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Poorly separable liquid/liquid mixtures often pose a major challenge in pharmaceutical extraction. In particular, compounds containing a low difference of density are difficult to separate using mixer-settler setups. The Zaiput device can prove an advantage over present liquid/liquid extractors. The research work involves characterizing the Zaiput apparatus for liquid/liquid extraction of pharmaceutical compounds with the model solvent system toluene-water. The extraction efficiency was investigated for a solvent mixture containing toluene-acetone-water residence times. To evaluate the separation success, the outlet’s concentrations are measured via IR spectroscopy., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

Im Rahmen dieser Arbeit wurden bereits etablierte NHC-Gold(I)-Komplexe synthetisiert und ihre Eigenschaften in biologisch relevanten Matrizes untersucht. Dazu wurde eine HPLC-MS Methode entwickelt, die in der Lage ist, den Hauptkomplex von potentiellen Abbauprodukten und Metaboliten zu trennen. Das chromatographische Verfahren ermöglichte dabei neben einer qualitativen Betrachtung eine einfache Quantifizierung der untersuchten Komplexe. Zunächst wurde die Stabilität in organischen Lösungsmitteln, wässrigen Lösungsmitteln und in saurer Lösung untersucht. Dabei konnte eine Instabilität in wässrigem Milieu gezeigt werden, die mit abnehmenden pH-Wert verstärkt wurde. Entstandene Abbauprodukte konnten identifiziert und quantifiziert werden. Weitere Untersuchungen mit Schwefelverbindungen als Reaktionspartnern ergaben weiteren Aufschluss über das Verhalten von Gold(I)-Komplexen in biologischem Umfeld. Die Experimente zeigten Ligandenaustauschreaktionen, welche in Form von Reaktionsgleichungen abgeleitet wurden. Um das Verständnis für das Verhalten von Goldkomplexen in biologischem Umfeld weiter zu vertiefen, wurden Stabilitätsuntersuchungen in der Zellkultur durchgeführt. Dazu wurde eine Zellextraktionsmethode entwickelt, die den intakten Komplex, Abbauprodukte und potentielle Metabolite extrahieren soll. Die anschließende Analytik wurde erneut mit der HPLC-MS durchgeführt. Zusätzlich wurden Zellaufnahmestudien mit einem Atomabsorptionsspektrometer durchgeführt. Da es sich bei den untersuchten Verbindungen um potentielle Zytostatika handelt, wurden diese in einem Xenograft-Modell auf ihre antitumorale Wirkung und Verträglichkeit untersucht., In this work, already established NHC-gold(I) complexes were synthesized and their properties in biologically relevant matrices were investigated. For this purpose, an HPLC-MS method capable of separating the main complex from potential degradation products and metabolites was developed. The chromatographic method thereby enabled an easy quantification of the investigated complexes in addition to a qualitative analysis. First, the stability in organic solvents, aqueous solvents and in acidic solution was investigated. An instability in aqueous medium could be shown, which was enhanced with a decreasing pH value. Formed degradation products could be identified and quantified. Further studies using sulfur compounds as reaction partners provided further insight into the behavior of gold(I) complexes in biological environments. The experiments revealed ligand exchange reactions, which were derived in the form of reaction equations. To further understand the behavior of gold complexes in biological environment, stability studies were carried out in cell culture. For this purpose, a cell extraction method was developed to extract the intact complex, degradation products and potential metabolites. Subsequent analysis was again performed using HPLC-MS. In addition, cell uptake studies were performed using an atomic absorption spectrometer. Since the compounds studied are potential cytostatic agents, they were evaluated for their antitumor activity and tolerability in a xenograft model.

The poor solubility of a large number of active pharmaceutical ingredients (API) is a major challenge in pharmaceutical research. Therefore, the extrusion of amorphous solid dispersions (ASDs) is one promising approach to enhance the dissolution rate by molecularly dissolve the API in an amorphous carrier polymer. During ASD development, crucial parameters as the dissolution and the decomposition need to be monitored. Within this study, a small scale twin screw extruder was coupled with special ColVisTec UV-Vis probes that are characterized by their small dimensions. This setup enables a systematic formulation development and optimization based on in-line monitoring of crucial parameters using small amounts of material., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

With the advances in sensor technology, big data, and artificial intelligence, unobtrusive in-home health monitoring has been a research focus for decades. Following up our research on smart vehicles, within the framework of unobtrusive health monitoring in private spaces, this work attempts to provide a guide to current sensor technology for unobtrusive in-home monitoring by a literature review of the state of the art and to answer, in particular, the questions: (1) What types of sensors can be used for unobtrusive in-home health data acquisition? (2) Where should the sensors be placed? (3) What data can be monitored in a smart home? (4) How can the obtained data support the monitoring functions? We conducted a retrospective literature review and summarized the state-of-the-art research on leveraging sensor technology for unobtrusive in-home health monitoring. For structured analysis, we developed a four-category terminology (location, unobtrusive sensor, data, and monitoring functions). We acquired 912 unique articles from four relevant databases (ACM Digital Lib, IEEE Xplore, PubMed, and Scopus) and screened them for relevance, resulting in n=55 papers analyzed in a structured manner using the terminology. The results delivered 25 types of sensors (motion sensor, contact sensor, pressure sensor, electrical current sensor, etc.) that can be deployed within rooms, static facilities, or electric appliances in an ambient way. While behavioral data (e.g., presence (n=38), time spent on activities (n=18)) can be acquired effortlessly, physiological parameters (e.g., heart rate, respiratory rate) are measurable on a limited scale (n=5). Behavioral data contribute to functional monitoring. Emergency monitoring can be built up on behavioral and environmental data. Acquired physiological parameters allow reasonable monitoring of physiological functions to a limited extent. Environmental data and behavioral data also detect safety and security abnormalities. Social interaction monitoring relies mainly on direct monitoring of tools of communication (smartphone; computer). In summary, convincing proof of a clear effect of these monitoring functions on clinical outcome with a large sample size and long-term monitoring is still lacking.

Spherical agglomeration (SA) is a key crystal shape modification technique commonly used to improve processability and physical properties of active pharmaceutical ingredients (APIs), such as flowability and dissolution. SA involves the introduction of a bridging liquid in the form of droplets into a suspension of API crystals. A novel approach was successfully implemented in this study by adopting a membrane emulsification (ME) system to control the size of bridging liquid droplets and spherical agglomerates. The study investigated ways to achieve the critical quality attributes of the spherical agglomerates by varying operating conditions and design parameters, including mixing rate, bridging liquid injection rate and membrane pore diameter. Antisolvent crystallization of benzoic acid in ethanol (solvent) and water (antisolvent) in presence of toluene as a bridging liquid was used as a case study. Spherical agglomerates of benzoic acid with enhanced quality attributes and mean size of 300 μm were obtained using a membrane pore size of 18 μm, a bridging liquid flow rate of 0.025 mL/min and mixing rate in the range of 800 RPM and 1000 RPM., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

Mixing is a crucial parameter for a wide range of processes in bio-/chemical engineering. In multiphase flows, the aim is a high and uniform mixing effect throughout the reaction volume without negative effects on the discrete phase. These are usually caused by the hydromechanical stress exerted by the mixing element. The present study introduces a novel reactor concept to overcome the use of fast rotating, locally acting mixing elements common in stirred tank reactors. Therefore, the reactor is designed of a flat, horizontally mounted cylinder, which enables mixing of the enclosed volume by its rotation around the central axis. To evaluate the mixing performance, a colorimetric method for mixing time determination is presented and the spatial evolution of mixing kinetics visualized. In addition, the method reveals the dependencies of the mixing time on the rotational speed and filling volume., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

Two-phase biocatalysis is commonly used for the bioconversion of water-insoluble chemical and pharmaceutical compounds. In research laboratories, such processes often take place in flasks and are driven and limited by slow diffusive processes. In this work, we developed droplet-based microfluidic systems and used them to perform testosterone dehydrogenation. The use of microdroplets as reaction vessels offers the advantage of greatly improved diffusion rates due to significantly increased surface-to-volume ratio. We found that indeed reaction time was reduced from tens of minutes to tens of seconds without sacrificing conversion efficiency. This demonstrates the potential of droplet-based two-phase biocatalysis., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

Apoptosis is a series of complex biochemical processes of programmed cell death which occurs in multicellular organisms. This mechanism must be highly regulated and controlled; otherwise, harmful illness such as cancer and autoimmune diseases may occur. Apoptosis is a key mechanism during a typical viral immune response.In the course of an infection, this mechanism eliminates infected cells and by that,limits the resources for viral replication. Recently, regulation of the death recep-tor pathway by cFLIP, a key regulator of signaling complexes downstream of death receptors, was targeted experimentally to study the role of apoptosis in influenza A virus (IAV) infection. By increasing the expression level of cFLIP in transgenic mice, a higher accumulation of Natural Killer (NK) cells and a higher viral load in comparison to wild type(WT) mice were observed. However, the viral load was similar in both mice after 10 days. Infection and functional impairment of NK cells was assumed to be responsible for the observed results. In order to validate this hypothesis, a mathematical model was used to quantify the role of apoptosis medi-ated cell death in NK cells during the viral clearance. The model contains variables and parameters that represent population and interactions between healthy and in-fected epithelial cells, influenza A virions, healthy and infected NK cells. Several simplifications were made in order to better estimate the critical parameters of the model. Simulation results showed that infection of NK cells with a longer lifespan alone cannot explain the observed high viral load. However, by increasing the viral production of infected NK cells, the viral load data was fitted. The lack of caspase3, due to the over-expression of cFLIP, may contribute to higher viral production inNK cells. Furthermore, a shorter lifespan of healthy NK cells was predicted, which may be related to biological exhaustion due to higher infection level. The posteriors and the correlation analysis of the parameters showed that parameters associated with lifespan and the viral production of infected NK cells are linearly correlated., Die Apoptose ist ein komplexer biochemischer Prozess des programmierten Zelltod,die in mehrzelligen Organismen auftritt. Dieser Mechanismus muss eng kontrolliert werden, sonst können schädliche Krankheiten auftreten, so wie z.B. Krebs und Autoimmunerkrankungen. Während einer typischen viralen Immunantwort, werden infizierte Zellen durch Apoptose eliminiert, so dass die Ressourcen für die Virus-replikation begrenzt werden. cFLIP ist ein Inhibitor der Todesrezeptor-gesteuerten Apoptose. Die Regulation des extrinsischen Signalwegs durch cFLIP wurde erforscht um die Rolle der Apoptose während Influenza A Infektionen zu untersuchen. Hierfür wurden transgene Mäuse, die cFLIP in allen hämatopowerten Zellen exprimieren,mit dem Influenza A-Virus infiziert. Diese transgenen Mäuse haben mehr natürliche Killerzellen (NK) und höhere Viruslast aufgewiesen. Die Viruslast nach 10 Tagen waren ähnlich bei transgenen und Wildtyp (WT) Mäusen. Tafrishi argumentierte, dass die infizierten NK-Zellen und ihre Funktionsstörung verantwortlich für die Ergebnissen sind. Hierin wurde ein mathematisches Modell verwendet, um diese Hypothese zu validieren und die Rolle der Apoptose in NK Zellen durch Influenza A-Infektionen zu quantifizieren. Die Modellvariablen und Parameter darstellen die Population und Wechselwirkungen beidem Influenza A-Virus, gesunden und infizierten Epithelzellen und gesunden und infizierten NK-Zellen. Die Simulationsergebnissen demonstrieren, dass infizierte NK-Zellen, die eine längere Lebensdauer haben, nicht der Grund für die erhöhte Viruslast sind. Stattdessen ist eine höhere Virusproduktion von der infizierten NK-Zellen notwendig, um die Viruslast Daten anzupassen.Deshalb kann das Fehlen von Caspase 3, aufgrund der Überexpression von cFLIP,zu einer höheren Virusproduktion in den NK-Zellen beitragen. Darüber hinaus hat die Datenanpassung eine kürzere Lebensdauer der gesunden NK-Zellen vorhergesagt,aufgrund der biologischen Erschöpfung. Korrelationsanalysen der Parameter zeigten,dass die Lebensdauer und die Virusproduktion der infizierten NK-Zellen linear korreliert sind.

