Establishment of an in vitro test strategy for orally inhaled drug products

This thesis describes and evaluates three animal experiment free approaches for the safety and efficacy assessment of pulmonary drug formulation in the pre-clinical phase. In the first approach the complex cell culture system MucilAir™ was exposed to pollen through the Vitrocell® Powder Chamber to simulate in vitro an allergic reaction and predict drug safety. However, cytokine response was not sufficient, and the setup has to be optimized for safety studies. Two in vitro setups were further developed for estimating drug safety and inflammatory response. For safety studies, an in vitro model should predict human data by using approved FDA excipient concentrations. From these excipients the in vitro IC50 was determined with pulmonary cell lines and ranked in the thus established SAFE classification. Consequently, SAFE allows for predicting human safety data by in vitro testing. For estimating potential inflammatory reactions located in the respiratory area, the alveolar cell line hAELVi was exposed to the pro-inflammatory cytokines TNF-α and IFN-γ. Similar to an inflammatory in vivo reaction, a loss of epithelial barrier function was observed. Furthermore, the anti-inflammatory response to hydrocortisone of the stimulated model underlines its po-tential for in vitro drug testing. The described in vitro models are a promising tool for replacing animal experiments, whereby their standardization and validation are future challenges according to recognized guidelinesDiese Arbeit beschreibt und bewertet drei tierversuchsfreie Ansätze zur Sicherheits- und Wirk-samkeitsbewertung von Inhalativa in der präklinischen Phase. Im ersten Ansatz wurde das Zellsystem MucilAir™ durch die Vitrocell® Powder Chamber Pol-len ausgesetzt, um in vitro eine allergische Reaktion zu simulieren. Die Zytokinreaktion war jedoch nicht ausreichend und der Aufbau muss für Sicherheitsstudien optimiert werden. Folg-lich wurden zwei in vitro Modelle entwickelt, um die Arzneimittelsicherheit und Entzündungs-reaktion abzuschätzen. Für eine Sicherheitsbewertung sollte ein in vitro Modell Humandaten unter Verwendung von FDA-Hilfsstoffkonzentrationen vorhersagen. Aus diesen Hilfsstoffen wurde der in vitro IC50 Wert für Lungenzellen bestimmt und in die etablierte SAFE-Klassifi-kation eingestuft. SAFE ermöglicht so die Vorhersage von Humandaten durch in vitro Tests. Zur Bewertung von Entzündungen wurden hAELVi Zellen den Zytokinen TNF-α und IFN-γ ausgesetzt. Ähnlich wie bei einer entzündlichen in vivo Reaktion wurde ein Verlust der epithe-lialen Barrierefunktion beobachtet. Darüber hinaus unterstreicht die entzündungshemmende Reaktion des Modells auf Hydrocortison sein Potenzial für eine in vitro Sicherheitsbewertungen von potenziellen Wirkstoffen. Die etablierten in vitro Modelle bieten einen vielversprechenden Ansatz, um Tierversuche zu ersetzen, wobei ihre Standardisierung und Validierung künftige Herausforderungen zur Auf-nahme in anerkannten Richtlinien darstellen

Conflict inside the European Commission is a key factor in shaping EU legislation

Why does the European Commission at times propose legislative drafts that provoke Member State opposition, that introduce strikingly high or low standards, or that actively contradict each other? Miriam Hartlapp, Julia Metz and Christian Rauh present findings from a study of 48 legislative drafting processes. They argue that while the Commission is often thought of as a unified actor, there is substantial disagreement within the Commission over the nature of legislative proposals. They write that studying this conflict is key to understanding the policies the institution proposes for Europe

How external interests enter the European Commission: mechanisms at play in legislative position formation

