Pruning at the cerebellar climbing fibre synapse: synaptic efficacy and glial involvement

Pruning, the elimination of excess synapses is a phenomenon of fundamental importance for correct wiring of the central nervous system. The establishment of the cerebellar climbing fiber (CF)-to-Purkinje cell (PC) synapse provides a suitable model to study pruning and pruning-relevant processes during early postnatal development. Until now the role of microglia in pruning remain under intense investigation. Here, we analyzed migration of microglia into the cerebellar cortex during early postnatal development and their possible contribution to the elimination of CF-to-PC synapses. Microglia enrich in the Purkinje cell layer at pruning-relevant time points giving rise to the possibility that microglia are actively involved in synaptic pruning. We investigated the contribution of microglial fractalkine (CX3CR1) signaling during postnatal development using genetic ablation of the CX3CR1 receptor and an in–depth histological analysis of the cerebellar cortex.:1 Introduction 6 1.1 Origin of microglia 6 1.2 Synaptic refinement 7 1.3 Fractalkine Receptor CX3CR1 9 1.4 Climbing fiber maturation and PCL development 10 1.5 Aim of the study 12 2 Materials and Methods 13 2.1 Materials 13 2.1.1 General material 13 2.1.1.1.1 Hardware 13 2.1.1.1.2 Consumable supplies 13 2.1.2 Chemicals 14 2.1.3 Solutions 14 2.1.4 Animals 14 2.1.5 Primary Antibodies 15 2.1.6 Secondary Antibodies 15 2.1.7 Software 15 2.2 Methods 15 2.2.1 Genotyping 15 2.2.2 Fixation and cryopreservation 16 2.2.3 Fluorescence Immunohistochemistry 16 2.2.4 Quantification of Microglia in the Cerebellum 16 2.2.5 Assessment of VGluT2 in the cerebellum 18 2.2.6 Statistical Analysis 19 3 Results 20 3.1 Postnatal enrichment of microglia cells in the cerebellar Purkinje cell layer 20 3.2 Microglial proximity to Climbing fibers 22 3.3 Population of the granular und molecular layer of CX3CR1 knock-out mice during early postnatal development 23 3.4 Influence of CX3CR1 knock-out on microglial morphology 25 3.5 Influence of CX3CR1 deletion on the VGluT2 expression during postnatal development 29 4 Discussion 31 4.1 Role of microglia in the developing cerebellum 31 4.2 CX3Cr1 Signaling and influence on microglial motility and morphology 32 4.3 CX3CR1 signaling and synaptic pruning 33 4.4 CX3CR1 signaling and formation of functional synapses 34 4.5 Correlation of immunohistological data with electrophysiological findings 35 5 Summary and conclusion 36 6 Zusammenfassung der Arbeit 38 7 References 41 8 Erklärung über die eigenständige Abfassung der Arbeit 49 9 Publications 5

Regulation of the Pregnane X Receptor Signaling Pathway

Liver-enriched nuclear receptors (NRs) collectively function as metabolic and toxicological `sensors' that mediate liver-specific gene-activation in mammals. NR-mediated gene-environment interaction regulates important steps in the hepatic uptake, metabolism and excretion of glucose, fatty acids, lipoproteins, cholesterol, bile acids, and xenobiotics. While it is well-recognized that ligand-binding is the primary mechanism behind activation of NRs, recent research is revealing that multiple signal transduction pathways modulate NR-function in liver. The interface between specific signal transduction pathways and NRs helps to determine their overall responsiveness to various environmental and physiological stimuli. The pregnane x receptor (PXR, NR1I2) was identified in 1998 as a member of the NR superfamily of ligand-activated transcription factors. PXR is activated by a broad range of lipophilic compounds in a species-specific manner. The primary function ascribed to PXR is the homeostatic control of steroids, bile acids, and xenobiotics. This function is mediated through PXR's ability to coordinately activate gene expression and regulate the subsequent activity of phase I and phase II metabolic enzymes, as well as several membrane transporter proteins. While PXR likely evolved primarily to protect the liver from toxic assault, its activation also represents the molecular basis for an important class of drug-drug, herb-drug, and food-drug interactions. While ligand binding is the primary mode of PXR activation, several signal transduction pathways interface with the PXR protein to determine its overall responsiveness to environmental stimuli. Multiple signaling pathways modulate the activity of PXR, likely through direct alteration of the phosphorylation status of the receptor and its protein cofactors. Therefore, specific combinations of ligand binding and cell signaling pathways affect PXR-mediated gene activation and determine the overall biological response. This dissertation contributes to the molecular understanding of the regulation of PXR by novel agonists, cAMP-dependent protein kinase (PKA) signaling, and phosphorylation. The results presented here were primarily obtained from mouse and tissue culture systems. This dissertation identifies Tian Xian, a traditional Chinese herbal anti-cancer remedy, as a novel PXR activator. This evidence suggests that Tian Xian should be used cautiously by cancer patients taking chemotherapy due to its potential to increase the metabolism of co-administered medications. In addition, data presented here show that activation of PKA signaling modulates PXR activity in a species-specific manner. It is further revealed that PXR exists as phospho-protein in vivo and that the activation of PKA signaling modulates the phospho-threonine status of PXR. Finally, the potential phosphorylation sites within the PXR protein are identified. These phosphorylation sites are characterized, using a phosphomimetic and phospho-deficient site-directed mutagenesis based approach, based on their ability to modulate PXR activity. Taken together, the work presented in this dissertation contributes to understanding the interface between ligands, signal transduction pathways and PXR activity, which is critical for the development of safe and effective therapeutic strategies

