The influence of the ratio of knock-in and homologous sequence on the accuracy of HR 142 4.8.7 Alternatives to double-stranded donor DNA 143 4.9 Endogenous gene tagging with IDPV donor DNA transduction 145 4.9.1 Alternative approaches for endogenous protein tagging 146 5. Conclusion 148 6. Summary 150 6.1 Summary 150 6.2 Zusammenfassung 153 7. Supplementary 157 8. References 159 9. Appendices 182 9.1 Indices 182 9.1.1 Abbreviations 182 9.1.2 Index of Figures 185 9.1.3 Index of Tables 187 9.2 Curriculum Vitae 188 9.3 Publication Record 189 9.4 Congress Contributions 189 9.5 Patent Applications 189 10. Statement of Authorship 19

Automatisierte Abgrenzung von Innenbereichen einschließlich Ergebnisevaluierung - Grundlage für ein Siedlungsflächenmonitoring

Der steigende Trend zur Urbanisierung stellt sowohl auf lokaler als auch auf globaler Ebene eine beträchtliche Herausforderung dar. Bis zum Jahr 2050 werden beinahe 7 von 10 Menschen in städtischen Gebieten leben, was einer Verdopplung der städtischen Bevölkerung gleichkommt. Die Schwierigkeit liegt darin, dass städtische Regionen momentan im Durchschnitt doppelt so schnell expandieren wie ihre Bevölkerung wächst. Man erwartet, dass das Anwachsen der städtischen Bevölkerung zusammen mit der wirtschaftlichen Entwicklung in den nächsten drei Jahrzehnten weltweit zu einer zusätzlichen bebauten Fläche von etwa 1,2 Millionen km2 führen wird. Neben negativen Auswirkungen wie dem Verlust von fruchtbaren Böden und Biodiversität durch Versiegelung und Fragmentierung, bieten Urbanisierungsprozesse auch Chancen für eine nachhaltigere Entwicklung. Daher ist es wichtig, diesen Prozess zu steuern, um die negativen Auswirkungen zu minimieren. Die Steuerung der Siedlungsentwicklung erfordert verlässliche Datengrundlagen. Die derzeitige Datenlage in Deutschland beruht jedoch eher auf Annahmen und Schätzungen denn empirischen Erhebungen. Ursache hierfür ist die Erfassung von Nutzungsänderungen aggregiert auf administrative Einheiten und deren Zusammenfassung in einer Nutzungsklasse, der Siedlungs- und Verkehrsfläche. Damit können weder kleinräumige Flächenänderungen noch die Qualität der Änderungen hinreichend bestimmt werden. In Wissenschaft, Stadt- und Regionalplanung besteht daher gleichermaßen Bedarf an aktuellen, homogenen, aussagekräftigen und ökonomisch generierten Informationen zur Siedlungsentwicklung. Im Rahmen dieser Arbeit wurde ein Abgrenzungsverfahren zur Erstellung detaillierter Siedlungsabgrenzungen, insbesondere des Innenbereichs im baurechtlichen Sinne, entwickelt. Um eine möglichst breite Anwendbarkeit zu gewährleisten, wurden als Eingangsdaten ausschließlich Gebäudegrundrisse und allgemein verfügbare topographische Daten verwendet. Nach einer Partitionierung des Datensatzes erfolgte eine semantische Filterung und Aggregation über einen dichtegesteuerten Clusteralgorithmus. Eine besondere Herausforderung stellte dabei die Berücksichtigung planerischer und rechtlicher Aspekte in Deutschland dar. Die Anwendung der Methode erfolgte in Untersuchungsgebieten in Frankfurt am Main, in der Region Hannover und im ländlichen Raum Brandenburgs. Die Abgrenzungsergebnisse wurden mit Hilfe von GIS-Analysen und Expertenbefragungen evaluiert. Die Ergebnisse zeigen, dass die Methode in der Lage ist, sowohl ländliche Gebiete als auch Großstädte und Ballungsräume mit einer Genauigkeit von 75 bis 94 % im Vergleich zu Expertenabgrenzungen abzubilden. Insbesondere Gebiete mit Wohn- und Mischbebauung liefern sehr gute Ergebnisse. Optimierungspotenzial