Die Morphologie der Buchlungen von Arachniden (Arachnida Lamarck, 1801 – Arthropoda, Chelicerata) wurde in der vorliegenden Dissertation einer streng vergleichenden Analyse unterzogen, welche mit Hilfe moderner Methoden eine neue Sicht auf die Phylogenie der Arachniden eröffnet. Aus dem Vergleich mit den potentiellen Schwestergruppen (Xiphosura, Eurypterida) und mit Skorpionsfossilien erweist sich der einmalige Landgang eines gemeinsamen Vorfahrens aller rezent terrestrischen Arachniden. Buchlungen-Daten von 200 rezenten + 2 fossilen Skorpionen, 16 Geißelspinnen (Amblypygi), 17 Geißelskorpionen (Thelyphonida), einem Schizomiden (Schizomida), einem ausgestorbenen Trigonotarbiden (Trigonotarbida) und der Außengruppe, den rezenten Pfeilschwanzkrebsen (Xiphosura), wurden zu einem illustrierten Katalog zusammengestellt. Die unüberschaubare Vielfalt der oft graduell variierenden Strukturen macht die eindeutige Definition der Merkmale und auftretenden Merkmalszustände notwendig. Es wurden folgende 5 Merkmale definiert: (1) die Oberflächen der Atemlamellen, (2) der Lamellenrand, (3) der posteriore Stigmenrand, (4) der anteriore Stigmenrand und (5) die Wand des Atriums. Hierbei tragen die Merkmale 1-3 mit ihrer unerwarteten Fülle innerhalb der Skorpione maßgeblich zur Aufklärung ihrer Verwandtschaftsverhältnisse bei. Ähnliches wird von den Merkmalen 4 und 5 bezogen auf die Uropygi s. lat. vermutet. Ein sechstes Merkmal (Sensillen/Poren) wurde definiert, doch nur vereinzelt erfasst. Raster-Elektronenmikroskopie diente der Erfassung der cuticulären Feinstruktur der Buchlungen. Die Struktur der Buchlungen im Gesamten wurde mittels histologischer Schnittserien und mit auf µCT basierenden 3D-Rekonstruktionen untersucht. Für die Untersuchung von Trigonotarbiden-Fossilien wurde eine Methode entwickelt, die 3D-Rekonstruktion aus Sequenzen von Mikrofotos in unterschiedlichen Fokusebenen generiert. / Strict comparative analyses of the morphology of arachnid book lungs (Arachnida Lamarck, 1801 - Arthropoda: Chelicerata) were carried out in the present doctoral thesis using modern methods, resulting in a new perspective on arachnid phylogeny. Comparisons with potential aquatic sistergroups (Xiphosura and/or Eurypterida) and fossil scorpions give rise to the hypothesis of the unique terrestrialization of ancestors, which is common for all arachnids. Data from 200 Recent + 2 extinct scorpions, 16 whip spiders (Amblypygi), 17 whip scorpions (Thely-phonida), 1 schizomid (Schizomida), 1 extinct trigonotarbid (Trigonotarbida) and 1 outgroup - horseshoe crab (Xiphosura) are assembled into an illustrated catalogue of arachnid book lungs. Following the observations of these gradually differing cuticular structures the vast variation of book-lung fine structure across Arachnida requires unequivocal definition of characters and character states. Five characters are defined, which are assigned to distinct homologous book-lung structures: (1) the surface structure of the respiratory lamellae, (2) structure of the distal edges of the lamellae, (3) the posterior edge of the spiracle, (4) the anterior edge of the spiracle and (5) the structure of the wall lining the atrial chamber. Especially the book-lung characters 1-3 contain unexpected information, which helps resolve relationships within Scorpiones to a high degree, and characters 4 and 5 are of considerable importance for Uropygi s. lat. One sixth character (sensilla/pores) is mentioned, but sporadically examined. However, cuticular book-lung fine structure is studied using SEM, the gross morphology is reassessed using histological sectioning and 3D-reconstructions based on µCT. For investigations on trigonotabid fossils a new method yielding 3D-reconstructions from stacks of subsequent focal layers was developed.