Betriebliche Aus- und Weiterbildung in den süddeutschen Metropolregionen

Die Wirtschaftskraft und Wettbewerbsfähigkeit einer Region hängen gerade vor dem Hintergrund des demographischen Wandels und dem daraus folgenden Fachkräftemangel ganz wesentlich von der Qualifikation der Beschäftigten sowie der Bevölkerungsstruktur und -entwicklung ab. Der Forschungsbericht untersucht anhand verschiedener Datenquellen die Entwicklung und Qualifikation der Bevölkerung sowie die Aus- und Weiterbildung der Beschäftigten in den vier süddeutschen Metropolregionen Nürnberg, München, Stuttgart und Frankfurt/Rhein-Main. Auch wenn sich die Strukturen der Metropolregionen recht deutlich voneinander unterscheiden, so wird offensichtlich, dass insbesondere die Metropolregion Nürnberg, auf der ein besonderer Focus in dieser Arbeit gelegt wird, sehr stark von den Folgen des demographischen Wandels betroffen ist. Dennoch bieten die vorhandenen Stärken der Metropolregion, wie beispielsweise bei der betrieblichen Berufsausbildung, eine gute Ausgangsposition, wie dem zukünftigen Fachkräftemangel begegnet werden kann. Desweiteren können ungenutzte Potentiale durch einen guten Mix der verschiedenen Formen der beruflichen Bildung erschlossen werden.Economic power and competiveness of a region depend to a great extent on the qualification of employees and on the demographic structure and the development of the population, especially because of demographic change and skilled worker shortage in future. This paper analyses the qualification and development of the population as well as the vocational training and further training of employees within the four metropolitan areas Nuremberg, Munich, Stuttgart and Frankfurt/Rhein-Main in southern Germany. Even though the structures are quite different among the metropolitan areas, especially the metropolitan area of Nuremberg will be affected quite hard by the demographic change. However this metropolitan area is very active in apprenticeship training and therefore has a good basis for preventing skilled worker shortage in future. By using different forms of further education it is also possible to strengthen the competiveness of the metropolitan area in future

“I Knew the Mechanics of Hip”: Fashioning Bodies in the Works of Joni Mitchell, 1968 - 1979

Since the late 1960s, Joni Mitchell has been best known for her remarkable music: her voice, compelling lyrics, and unusual guitar tunings, all of which have conveyed sentiments of her generation and have remained relevant to later generations. Although her lyrics and album covers are replete with references to fashion, textiles, dress, and appearance, they have not been critically analyzed in relation to her status as a cultural icon. The fashion industry is clearly aware of this status, as evidenced, for example, by her inclusion in a 2015 Yves St. Laurent campaign. In our study, we ask: (1) What does Mitchell’s music convey about fashion, textiles, dress, and appearance? (2) What fashion philosophies and themes are found in her vocal and visual arts? To answer these questions, we completed a content analysis of Mitchell’s lyrics, alongside a visual analysis of her album art. By studying the period of 1968 to 1979, our research captures the epoch of her greatest radio influence and record sales, and identifies the messages and fashion philosophies threaded throughout her songs and artwork

Betriebliche Aus- und Weiterbildung in den süddeutschen Metropolregionen (In-house training and further training in Southern German metropolitan areas)