"The European Commission, Europe's central supranational bureaucracy, is often depicted as a playground for external interests. The scholarly literature offers a range of sensible explanations how the Commission position is bound by national, organised societal, or party political interests. Despite substantial contributions, the empirical approach supporting such arguments often focuses on the mere comparison between a particular external interest and the legislative outcome. In this paper, we aim to add a more process-based, mechanismic perspective by empirically focussing the question how certain external interest found their way into a Commission position as captured by a legislative proposal. Against the background of a larger research project which analyses 48 position formation processes in the Commission on the basis of 133 in-depth interviews with participating officials, this paper presents mechanisms linking external interest to the final Commission position and that are transferable across specific drafting processes and across different policy fields." (author's abstract)"Die europäische Kommission, zentraler bürokratischer Akteur der Europäischen Union, wird oft als Spielwiese externer Interessen dargestellt. Die politikwissenschaftliche Literatur bietet vor diesem Hintergrund eine Reihe sinnvoller Erklärungen an, wie die Kommission bei ihrer Positionsbildung durch nationale, gesellschaftlich-organisierte oder parteipolitische Interessen beeinflusst ist. Trotz beachtlicher Forschungsbeiträge, beschränken sich empirische Arbeiten, die dieses Argument untermauern allerdings häufig auf den bloßen Vergleich eines bestimmten externen Interesses und dem Inhalt des letztlich von der Kommission vorgeschlagenen Gesetzes. Mit diesem Papier zielen wir darauf ab, diese Literatur um einen eher prozess- und mechanismus-orientierten Ansatz zu ergänzen indem wir empirisch der Frage nachgehen, wie externe Interessen ihren Weg in einen legislativen Kommissionsvorschlag gefunden haben. Wir greifen dabei auf ein breiteres Forschungsprojekts zurück, das 48 Politikformulierungsprozesse innerhalb der Kommission auf der Basis von 133 Interviews mit beteiligten Kommissionsbeamten analysiert. Auf dieser Grundlage lassen sich eine Reihe von Mechanismen identifizieren, die aufklären, wie externe Interessen mit der finalen Kommissionsposition verbunden sind, und die gleichzeitig über einzelne Prozesse und Politikfelder transferierbar sind." (Autorenreferat

Made in Brussels: wie externe Interessen ihren Weg in die Politikformulierung der EU-Kommission finden

"Die EU-Kommission als zentrale supranationale Bürokratie der Europäischen Union wird häufig als Spielwiese externer Interessen dargestellt. In der Tat werden mittels bestimmter Mechanismen externe Interessen an die abschließende Position der Kommission zu EUGesetzesvorhaben gekoppelt. Die Analyse zeigt, wie organisierte gesellschaftliche oder parteipolitische Interessen ihren Weg in Gesetzesinitiativen der Kommission finden: etwa durch die Antizipation der Mehrheit im Rat oder im Europäischen Parlament, durch den Transfer bestehender (nationaler) Politiklösungen, durch eine Art Gütertausch von Einfluss gegen Legitimität oder Expertise oder durch ideologische Fürsprache." [Autorenreferat]"The European Commission, Europe’s central supranational bureaucracy, is often depicted as a playground for external interests. Indeed, certain mechanisms link external interests to the final Commission position. Research shows how national organized societal or party political interests find their way into a Commission initiative: by anticipating the Council or European Parliament shadow, by transfer of (national) templates, by exchange of resources (legitimacy and expertise) or by ideological advocacy." [author's abstract

Inflammatory bowel disease addressed by Caco-2 and monocyte-derived macrophages : an opportunity for an in vitro drug screening assay

Infammatory bowel disease (IBD) is a widespread disease, afecting a growing demographic. The treatment of chronic infammation located in the GI-tract is dependent on the severity; therefore, the IBD treatment pyramid is commonly applied. Animal experimentation plays a key role for novel IBD drug development; nevertheless, it is ethically questionable and limited in its throughput. Reliable and valid in vitro assays ofer the opportunity to overcome these limitations. We combined Caco-2 with monocyte-derived macrophages and exposed them to known drugs, targeting an in vitro-in vivo correlation (IVIVC) with a focus on the severity level and its related drug candidate. This co-culture assay addresses namely the intestinal barrier and the immune response in IBD. The drug efcacy was analyzed by an LPS-infammation of the co-culture and drug exposure according to the IBD treatment pyramid. Efcacy was defned as the range between LPS control (0%) and untreated co-culture (100%) independent of the investigated read-out (TEER, Papp, cytokine release: IL-6, IL-8, IL-10, TNF-α). The release of IL-6, IL-8, and TNF-α was identifed as an appropriate readout for a fast drug screening (“yes–no response”). TEER showed a remarkable IVIVC correlation to the human treatment pyramid (5-ASA, Prednisolone, 6-mercaptopurine, and infiximab) with an R2 of 0.68. Similar to the description of an adverse outcome pathway (AOP) framework, we advocate establishing an “Efcacy Outcome Pathways (EOPs)” framework for drug efcacy assays. The in vitro assay ofers an easy and scalable method for IBD drug screening with a focus on human data, which requires further validation

Fundamental challenges in designing a collaborative travel app

The growing capabilities of smartphones have opened up new opportunities for travel coordination and transport is a fertile area for app development. One stream of development is apps that enable collaborative travel, either in the form of lift sharing or collaborative shopping, but despite growing interest from governmental agencies, there is little evidence of the efficacy of such apps. Based on trials of purpose built travel collaboration apps, deployed in tourism, urban and rural residential communities, and logistics, this paper analyses the fundamental challenges facing users adopting such travel apps. The findings suggest that transport practitioners, policy makers and app developers need to better understand the challenges associated with attracting users, the use of incentives and the types of communities most appropriate to implement collaborative travel concepts using such approaches. Also, how the users’ sense of time pressure and the issues around reciprocal exchange can impact on their long-term success and wider adoption