Secondary nucleation is widely present in crystallisation processes and it is often relied upon to attain desirable critical quality attributes of crystalline products, such as polymorphic form and crystal size distribution. This is particularly the case in continuous crystallisation, where a good control of secondary nucleation can be crucial in order to achieve and maintain steady state operation. This work utilises rapid, small-scale experiments in agitated vials with in-situ imaging for crystal counting and sizing (using the Crystalline platform), to quantify nucleation and crystal growth kinetics of α-glycine across a range of supersaturations in aqueous solutions under isothermal conditions. Both seeded and unseeded crystallisation experiments were conducted. It was found that secondary nucleation and crystal growth rates determined from the same vials show a close correlation across the whole range of supersaturations investigated, which suggest a close relationship between the mechanisms of shear induced secondary nucleation and crystal growth in this system., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

The cryopreservation of cells and tissue engineered constructs is determined by multiple factors, such as possible cellular damages due to mass and heat transfer during the freezing and thawing process. It is assumed that the utilization of cryobags improves the heat transfer during freezing and thawing and is therefore superior in comparison to currently applied cryovials and multiwell plates. Therefore, we have analysed the cryopreservation of cell-seeded electrospun polycaprolactone/polylactide scaffolds within cryobags. Additionally, the performance of four different cryoprotective agents was analysed, which included 10% (v/v) dimethyl sulfoxide and 10% (v/v) ethylene glycol as separately applied agents as well as their combination with 0.3 M sucrose. The samples were frozen in in-house made cryobags with a controlled rate freezer. The cell viability was analysed by fluorescence microscopy before freezing and after thawing. The results show, that the application of cryobags in combination with 10% (v/v) ethylene glycol can have a positive impact onto the cell viability., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

Jahrbuch der Braunschweigischen Wissenschaftlichen Gesellschaft, vol. 2020Jahrbuch 2020 der Braunschweigischen Wissenschaftlichen Gesellschaft, vol. 2020, p. 170

Without any doubt the young technology of additive manufacturing, including 2D and 3D printing, has been changing the world. A wide spectrum of objects in medical and pharmaceutical field can already be successfully printed. This ranges from a detailed figure in the micrometer regime to a full size residential house unit made from concrete [Sakin et al., 2017; Doherty et al., 2020]. In the pharmaceutical sector, additive manufacturing offers the option of medication customization [Pravin et al., 2018]. Integrating this new technology has several advantages over the current, well-established but outdated “one-size-fits-all” approach. Besides avoiding medication errors, printing tailored oral dosage forms is financially attractive for small scale on-demand production [Awad et al., 2018]. The development of dosage forms, production of sample batches and modification of samples can be done with little effort compared to generic powder-pressed pills which require heavy pharmaceutical machinery [Awad et al., 2018; Dachtler et al., 2020]. To further progress and to create the legal framework for the approval of additively manufactured drugs it requires a paradigm change of stakeholders who have the power to make decisions and changes [Horst et al., 2019]., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

Amorphous solid dispersions (ASDs) of a poorly water-soluble active pharmaceutical ingredient (API) in a polymer matrix can enhance the water solubility and improve the bioavailability of the API. Many ASD products are kinetically stabilized, and the inhibition of crystallization of an API within and beyond shelf life is still a matter of debate, since the formation of crystals may impact bioavailability. We present the first literature examples of ASD long-term stability studies over up to 25 years under ambient storage conditions where no API crystallization was observed. Additionally, a risk assessment and mitigation strategy of API crystallization in packaged ASD drug products (DP) is outlined. The risk of shelf-life crystallization and the respective mitigation steps are assigned for different DP development scenarios. Ultimately, the physical stability of ASD DPs during shelf-life storage is modeled by quantifying crystal growth kinetics by transmission Raman spectroscopy (TRS), modeling the impact of water sorption on the glass-transition temperature of the ASD, and predicting the moisture uptake by the packaged ASD DP during storage. This approach is applied to an ASD of an AbbVie-internal compound showing stability beyond the anticipated shelf-life., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

In December 2019, a series of pneumonia of unknown cause emerged in Wuhan, China, which developed into a global pandemic with more than 200 million cases and 4 million deaths worldwide as of August 2021. The enveloped RNA betacoronavirus which was found to cause the pandemic was named severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Academia and industry started with non-clinical and clinical development of several SARS-CoV-2 vaccine candidates immediately and results were achieved enormously fast. As an example, BioNTech and Pfizer got their joint SARS-CoV-2 vaccine approved less than eight months after trials started, while under normal circumstances vaccine development takes up to 10 – 15 years. In the case of the SARS-CoV-2 vaccine the extremely fast turnaround time from start of clinical trials to approval of the first vaccine candidates was driven by the urgent medical need and made possible by the mobilisation of enormous financial resources from the private and public sectors, as well as manpower and effort and the willingness of regulators to speed up timelines and approval processes. Looking at the extremely fast and successful development of several SARS-CoV-2 vaccine candidates, it is difficult to understand from a layman’s perspective why for others diseases like Human Immunodeficiency Virus/ Acquired Immunodeficiency Syndrome (HIV/AIDS) and tuberculosis (TB), which are among the top ten causes of death worldwide, development of effective vaccines takes so long. More than 30 years after HIV has been discovered, there is still no vaccine available for prevention of HIV infection. Development of several vaccine candidates has failed in clinical trials. The difficulties stem in large part from HIV’s unusual characteristics of replicating and mutating rapidly and high sequence diversity, its ability of immune system evasion and the creation of a persistent viral reservoir within the host. Development of the novel TB vaccine candidate VPM1002, which is designed to be a safe and effective vaccine for prevention of TB infection and TB recurrence, is an example of classical vaccine development. VPM1002 was invented in Germany, went through successful non-clinical development and is currently being tested in phase III clinical trial. The achievements during the last year fast development of SARS-CoV-2 vaccines during the last year have proven that in case of a pandemic situation and an urgent medical need, fast vaccine development is possible. The achievements were reached with high financial commitment and regulators have shown willingness and engagement to accelerate timelines for review and approval processes. It would be desirable if further vaccine developments could benefit from these efforts., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

Polymerized Ionic Liquids (PILs) are hydrogels synthesized from ionic liquids. They combine the positive characteristics of ionic liquids and polymers which is why they have a wide range of applications, e.g. in the pharmaceutical sector. They are characterized by properties such as high mechanical strength, biocompatibility, and favorable swelling behavior. In this study, the extremely high mechanical strength of novel polymerized ionic liquids (PILs)-based hydrogels was confirmed by mechanical tests. The swelling behavior was studied of hydrogels based on 1-Vinyl-3-alkylimidazolium bromide with different crosslinkers in various solvents – aqueous and organic., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

Bei der Entwicklung Digitaler Gesundheitsanwendungen (DiGA) sind eine Reihe rechtlicher, finanzieller und qualitativer Rahmenbedingungen zu berücksichtigen. Hinzu kommen organisatorische und technische Anforderungen zur Einbindung einer DiGA in Versorgungsprozesse. In Rehabilitationsprozessen spielt die Therapieadhärenz der Patienten eine wesentliche Rolle. Wie bereits vergangene Studien gezeigt haben, können diese durch den Einsatz von Spiel-Design-Elementen gesteigert werden. Die Auswahl, Kombination und das Design einzelner Spiele-Komponenten wirkt jedoch oftmals unüberlegt. Das Ziel dieser Dissertation ist es, einen ganzheitlichen Ansatz zur Entwicklung und Verbesserung von DiGA für die Rehabilitation zum Zweck der Adhärenzsteigerung durch Gamification zu erstellen und in der Praxis zu erproben. Um den Realitätsbezug des Ansatzes zu sichern, wurde das Krankheitsbild der Schulterläsionen als Anwendungsbeispiel genutzt. In acht Ebenen erfolgte die systematische Entwicklung des GISMOR-Ansatzes – ‘Gamification Increasing Motivation for Rehabilitation’. GISMOR unterstützt sowohl bei der Identifikation potentieller Adhärenzfaktoren (Patientenebene) als auch bei der adäquaten auf die Zielgruppe ausgerichteten Auswahl, Kombination und Umsetzung von Spiel-Design-Elementen (Gamificationebene). Dabei legt GISMOR besonderen Wert auf die Interaktion zwischen der DiGA und dem Patienten und stellt damit nicht nur die interaktive und visuelle Gestaltung des User Interface in den Vordergrund, sondern auch die resultierende User Experience (Interaktionsebene). Darüber hinaus gibt GISMOR den rechtlichen, finanziellen und qualitativen Rahmen für die Umsetzung abrechnungsfähiger DiGA vor (Dienstleistungsebene). Für den erfolgreichen Einsatz von DiGA liefert GISMOR weiterhin Anhaltspunkte zur Einbindung dieser in Rehabilitationsprozesse (Prozessebene) sowie zur Integration in transinstitutionelle Informationssystemarchitekturen (Informationssystemebene). Über den Anwendungsleitfaden zu GISMOR wird der entwickelte Ansatz schließlich in der Praxis einsetzbar. Als eigenständiges Dokument dient der Anwendungsleitfaden Softwareentwicklern, Medizininformatikern und Gesundheitsversorgern als Handlungshilfe zur Entwicklung und Verbesserung von DiGA sowie Medizinischen Assistenzsystemen. Am Beispiel des Telerehabilitationssystems AGT-Reha konnte dabei gezeigt werden, dass GISMOR sowohl für die Analyse als auch Ausarbeitung konkreter Maßnahmen zur Verbesserung einer DiGA geeignet ist., A number of legal, financial, and qualitative constraints must be considered in developing operational and billable digital health applications (DiGA). Furthermore, there are organizational and technical requirements for integrating DiGAs into care processes. In rehabilitation processes, patients' motivation and adherence are essential factors as well. Past studies have already shown that these can be increased through the use of game design elements. However, the selection, combination, and design of game components often seems rash. The objective of this thesis is to design and evaluate a holistic approach for developing and improving DiGAs in rehabilitation for the purpose of increasing motivation and adherence through gamification. Shoulder lesions were used as a consistent application example. The systematic development of GISMOR – ‘Gamification Increasing Motivation for Rehabilitation’ – was done in eight tiers. GISMOR supports both the identification of potential adherence factors (patient tier) and the adequate selection, combination, and implementation of game design elements aligned with the target group (gamification tier). GISMOR emphasizes the interaction between a DiGA and the patient, thus focusing not only on the interactive and visual design of the user interface, but also on the resulting user experience (interaction tier). In addition, GISMOR provides a legal, financial and qualitative framework for the implementation of billable DiGAs (service tier). For the successful use of DiGAs, GISMOR continues to provide guidance on how to embed them in rehabilitation processes (process tier) and how to integrate them into transinstitutional information system architectures (information system tier). By means of the GISMOR guide for use, the developed approach will finally be applicable in practice. As a stand-alone document, the User Guide serves software developers, medical informaticians, and healthcare providers as a reference for action in the development and improvement of DiGAs and Medical Assistance Systems. Using the telerehabilitation system AGT-Reha as an example, it could be shown that GISMOR is suitable for both the analysis and the development of concrete measures for the improvement of a DiGA.

