Autologous coculture of primary human alveolar epithelial cells and macrophages for evaluating the safety and efficacy of novel inhalation pharmaceuticals

Animal experiments are general practice to study safety and to estimate the efficacy of orally inhaled drugs in preclinical trials. The relevance of these data for humans is questionable. The aim of this thesis is the development of a relevant in vitro model of the air-blood barrier that can better predict the human in vivo situation. The first chapter describes how particle uptake by alveolar macrophages can be visualized in vitro by correlative light and electron microscopy (CLEM). The second chapter focuses on the development of a cellular in vitro model addressing the alveolar space. The human alveolar autologous coculture consists of primary alveolar type I-like pneumocytes cocultured with primary alveolar macrophages from the same human donor. The model demonstrated its use to investigate cell-particle inreractions at the air-liquid interface. Only macrophages engulfed foreign particles in the in vitro model visualized by CLEM. The ability of the autologous coculture to mimic inflammatory processes in the lung is the focus of the third chapter. The in vitro model showed a typical interleukin release of inflammatory markers after stimulation with lipopolysaccharides. A subsequent treatment with IL-10 loaded particles counterregulated the inflammation.Tierversuche sind gängige Praxis um Unbedenklichkeit oder Wirksamkeit von Inhalanda in präklinischen Studien zu untersuchen. Die Relevanz der durch Tierversuche generierten Daten für den Menschen wird zunehmend in Frage gestellt. Das Ziel dieser Arbeit ist deshalb die Entwicklung eines für den Menschen relevanten in vitro Modells der Blut-Luft-Schranke zur genaueren Vorhersage der humanen in vivo Situation. Im ersten Kapitel wird beschrieben, wie die Aufnahme von Partikeln in Alveolarmakrophagen mit Korrelativer Licht- und Elektronenmikroskopie (CLEM) in vitro visualisiert werden kann. Das zweite Kapitel legt den Fokus auf die Entwicklung eines zellulären alveolaren in vitro Modells. Die humane autologe Kokultur besteht aus primären alveolaren Typ-I ähnlichen Pneumozyten, welche mit primären Alveolarmakrophagen des gleichen Spenders kokultiviert werden. Das Modell erwies sich als geeignet, um Zell-Partikel-Interaktionen zu untersuchen, wobei die Partikel direkt aus der Luft auf die Zelloberfläche abgeschieden wurden. Mittels CLEM konnte gezeigt werden, dass es lediglich die Makrophagen sind, die die Fremdpartikel aufnehmen. Der Fokus des dritten Kapitels liegt auf der Fähigkeit der autologen Kokultur entzündliche Prozesse nachzuahmen. Das in vitro Modell zeigte eine typische Freisetzung entzündlicher Marker nach Stimulation mit Lipopolysacchariden. Eine anschließende Behandlung mit IL-10 beladenen Partikeln führte zu einer anti-inflammatorischen Wirkung

The Relationship Between Stress, Social Problem Solving, and Psychological Distress in Mothers of Daughters with Eating Disorders

Mothers of daughters with eating disorders are at risk for experiencing high levels of psychological distress, which can have negative implications for mothers themselves and the children under their care. Coping strategies that effectively manage the stress of the caregiving role have been found to reduce psychological distress, but the literature does not clearly define the type of coping that is best indicated for this population of caregivers. Coping via social problem solving has been found to predict less psychological distress and moderate the stressdistress relationship in many caregiver populations. The relationship between stress, coping via social problem solving, and psychological distress was explored; it was hypothesized that stress related to being a mother of a daughter with an eating disorder and social problem solving would predict the level of experienced psychological distress, and that social problem-solving skills would moderate the stress-distress relationship. The present study was able to replicate the finding that mothers of daughters with eating disorders are highly distressed, and was unsuccessful at finding social problem-solving skills to predict psychological distress or moderate the relationship between stress and distress. Implications of these findings are discussed

Qualitative Meta-analysis on Three Aspects of the Federal Budget: The Process, Politics and Policies