"Economic power and competiveness of a region depend to a great extent on the qualification of employees and on the demographic structure and the development of the population, especially because of demographic change and skilled worker shortage in future. This paper analyses the qualification and development of the population as well as the vocational training and further training of employees within the four metropolitan areas Nuremberg, Munich, Stuttgart and Frankfurt/Rhein-Main in southern Germany. Even though the structures are quite different among the metropolitan areas, especially the metropolitan area of Nuremberg will be affected quite hard by the demographic change. However this metropolitan area is very active in apprenticeship training and therefore has a good basis for preventing skilled worker shortage in future. By using different forms of further education it is also possible to strengthen the competiveness of the metropolitan area in future." (Author's abstract, IAB-Doku) ((en))Metropolregion, demografischer Wandel, Wirtschaftsstruktur, Qualifikationserwerb, Qualifikationsstruktur, betriebliche Berufsausbildung, betriebliche Weiterbildung, Hochschulbildung, Ausbildungsquote, Ausbildungsplatzangebot, IAB-Betriebspanel, Süddeutschland, Rhein-Main-Gebiet, München, Nürnberg, Stuttgart, Baden-Württemberg, Bayern, Bundesrepublik Deutschland

Live donor study – implications of kidney donation on cardiovascular risk with a focus on lipid parameters including lipoprotein a

In this prospective observational cohort study, we evaluate the change in cardiovascular risk parameters, with a focus on lipids, in live kidney donors 1 year post donation. Body mass index, systolic/diastolic blood pressure, kidney function (chromium-51 ethylenediaminetetraacetic acid estimated glomerular filtration) and lipid parameters were measured at baseline and 1 year. Data on 87 live kidney donors were collected. Body mass index increased from 26.5 ± 2.7 pre to 27.4 ± 3.0 kg/m post donation (p < 0.0001). Chromium-51 ethylenediaminetetraacetic acid estimated glomerular filtration decreased from 111.8 ± 20.0 pre to 72.1 ± 13.1 mL/min/1.73 m post donation (p < 0.0001). Serum triglyceride levels increased from 0.8 (interquartile range 0.6–1.3) pre to 1.0 mmol/L (interquartile range 0.7–1.6) post donation (p = 0.0004). Statin use increased from 11.5% pre to 21% post donation (p < 0.005). Low-density lipoprotein remained stable, and other lipids (high-density lipoprotein, apolipoprotein B and lipoprotein a) did not change post donation

Undisturbed climbing fiber pruning in the cerebellar cortex of CX3CR1-deficient mice

Pruning, the elimination of excess synapses is a phenomenon of fundamental importance for correct wiring of the central nervous system. The establishment of the cerebellar climbing fiber (CF)-to-Purkinje cell (PC) synapse provides a suitable model to study pruning and pruning-relevant processes during early postnatal development. Until now, the role of microglia in pruning remains under intense investigation. Here, we analyzed migration of microglia into the cerebellar cortex during early postnatal development and their possible contribution to the elimination of CF-to-PC synapses. Microglia enrich in the PC layer at pruning-relevant time points giving rise to the possibility that microglia are actively involved in synaptic pruning. We investigated the contribution of microglial fractalkine (CX3CR1) signaling during postnatal development using genetic ablation of the CX3CR1 receptor and an in-depth histological analysis of the cerebellar cortex. We found an aberrant migration of microglia into the granule and the molecular layer. By electrophysiological analysis, we show that defective fractalkine signaling and the associated migration deficits neither affect the pruning of excess CFs nor the development of functional parallel fiber and inhibitory synapses with PCs. These findings indicate that CX3CR1 signaling is not mandatory for correct cerebellar circuit formation. Main Points - Ablation of CX3CR1 results in a transient migration defect in cerebellar microglia. - CX3CR1 is not required for functional pruning of cerebellar climbing fibers. - Functional inhibitory and parallel fiber synapse development with Purkinje cells is undisturbed in CX3CR1-deficient mice