Encapsulation of clinically relevant cells, such as multipotent stromal cells (MSCs), in three-dimensional (3D) core-shell alginate structures is a promising method for the treatment of various diseases as well as drug testing and development. By utilizing the alginate capsules, drugs and active substances can be delivered specifically to the desired place within the patients’ body. The aim of this work was to investigate the effect of cryopreservation on the integrity of cell-free coaxial alginate capsules after thawing using slow freezing to ensure their long-term storage. Alginate capsules were produced by electro-spraying and cryopreserved using different cryoprotective agents (CPAs), incubation time and thawing temperature. The results suggest that the capsules loaded with 10% (v/v) dimethyl sulfoxide (DMSO) and 0.3 M sucrose and using high thawing rates demonstrated a beneficial effect on the integrity of capsules, as compared to 10% (v/v) DMSO alone and applying low thawing rates. This study is a further development towards the application of cryopreservation for long-term storage and the emerging cell-based medicine., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

Mit freundlicher Genehmigung des DAV - Deutscher Apotheker Verlag und der Deutschen Gesellschaft für Geschichte der Pharmazie., Geschichte der Pharmazie: Zeitschrift der Deutschen Gesellschaft für Geschichte der Pharmazie e.V., vol. 73, no. 4, p. 61

Jahrbuch der Braunschweigischen Wissenschaftlichen Gesellschaft, vol. 2020Jahrbuch 2020 der Braunschweigischen Wissenschaftlichen Gesellschaft, vol. 2020, p. 99

Mit freundlicher Genehmigung des DAV - Deutscher Apotheker Verlag und der Deutschen Gesellschaft für Geschichte der Pharmazie., Geschichte der Pharmazie: Zeitschrift der Deutschen Gesellschaft für Geschichte der Pharmazie e.V., vol. 73, no. 3, p. 33

Mit freundlicher Genehmigung des DAV - Deutscher Apotheker Verlag und der Deutschen Gesellschaft für Geschichte der Pharmazie., Geschichte der Pharmazie: Zeitschrift der Deutschen Gesellschaft für Geschichte der Pharmazie e.V., vol. 73, no. 1/2, p. 1

Pharmaziehistorische Bibliographie PhB, vol. 29

Die Untersuchung des Herzens war jeher ein zentraler Punkt der ärztlichen Diagnostik. Mit der Weiterentwicklung diagnostischer Methoden und Verfahren, von der Echokardiographie bis hin zu modernen bildgebenden Verfahren ist das Wissen über die Funktion des Herzens weitergewachsen. Doch können EKG oder MRT die Bedürfnisse des mobilen Alltags durch Größe und Komplexität nicht erfüllen. Die Ballistokardiographie (BCG) bietet hier eine potenzielle diagnostische Unterstützung. BCG als Methode zur Signalaufzeichnung von Schwingungen an der Körperoberfläche, um Rückschlüsse auf die Physiologie des kardiovaskulären Systems zu ziehen, bietet hier grundsätzliche neue Möglichkeiten. Diese Form der Diagnose, im Besonderen auch der Spezialfall der Seismokardiographie (SCG) lässt sich über kostengünstige, leichte, kleine und energieeffiziente Sensoren realisieren. Hierbei ist die Nutzung von digitalen Accelerometer eine Chance für die Weiterentwicklung der Herzdiagnostik. In dieser Arbeit wird die Untersuchung der Einsetzbarkeit kostengünstiger, digitaler 3D-Accelerometer untersucht. Dabei wird in verschiedenen Iterationen und Variationen ein vollumfängliches Messgesamtsystem für den potenziellen diagnostischen Einsatz in prototypenhafter Form konzipiert, erstellt und in verschiedenen Umgebungen und Settings, unter anderem an herzgesunden Erwachsenen und Kindern sowie auch an Herzinsuffizienten, untersucht. Hierbei stehen sowohl die Eruierung geeigneter Messpositionen, die Prüfung von Erkrankungen, welche sich aufgrund ihrer bekannten Symptome für die BCG/SCG-Messung eignen, als auch die Erarbeitung von Systemeigenschaften mit negativen Einfluss auf die Signalgüte, im Vordergrund. Nicht zuletzt wird die Herzinsuffizienz für den diagnostischen Einsatz untersucht und ein potenzielles Messsystem vorgestellt und ein Ausblick auf die klinische Nutzung gegeben. Als Fazit der beschriebenen Arbeit zeigt sich, dass die dargelegten Ergebnisse einen vollständigen Aufbau eines geeigneten Gesamtsystems in HW und SW ermöglichen. Zudem werden sowohl vorteilhafte Eigenschaften detailliert und umfassend beschrieben, wie zum Beispiel die Messposition oder Messumstände einer qualifizierten BCG/SCG-Messung. Mit den Schilderungen, Ergebnissen und Darstellungen der ausgewählten Analysen wird dem Leser eine Basis für zukünftige Messungen und Auswertungen sowie eine Unterstützung bei der Planung von Studien für die Erhebung von BCG- und SCG-Signalen zur Verfügung gestellt., The examination of the heart has always been a central point of medical diagnosis. With the further development of diagnostic methods and procedures, from echocardiography to modern imaging procedures, knowledge about the function of the heart has increased. However, due to their size and complexity, EKG or MRI cannot meet the needs of everyday mobile life. Ballistocardiography (BCG) offers potential diagnostic support here. BCG as a method for signal recording of vibrations on the body surface in order to draw conclusions about the physiology of the cardiovascular system offers fundamentally new possibilities here. This form of diagnosis, in particular the special case of seismocardiography (SCG), can be implemented using inexpensive, lightweight, small and energy-efficient sensors. The use of digital accelerometers is an opportunity for the further development of cardiac diagnostics. In this thesis, the investigation of the applicability of inexpensive, digital 3D-accelerometers is investigated. In different iterations and variations, a comprehensive overall measurement system for potential diagnostic use in prototype form is designed, created and examined in different environments and settings, including adults and children with healthy heart as well as heart failure. The focus here is on finding suitable measurement positions, testing diseases that are suitable for BCG/SCG measurement due to their known symptoms, and developing system properties with a negative impact on the signal quality. Finally yet importantly, heart failure is examined for diagnostic use, a potential measurement system is presented and an outlook on clinical use is given. The conclusion of the work described shows that the results presented enable a complete construction of a suitable overall system in HW and SW. In addition, advantageous properties are described in detail and comprehensively, such as the measurement position or measurement conditions of a qualified BCG/SCG measurement. The descriptions, results and representations of the selected analyzes provide the reader with a basis for future measurements and evaluations as well as support in planning studies for the collection of BCG and SCG signals.

Two-photon polymerization(2PP) is a process for three-dimensional (3D) micro-and Nano structuring based on the locally controlled curing of liquid precursors (light-sensitive resins) by photochemical triggered polymerization. In this decade, will be hearing a lot about this technic being applied to pharmaceutical applications like fabricating 3D microchannels for nanoparticle precipitation, nano-porous membranes and scaffolds for cell culturing, biomimetic organ-on-chip systems. This paper presents 2pp applied microsystems for continuously generating lipid nanoparticles which are one of the most important drug carrier system. The most important advantages of 2pp is manufacturing 3D shapes that is not possible with lithographic micro and nano fabrication technologies. Also, it will be shown how 2pp fabricated microchannel can be integrated with continuous size measurement by flowDLS for the feed-back controlled generation of nanoparticles., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

Jahrbuch der Braunschweigischen Wissenschaftlichen Gesellschaft, vol. 2020Jahrbuch 2020 der Braunschweigischen Wissenschaftlichen Gesellschaft, vol. 2020, p. 140

In this study, a novel approach was developed for determining the density of compacts using ultraviolet–visible spectrophotometry. The assumption within this context was that a change in density affects the corresponding color information of the compact. From the obtained spectra of the visible range, the color information of the compact was calculated which turned out to be directly proportional to the density of the compact. In comparison, the obtained spectra were analyzed using partial least square regression. The results of this study showed that both methods could be used predicting the density of a compact from the corresponding visible spectrum at identical accuracy. In contrast to the partial least square regression, the correlation of the color information as a direct output parameter of the spectrophotometer with the density required no excessive data pre-processing. Subsequently, the easier and faster data processing of the color information over the partial least square regression, conceives using this novel approach as potential process analytical technology tool for implementation into a compression process e.g., tableting or roller compaction., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

Dermatoses are skin diseases caused by lowered level of lipids in the intercellular lipid matrix of skin’s uppermost layer stratum corneum. A promising therapy is the application of liposomally formulated skin lipids (ceramides, cholesterol, fatty acids). These so called cerosomes proved very effective in disrupted skin barrier repair; however, they aim primarily on a mechanical surface restoration. For this reason an interesting idea is to combine the cerosomes and anti-inflammatory drugs (such as hydrocortisone) used for the treatment of pathophysiological processes in the skin. Unfortunately, liposomes generally show low drug encapsulation rates, hence they would not be very effective carriers. On the other hand, great encapsulation effectivity can be found in lipid nanocapsules (LNCs). We therefore aimed to create a combined system cerosomes-LNCs that would ensure the skin barrier restoration thanks to the ceramides and simultaneously effectively deliver hydrocortisone as a representative of anti-inflammatory drugs into lower layers of the skin. In this work, we characterized the formulations by dynamic light scattering and transmission electron microscopy to assess their size and morphology., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

Die Hämoglobinkonzentration in Blut ist einer der am häufigsten gemessenen Parameter. Für Hämoglobin ist die am häufigsten eingesetzte Messmethode die Cyanmethämoglobinmethode. Diese ist nicht auf das internationale Einheitensystem rückgeführt. Die Entwicklung von Verfahren, die eine metrologische Rückführbarkeit gewährleisten, ist in den letzten stark in den Vordergrund der Aktivitäten nationaler Metrologieinstitute gerückt. In dieser Arbeit wurde daher versucht einen vergleichbaren Immunoassay, basierend auf der oberflächenverstärkten Raman-Spektroskopie zu entwickeln. Die Kombination mit der Isotopenverdünnung (ID) ermöglicht präzise Messungen in selbst kleinsten Konzentrationsbereichen. Die Detektion des Hämoglobins erfolgt indirekt durch die Verwendung des ramanaktiven Markers 5,5′-Dithiobis-2-nitrobenzoesäure (DTNB) welcher auf Goldnanopartikeln mit einem Durchmesser von 40 nm immobilisiert wurde. Neben DTNB in nativer Form wurde für die Durchführung der ID ebenfalls ein isotopenangereichertes Analogon benötigt. Für einen erfolgreichen Immunoassay wurden weitere Funktionalisierungen der AuNP mit monoklonalen Antikörpern und einem zertifizierten Hämoglobin mit bekannter Konzentration durchgeführt. Die Funktionalisierungen der AuNP wurden eingehend mit Verfahren wie der UV/Vis-Spektroskopie, der Agarose-Gelelektrophorese und der Röntgenkleinwinkelstreuung untersucht. Der Nachweis der Spezifizität von monoklonalen sowie polyklonalen Antikörpern für Hämoglobin wurde mittels Gelelektrophorese überprüft. Für die Realisierung der ID wurden die AuNP entweder mit DTNB (+ Antikörper, + Hämoglobin) oder mit DTNB* (+ Antikörper, + Hämoglobin) in bestimmten äquidistanten Verhältnissen gemischt. Das höhere molekulare Gewicht des führt zu einer Verschiebung der Raman-Spektren, die in Kombination mit den Mischungsverhältnissen eine Bestimmung der (unbekannten) Proteinkonzentration ermöglicht. Durch die Kenntnis der für die Mischungsverhältnisse verwendeten Mengen (Nanopartikel, DTNB und Hämoglobin) kann eine Berechnung der unbekannten Konzentration erfolgen. Es wurden verschiedene Oberflächen wie Cellulosenitrat-Membranen und Glaswafer zum Immobilisieren getestet. Glaswafer zeigten sich als zielführend und ermöglichten die Erstellung eines Testformates des SERS-Immunoassays. Aufgrund der Problematik einer Aggregation der Partikel kann das Testformat des ID-SERS-Immunoassays nur eingeschränkt verwendet werden., Haemoglobin is one of the most frequently measured parameters in laboratory medicine. The only approved method for the detection of haemoglobin concentration the cyanmethaemoglobin method is not traceable to the SI. In general, there are severe problems in comparability of results in clinical laboratories and they can vary depending on the used testing kit. Raising from the problem of this lacking comparability and traceability the scope of this research was the development of a SERS-Immunoassay in combination with the Isotope Dilution approach which, is capable of an SI traceable quantification of haemoglobin. The detection of haemoglobin was performed indirect due to the usage of the strong Raman active marker DTNB. The aimed quantification of haemoglobin required additionally an isotopic enriched analogue of this marker. The used spherical goldnanoparticles with a size of 40 nm were functionalized with DTNB and its isotopologue and specific antibodies targeted against haemoglobin. The functionalization could be proved with different methods like UV/Vis, SAXS and agarose gel-electrophoresis. The specificity between antibodies and haemoglobin was successfully tested with Western blotting. This nanoparticle antibody complex was able to specific bind the haemoglobin and could be immobilized on a functionalized nitrocellulose membrane. The nitrocellulose-based approach appeared not to be suitable for a further development of the Immunoassay and an alternative was chosen. Due to these reasons an alternative approach with modified glass slides was executed. The alternative approach managed to minimize potential nanoparticle aggregation. With the achieved spectra a first try for a successful calibration model was made but with some limitations of the model itself. There are noticeable differences between the model based on the nitrocellulose membrane and the slides. Despite the influences of the nanoparticles, the ID-SERS-Immunoassay can be the basis for a further improvement of the quantification model for haemoglobin.