Laterale Gentransfer wurde zuerst von Schwartz und Dayhoff (1978) entdeckt, die es aber als eine Exzentrizität werteten und als solche ignorierten. Später, als mehrere DNS- und Eiweißsequenzen sequenziert und raffiniertere Phylogenien rekonstruiert wurden, hat die Rolle an Relevanz gewonnen, die der laterale (oder horizontale) Gentransfer in der evolutionären Geschichte von lebendigen Organismen gespielt hat. Außerdem existiert auch zwischen Endosymbionten und Zellkernen statt. Ich habe ein theoretisches Modell entwickelt, das den lateralen Gentransfer zwischen Endosymbionten und dem Zellkern repräsentiert. Das Modell erforscht die Bedeutung des Fehlens von Rekombination in den Organellen (Muller’s Ratchet) sowie Abweichungen von Muller’s Ratchet in Form der non-symmetrical homologous recombination in Gentransfermechanismen. Ich habe zum einen Zellkern-Inkompatibilitäten, die aus der Übertragung eines Gens resultieren, und zum anderen Zyto- und Zellkern-Inkompatibilitäten zwischen den mutierten endosymbiotischen Genomen und dem modifizierten Zellenkern untersucht. Die Ergebnisse zeigen, dass unter bestimmten Bedingungen die Existenz oder Nicht-Existenz von Rekombination die gleiche Wirkung haben können. Es zeigte sich auch, dass Rekombination, wenn sie vorkommt und wenn sie nicht symmetrisch ist, starke Auswirkungen auf die Allelenfrequenz einer Population haben kann. Es wurde auch klar, dass es eine starke Beziehung zwischen dem Zellkern und endosymbiotischen Genomen gibt, und dass das evolutionäre Schicksal des einen größtenteils von den evolutionären Kräften abhängig ist, die das andere beeinflussen. Wenn man Zellkern- und Cyto-Zellkerninkompatibilitäten in das Modell einführt, dann zeigen die Ergebnisse, dass die Inkompatibilitäten, die der laterale Gentransfer produziert hat, möglicherweise eine ähnliche Rolle im Speziationsmechanismus spielen könnten wie die Inkompatibilitäten zwischen Mitochondrien und Zellkernen in verschiedenen Nasonia-Arten. / Lateral gene transfer has played a key role in the evolution of living beings. This process was first acknowledged in 1978 by Schwartz and Dayhoff but considered a relatively infrequent eccentricity and ignored. Later on, as DNA and protein sequences accumulated and more refined phylogenies were reconstructed, the contribution of lateral (or horizontal) gene transfer to the evolutionary history of living organisms gained relevance. Besides, gene transfer is known to occur not only between independent organisms but also, and more frequently between endosymbionts including eukaryotic organelles. I developed a theoretical model to study the lateral gene transfer process between cell organelles (but extendible to other endosymbionts) and the cell nucleus. The model explores the role of the lack of recombination in the organelles (Muller''s ratchet) as well as deviations from Muller''s ratchet in the form of non-symmetrical homologous recombination in relation with the gene transfer process. Also, nuclear incompatibilities resulting from the inclusion of a transferred gene, and cyto-nuclear incompatibilities between the mutant endosymbiotic genomes and the modified nuclear genome are investigated. The results obtained show that under certain circumstances the existence recombination or its non-existence produce the same results, and that deviations from symmetry in the recombination process might have important effects on the frequency of different alleles. It is also clear that there is a strong relation between nuclear and endosymbiotic genomes, and that the evolutionary fate of one largely depends on the forces affecting the other. When nuclear and cyto-nuclear incompatibilities are introduced in the model, the results show that lateral gene transfer-induced incompatibilities could potentially play a role in the speciation process similar to the one produced by mitochondria in the Nasonia species.

In der eukaryotischen Ontogenese ist die Etablierung differenzierungsspezifischer Genexpression eng an die Unterteilung des Genoms in funktionell getrennte Domänen gekoppelt. Solche Domänen lassen entweder erhöhte transkriptionelle Aktivität zu oder unterdrücken sie und werden Eu- bzw. Heterochromatin genannt. Heterochromatin enthält spezielle Proteine, die zur Ausbildung dieser repressiven Chromatinstruktur beitragen. Eine der Hauptfragen in der Heterochromatinbiologie ist, wie solche Proteine rekrutiert werden. Dieser Prozess ist entscheidend damit einzelnde Regionen im Genom koordiniert zeit- und ortsabhängig reprimiert werden können. In Saccharomyces cerevisiae entsteht Hetero-chromatin an den silent-mating-type Loci HMRa und HMLalpha durch die zielgerichtete Rekrutierung des Sir-Komplexes über eine Gruppe von Proteinen, die an sogenannte silencer-DNA Sequenzen binden. In diese Arbeit wird gezeigt, daß das Protein Sum1, bisher bekannt als Repressor meiotischer Gene im vegetativen Zellzyklus, als Heterochromatin-Rekrutierungsfaktor für HMLalpha fungiert. Sum1 konnte in vitro und in vivo an HMLalpha über ein funktionelles Element innerhalb des HML-E silencers binden und die Deletion von SUM1 verursachte einen Verlust von Repression an HMLalpha. SUM1 beeinflußte außerdem die Fähigkeit von HML-E als Replikationsstartpunkt (origin) zu agieren, was eine Rolle von Sum1 in der Replikation nahelegt. Die Beobachtung, daß orc2-1 und orc5-1 mit sum1Delta synthetisch lethal waren und daß cdc6-1, cdc7-1 oder cdc45-1 mit sum1Delta einen synthetischen Wachstumsdefekt aufwiesen unterstützt die Vermutung, daß