Chemie im Wandel

CHEMIE IM WANDEL Chemie im Wandel / Münnich, Paul (CC BY-NC-SA) ( -

Cold Atmospheric Plasma Promotes the Immunoreactivity of Granulocytes In Vitro

Cold atmospheric plasma (CAP) reduces bacteria and interacts with tissues and cells, thus improving wound healing. The CAP-related induction of neutrophils was recently described in stained sections of wound tissue in mice. Consequently, this study aimed to examine the functionality of human polymorphonuclear cells (PMN)/granulocytes through either a plasma-treated solution (PTS) or the direct CAP treatment with different plasma modes and treatment durations. PTS analysis yielded mode-dependent differences in the production of reactive oxygen species (ROS) and reactive nitrogen species (RNS) after CAP treatment. Live-cell imaging did not show any chemo-attractive or NETosis-inducing effect on PMNs treated with PTS. The time to maximum ROS production (TmaxROS) in PMNs was reduced by PTS and direct CAP treatment. PMNs directly treated with CAP showed an altered cell migration dependent on the treatment duration as well as decreased TmaxROS without inducing apoptosis. Additionally, flow cytometry showed enhanced integrin and selectin expression, as a marker of activation, on PMN surfaces. In conclusion, the modification of PMN immunoreactivity may be a main supporting mechanism for CAP-induced improvement in wound healing

Rebound After Fingolimod and a Single Daclizumab Injection in a Patient Retrospectively Diagnosed With NMO Spectrum Disorder—MRI Apparent Diffusion Coefficient Maps in Differential Diagnosis of Demyelinating CNS Disorders

Objective: Differential diagnosis of neuromyelitis optica spectrum disorders (NMOSD) and multiple sclerosis (MS) or mimics can be challenging, especially in patients with atypical presentations and negative serostatus for aquaporin-4 antibodies (AQP4-Ab). This brief research report describes magnetic resonance imaging (MRI) findings focusing on quantitative apparent diffusion coefficient (ADC) histogram analysis as a potential tool to differentiate NMOSD from MS.Methods: Longitudinal MRI data obtained during routine clinical examinations were retrospectively analyzed in a patient with histologically determined cerebral NMOSD, a patient with an acute tumefactive MS lesion, and a patient with ischemic stroke. Histogram analyses of ADC maps were evaluated.Results: A patient diagnosed with MS experienced a severe rebound after fingolimod withdrawal and a single daclizumab injection. Cerebral NMOSD manifestation was confirmed by brain biopsy. However, the patient did not fulfill consensus criteria for NMOSD and was AQP4-Ab negative. Comparison of ADC histogram analyses of this patient with those from a patient with MS and one with ischemic stroke revealed differential ADC characteristics: namely a more pronounced and prolonged ADC leftward shift in inflammatory than in ischemic pathology, even more accentuated in NMOSD versus MS.Conclusion: ADC map histograms and ADC threshold values for different conditions may be useful for differentiation of large inflammatory brain lesions and further studies are merited

Towards More Predictive, Physiological and Animal-free In Vitro Models: Advances in Cell and Tissue Culture 2020 Conference Proceedings

Experimental systems that faithfully replicate human physiology at cellular, tissue and organ level are crucial to the development of efficacious and safe therapies with high success rates and low cost. The development of such systems is challenging and requires skills, expertise and inputs from a diverse range of experts, such as biologists, physicists, engineers, clinicians and regulatory bodies. Kirkstall Limited, a biotechnology company based in York, UK, organised the annual conference, Advances in Cell and Tissue Culture (ACTC), which brought together people having a variety of expertise and interests, to present and discuss the latest developments in the field of cell and tissue culture and in vitro modelling. The conference has also been influential in engaging animal welfare organisations in the promotion of research, collaborative projects and funding opportunities. This report describes the proceedings of the latest ACTC conference, which was held virtually on 30th September and 1st October 2020, and included sessions on in vitro models in the following areas: advanced skin and respiratory models, neurological disease, cancer research, advanced models including 3-D, fluid flow and co-cultures, diabetes and other age-related disorders, and animal-free research. The roundtable session on the second day was very interactive and drew huge interest, with intriguing discussion taking place among all participants on the theme of replacement of animal models of disease