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Benedikt Deuringer,1 Carmen Härdtner,2 Katja Krebs,2 Ralf Thomann,3 Martin Holzer,1 Ingo Hilgendorf,2,4,* Regine Süss1,* 1Department of Pharmaceutical Technology and Biopharmacy, Institute of Pharmaceutical Sciences, University of Freiburg, Freiburg, 79104, Germany; 2Department of Cardiology and Angiology, University Heart Center Freiburg-Bad Krozingen and Faculty of Medicine, University of Freiburg, Freiburg, 79106, Germany; 3FMF Materials Research Center, University of Freiburg, Freiburg, 79104, Germany; 4Institute for Experimental Cardiovascular Medicine, University Heart Center Freiburg-Bad Krozingen and Faculty of Medicine, University of Freiburg, Freiburg, 79110, Germany*These authors contributed equally to this workCorrespondence: Benedikt Deuringer, Pharmaceutical Technology and Biopharmacy, Sonnenstraße 5, Freiburg, 79104, Germany, Tel +49 761 203 6329, Fax +49 761 203 6326, Email bdeuringer@pharmazeutischetechnologie.dePurpose: The conventional techniques for the preparation of reconstituted high-density lipoprotein (rHDL) are hampered by long process times, the need for large amounts of starting material, and harsh preparation conditions. Here, we present a novel rHDL preparation method to overcome these challenges. Furthermore, we propose a dual mode of action for rHDL loaded with the immunosuppressant drug everolimus (Eve-rHDL) in the context of atherosclerosis and cardiovascular disease.Methods: We use dual centrifugation for rHDL nanoparticle preparation and characterize the physicochemical properties by NS-TEM, N-PAGE, DLS, AF4, and HPLC. In addition, we determine the biological efficacy in human and murine cell culture with regard to cellular uptake, cholesterol efflux, and proliferation.Results: We confirm the characteristic particle size of 10 nm, discoidal morphology, and chemical composition of the rHDL preparations and identify dual centrifugation as an ideal method for cost-effective aseptic rHDL manufacturing. rHDL can be prepared in approx. 1.5 h with batch sizes as little as 89 μL. Moreover, we demonstrate the cholesterol efflux capacity and anti-proliferative activity of Eve-rHDL in vitro. The anti-proliferative effects were comparable to free Eve, thus confirming the suitability of rHDL as a capable drug delivery vehicle.Conclusion: Eve-rHDL shows great efficacy in vitro and may further be employed to target atherosclerotic plaques in vivo. Highly effective anti-atherosclerotic therapy might be feasible by reducing both inflammatory- and lipid burden of the plaques. Dual centrifugation is an ideal technique for the efficient application of the rHDL platform in cardiovascular disease and beyond.Keywords: atherosclerosis, cholesterol efflux, drug delivery, everolimus, proliferation, protein-conjugate

This article explores and extends the capacity of the framework of inclusion and exclusion as adhered to and cultivated by the core of the field of adaptation studies. Investigating the focus on the textual, this article is a consideration of the role and position of the textual in the discourse of adaptation studies before considering three different types of adaptation in performance in Victorian Music Hall where the adaptation of the textual as well as the non-textual form a common base for a variety, if not majority, of performances. Considering popular performance outside of the confines of the theatre offers examples of adaptations based on textualities and those beyond textualities as well as a combination of the two. Examples analysed include the singer Marie Lloyd, who is probably most widely known as a Music Hall star outside of the confines of theatre history, the magician William Elsworth Robinson aka Chung Ling Soo and monkey-man Harvey Teasdale. Such opening-up of adaptation studies’ textual focus will subsequently allow a flexing, a keeping supple of any framework of inclusion and exclusion constructed.

Aims P-selectin is an activatable adhesion molecule on platelets promoting platelet aggregation, and platelet–leukocyte complex (PLC) formation. Increased numbers of PLC are circulating in the blood of patients shortly after acute myocardial infarction and predict adverse outcomes. These correlations led to speculations about whether PLC may represent novel therapeutic targets. We therefore set out to elucidate the pathomechanistic relevance of PLC in myocardial ischemia and reperfusion injury. Methods and results By generating P-selectin deficient bone marrow chimeric mice, the post-myocardial infarction surge in PLC numbers in blood was prevented. Yet, intravital microscopy, flow cytometry and immunohistochemical staining, echocardiography, and gene expression profiling showed unequivocally that leukocyte adhesion to the vessel wall, leukocyte infiltration, and myocardial damage post-infarction were not altered in response to the lack in PLC. Conclusion We conclude that myocardial infarction associated sterile inflammation triggers PLC formation, reminiscent of conserved immunothrombotic responses, but without PLC influencing myocardial ischemia and reperfusion injury in return. Our experimental data do not support a therapeutic concept of selectively targeting PLC formation in myocardial infarction.

Statins induce plaque regression characterized by reduced macrophage content in humans, but the underlying mechanisms remain speculative. Studying the translational APOE*3-Leiden.CETP mouse model with a humanized lipoprotein metabolism, we find that systemic cholesterol lowering by oral atorvastatin or dietary restriction inhibits monocyte infiltration, and reverses macrophage accumulation in atherosclerotic plaques. Contrary to current believes, none of (1) reduced monocyte influx (studied by cell fate mapping in thorax-shielded irradiation bone marrow chimeras), (2) enhanced macrophage egress (studied by fluorescent bead labeling and transfer), or (3) atorvastatin accumulation in murine or human plaque (assessed by mass spectrometry) could adequately account for the observed loss in macrophage content in plaques that undergo phenotypic regression. Instead, suppression of local proliferation of macrophages dominates phenotypic plaque regression in response to cholesterol lowering: the lower the levels of serum LDL-cholesterol and lipid contents in murine aortic and human carotid artery plaques, the lower the rates of in situ macrophage proliferation. Our study identifies macrophage proliferation as the predominant turnover determinant and an attractive target for inducing plaque regression. Supplementary Information The online version contains supplementary material available at 10.1007/s00395-020-00838-4.