Over the last few years, policymakers and the media have paid special attention to the federal budget, deficits and the national debt as the recent recession plunged the nation into the red. In response, some called for fiscal tightening while others simultaneously advocated for further stimulus spending to accelerate the recovery. Beyond the debate on how to respond to the short-term deficits caused by the recession and a sluggish recovery are discussions of looming budgetary struggles. The medium to long-term budget concerns are driven primarily by the United States’ rapidly aging society and rising health care costs. These two factors are of chief concern because Medicare, Medicaid, and Social Security comprise a large portion of annual federal spending. Some respond to these warnings by emphasizing the unreliability that inevitably results when one attempts to project into the future. Others counter that the recent recession has led to historically high levels of debt and the United States must proactively reduce the level of debt relative to the economy in order to provide flexibility for additional debt accumulation should these programs or another unforeseen event strain the nation’s finances

God, self, humanity, Earth: Christian ecological ethics in local contexts

The complex issues of environmental justice require a contextualized approach, analyzing the intricacy of global ecological issues and studying local communities in which ecological issues are entangled with other concerns, such as ensuring economic survival and protecting the viability of local communities. Such contextual analysis is needed to guide Christians in making ethical decisions. This dissertation utilizes an ethnographic case study of hydraulic fracturing (fracking) in two local communities in Northeastern Pennsylvania to describe and analyze the global and local tensions created by the social, economic, and environmental effects of these practices. The description and analysis of the case study contributes to the development of a contextual Christian ecological ethical framework that is forged at the intersection of lived experience and Christian theological ethical principles and practices. Lutheran and ecofeminist theological perspectives provide a framework for a contextualized Christian ethic that identifies and negotiates through the economic and ecological tensions present for individuals and communities facing ecological injustice, resulting in the development of ethical principles that enable practical action in establishing justice within communities. By reshaping the dialectical method found in Martin Luther’s theology, feminist intersectional theory, and middle-axiom ethical approaches, this dissertation argues that a contextualized ethical framework can be employed in local situations of ecological injustice for a more realistic and potentially transformative outcome. Specifically, I engage the coram relational model found in Luther’s writing, as described by Gerhard Ebeling and interpreted by Lutheran feminist theologian Caryn D. Riswold. My theological ethical approach is to engage people in dialectical reflection on their relationships with God, themselves as individuals, humanity, and the Earth. Through investigating these relationships, individuals come to understand the self as a complex manifestation of identities that influence the ethical decision-making process. Based on the self-knowledge gained in these reflections, community members can participate in dialogue to develop ethical principles in the form of middle axioms that can guide their community as a whole toward practical action. The case study analysis reveals how dialectical reflection and the development of middle axioms build upon each other, enabling practical ethical action in communities facing ecological and economic injustice

Mitochondria-encoded genes contribute to evolution of heat and cold tolerance in yeast

Genetic analysis of phenotypic differences between species is typically limited to interfertile species. Here, we conducted a genome-wide noncomplementation screen to identify genes that contribute to a major difference in thermal growth profile between two reproductively isolated yeast species, Saccharomyces cerevisiae and Saccharomyces uvarum. The screen identified only a single nuclear-encoded gene with a moderate effect on heat tolerance, but, in contrast, revealed a large effect of mitochondrial DNA (mitotype) on both heat and cold tolerance. Recombinant mitotypes indicate that multiple genes contribute to thermal divergence, and we show that protein divergence in COX1 affects both heat and cold tolerance. Our results point to the yeast mitochondrial genome as an evolutionary hotspot for thermal divergence.This work was supported by the NIH (grant GM080669) to J.C.F. Additional support to C.T.H. was provided by the USDA National Institute of Food and Agriculture (Hatch project 1003258), the National Science Foundation (DEB-1253634), and the DOE Great Lakes Bioenergy Research Center (DOE BER Office of Science DE-SC0018409 and DE-FC02-07ER64494 to T. J. Donohue). C.T.H. is a Pew Scholar in the Biomedical Sciences and a Vilas Faculty Early Career Investigator, supported by the Pew Charitable Trusts and the Vilas Trust Estate, respectively. D.P. is a Marie Sklodowska-Curie fellow of the European Union’s Horizon 2020 research and innovation programme (grant agreement no. 747775).Peer reviewe