SUM1 eine globale Rolle in der Replikationsinitiation besitzt. In einer genomweiten Suche wurden ARS Elemente gefunden, die sowohl Sum1 als auch ORC rekrutieren. Dabei konnte gezeigt werden, daß die Replikationsaktivität dieser ARS Elemente von Sum1 bzw. Sum1 Bindungsstellen abhängig war. Als Repressor von meiosespezifischen Genen interagiert Sum1 oft mit der Histondeacetylase Hst1. In diesem Zusammenhang konnte gezeigt werden, daß SUM1-regulierte origins ebenfalls HST1 zur vollen Aktivität benötigten. Zusammenfassend schlagen wir Sum1 als neuartigen Modulator für die Replikationsinitiation an einer Untergruppe chromosomaler Replikationsstartpunkte vor. / The division of eukaryotic chromatin into functionally distinct domains is critical to implement gene expression programs that drive the development of multicellular organisms. Regions termed euchromatin exist in the genome that are generally conducive to transcription, whereas heterochromatin contains specialized chromatin binding proteins that repress transcription in these regions. A central question in heterochromatin biology is how the heterochromatin factors are targeted to specific genomic regions, a process that is crucial to ensure that the designated domains, and only they, are repressed in the appropriate spatial and temporal fashion. In Saccharomyces cerevisiae heterochromatinization at the silent mating-type loci HMRa and HMLalpha is achieved by targeting the Sir complex to these regions via a set of anchor proteins that bind to the silencers. Here, we have identified a novel heterochromatin targeting factor for HMLalpha, the protein Sum1, a repressor of meiotic genes during vegetative growth. Sum1 bound both in vitro and in vivo to HMLalpha via a functional element within the HML-E silencer, and deletion of SUM1 caused HMLalpha derepression. Significantly SUM1 was also required for origin activity of HML-E, suggesting a role of Sum1 in replication initiation. Our observations of a synthetic lethality between orc2-1 or orc5-1 and sum1Delta as well as a synthetic growth defect of cdc6-1, cdc7-1 and cdc45-1 with sum1Delta support the notion that SUM1 has a global role in replication initiation. In a genome-wide search for Sum1-regulated origins, we identified a set of autonomous replicative sequences (ARS elements) that bound both the origin recognition complex and Sum1. Full initiation activity of these origins required Sum1, and their origin activity was decreased upon removal of the Sum1 binding site. In its role as a repressor of meiosis specific genes, Sum1 often works in concert with the histone deacetylase Hst1. We found that SUM1-regulated origins also required HST1 for full activity. Taken together we propose that Sum1 is a novel replication initiation modulator for a subset of chromosomal origins.

Submitted by JOSIANE SANTOS DE OLIVEIRA (josianeso) on 2018-09-24T12:37:05Z No. of bitstreams: 1 Tatiana Dilly Dexheimer_.pdf: 564022 bytes, checksum: dc30c0b4e680aa78777ee81a76700bb5 (MD5) / Made available in DSpace on 2018-09-24T12:37:05Z (GMT). No. of bitstreams: 1 Tatiana Dilly Dexheimer_.pdf: 564022 bytes, checksum: dc30c0b4e680aa78777ee81a76700bb5 (MD5) Previous issue date: 2018-06-19 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Esta dissertação de mestrado é um subprojeto de um projeto maior, intitulado “A mobilização do saber e do fazer: episteme e deonticidade na fala-em-interação institucional e na conversa cotidiana”. (OSTERMANN, 2016). Investigamos o tema de respostas não-conformativas no português brasileiro como resposta a perguntas polares e perguntas abertas do tipo específicas. O contexto de pesquisa é um hospital do SUS, localizado no sul do Brasil, e as interações gravadas consistem em aconselhamentos genéticos, ecografias obstétricas e morfológicas e ecocardiografias. A análise dos dados é realizada com base no aparato teórico-metodológico da Análise da Conversa. (SACKS; SCHEGLOFF; JEFFERSON, 1974). O objetivo desta dissertação é analisar e descrever as ações e implicações das respostas não-conformativas fornecidas por médicos/as a gestantes de médio e alto risco. Os resultados da investigação mostram que, no contexto investigado, as respostas não-conformativas apresentam características similares as respostas não-conformativas transformativas, indiretas e oracionais descritas em estudos anteriores em outras línguas. (STIVERS, HAYASHI, 2010; WALKER; DREW; LOCAL, 2011; FOX; THOMSPON, 2011). Evidenciamos, contudo, que as respostas transformativas de especificação e substituição geram trabalho interacional maior por parte do respondente, assim como as respostas transformativas de foco e pressuposição. Além disso, identificamos um tipo de resposta não-conformativa não descrita anteriormente na literatura: respostas transformativas que postergam o provimento da informação. Esse tipo de resposta foi identificado tanto como resposta para perguntas polares quanto para perguntas abertas do tipo específicas. Observamos que esse tipo de resposta ocorre em momentos que se revelam como interacionalmente ‘delicados’, tais como, comunicação de más notícias, de risco de morte fetal e de riscos decorrentes de procedimentos. Concluímos que as respostas transformativas que postergam o provimento da informação são uma forma de ajustar a fala levando em consideração o interlocutor e a potencial delicadeza das ações envolvidas naquela sequência interacional. Por fim, entendemos que as respostas transformativas permitem ao interagente resistir às imposições da