bieten dagegen Flächen mit Sondernutzungen oder stark heterogene Siedlungsbereiche. Das Verfahren stellt somit ein wertvolles Instrument zur Generierung von Grundlagendaten für zukünftige Studien oder Fachanwendungen dar. Die automatisiert generierten Abgrenzungen können Planungsaufgaben auf verschiedenen räumlichen Ebenen unterstützen, sie bilden die Grundlage für die Bestimmung von Innenentwicklungspotenzialen bzw. -indikatoren und eignen sich außerdem für Untersuchungen zur Zersiedelung.:Übereinstimmungserklärung Danksagung Zusammenfassung Abstract Abkürzungsverzeichnis Abbildungsverzeichnis Tabellenverzeichnis 1. Einleitung 1 1.1. Reduzierung der Flächeninanspruchnahme als politisches Ziel 1.2. Funktion des Siedlungsflächenmonitoring 1.3. Problemstellung 1.4. Zielstellung und Forschungsfragen 1.5. Aufbau der Arbeit 2. Forschungsansätze für automatisierte Siedlungsabgrenzungen 2.1. Datenquellen 2.2. Methoden 2.3. Bewertung bisheriger Ansätze 3. Begriffliche und konzeptionelle Grundlagen 3.1. Im Zusammenhang bebauter Ortsteil 3.1.1. Vorhaben 3.1.2. Bebauungszusammenhang 3.1.3. Ortsteil 3.2. Baulücke 3.3. Außenbereich im Innenbereich 3.4. Innenbereichssatzungen 3.4.1. Klarstellungssatzung 3.4.2. Entwicklungssatzung 3.4.3. Einbeziehungs- oder Ergänzungssatzung 3.5. Außenbereich 4. Verfahrensentwicklung 4.1. Allgemeiner methodischer Ansatz 4.2. Eingangsdaten 4.3. Anforderungen 4.4. Methodik 4.4.1. Partitionierung 4.4.2. Erstellung von Baublockgeometrien 4.4.3. Ermittlung Überbauungsgrad 4.4.4. Filterung 4.4.5. Identifizierung dicht bebauter Blöcke 4.4.6. Aggregation mittels minimalen Spannbaum 4.4.7. Nachbearbeitung 4.5. Parametrisierung des Verfahrens 4.5.1. Eingangsdaten für Innenbereichsabgrenzung 4.5.2. Objektauswahl für die Filterung 4.5.3. Schwellwert für dicht bebaute Blöcke 4.5.4. Größe von Baulücken und Freiflächen 4.5.5. Größe von Außenbereichsflächen im Innenbereich 4.5.6. Mindestanzahl von Gebäuden und Mindestgröße eines Ortsteils 4.5.7. Abgrenzung der Innenbereichsflächen vom Außenbereich 4.6. Programmtechnische Umsetzung 5. Evaluationsmethodik 5.1. Vorüberlegungen 5.1.1. Ziel der Bewertung 5.1.2. Definition von Qualität 5.1.3. Kontext der Bewertung 5.1.4. Qualitative und quantitative Methoden zur Bewertung von Geometrien 5.2. Empirische Bestimmung der Abgrenzungsqualität 5.3. Bestimmung der Abgrenzungsqualität mittels Expertenbefragung 5.3.1. Umsetzung der Datenerhebung mittels Fragebogen 5.3.2. Datenerhebungstechnik 5.3.3. Erfassen und Auswerten von Daten 5.3.4. Stichprobenbildung für Experteninterviews 5.3.5. Bewertung der Innenbereichsgeometrien 5.3.6. Mögliche Fehlerquellen von Umfragen 5.3.7. Gütekriterien 6. Ergebnisse 6.1. Vergleich der Abgrenzungsergebnisse mittels Referenzdaten 6.1.1. Untersuchungsgebiete und verwendete Daten 6.1.2. Auswertung GIS-Analyse 6.2. Auswertung der Befragung und Experteneinschätzung 6.2.1. Vorerfahrungen der Teilnehmenden 6.2.2. Allgemeine Fragen zum Thema Innenbereich 6.2.3. Beurteilung der Karten 6.2.4. Auswertung der Bewertungen der Karten nach Flächennutzung 6.2.5. Bewertung aller Abgrenzungen im Untersuchungsgebiet 6.2.6. Urteile zu Einsatzmöglichkeiten des Verfahrens 7. Diskussion 7.1. Stärken und Schwächen des Verfahrens 7.2. Beurteilung der quantitativen Bewertungsmethode 7.2.1. Einfluss der Siedlungsstruktur auf das Abgrenzungsergebnis 7.2.2. Umgang mit unvollständigen Referenzabgrenzungen 7.3. Bewertung der Expertenbefragung 7.4. Herausforderungen der Innenbereichsabgrenzung 7.4.1. Wohnbebauung 