Malaria ist eine der häufigsten und gefährlichsten Infektionskrankheiten weltweit, die durch Parasiten der Gattung Plasmodium ausgelöst wird. Aufgrund der zunehmenden Resistenzen der Erreger gegen die derzeit eingesetzten Arzneimittel werden zusätzliche antiplasmodiale Wirkstoffe mit neuen Wirkungsmechanismen benötigt. In vorangehenden Untersuchungen identifizierten FUGEL und MASCH die Substanzklasse der 2-Acyl-4-arylthieno[2,3-b]pyridine als Hemmstoffe des plasmodialen Enzyms PfGSK-3. Einzelne Vertreter dieser Wirkstoffklasse zeigten auch antiplasmodiale Aktivität in einstellig mikromolaren Konzentrationen. Analoge Carbonsäureamide erwiesen sich jedoch als inaktiv gegenüber der postulierten biologischen Zielstruktur PfGSK-3. Neuere Untersuchungen ergaben jedoch, dass auch 42 ein potenter antiplasmodialer Wirkstoff ist. Die hier vorgelegte Arbeit war dem Design und der Synthese von Analoga der Verbindung 42 gewidmet, um Struktur-Eigenschafts- sowie Struktur-Wirkungsbeziehungen aufzustellen. Für die Herstellung neuer Strukturen wurden literaturbekannte Synthesesequenzen abgewandelt, wobei die Thieno[2,3-b]pyridin-Grundstruktur jeweils durch eine Thorpe-Ziegler-Zyklisierung als Schlüsselschritt gebildet wurde. Die durchgeführten biologischen Testungen ergaben, dass in dieser Substanzklasse am Aromaten in 4-Position die ortho-Chlor-Substitution verzichtbar ist und stattdessen eine meta-Methylgruppe besonders gute antiplasmodiale Aktivitäten ergibt. Die Cyan- und Aminogruppe in 5- bzw. 6-Position am Thieno[2,3-b]pyridin kann durch aliphatische Reste oder Carbonsäurestrukturen ersetzt werden. Strukturen wie 42 und 46f zeigen zwar deutliche antiplasmodiale Aktivität, hemmen jedoch nicht das plasmodiale Enzym PfGSK-3. Wird der aromatische Substituent am Amid-Stickstoffatom gegen einen aliphatischen Rest ersetzt, kehren sich diese Eigenschaften um. So ist das N-Methylamid 47k ein PfGSK-3-Inhibitor, zeigt aber keine nennenswerte antiplasmodiale Aktivität. Ebenso wie die Verbindungen 42, 46f und 47k zeigt der Carbonsäureester 46h antiplasmodiale Aktivität bei schlechter Wasserlöslichkeit. Die entsprechende Carbonsäure 46i ist dagegen deutlich besser wasserlöslich, weist jedoch keine antiplasmodiale Aktivität mehr auf. Stabilitätsuntersuchungen ergaben außerdem, dass sich die potenteste Verbindung 46f unter Lichtbestrahlung in gelöster Form über einen Zeitraum von einigen Tagen zersetzt, unter Lichtausschluss jedoch über längere Zeit stabil ist., Parasites of the genus Plasmodium induce malaria, one of the most common and dangerous infectious diseases worldwide. Due to an increasing resistance of the pathogens to currently used drugs, additional antiplasmodial agents with new mechanisms of action are needed. In previous studies, FUGEL and MASCH identified the substance class of 2-acyl-4-arylthieno[2,3-b]pyridines as inhibitors of the plasmodial enzyme PfGSK-3. Individual representatives of this substance class also showed antiplasmodial activity in single-digit micromolar concentrations. Analogous carboxylic acid amides, however, proved to be inactive towards the postulated biological target structure PfGSK-3. Yet, recent investigations have shown that 42 is also a potent antiplasmodial agent. The work presented here was dedicated to the design and synthesis of analogues of compound 42 to establish structure-property and structure-activity relationships. For the preparation of new structures, literature-known syntheses sequences were modified, whereby the thieno[2,3-b]pyridin basic structure was formed by a Thorpe-Ziegler cyclization as a key step in each case. Biological tests showed that in this substance class ortho-chlorine substitution on the aromatic substituent in the 4-position is dispensable and instead a meta-methyl group shows particularly good antiplasmodial activities. The cyano and amino groups in the 5-position and 6-position on thieno[2,3-b]pyridine-scaffold can be replaced by aliphatic residues or carboxylic acid structures. Structures such as 42 and 46f show clear antiplasmodial activity, but do not inhibit the plasmodial enzyme PfGSK-3. By replacing the aromatic substituent on the amide nitrogen atom with aliphatic residues, these properties are reversed. For example, N-methylamide 47k is a PfGSK-3 inhibitor but does not exhibit significant antiplasmodial activity. Like the compounds 42, 46f and 47k, the carboxylic acid ester 46h shows antiplasmodial activity along with poor water solubility. The corresponding carboxylic acid 46i, on the other hand, is much more water soluble, but no longer exhibits antiplasmodial activity. Stability tests also showed that the most potent compound 46f decomposes in dissolved form under light irradiation over a period of several days but is stable over a longer period under exclusion of light.

Chondroitin sulfate A was covalently immobilized onto a monolithic silica epoxy column involving a Schiff base formation in the presence of ethylenediamine as a spacer and evaluated in terms of its selectivity in enantioseparation. The obtained column was utilized as a chiral stationary phase in enantioseparation of amlodipine and verapamil using a mobile phase consisting of 50 mM phosphate buffer pH 3.5 and UV detection. Sample dilution by organic solvents (preferably 25% v/v acetonitrile-aqueous solution) was applied to achieve baseline enantioresolution (Rs &gt, 3.0) of the individual drug models within 7 min, an excellent linearity (R2 = 0.999) and an interday repeatability of 1.1% to 1.8% RSD. The performance of the immobilized column for quantification of racemate in commercial tablets showed a recovery of 86&ndash, 98% from tablet matrices. Computational modeling by molecular docking was employed to investigate the feasible complexes between enantiomers and the chiral selector.

In situ tissue engineering requires to functionalize the surfaces of implants, to achieve better biocompatibility or to release pharmaceuticals. Often a burst release of such pharmaceuticals takes place and the surface functionality is gone rapidly. Casein-micelles (M-Cas) offer unique release capabilities for hydrophobic and hydrophilic substances. In this work, M-Cas were reconstructed and used as release system for curcumin as a model drug. Modified electrospun polycaprolactone (PCL)-fiber mats were coated with chitosan / tripolyphosphate nanoparticles (CS/TPP-NP) and functionalized with the reconstructed M-Cas. [1] CS/TPP-NPs and Na-caseinate (Cas) were used as blocking layer to slow down the release of curcumin loaded micelles. The layers were investigated by surface zeta-potential (ZP). The release of the curcumin from surfaces was possible and the blocking layers had a significant influence on the kinetics., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

Solid triglyceride nanoparticles are under intensive investigation as drug delivery systems. In the solid state, triglycerides exist in three polymorphic modifications which differ from each other in their physicochemical properties. Thus, it is particularly important to know the influence of parameters affecting the polymorphic behavior of the solid triglyceride nanoparticles to get a stable product with defined physicochemical properties over an appropriate storage time. This study investigated the effect of subsequent addition of SDS to tripalmitin nanoparticles, which had been stabilized with partially hydrolyzed poly(vinyl alcohol) (PVA). The polymorphic behavior was characterized with DSC and X-ray diffraction over a storage time of 28 d at 20 °C. A faster transition into more stable polymorphic modifications was observed after the addition of SDS to the nanoparticles., SPhERe Proceedings: 4th International Symposium on Pharmaceutical Engineering Research

All medical devices and implants are made of synthetic or natural, but usually not endogenous, materials. Implantation of such devices, into living tissue, is therefore subject to the risk of nosocomial infections, biofilm formation and may cause implant-associated infections. Microorganisms (including bacteria) that grow in biofilms and cause chronic inflammation are the typical source of these infections. Subsequently, they can lead to implant failure, requiring further surgical treatments. To prevent this, this study investigated and presented enzymatically degradable drug delivery systems that enable encapsulation and targeted release of an antibacterial agent triggered by an infection. To achieve this, the dissertation focused on the fabrication, characterization of suitable nanocarriers and their immobilization on implant surfaces such as titanium. To build a stable and degradable encapsulation system, different steps were established: 1) the integration of an enzyme-labile model peptide into two different polymer-based systems 2) generation of particles 3) coating of titanium surfaces with the particles 4) investigation of stability and degradability of those systems 5) the release of ciprofloxacin as a model substance. For this, two approaches have been tested. The first approach included the synthesis of chitosan‐g‐[peptide‐poly‐L‐caprolactone] and its self‐assembly into polymeric vesicles by the solvent shift method. For the second approach, nanogels dispersions were prepared by ionotropic gelation of the alginate with the poly-L-lysine, which was conjugated with ciprofloxacin via a copper-free 1,3-dipolar cycloaddition (click reaction)., Alle medizinischen Geräte und Implantate bestehen aus synthetischen oder natürlichen, jedoch in der Regel aus nicht endogenen Materialien. Die Implantation solcher Geräte in lebendes Gewebe unterliegt daher dem Risiko nosokomialer Infektionen und der Bildung von Biofilmen und kann implantatassoziierte Infektionen verursachen. Mikroorganismen (einschließlich Bakterien), die in Biofilmen wachsen und chronische Entzündungen verursachen, sind die typische Quelle dieser Infektionen. Anschließend können sie zu einem Implantatversagen führen, was weitere chirurgische Behandlungen erforderlich macht. Um dies zu verhindern, untersuchte und ergab diese Studie enzymatisch abbaubare Arzneimittelabgabesysteme, die die Einkapselung und gezielte Freisetzung eines durch eine Infektion ausgelösten antibakteriellen Mittels ermöglichen. Um dies zu erreichen, konzentrierte sich die Dissertation auf die Herstellung und Charakterisierung geeigneter Nanoträger sowie deren Immobilisierung auf Implantatoberflächen wie Titan. Um ein stabiles und abbaubares Einkapselungssystem aufzubauen, wurden verschiedene Schritte festgelegt: 1) Integration eines enzymlabilen Modellpeptids in zwei verschiedene Systeme auf Polymerbasis 2) Erzeugung von Partikeln 3) Beschichtung von Titanoberflächen mit den Partikeln 4) Untersuchung von Stabilität und Abbaubarkeit dieser Systeme 5) Freisetzung von Ciprofloxacin als Modellsubstanz. Hierzu wurden zwei Ansätze getestet. Der erste Ansatz umfasste die Synthese von Chitosan-g-[Peptid-Poly-ε-Caprolacton] und dessen Selbstorganisation zu polymeren Vesikeln durch das Lösungsmittel-Verschiebungsverfahren. Für den zweiten Ansatz wurden Nanogeldispersionen durch ionotrope Gelierung des Alginats mit dem Poly-L-Lysin hergestellt, das über eine kupferfreie 1,3-dipolare Cycloaddition (Klickreaktion) mit Ciprofloxacin konjugiert wurde.