Unearthing Franklinton's Potential: Cultivating a Vibrant Foodscape

This presentation will highlight some results of a three-year endeavor with Franklinton Farms to improve healthy food access through a neighborhood community supported agriculture (CSA) program, strengthen community self-reliance and inspire community empowerment through cooking classes and workshops on nutrition, food processing, and leadership, engage residents in different parts of the food system, preserve land for urban agriculture, and create inviting urban farm spaces. The Franklinton neighborhood (Census Tracts 30, 32, 40, 42, 43, 50, 51) is an impoverished urban Appalachian neighborhood of 10,000 people on the near-west side of Columbus, Ohio plagued by low educational attainment, drugs, housing decay, historical disinvestment, and consequential obesity, food insecurity, chronic diet-related disease, and high infant mortality rates. 1-4 Health and wellness are a serious problem in the neighborhood as access to adequate health care and a healthy food environment is difficult to realize for residents. An Ohio State Food Mapping Team (FMT) surveyed 74 Franklinton residents in 2014. Of those who were interviewed, 44.4 percent reported high blood pressure, 34.7 percent reported high cholesterol, 18.1 percent reported pre-diabetes, and 13.5 percent reported Type II Diabetes. Underlying many of these issues are the high rates of poverty and high rates of food insecurity. The FMT found that 60.5 percent of all participants were food insecure (low and very low food secure combined); and 31 percent were very low food secure. Access to healthy, fresh food is hampered by the lack of a full-service grocery store in or near Franklinton. At least 30 percent of households do not have a vehicle. Restricted access to transportation means that residents buy their food at corner stores (49 percent), fast food outlets (77 percent), and dollar stores (58 percent). FMT survey participants stated that nutrition (89 percent) and price (91 percent) were important aspects of their food purchasing decisions, while 38 percent were not satisfied with being able to access the food they wanted and did not find it easy to find fresh produce. Barriers to food choices included price, transportation, and distance to the food store. This presentation will highlight the results of the CSA portion of the project, in addition to challenges experience and opportunities for learning if other groups are interested in such a project. We have had more than 100 households participate in the CSA and many have participated in our pre- and post-season surveys. Specifically, we will share about participants’ (including the 50 percent who are low-income, paying half price) experiences using quantitative and qualitative data related to food security, dietary patterns, sense of community, and food access. Additional partners have included the Ohio State College of Social Work, AmeriCorps VISTA, Mount Carmel Healthy Living Center, and Local Matters.AUTHOR AFFILIATION: Michelle Kaiser, associate professor, Ohio State College of Social Work, [email protected] (Corresponding Author); Nick Stanich, executive director, Franklinton FarmsThe authors will highlight some results of a three-year endeavor with Franklinton Farms to improve healthy food access through a neighborhood Community Supported Agriculture (CSA) program, strengthen community self-reliance, and inspire community empowerment through cooking classes and workshops on nutrition, food processing, and leadership, engage residents in different parts of the food system, preserve land for urban agriculture, and create inviting urban farm spaces. More than 100 households have participated in the CSA, and many have participated in our pre- and post-season surveys. Specifically, we will share about participants’ (including the 50 percent who are low-income, paying half price) experiences using quantitative and qualitative data related to food security, dietary patterns, sense of community, and food access

PARCS: A Safety Net Community-Based Fitness Center for Low-Income Adults

Background: Physical activity (PA) and fitness are critical to maintaining health and avoiding chronic disease. Limited access to fitness facilities in low-income urban areas has been identified as a contributor to low PA participation and poor fitness. Objectives: This research describes community-based fitness centers established for adults living in low-income, urban communities and characterizes a sample of its members. Methods: The community identified a need for physical fitness opportunities to improve residents’ health. Three community high schools were host sites. Resources were combined to renovate and staff facilities, acquire equipment, and refer patients to exercise. The study sample included 170 members older than age 18 who completed demographic, exercise self-efficacy, and quality of life surveys and a fitness evaluation. Neighborhood-level U.S. Census data were obtained for comparison. Results: The community-based fitness centers resulted from university, public school, and hospital partnerships offering safe, accessible, and affordable exercise opportunities. The study sample mean body mass index was 35 + 7.6 kg/m2 (class II obesity), mean age was 50 ± 12.5 years, 66% were Black, 72% were female, 66% completed some college or greater, and 71% had an annual household income of less than $25,000 and supported 2.2 dependents. Participants had moderate confidence for exercise participation and low fitness levels. When compared with census data, participants were representative of their communities. Conclusion: This observational study reveals a need for affordable fitness centers for low-income adults. We demonstrate a model where communities and organizations strategically leverage resources to address disparities in physical fitness and health

Towards planning and control in cognitive factories - A generic model including learning effects and knowledge transfer across system entities

Cognitive abilities allow robots to learn and reason from their environment. The gained knowledge can then be incorporated into the robot’s actions which in turn affect the environment. Therefore, a cognitive robot is no longer a static system that performs actions based on a pre-defined set of rules but a complex entity that dynamically adjusts over time. With this, challenges arise for production systems that need to observe and ideally anticipate the cognitive robot’s behavior. Often, digital twins are employed to test and optimize production control systems. This paper presents a generic approach to characterize, model and simulate learning processes and formalized knowledge in hybrid production systems assuming different station types with learning effects. Thereby, quantitative and qualitative learning processes are mapped including knowledge sharing and transfer across entities. A modular and parameterizable design enables the adjustment to different use cases. Eventually, the model is instantiated as a digital twin of a real production system for product disassembly employing cognitive-autonomous robots among human operators and rigidly automated machines. The model shows great potential to be integrated into test beds for planning and control systems of cognitive factories