pergunta e controlar, também, a pauta da interação. / This master’s dissertation consists of a subproject of a larger study, entitled “A mobilização do saber e do fazer: episteme e deonticidade na fala-em-interação institucional e na conversa cotidiana”. (OSTERMANN, 2016). It investigates non-conformative answers in Brazilian Portuguese to polar questions and wh-questions, more specifically, specifying questions. All the consultations audio recorded were held at a public health system hospital located in southern Brazil and the interactions consist of genetic counseling, morphological and fetal ultrasound and echocardiography. The data analysis is based on a conversational analytical perspective. (SACKS; SCHEGLOFF; JEFFERSON, 1974). The objective of this dissertation is to analyze and describe the actions and implications of non-conformative answers provided by doctors to moderate and high pregnancy women. The results indicate that non-conformative answers and transformative, indirect and clausal responses present similar characteristics. (STIVERS, HAYASHI, 2010; WALKER; DREW; LOCAL, 2011; FOX; THOMSPON, 2011). However, transformative answers of specification and replacement engender greater interactive work for the respondent, as well as transformative answers of focus and presupposition. Additionally, we have identified a kind of transformative answer that has not been described in the literature yet: transformative answers that postpone the provision of the information. These kind of answers have been identified as answers to polar questions as well as wh-questions on the kind of specifying questions. We have observed that these answers occur in moments that reveal to be interactionally delicate, for instance, delivery of bad news, fetal death risk, and risks arising from medical procedures. We have concluded that transformative answers that postpone the provision of the information are a way to adjust the talk taking into consideration the interlocutor and the potential delicacy of the actions involved in that interactional sequence. Finally, we understand that transformative answers allow the interlocutor to resist to the constraints of the questions and also control the agenda of the interaction.

HIV, syphilis, malaria, and anaemia are major causes of adverse pregnancy outcomes in sub-Saharan Africa (SSA). Despite global and national policies advocating for screening of these conditions, only HIV testing has achieved good coverage, precluding early detection and appropriate management in pregnancy. Rapid pointof-care tests (POCTs) provide an opportunity to integrate diagnosis and provide timely treatment of these conditions in rural antenatal care (ANC) settings. After an introductory chapter, a review of the literature on these four conditions in pregnancy is presented with a focus on SSA. The thesis then shifts attention to Kenya, a country that embodies many of the disease challenges and health system characteristics of the region. Kenyan ANC policy recommends testing for HIV, syphilis and anaemia and preventive strategies for malaria. The following chapters are comprised of three linked studies conducted in western Kenya, that use different methods to progressively investigate the implementation success of integrated point-of-care testing (POCT) for HIV, syphilis, malaria and anaemia at seven peripheral dispensaries. Baseline data confirmed that testing requirements for syphilis, malaria and anaemia are not currently met at dispensary level. We implemented an intervention where test kits were supplied and training plus supervision were provided to enable healthcare workers to conduct integrated POCT for pregnant women. Adoption and fidelity were measured quantitatively using exit interviews, antenatal registers and proficiency scores (Study 1: Integrating point-of-care testing (POCT) for HIV, syphilis, malaria and anaemia in antenatal care at dispensary level in western Kenya: an implementation study) while acceptability, appropriateness and feasibility were assessed qualitatively (Study 2: Exploring healthcare workers and pregnant women’s perspectives on appropriateness, acceptability and feasibility of integrating point-of care testing: A qualitative study). Our findings show that the innovation was highly adopted, meaning almost all pregnant women received the essential tests. This was supported by the qualitative findings where healthcare workers and pregnant women found the innovation acceptable and appropriate. However, fidelity to clinical management guidelines can still be improved. Our qualitative findings provide some explanation for these gaps. One common sentiment among interviews with healthcare workers was that workload was perceived to be a barrier to providing quality care. We explored this further with discrete-event simulation modelling (Study 3: Investigating the operational impact of integrating HIV, syphilis, malaria and anaemia point-of-care testing in antenatal care clinics in western Kenya: a discrete event simulation model) and found the healthcare workers were actually under-utilized. This suggests that nurses should, in theory, have sufficient time to deliver essential ANC services. While integrating POCT addresses one gap, additional interventions to support and supervise healthcare workers are needed to ensure appropriate and high quality of care. An integrated approach to health systems strengthening and more investment in implementation and translation research using multi-methods are needed.