Analytical-method development based on design of experiment has been applied for optimizing the enantioseparation of amlodipine by chiral capillary electrokinetic chromatography using maltodextrin as the chiral selector. The effect of different factors on the enantioresolution quality was screened. Three separation factors, namely maltodextrin concentration, pH of the background electrolyte and applied voltage were selected as independent variables. The number of experiments was reduced while maximizing the information content using design of experiment. Based on a full-quadratic design that included three variables on three levels, the total design space could be reduced to fifteen factor combinations using a D-optimal algorithm. The aim of the experiment was to find the optimal factor combinations with respect to resolution. The maltodextrin concentration (7.5–10% w/v) demonstrated the strongest effect on the resolution followed by pH (2–4) of the background electrolyte and the applied voltage (15–20 kV). An increase in the maltodextrin concentration was found to result in a greater stereoselectivity, represented by the higher resolution values (Rs ≥ 1.5). The separation conditions in the proposed method were feasible to be adjusted within the applied range with an acceptable resolution.

Bone morphogenetic protein 2 (BMP-2) has a high tendency to aggregate at physiological pH and physiological ionic strength, which can complicate the development of growth factor delivery systems. The aggregation behavior in differently concentrated BMP-2 solutions was investigated using dynamic and static light scattering. It was found that at higher concentrations larger aggregates are formed, whose size decreases again with increasing dilution. A solubilizing effect and therefore less aggregation was observed upon the addition of albumin. Imaged capillary isoelectric focusing and the simulation of the surface charges of BMP-2 were used to find a possible explanation for the unusually low solubility of BMP-2 at physiological pH. In addition to hydrophobic interactions, attractive electrostatic interactions might be decisive in the aggregation of BMP-2 due to the particular distribution of surface charges. These results help to better understand the solubility behavior of BMP-2 and thus support future pharmaceutical research and the development of new strategies for the augmentation of bone healing.

Eine gesunde Cornea ist essenziell wichtig für das Sehvermögen. Die Transplantation einer Spenderhornhaut kann bei einer Vielzahl von Erkrankungen der Augenoberfläche das Sehvermögen wiederherstellen. Da die Verfügbarkeit der Spenderhornhäute nicht ausreicht, um den Bedarf zu decken, eröffnet die In-vitro-Rekonstruktion, das sogenannte Tissue Engineering, vielversprechende Möglichkeiten. In dieser Arbeit wurden primäre, corneale Stromazellen aus humanen Hornhäuten isoliert und in-vitro über mehrere Wochen kultiviert. Der Einfluss verschiedener Kultivierungsbedingungen auf die Bildung eines zusammenhängenden, von der Kultivierungsoberfläche ablösbaren Zellsheets, bestehend aus Zellen und extrazellulärer Matrix (EZM) wurde untersucht. Die Zellen wurden in unterschiedlichen Zelleinsaatdichten und mit verschiedenen Konzentrationen an Mediensupplementen wie Vitamin C, fetalem bovinem Serum und L-Glutamin kultiviert. Des Weiteren wurde der Einfluss von Kollagenbestandteilen (Glycin, L-Prolin, L-4-Hydroxyprolin) und der Einfluss von TGF-β1, bFGF, IGF-2, PDGF-BB und Insulin auf die Proliferation, die Kollagen- sowie die Glykosaminoglykansynthese der Zellen untersucht. Zur besseren Standardisierbarkeit wurde außerdem ein serumfreies Kultivierungsmedium entwickelt. Die entstehenden Zellsheets wurden hinsichtlich ihrer Lichttransmission, Ultrastruktur und biomechanischen Eigenschaften charakterisiert. Die Synthese EZM wurde durch die Kultivierung der Zellen mit Insulin, TGF-β1 und Vitamin C signifikant erhöht. Die Zellsheets wiesen eine hohe Lichttransmission, geeignete Materialeigenschaften und teilweise Ähnlichkeiten zur Ultrastruktur der humanen Hornhaut auf. Die EZM bestand, wie das corneale Stroma, vor allem aus Kollagen Typ I und zu kleineren Anteilen aus Kollagen Typ V und Typ III. Die für das humane Stroma charakteristische Anordnung der Kollagenfibrillen, parallel zueinander und in Lamellen übereinander, konnte in einigen Bereichen der Zellsheets gefunden werden. Eine serumfreie Kultivierung der Zellen mit TGF-β1-haltigem Medium resultierte in einem zusammenhängenden Zellsheet, während die Kultivierung ohne TGF-β1 zu fragilen Sheets führte. Die Arbeit beschreibt die Entwicklung eines im Querschnitt in-vivo-ähnlichen Stroma-Konstrukts durch Übereinanderstapeln der Zellsheets. Hier scheint eine serumfreie Kultivierung der Zellsheets übereinander in einer Zellkrone (zwei Edelstahlzylinder, zwischen denen das Konstrukt eingespannt wird) vielversprechend zu sein., A healthy cornea is essential for vision. The transplantation of a donor cornea can restore vision in a variety of ocular surface diseases. Since the availability of donor corneas is insufficient to meet the demand, in vitro reconstruction, so-called tissue engineering, provides promising possibilities. In this work, primary corneal stroma cells were isolated from human corneas and cultivated in-vitro over several weeks. The influence of different cultivation conditions on the formation of a coherent cell sheet consisting of cells and extracellular matrix (ECM), which can be detached from the cultivation surface, was investigated. The cells were cultivated at different cell seed densities and with different concentrations of media supplements such as vitamin C, fetal bovine serum and L-glutamine. Furthermore, the influence of collagen components (glycine, L-proline, L-4-hydroxyproline) and the influence of TGF-β1, bFGF, IGF-2, PDGF-BB and insulin on cell proliferation, collagen and glycosaminoglycan synthesis was investigated. In addition, a serum-free culture medium was developed for better standardization. The resulting cell sheets were characterized with respect to light transmission, ultrastructure and biomechanical properties. The synthesis of ECM was significantly increased by culturing the cells with insulin, TGF-β1 and vitamin C. The cell sheets showed high light transmission, suitable material properties and partial similarities to the ultrastructure of the human cornea. The ECM, like the corneal stroma, consisted mainly of collagen type I and to smaller parts of collagen type V and type III. The arrangement of collagen fibrils, parallel to each other and in lamellae on top of each other, which is characteristic for the human stroma, could be found in some areas of the cell sheets. Serum-free cultivation of the cells with TGF-β1-containing medium resulted in a coherent cell sheet, while serum-free cultivation without TGF-β1 resulted in fragile sheets. Finally, the work describes the development of a cross-sectional in-vivo-like stroma construct by stacking the single cell sheets on top of each other. Here, a serum-free cultivation of the cell sheets on top of each other in a cell crown (two stainless steel cylinders between which the construct is clamped) seems promising.

Nowadays, vaccination represents the most effective medical intervention to control and prevent the spreading of infectious diseases in the population. Nonetheless, the emergence of new pathogens, the logistic constraints for the vaccine storage and the safety concerns regarding the use of whole pathogens, as well as the rising wave of skepticism among the population toward vaccination, have led to the rebound of some almost eradicated pathogens and delayed the success to control some other diseases like influenza infection. Hence, the development of non-invasive needle-free vaccination strategies combined with subunit vaccines able to stimulate protective immune responses is required. In this thesis, we have confirmed the potential of the transfollicular (TF) route for the delivery and the generation of a protective immune response using inactivated influenza viruses. Moreover, by investigating the early innate immune events occurring in the skin after TF immunization using polymeric nanocarriers, we have shown that the nanocarrier is needed for the effective delivery and uptake of the vaccine antigen. In addition, the co-formulation of the nanoparticles with either of Bis-(3´, 5´)-cyclic dimeric adenosine monophosphate (c-di-AMP) or pegylated aGalactosylceramide (aGCMPEG) adjuvants, help to tailor the innate immune response by acting on specific skin dendritic cells and therefore participate in the induction of a differentially polarized adaptive immune response. Altogether, this study has revealed not only the potential of the TF route for vaccine delivery but also that the combination of this route with adjuvanted polymeric nanoparticles is able to induce a protective immune response against live influenza virus challenge., Impfungen stellen die effizienteste Erfindung in der Medizin dar, um Infektionskrankheiten zu kontrollieren und ihre Ausbreitung zu verhindern bzw. einzudämmen. Nichtsdestotrotz führen neu auftretende Krankheiten, Sicherheitsbedenken bei der Verwendung von Lebendimpfstoffen, sowie logistische Probleme bei der Lagerung und Einhaltung von Kühlketten und nicht zuletzt die zunehmende Impfmüdigkeit dazu, dass bereits unter Kontrolle oder gar ausgerottet geglaubte Pathogene wieder neu erstarken. Aus diesen Gründen ist es notwendig, neue nadelfreie nicht-invasive Impfstrategien zu entwickeln, die sich durch ein erhöhtes Sicherheitsprofil bei gleichzeitig hoher Effektivität kennzeichnen. In der vorliegenden Arbeit konnten wir bestätigen, dass sich die transfollikuläre (TF) Impfroute dazu eignet, schützende Immunantworten gegen das Influenzavirus zu stimulieren. Darüber hinaus konnte durch Analyse der unmittelbar nach TF Immunisierung ablaufenden Prozesse des angeborenen Immunsystems in der Haut gezeigt werden, dass Nanopartikel für diese Impfstrategie notwendig sind, um einen effektiven Transport des Impfstoffantigens zu den, sowie dessen Verarbeitung durch die Immunzellen zu gewährleisten. Ferner kann durch Zugabe der Adjuvantskandidaten Bis-(3´,5´)-zyklisches dimerisches Adenosinmonophosphat (c-di-AMP) und dem pegylierten αGalactosylceramid (αGCMPEG) zu dem Polymer-basierten Nanotransporter die frühe Immunantwort derart modifiziert werden, dass die erworbene Immunantwort unterschiedlich polarisiert verläuft. Die vorliegende Arbeit hat somit nicht nur das Potential der TF-Impfstrategie als Alternative zur klassischen Impfung überhaupt deutlich gemacht, sondern auch dargelegt, dass diese Route bei Verwendung von Nanotransportern in Verbindung mit Adjuvantien ebenfalls für Subunit-Impfstoffe geeignet ist, um z. B. eine schützende Immunantwort gegen ein Influenzavirus zu stimulieren.