Background Guidelines recommend cholesterol lowering for primary and secondary prevention of cardiovascular disease. While lipid lowering has been reported to induce plaque regression, the underlying mechanisms have remained speculative. Purpose We hypothesize that lipid uptake triggers local macrophage proliferation in the plaque, and conversely, statin treatment inhibits local macrophage proliferation leading to plaque regression. Methods Mixed bone marrow chimeras were generated in LDLR−/− mice reconstituted with wild type and scavenger receptor deficient or cholesterol exporter deficient bone marrow cells to study cell autonomous effects on macrophage proliferation. APOE*3-Leiden.huCETP mice with established atherosclerosis were randomized to three groups: Continued cholesterol diet, cholesterol diet supplemented with 0.01% atorvastatin, and cholesterol free diet for 4 weeks to study mechanisms of plaque regression. Results Proliferation of scavenger receptor A and CD36 deficient macrophages with impaired lipid uptake was reduced by 30–50% in the plaque, while ABCA1/ABCG1 exporter deficiency resulted in cholesterol overloading and apoptosis. Oral atorvastatin treatment decreased total plasma cholesterol levels by 50% to the same extend as cholesterol free diet feeding in APOE*3-Leiden.huCETP. Cholesterol lowering resulted in a 50% reduction in local macrophage proliferation and plaque regression with reduced macrophage and lipid contents and increased collagen. GFP bone marrow reconstitution of APOE*3-Leiden.huCETP mice in which the aortas were shielded from irradiation showed infiltrating monocytes to contribute only 11% to the plaque macrophage pool during plaque progression, thereby underscoring the relevance of targeting macrophage proliferation for plaque regression. Finally, rates of macrophage proliferation in human carotid artery plaques correlated with serum LDL-cholesterol levels, in line with our experimental studies. Conclusion Foam cell formation in atherosclerotic plaques triggers their proliferation. Targeting macrophage proliferation leads to plaque regression.

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Als eine Form der Zusammenarbeit zwischen pädagogischen Fachkräften und Eltern wurden im Rahmen des Projekts „Schulreifes Kind“ sogenannte „Runde Tische“ eingeführt. Hierbei handelt es sich um Treffen, bei denen gemeinsam die Entwicklung einzelner Kinder betrachtet und mögliche Förderinhalte besprochen werden. In der vorliegenden Arbeit wird anhand einer Fragebogenuntersuchung von N = 516 beteiligten Personen eine Evaluation der „Runden Tische“ vorgenommen. Die Ergebnisse weisen auf eine gute Zusammenarbeit zwischen pädagogischen Fachkräften und Eltern hin, wobei vor allem seitens der pädagogischen Fachkräfte der Wunsch eines noch intensiveren Austauschs deutlich wird. Weiterhin besteht Einigkeit darüber, dass die „Runden Tische“ eine gute Möglichkeit darstellen die Eltern enger in vorschulische Förderentscheidungen einzubinden. Als besonders positiv wird herausgestellt, dass es bei den Treffen zu einem gezielten Austausch über die Kinder aus den unterschiedlichsten Blickwinkeln kommt.

Im Rahmen einer längsschnittlichen Evaluation des Baden-Württemberger Modellprojektes „Schulreifes Kind“ wurden mehr als 900 Kinder des Einschulungsjahrgangs 2009 zu drei Messzeitpunkten vor Schuleintritt und zwei weiteren nach Schuleintritt untersucht. Die hier vorgelegten Datenanalysen wurden zur Klärung der folgenden drei Fragen genutzt: (1) Führen die kompensatorischen Fördermaßnahmen zur Verbesserung schulrelevanter Fertigkeiten bis zum Ende der Kindergartenzeit? (2) Kommt es infolge des Projektes zu einer höheren Einschulungswahrscheinlichkeit? (3) Zeigen sich mittelfristige Wirkungen auf den Erwerb schulischer Fertigkeiten? Die Analysen zeigen, dass die ersten zwei Fragen mit „ja“ beantwortet werden können. Die Frage nach mittelfristigen Transferwirkungen auf den Erwerb schulischer Fertigkeiten muss dagegen zunächst unbeantwortet bleiben. Insgesamt zeigt sich ein vergleichsweise positives Wirkungsmuster der Zusatzförderung im letzten Kindergartenjahr.

This paper is a dialogue between the artist and the analyst in the examination of a digital video art work as a piece of practice as research. The paper identifies and addresses the significant research questions that have arisen out of precisely such a dialogue about work in progress. Rather than focusing on the completed installation as a piece of research, this paper emphasises the artistic practice as experience and offers a concurrent academic debate, which complements the art work rather than substitute it as a research outcome. By embracing such plurality inherent in artistic practice as research, this article examines questions of authenticity of voice, narrative abstraction and interpretation and the effect low budget constraints have on film practice. Rather than offering certain conclusions, this paper is to be one in a series, discussing the research practice at various points in the artistic process.