7.4.2. Kleingartensiedlungen 7.4.3. Wochenendhaussiedlungen 7.4.4. Industrie- und Gewerbegebiete 7.4.5. Tagebau 7.4.6. Landwirtschaftliche Betriebe 7.4.7. Streusiedlungen und Einzelgehöfte 7.4.8. Großflächige Photovoltaikanlagen 7.4.9. Kasernen 7.5. Fehlerbetrachtung 7.6. Vergleich mit anderen Studien 7.7. Anwendung und Übertragbarkeit 8. Fazit und Ausblick 8.1. Kriterien für die Innenbereichsabgrenzung und Aspekte bei der Abgrenzung durch Experten 8.2. Ansatz, Voraussetzungen und Vorgehen beim automatisierten Abgrenzen von Innenbereichen 8.3. Einfluss Struktur der Siedlungen auf die Abgrenzungsqualität 8.4. Anwendungsfelder einer Innenbereichsabgrenzung 8.5. Forschungsbedarf A. Anhang A.1. Befragungsergebnisse A.1.1. Einführungstext zur Befragung A.1.2. Hintergrund der Teilnehmenden A.1.3. Grundlagen Innenbereich A.1.4. Bewertung der Karten A.1.5. Bewertung der Abgrenzung in der Gesamtheit A.2. Detailauswertung nach Flächennutzungen A.3. Quellcode A.3.1. Skripte IB-Tool A.3.2. Skript qualitative Bewertung A.3.3. Skript Auswertung Befragung A.3.4. Zufallsfunktion für Flächenauswahl A.4. Geodaten Literaturverzeichni

Desecularisation of the State and Sacred Secularism: Politics and Religion in Mexico within the Latin-American Context

Recent political conflicts have highlighted the influence of religious actors and organisations in the public spheres of Latin American countries. The Pentecostal Evangelical movement in Colombia was crucial to the success of the ‘No’ campaign in the 2016 plebiscite, in which citizens rejected the peace agreement between the government and the Revolutionary Armed Forces of Colombia (FARC). In Brazil, evangelical congregations played a central role during the 2016 impeachment of President Dilma Rousseff, and in the subsequent rise of Jair Bolsonaro. In Bolivia, evangelical leaders and conservative elements of the Catholic Church alike supported the 2019 coup against Evo Morales. In Mexico, the 2018 rise of left-wing President Andrés Manuel López Obrador has been accompanied by criticism of his proximity to religious actors, and his moralising political rhetoric. Some authors have even described the Mexican leader as a politician with messianic overtones. Against this background, it is worth asking what the implications of the recent convergence between politics and religion in Latin America are. To answer this question, we must avoid the oversimplification of suggesting a singular process of religious advance in Latin American societies. It is important to instead highlight the complex interaction of: 1) the process of secularisation (involving both secularism and secularity) in the region, 2) the trend towards pluralisation of the religious field, 3) the concurrence of counter-secular expressions in the public space, 4) and the occurrence of conflict in the political arena. Although secularisation in Latin America historically emerged as a process of distinction of the political sphere, I argue here that it is currently expressed as a democratic ideal through the process of the dispersal in society of certain secular notions favouring state autonomy, especially in those countries that maintain the secularism legally established in the nineteenth century. My approach raises the question of how the boundaries between religion and the state in Mexico have been defined historically, and what the current status of this differentiation is. I also advance the analytical notion of sacred secularism, as a principle and expectation in the public space