In Deutschland dürfen Reizstoffsprühgeräte (RSG) nur mit Zulassung hergestellt und verkauft werden, wenn sie als Selbstverteidigungsmittel gegen Menschen gedacht sind. Nach der Änderung des Waffenrechts im Jahr 2003 hat die Fraunhofer-Gesellschaft die Zuständigkeit der Prüfung der Reizstoffsprühgeräte abgegeben. Es gab seitdem keine Richtlinien für die Messeinrichtungen. Aktuell ist die Physikalisch-Technische Bundesanstalt für die Zulassung zuständig. Im ersten Teil dieser Arbeit wurde eine Anforderung für Reizstoffsprühgeräte erstellt. Sie ermöglicht neue Zulassungen von RSG durch die PTB. Das Prüfverfahren wurde getestet. Der zweite Teil der Arbeit fokussiert sich auf die Untersuchung der Gefahr des Herzkammerflimmerns von Elektroimpulsgeräten (EIG) und deren neue Bewertungsverfahren. EIG gehören zu den bekanntesten Personenschutzgeräten und dürfen in Deutschland verkauft werden. Sie übertragen wie ein TASER© Impulse in den Körper der Zielperson. Die Wirkung von Elektroimpulsgeräten soll die direkte oder indirekte Erregung einer großen Masse von Zellen im Körper durch den Strom hervorrufen. Zu den erregbaren Zellen zählen die Nervenzellen und die Muskelzellen. Die Gefährdung eines EIG wird auch von seinem Strom verursacht. Ein anatomisches menschliches CAD-Körpermodell wurde nach der Anforderung dieser Arbeit angepasst. Das Modell wurde in einer Finite-Elemente-Methode-Modellierung (FEM) eingesetzt. Mit der Modellierung kann die Stromverteilung im Körper berechnet werden, welche als Mittel der Bewertung der Gefährlichkeit eines möglichen vom EIG verursachten Herzkammerflimmerns verwendet wird. Um das computerbasierte Modell zu erstellen, werden die elektrischen Eigenschaften des menschlichen Körpers benötigt. Die Leitfähigkeit von Muskelgewebe wurde im Rahmen dieser Arbeit im Hochspannungsbereich gemessen. Die Feldstärkeabhängigkeit der Leitfähigkeit wurde nachgewiesen. Deren gemessene Kennlinie wurde zum Aufbau des Gleichungssystems in das FEM-Modell eingesetzt. Drei physikalische Modelle wurden in dieser Arbeit verglichen, nämlich ein elektrisches quasistatisches Modell, ein frequenzabhängiges Modell und ein Modell mit einer feldstärkeabhängigen Leitfähigkeit. Die Kenngrößen Ausgangsspannung, Elektrodenabstand und Elektrodendurch-messer eines EIG führten über das FEM-Modell zu einer Aussage der Stromdichte auf der Herzoberfläche. Das Modell mit einer feldstärkeabhängigen Leitfähigkeit wurde zusammen mit einem Nervenzellenmodell als das Berechnungstool in dem neuen Bewertungsverfahren verwendet. Das neue Verfahren kann auch die Wirksamkeit von EIG evaluieren. Um die zukünftige komplizierte Berechnung mit dem FEM-Modell zu vermeiden, wurde eine Lookup-Tabelle erstellt., In Germany, irritant gas spraying devices (RSG) may only be manufactured and sold with approval if they are intended as a self-defense device against humans. Following the amendment of the law in 2003, the Fraunhofer-Gesellschaft has given up responsibility for testing irritant gas spraying devices. Since then, there have been no guidelines for the measuring devices. Currently, the Physikalisch-Technische Bundesanstalt (PTB) is responsible for the approval. In the first part of this work, a requirement for irritant gas spraying devices has been established. It enables new approvals of RSG by the PTB. The test method was established. The second part of the thesis focuses on the investigation of the risk of ventricular fibrillation of electric shock devices (EIG) and their new evaluation procedures. EIG are among the most well-known personal defense equipment and can be sold in Germany. They transmit impulses into the body of the target person like a TASER©. The effect of EIG is intended to cause the direct or indirect stimulation of a large mass of cells especially nerve cells and muscle cells in the body through the current. The threat of an EIG is caused by its current. An anatomical CAD human model was adapted to the requirement of this work. The model was used in a finite element method modeling (FEM). This modeling can be applied to calculate the distribution of current in the body, which is used as a means of assessing the danger of any potential ventricular fibrillation caused by the EIG. To create the computer-based model, the electrical properties of the human body are required. The conductivity of muscle tissue was measured in the context of this work in the high voltage range. The field strength dependence of the conductivity was detected. The measured characteristic curve was used to build the equation system in the FEM model. Three physical models were compared in this work: an electrical quasi-static model, a frequency-dependent model and a model with a field-strength-dependent conductivity. The output voltage, the electrode spacing and the electrode diameter of an EIG led to a statement of the current density on the heart surface using the FEM model. The model with a field-strength-dependent conductivity was used together with a neuron model as the calculation tool in the new test method. The new method can also evaluate the effectiveness of EIG. To avoid a complicated calculation with the FEM model in the future, a lookup table has been created., Berichte aus dem Institut für Elektrische Messtechnik und Grundlagen der Elektrotechnik, vol. 69

Chronic hepatotropic viral infections represent a major challenge in current medicine. Chronic infections are a consequence of dysfunctional CD8+ T cell responses which impair effective viral clearance. Dysfunction in CD8+ T cells, as a result of T cell exhaustion, lift the break on viral replication, thereby promoting the development of liver diseases. Currently, immunomodulatory strategies are being explored as an attractive option to reinvigorate exhausted T cells in chronic infection as well as in cancer. However, the dynamic regulation of exhausted T cells as well as the mechanisms enforcing reinvigoration of exhausted T cells in the liver are not clearly defined. In this study, an OvaXCre transgenic mouse model characterized by antigen expression specifically in the liver was used to elucidate the dynamic differentiation and reinvigoration of exhausted T cells in the liver. The expression of antigen in a fraction of hepatocytes mimics persisting antigen situation in chronic liver infections. In this regard, it was observed that host conditioning with CpG oligodeoxynucleotides (CpG ODN), which modulates the liver environment, translates into eradication of chronic Ova-antigen from the liver by the action of cytotoxic T cells, probably as a result of exhausted T cell reinvigoration. Interestingly, antigen clearance was accompanied with an extreme loss of antigen specific T cells and T cell stemness. In depth characterization of exhausted T cells in liver revealed two distinct T cell subsets, identified by presence or absence of the chemokine receptor CXCR5. CXCR5+ and CXCR5- T cells exhibit profound differences in cytotoxic, effector and memory functions, with all these functional capacities skewed towards CXCR5+ T cells in the liver. The CXCR5+ T cells are follicular-like and liver resident cells by virtue of upregulation of the transcription factors Bcl6 and LFA-1. Robust and efficient mitochondria activities were manifested in CXCR5+ T cells, which was in contrast to CXCR5- T cells. Importantly, CXCR5+ T cells demonstrated a significant nutrient acquisition advantage and show enhanced recall responses and self-maintenance in vivo. In sum, these data show that CpG ODN immunomodulation reinvigorates exhausted T cells. While heterogeneity exists in the intrahepatic pool of exhausted T cells, the CXCR5+ T cell subset plays a major role in eliminating chronic antigen expression in liver. Therefore, this subset may be of interest for the development of novel immune based therapeutic strategies against chronic liver infections., Chronische hepatotrope Virusinfektionen stellen in der heutigen Medizin eine große Herausforderung dar. Dies ist eine Folge der dysfunktionalen CD8+ T-Zell-Reaktionen, die als T-Zell-Erschöpfung bekannt sind und die effektive virale Beseitigung beeinträchtigen. Gegenwärtig werden immunmodulatorische Strategien zur Wiederbelebung erschöpfter T-Zellen bei chronischen Infektionen und Krebs erforscht. Die dynamische Regulation erschöpfter T-Zellen und die Mechanismen, die die Wiederbelebung erschöpfter T-Zellen in der Leber verstärken, sind jedoch nicht klar definiert. In dieser Studie wurde ein transgenes OvaXCre-Mausmodell verwendet, das durch die Expression von Ova-Antigen in der Leber charakterisiert ist, um die dynamische Differenzierung und die Wiederbelebung erschöpfter T-Zellen aufzuklären. Das in Hepatozyten exprimierte Antigen imitiert die persistierende Antigensituation bei chronischen Leberinfektionen. Es wurde beobachtet, dass die Wirtskonditionierung mit CpG-Oligodesoxynukleotiden, die das Lebermilieu moduliert, durch die Wirkung zytotoxischer T-Zellen zu einer chronischen Erythrozyten-Antigen-Eradikation führt, wahrscheinlich als Folge der Wiederbelebung erschöpfter T-Zellen. Interessanterweise ging die Antigen-Beseitigung mit dem Verlust von antigenspezifischen T-Zellen und T-Zell-Stammzellen einher. Die Charakterisierung erschöpfter T-Zellen in der Leber ergab zwei verschiedene T-Zell-Untergruppen, die durch das Vorhandensein oder Fehlen des Chemokinrezeptors CXCR5 identifiziert wurden. CXCR5+ und CXCR5- T-Zellen weisen tiefgreifende Unterschiede in zytotoxischen, Effektor- und Gedächtnisfunktionen auf, wobei alle diese funktionellen Kapazitäten verstärkt in CXCR5+ T-Zellen in der Leber sind. Es wurde beobachtet, dass die CXCR5+ T-Zellen follikelähnliche und leberresidente Zellen sind. Zudem manifestierten sich robuste und effiziente Mitochondrienaktivitäten in CXCR5+ T-Zellen. Wichtig ist, dass die CXCR5+ T-Zellen einen signifikanten Vorteil bei der Nährstoffbeschaffung zeigten und in vivo erhöhte Rückrufreaktionen und Selbsterhaltung zeigten. Zusammenfassend zeigen diese Daten, dass die CpG-ODN-Immunmodulation erschöpfte T-Zellen wiederbelebt. Während im intrahepatischen Pool erschöpfter T-Zellen Heterogenität besteht, spielt die CXCR5+ T-Zell-Untergruppe eine wichtige Rolle bei der Eliminierung der chronischen Antigenexpression in der Leber. Daher könnte diese Untergruppe für die Entwicklung neuer immunbasierter therapeutischer Strategien gegen chronische Leberinfektionen von Interesse sein.