Diese Habilitationsschrift konzentriert sich auf die Evolution osteologischer Korrelate, anhand derer Rückschlüsse auf die Struktur der Haut sowie die Art der Atmung und der Nahrungsaufnahme früher Tetrapoden gezogen werden können. Die äußere Skulptur der Hautknochen früher Tetrapoden trug zur Konsolidierung der darüber liegenden Dermis bei; dies und die zahlreichen Hautverknöcherungen deuten darauf hin, dass Hautatmung im Gegensatz zu heutigen Lissamphibien keinen wesentlichen Teil des Gasaustausches ausmachen konnte. Es wird die Hypothese aufgestellt, dass eine weitere Funktion der Knochenskulptur in der Pufferung von CO2-induzierter Azidose bestand, wenn sich die Tiere an Land aufhielten. Mehrere Gruppen früher Tetrapoden entwickelten unabhängig voneinander Osteoderme im Rumpf, die unter anderem die Wirbelsäule bei der Fortbewegung an Land unterstützen konnten oder als Kalzium-Reservoir dienten. Die Veränderungen in der Morphologie, Histologie und Anordnung der Dermalschuppen während des Fisch-Tetrapoden-Überganges erfolgte etwa simultan zur Entwicklung der Tetrapodenextremität und ermöglichte eine größere Flexibilität des Körpers und der Beine. Der Bau des Kiemenskeletts ihrer Fischvorfahren wurde bei einer unerwartet großen Anzahl früher Tetrapoden beibehalten. Dies zeigt, dass viele frühe Tetrapoden sowohl der Stamm- als auch der Kronengruppe als Adulte über innere Kiemen atmeten, während ihre Larven äußere Kiemen besaßen. Das Kiemenskelett und die Rekonstruktion assoziierter Muskeln zeigen, dass viele aquatische frühe Tetrapoden ihre Nahrung durch akinetisches Saugschnappen erbeuteten. Trotz spärlicher Fossilüberlieferung gibt es Hinweise darauf, dass die ersten an Land fressenden Tetrapoden noch keine bewegliche Zunge besaßen. Ein ontogenetischer Umbau des larvalen, kiementragenden Kiemenskelets in einen zungenstützenden Apparat kann bei frühen Tetrapoden nur in Ausnahmefällen belegt werden. / This habilitation thesis focuses on the evolution of skeletal correlates from which the integumentary structure and the mode of breathing and feeding in early tetrapods can be inferred. Sculpture on the external surface of dermal bones contributed to the consolidation of a dense integument; this and numerous dermal ossifications render substantial cutaneous respiration as in extant lissamphibians unlikely. It is hypothesised that a further function of dermal bone sculpture was to buffer CO2-induced acidosis while the animals were on land. Numerous early tetrapods independently developed osteoderms in the trunk that may have served among other things for support of the vertebral column in terrestrial locomotion or as a physiological calcium reservoir. The alterations in morphology, histology and arrangement of dermal scales during the fish-to-tetrapod transition occurred at roughly the same time when digits appeared and enabled greater flexibility of body and limbs. The structure of the hyobranchium of their fish-like ancestors was retained in an unexpected large number of early tetrapods, showing that many early tetrapods of both the stem- and crown-group breathed via the associated internal gills as adults, and via external gills as larvae. The hyobranchial apparatus and reconstruction of the associated musculature indicate that many aquatic early tetrapods fed by akinetic suction feeding. There is evidence that the earliest terrestrial tetrapods captured prey on land by jaw prehension rather than by tongue based feeding. Ontogenetic remodelling into a tongue supporting adult hyobranchium in early tetrapods can only be demonstrated in exceptional cases.

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