Um bereits in der frühen Entwicklungsphase die Durchlässigkeit potentieller Wirkstoffe durch die Blut-Hirn-Schranke abschätzen zu können, sind zuverlässige in vitro-Zellkulturmodelle erforderlich. Eine breite Anwendung findet hierbei die humane Zelllinie hCMEC/D3, wobei ihre mäßig restriktiven Barriereeigenschaften einen erheblichen Nachteil darstellen. Eine Erklärung für ihre geringe Barriereintegrität und der hohen Durchlässigkeit für Natriumfluoreszein und FITC-Dextran 4 kDa lieferte die immunzytochemische Untersuchung wichtiger TJ-Proteine. So waren Occludin, ZO-1 und Claudin-5 größtenteils zytosolisch lokalisiert und nicht wie erwartet intramembranös. Hiervon ausgehend wurden in der vorliegenden Arbeit verschiedene statische sowie dynamische Kultivierungsvariationen zur Verbesserung der Barrierefunktionalität analysiert. Bei der Untersuchung verschiedener Zellkulturmedien stellten sich die Medien ECGM MV-2, MCDB 131 und das serumfreie Medium hESFM neben dem Standardmedium EGM-2 am geeignetsten heraus. Allerdings zeigte keines der untersuchten Medien einen positiven Einfluss auf die endotheliale Barrierewirksamkeit. Auch verschiedene Beschichtungsmaterialien (Laminin, Kollagen, und Fibronektin), Medienzusätzen (Serum, Hydrocortison, Calcium und pCPT-cAMP kombiniert mit Ro-20-1724), Porengrößen (0,4, 1,0, 3,0 µm) und Ko-Kultivierungsvariationen (direkt und indirekt mit SVGmm) verbesserten die Barriereintegrität der hCMEC/D3 Zellen nicht. Zudem wurde der Einfluss einer dynamischen Kultivierung unter Anwendung des Zellkulturmodells DynaMiTES untersucht. Zunächst wurden die Kultivierungsbedingungen erfolgreich an das DynaMiTES-Design angepasst, indem die Aussaatrichtung von normal auf invers geändert wurde. Die Endothelzellen wurden bei einer Flussrate bis 3,33 ml/min für 3 h und bei 0,03 ml/min für 72 h ohne einen Verlust der Zellviabilität und der Zellzahl kultiviert. Jedoch blieben die Barriereeigenschaften der Endothelzellen unter allen untersuchten Flussraten unverändert. Die Ausbildung dichter Zell-Zell-Kontakte ist eine Voraussetzung für die restriktiven Eigenschaften der Blut-Hirn-Schranke. Die Ergebnisse verdeutlichen, dass die hCMEC/D3 Zelllinie, trotz der getesteten Kultivierungsvariationen, nicht für Permeationsstudien von niedermolekularen Arzneimittelkandidaten geeignet ist. Somit bleibt der Bedarf nach einer Zelllinie mit restriktiven Barriereeigenschaften weiterhin bestehen., Reliable in vitro cell culture models are required to estimate the brain accessibility in the early phase of drug development. For this purpose the human endothelial cell line hCMEC/D3 is widely used, but its moderately restrictive barrier properties are a considerable disadvantage. The low barrier integrity and high permeability for sodium fluorescein and FITC-dextran 4 kDa might be due to the incomplete formation of tight junction proteins. Thus, ZO-1, occludin and claudin-5 were mostly cytosolically localized and not intramembranously, as expected. Based on this, various static and dynamic cultivation variations were analyzed to improve barrier functionality of hCMEC/D3 cells regarding an in vivo-like blood-brain barrier model for drug permeation studies. Firstly, the effect of ten different cell culture media, including two serum-free ones, on barrier properties was investigated. Besides the standard medium EGM-2, the basal media ECGM MV-2, MCDB 131 and the serum-free medium hESFM seemed to be most suitable. Though, none of the tested cell culture media showed a positive effect on the barrier integrity of the endothelial cell line. Variations of different coating materials (laminin, collagen and fibronectin), media supplements (serum, hydrocortisone, calcium and pCPT-cAMP combined with Ro 20 1724), pore sizes (0.4, 1.0, 3.0 µm) and co-cultivation variations (directly and indirectly with SVGmm) did not improve the barrier integrity of the hCMEC/D3 cells. Furthermore, the cell line was cultivated under dynamic conditions using the cell culture model DynaMiTES. The cultivation conditions were successfully adapted to the DynaMiTES design by changing the seeding orientation from normal to inverse. The endothelial cells were cultivated for 3 h at a flow rate up to 3.33 ml/min and for 72 h at a flow rate of 0.03 ml/min without loss of cell viability and cell count. However, the barrier integrity of the endothelial cell line remained unchanged under all dynamic conditions. The formation of tight cell-cell contacts is essential for the restrictive properties of the blood-brain barrier. The results show that the hCMEC/D3 cell line is not suitable for permeation studies of small molecule drug candidates. Even after the tested cultivation variations the hCMEC/D3 model showed considerable limitations and therefore the need for an alternative cell line with restrictive barrier properties remains.

We designed a low-cost wrist-worn prototype for simultaneously measuring environmental, behavioral, and physiological domains of influencing factors in healthcare. Our prototype continuously monitors ambient elements (sound level, toxic gases, ultraviolet radiation, air pressure, temperature, and humidity), personal activity (motion tracking and body positioning using gyroscope, magnetometer, and accelerometer), and vital signs (skin temperature and heart rate). An innovative three-dimensional hardware, based on the multi-physical-layer approach is introduced. Using board-to-board connectors, several physical hardware layers are stacked on top of each other. All of these layers consist of integrated and/or add-on sensors to measure certain domain (environmental, behavioral, or physiological). The prototype includes centralized data processing, transmission, and visualization. Bi-directional communication is based on Bluetooth Low Energy (BLE) and can connect to smartphones as well as smart cars and smart homes for data analytic and adverse-event alerts. This study aims to develop a prototype for simultaneous monitoring of the all three areas for monitoring of workplaces and chronic obstructive pulmonary disease (COPD) patients with a concentration on technical development and validation rather than clinical investigation. We have implemented 6 prototypes which have been tested by 5 volunteers. We have asked the subjects to test the prototype in a daily routine in both indoor (workplaces and laboratories) and outdoor. We have not imposed any specific conditions for the tests. All presented data in this work are from the same prototype. Eleven sensors measure fifteen parameters from three domains. The prototype delivers the resolutions of 0.1 part per million (PPM) for air quality parameters, 1 dB, 1 index, and 1 &deg, C for sound pressure level, UV, and skin temperature, respectively. The battery operates for 12.5 h under the maximum sampling rates of sensors without recharging. The final expense does not exceed 133&euro, We validated all layers and tested the entire device with a 75 min recording. The results show the appropriate functionalities of the prototype for further development and investigations.

BackgroundGamification has become increasingly important both in research and in practice. Particularly in long-term care processes, such as rehabilitation, playful concepts are gaining in importance to increase motivation and adherence. In addition to neurological diseases, this also affects the treatment of patients with musculoskeletal diseases such as shoulder disorders. Although it would be important to assist patients during more than one rehabilitation phase, it is hypothesized that existing systems only support a single phase. It is also unclear which game design elements are currently used in this context and how they are combined to achieve optimal positive effects on motivation. ObjectiveThis scoping review aims to identify and analyze information and communication technologies that use game design elements to support the rehabilitation processes of patients with musculoskeletal diseases of the shoulder. The state of the art with regard to fields of application, game design elements, and motivation concepts will be determined. MethodsWe conducted a scoping review to identify relevant application systems. The search was performed in 3 literature databases: PubMed, IEEE Xplore, and Scopus. Following the PICO (population, intervention, comparison, outcome) framework, keywords and Medical Subject Headings for shoulder, rehabilitation, and gamification were derived to define a suitable search term. Two independent reviewers, a physical therapist and a medical informatician, completed the search as specified by the search strategy. There was no restriction on year of publication. Data synthesis was done by deductive-inductive coding based on qualitative content analysis. ResultsA total of 1994 articles were screened; 31 articles in English, published between 2006 and 2019, were included. Within, 27 application systems that support patients with musculoskeletal diseases of the shoulder in exercising, usually at home but also in inpatient or outpatient rehabilitation clinics, were described. Only 2 application systems carried out monitoring of adherence. Almost all were based on in-house developed software. The most frequently used game components were points, tasks, and avatars. More complex game components, such as collections and teams, were rarely used. When selecting game components, patient-specific characteristics, such as age and gender, were only considered in 2 application systems. Most were described as motivating, though an evaluation of motivational effects was usually not conducted. ConclusionsThere are only a few application systems supporting patients with musculoskeletal diseases of the shoulder in rehabilitation by using game design elements. Almost all application systems are exergames for supporting self-exercising. Application systems for multiple rehabilitation phases seem to be nonexistent. It is also evident that only a few complex game design elements are used. Patient-specific characteristic are generally neglected when selecting and implementing game components. Consequently, a holistic approach to enhance adherence to rehabilitation is required supporting patients during the entire rehabilitation process by providing motivational game design elements based on patient-specific characteristics.

Die in einem Hochdurchsatz-Verfahren identifizierten Wirkstoffe weisen oft eine hohe Molmasse und eine schlechte Wasserlöslichkeit auf. Im Laufe des Optimierungsprozesses hin zu einem klinischen Arzneistoffkandidaten wird meist die Molmasse weiter erhöht und dadurch die Wasserlöslichkeit noch stärker vermindert. Für die Anwendung als oral applizierbarer Arzneistoff ist jedoch eine ausreichende Wasserlöslichkeit eine notwendige Voraussetzung. Zur Verbesserung der Wasserlöslichkeit können ionisierbare Gruppen in die Moleküle eingebracht werden. Diese Strukturelemente bedingen jedoch oft weitere Nachteile. Beispielsweise interagieren basische Aminogruppen häufig mit hERG-Ionenkanälen, was zu unerwünschten kardialen Nebenwirkungen führen kann. Vor diesem Hintergrund wurde in der hier vorgelegten Arbeit untersucht, ob sich N-alkylierte Sulfamide als neutrale löslichkeitsverbessernde Gruppen (engl. neutral solubility improving groups, NeuSIGs) eignen. Als Beispielstrukturen für Wirkstoffe wurden dazu zwei Serien von Proteinkinase-Inhibitoren herangezogen: Erstens Anilinopyrimidine, die die Aurora-Kinase A hemmen und daher potentiell zur Entwicklung von Antitumorwirkstoffen dienen könnten. Zweitens wurde der etablierte Arzneistoff Gefitinib modifiziert, der als Hemmstoff der epidermalen Wachstumsfaktor-Rezeptorkinase (engl. epidermal growth factor receptor, EGFR) gegen bestimmte Formen von Lungenkrebs eingesetzt wird. An diesen Beispielen wurde untersucht: a) ob sich durch molecular docking-Studien vorhersagen lässt, an welchen Positionen in den Molekülen NeuSIGs eingeführt werden können, ohne die biologische Aktivität zu verringern, b) ob die so konzipierten Verbindungen verbesserte Wasserlöslichkeit aufweisen, c) ob Berechnungsmethoden zur Vorhersage der Löslichkeit auf der Grundlage von Strukturdaten bei diesen Substanzen realistische Werte ergeben. Theoretische Berechnungsmethoden prognostizierten für die so konzipierten Moleküle zwar niedrigere Lipophilie, jedoch nicht in jedem Fall bessere Löslichkeit in wässrigen Systemen. Die Synthese und experimentelle Untersuchung der Verbindungen bestätigte einerseits den Erhalt der biologischen Aktivität. Andererseits wurde keine generelle Verbesserung der Wasserlöslichkeit durch den Einbau von Sulfamidstrukturen beobachtet., New drugs are usually developed from compounds identified by high throughput screening (HTS) campaigns. These substances often display a high molecular weight and poor aqueous solubility. During the optimization process towards a clinical candidate, the molar mass is usually increased further, thereby reducing the aqueous solubility even more. However, sufficient aqueous solubility is a necessary prerequisite for use as an orally administered drug. Ionizable groups can be introduced into the molecules to improve the aqueous solubility. However, these structural elements often cause disadvantages. For example, basic amino groups may interact with hERG channels, a property leading to undesirable cardiac side effects. Against this background, the present study investigated whether N-alkylated sulfamides are suitable as neutral solubility improving groups (NeuSIGs). Two series of protein kinase inhibitors were used as example structures for active substances: First, anilinopyrimidines, which inhibit aurora kinase A and which may serve as starting points for the development of antitumor drugs. Secondly, the established drug gefitinib was modified, which is an inhibitor of the epidermal growth factor receptor kinase (EGFR) and is used as a drug against certain forms of lung cancer. These two compound classes were selected to investigate: a) whether molecular docking studies can predict at which positions in the molecules NeuSIGs can be introduced without reducing the biological activity b) whether the designed compounds display improved aqueous solubility c) whether calculation methods for predicting solubility based on structural data generate realistic values for these compounds. Theoretical calculation methods predicted lower lipophilicity for the molecules designed in this way, but not always better solubility in aqueous systems. On the one hand, the synthesis and experimental investigation of the compounds confirmed the preservation of the biological activity. On the other hand, no general improvement of the aqueous solubility by the incorporation of sulfamide elements was observed.

Malaria causes hundreds of thousands of deaths every year, making it one of the most dangerous infectious diseases worldwide. Because the pathogens have developed resistance against most of the established anti-malarial drugs, new antiplasmodial agents are urgently needed. In analogy to similar antiplasmodial ketones, 4-arylthieno[2,3-b]pyridine-2-carboxamides were synthesized by Thorpe-Ziegler reactions. In contrast to the related ketones, these carboxamides are only weak inhibitors of the plasmodial enzyme PfGSK-3 but the compounds nevertheless show strong antiparasitic activity. The most potent representatives inhibit the pathogens with IC50 values in the two-digit nanomolar range and exhibit high selectivity indices (&gt, 100).

The Process Analytical Technology initiative and Quality by Design paradigm have led to changes in the guidelines and views of how to develop drug manufacturing processes. On this occasion the concept of the design space, which describes the impact of process parameters and material attributes on the attributes of the product, was introduced in the ICH Q8 guideline. The way the design space is defined and can be presented for regulatory approval seems to be left to the applicants, among who at least a consensus on how to characterize the design space seems to have evolved. The large majority of design spaces described in publications seem to follow a “static” statistical experimentation and modeling approach. Given that temporal deviations in the process parameters (i.e., moving within the design space) are of a dynamic nature, static approaches might not suffice for the consideration of the implications of variations in the values of the process parameters. In this paper, different forms of design space representations are discussed and the current consensus is challenged, which in turn, establishes the need for a dynamic representation and characterization of the design space. Subsequently, selected approaches for a dynamic representation, characterization and validation which are proposed in the literature are discussed, also showcasing the opportunity to integrate the activities of process characterization, process monitoring and process control strategy development.

Gestational diabetes mellitus (GDM) is a complex metabolic disease involving genetic and environmental factors. Recent studies have underlined its heterogeneity, so it is reasonable to divide patients into subpopulations depending on whether an insulin secretion or sensitivity defect is predominant. Since testing for GDM is usually performed in the second trimester, misinterpretation of prediabetes as gestational diabetes may occur. As with type 2 diabetes (T2DM), rodent models are needed for both GDM and prediabetes, but few do exist. Here, we compared the metabolic changes in pregnant normal NMRI mice with those in New Zealand obese (NZO) mice. Male animals of this strain are an established model of T2DM, whereas female mice of this strain are protected from hyperglycemia and β‐cell death. We demonstrate that female NZO mice exhibited impaired glucose tolerance, preconceptional hyperinsulinemia, and hyperglucagonemia without any signs of manifest diabetes. The NZO model showed, compared with the NMRI control strain, a reduced proliferative response of the Langerhans islets during pregnancy (3.7 ± 0.4 vs. 7.2 ± 0.8% Ki‐67‐positive nuclei, p = .004). However, oral glucose tolerance tests revealed improved stimulation of insulin secretion in both strains. But this adaption was not sufficient to prevent impaired glucose tolerance in NZO mice compared with the NMRI control (p = .0002). Interestingly, glucose‐stimulated insulin secretion was blunted in isolated primary NZO islets in perifusion experiments. In summary, the NZO mouse reflects important characteristics of human GDM and prediabetes in pregnancy and serves as a model for subpopulations with early alterations in glucose metabolism and primary insulin secretion defect., The aim of this work was to evaluate metabolic changes during pregnancy within the polygenic NZO mouse model and if it is a suitable one for the human disease. NZO mice showed early alterations in glucose homeostasis and insulin secretion, but no overt diabetes. Interestingly, the preconceptionally impaired glucose tolerance did not deteriorate during gestation despite a proliferation defect of Langerhans islets.

Unobtrusive in-vehicle health monitoring has the potential to use the driving time to perform regular medical check-ups. This work intends to provide a guide to currently proposed sensor systems for in-vehicle monitoring and to answer, in particular, the questions: (1) Which sensors are suitable for in-vehicle data collection? (2) Where should the sensors be placed? (3) Which biosignals or vital signs can be monitored in the vehicle? (4) Which purposes can be supported with the health data? We reviewed retrospective literature systematically and summarized the up-to-date research on leveraging sensor technology for unobtrusive in-vehicle health monitoring. PubMed, IEEE Xplore, and Scopus delivered 959 articles. We firstly screened titles and abstracts for relevance. Thereafter, we assessed the entire articles. Finally, 46 papers were included and analyzed. A guide is provided to the currently proposed sensor systems. Through this guide, potential sensor information can be derived from the biomedical data needed for respective purposes. The suggested locations for the corresponding sensors are also linked. Fifteen types of sensors were found. Driver-centered locations, such as steering wheel, car seat, and windscreen, are frequently used for mounting unobtrusive sensors, through which some typical biosignals like heart rate and respiration rate are measured. To date, most research focuses on sensor technology development, and most application-driven research aims at driving safety. Health-oriented research on the medical use of sensor-derived physiological parameters is still of interest.

Viele neue pharmazeutische Wirkstoffe sind schlecht wasserlöslich. Lipidnanopartikel (LNP) stellen ein vielversprechendes Trägersystem für diese Wirkstoffe dar und können aufgrund ihrer geringen Teilchengröße direkt in die Blutbahn appliziert werden. Um die Einsatzmöglichkeiten dieser Trägersysteme bewerten zu können, sind Kenntnisse über ihr Wirkstofffreisetzungsverhalten wichtig. Die verwendeten Akzeptormedien sollten lipophile Kompartimente enthalten, um die physiologischen Bedingungen möglichst genau darzustellen. Allerdings erschwert die geringe Größe der Trägerpartikel die Trennung von Donor- und Akzeptorsystem in Freisetzungsuntersuchungen unter realitätsnahen Bedingungen erheblich, da sie im selben Größenbereich wie die lipophilen Akzeptorpartikel liegt. Diese Arbeit beschreibt die Entwicklung einer neuen Methode zur Bestimmung der Freisetzungs-eigenschaften von echten Wirkstoffen aus LNP unter realitätsnahen Bedingungen, ohne vorherige Trennung von Donor- und Akzeptorpartikel. Die Methode basiert auf der Messung der Kristallisationstemperatur (TKrist) von Trimyristin(TM)-Nanopartikeln mittels DSC. Die TKrist der LNP sinkt proportional mit der Menge an eingeschlossenem Arzneistoff und erhöht sich mit Wirkstofffreisetzung. TM-Nanoemulsionen, die mit den schwer wasserlöslichen Wirkstoffen Fenofibrat, Orlistat, Tocopherolacetat und Ubidecarenon beladen waren, wurden in drei Akzeptormedien mit steigender Annäherung an physiologische Bedingungen untersucht: in einer Rapsöl-Nanoemulsion, in Schweineserum und Schweineblut. Die Ergebnisse zeigten eine Korrelation der Freisetzungseigenschaften mit der Lipophilie der Wirkstoffe: je höher der logP-Wert, desto langsamer war die Freisetzung. Eine erhöhte Freisetzung in einer Rapsöl-Nanoemulsion im Vergleich zu Serum und Blut deutete darauf hin, dass das Ausmaß der Freisetzung von einem Verteilungsgleichgewicht bestimmt wurde. Bei der Methodenentwicklung wurde beobachtet, dass die Verdünnung von Poloxamer 188-stabilisierten TM-Nanoemulsionen mit Wasser zu einem Anstieg der TKrist führte, während sie bei anderen Emulgatoren wie Tyloxapol, Poloxamer 407, Saccharoselaurat und einer Lecithin/Glycocholat-Kombination unverändert blieb. Es wurde eine Veränderung der Partikelzusammensetzung und der Grenzflächenbelegung mit dem Emulgator vermutet und weitergehende Untersuchungen wiesen auf eine Abhängigkeit der TKrist von der Poloxamer 188-Konzentration an der Partikelgrenzfläche hin., Lipid nanoparticles are an interesting parenteral delivery system for poorly water-soluble drugs. In order to approach physiological conditions when conducting release studies from such systems, the release media should contain lipophilic acceptor compartments. In practice, drug release studies under such close to physiological conditions may be complicated by the small size of lipid nanoparticles, which is in the same range as that of the acceptor particles. This work describes a novel method for drug release measurements which works without separation of donor and acceptor particles. The technique is based on measuring the crystallization temperature (Tcryst) of poloxamer (pol) 188 -stabilized trimyristin (tm) nanoparticles by DSC. The Tcryst of the nanoparticles decreases proportionally with the amount of drug incorporated and thus increases as a result of drug release. Liquid tm-nanoparticles loaded with fenofibrate, orlistat, tocopherol acetate and ubidecarenone were studied in three release media with increasing comparability to physiological conditions: a rapeseed oil nanoemulsion, porcine serum and porcine blood. Using the new method, a correlation between release behavior and drug lipophilicity was observed: the higher the logP value of the drug, the slower the release. The extent of drug release was influenced by partition equilibrium as indicated by increased drug release in the rapeseed oil nanoemulsion compared to porcine serum and blood. Based on the method developed in this thesis, it is possible to investigate slow transfer processes from lipid emulsion systems. However, the applicability of this method is limited to supercooled tm donor nanoparticles and similar systems. During method development, it was observed that dilution of pol 188-stabilized tm nanoemulsions with water caused an increase in Tcryst while it remained the same with other emulsifiers like tyloxapol, pol 407, sucrose laurate and a lecithin/sodium glycocholate combination. Pol 188 was the only emulsifier used in a concentration below the critical micelle concentration (cmc). Presumably, the monomers were driven out of the nanoparticle interface to reach a new equilibrium after dilution with water. In contrast, when diluted with a solution of the same pol 188 concentration, monomers from the interface were no longer needed and no change in Tcryst was observed. Furthermore, it was shown that the pol 188 content of the system correlated with the Tcryst of tm nanoparticles.

Background Psychotherapy is highly effective and widely acknowledged for treating various mental disorders. Nevertheless, in terms of methods for teaching effective psychotherapeutic approaches and competencies, there has been a lack of investigation. Training and supervision are the main strategies for teaching therapist competencies, and standardized role-plays with simulated patients (i.e., trained individuals playing someone with a mental disorder) seem useful for evaluating training approaches. In medical education, this procedure is now internationally established. However, so far, little use has been made of standardized role-playing to evaluate training and supervision in the area of clinical psychology and psychotherapy. Methods In this study, standardized role-plays are used to evaluate methods for training and supervision. Central cognitive behavioral approaches for treating depression are taught in the training. The first experiment compares an active training approach (i.e., model learning) with a passive one (i.e., reading manual-based instructions). The second experiment compares a direct supervision technique (i.e., supervision based on video analysis) with an indirect one (i.e., supervision based on verbal reporting). In each experiment, 68 bachelor’s and master’s students of psychology will be randomly assigned to the experimental and control groups. Each student takes part in three role-plays (baseline, post and 3-month follow-up), which are all videotaped. Two independent raters assess therapist competencies in each role-play on the basis of a standardized competence scale. Discussion The research project aims to contribute to the development of specific training and supervision methods in order to improve psychotherapy training and patient care.