Dual inhibitors of microsomal prostaglandin E2 synthase-1 and 5-lipoxygenase as anti-inflammatory payloads in polymer-based nanoparticles

Originated from the enzymatic oxygenation of polyunsaturated fatty acids, lipid mediators (LM) conduct inflammatory and resolving responses of the innate immune system to restore homeostasis after infections or injuries1. In current pharmacotherapy the production of pro-inflammatory prostaglandins (PG) and leukotrienes (LT) is reduced by inhibition of cyclooxygenase (COX)-1 and -2 or 5-lipoxygenase (LOX), which ameliorates inflammation but also exerts severe side effects upon long-term therapy, because the mediators that are crucial for homeostasis are influenced or substrates are redirected to other metabolic pathways, resulting in dysregulation of the lipid mediator network2. Therefore, new pharmacological strategies to specifically modify the production of different lipid mediators are under constant development and the impact of drugs interfering with these metabolic pathways needs to be understood. This thesis reveales the necessity for new pharmacological options for intervention in inflammatory diseases and disclosed a promising new approach to effectively deliver potent dual inhibitors to the site of action via NPs that would otherwise fail to act due to high lipophilicity and overall poor bioavailability. The sophisticated mode of action of these dual inhibitors not only reduces pro-inflammatory mediators but can actively induce the resolution phase by promoting SPM formation. These findings could contribute to a paradigm shift in inflammatory therapy via the combination of novel smart molecules and nanomedicine

Label-Free Characterization of Macrophage Polarization Using Raman Spectroscopy

Macrophages are important cells of the innate immune system that play many different roles in host defense, a fact that is reflected by their polarization into many distinct subtypes. Depending on their function and phenotype, macrophages can be grossly classified into classically activated macrophages (pro-inflammatory M1 cells), alternatively activated macrophages (anti-inflammatory M2 cells), and non-activated cells (resting M0 cells). A fast, label-free and non-destructive characterization of macrophage phenotypes could be of importance for studying the contribution of the various subtypes to numerous pathologies. In this work, single cell Raman spectroscopic imaging was applied to visualize the characteristic phenotype as well as to discriminate between different human macrophage phenotypes without any label and in a non-destructive manner. Macrophages were derived by differentiation of peripheral blood monocytes of human healthy donors and differently treated to yield M0, M1 and M2 phenotypes, as confirmed by marker analysis using flow cytometry and fluorescence imaging. Raman images of chemically fixed cells of those three macrophage phenotypes were processed using chemometric methods of unmixing (N-FINDR) and discrimination (PCA-LDA). The discrimination models were validated using leave-one donor-out cross-validation. The results show that Raman imaging is able to discriminate between pro- and anti-inflammatory macrophage phenotypes with high accuracy in a non-invasive, non-destructive and label-free manner. The spectral differences observed can be explained by the biochemical characteristics of the different phenotypes

Archivierung und Zugang zu qualitativen Daten

Der vorliegende Band dokumentiert die Beiträge und Ergebnisse eines vom Rat für Sozial- und Wirtschaftsdaten (RatSWD) und dem Datenservicezentrum Qualiservice am 27. und 28. April 2018 an der Universität Bremen veranstalteten Workshops zum Thema "Archivierung und Zugang zu qualitativen Daten". Ziel des Workshops mit mehr als 50 Forschenden verschiedener sozial- und geisteswissenschaftlicher Disziplinen sowie Vertreterinnen und Vertretern von Forschungsdatenzentren war es, bisherige Erfahrungen zu evaluieren und Anforderungen für eine verbesserte Archivierungsinfrastruktur in der qualitativen Forschung zu benennen

Long-term durability of alumina ceramic heads in THA

Background: The optimal type of bearing for hip arthroplasty remains a matter of debate. Ceramic-on-polyethylene (CoP) bearings are frequently used in younger and more active patients to reduce wear and increase biocompatibility compared to Metal-on-Polyethylene (MoP) bearings. However, in comparison to metal heads, the fracture risk of ceramic heads is higher. In addition, ceramic head fractures pose a serious complication which often necessitates major revision surgery. To date, there are no long-term data (>20 years of follow-up) reporting fracture rates of the ceramic femoral heads in CoP bearings. The purpose of this research was to investigate long-term CoP fracture rate. Methods: We evaluated the clinical and radiographic results of 348 cementless THAs treated with 2nd generation Biolox® Al2O3 Ceramic-on-Polyethylene (CoP) bearings consecutively implanted between January 1985 and December 1989. The mean age at implantation was 57 years. The patients were followed for a minimum of 20 years. At the final 111 had died, and 5 were lost to follow-up. The cumulative incidence of ceramic head fractures in the long-term was estimated using a competing risk analysis. Results: The cumulative incidence of ceramic head fracture after 22-years was estimated with a competing risk analysis at 0.29% after 22-years (SE = 2.09%; 95% - CI: 0.03-1.5%). The radiographic analysis revealed no impending failures at final follow-up. Discussion/Conclusion: The fracture rate of second-generation ceramic heads using a CoP articulation remains very low into the third decade after cementless THA

Wear performance of ceramic-on-metal hip bearings.

Ceramic-on-metal (CoM) bearings are considered to be a promising alternative to polyethylene-based bearings or hard-on-hard bearings (Ceramic-on-Ceramic (CoC) and Metal-on-Metal (MoM)). Although, CoM shows lower wear rates than MoM, in-vitro wear testing of CoM shows widely varying results. This may be related to limitations of wear-measuring methods. Therefore, the aim of this study was to improve the gravimetric measurement technique and to test wear behaviour of CoM bearings compared to CoC bearings. Level walking according to ISO-14242 was simulated for four CoM and four CoC bearings. Prior to simulation, errors in measurement of gravimetric wear were detected and improvements in measurement technique incorporated. The results showed no differences in mean wear rates between CoM and CoC bearings. However, the CoM bearings showed wear results over a wide range of wear performance. High reliability of wear results was recorded for the CoC bearings. Material transfer was observed on the ceramic heads of the CoM bearings. Therefore, for level walking a partial mixed or boundary lubrication has to be assumed for this type of bearing. CoM is a highly sensitive wear-couple. The reasons for the observed behaviour cannot be clarified from this study. Simulator studies have to be considered as an ideal loading condition. Therefore, high variations in wear rates as seen in this study, even at low levels, may have an adverse effect on the in-vivo wear behavior. Careful clinical use may be advisable until the reasons for the variation are fully clarified and understood

Wear testing of moderate activities of daily living using in vivo measured knee joint loading.

Resumption of daily living activities is a basic expectation for patients provided with total knee replacements. However, there is a lack of knowledge regarding the impact of different activities on the wear performance. In this study the wear performance under application of different daily activities has been analyzed. In vivo load data for walking, walking downstairs/upstairs, sitting down/standing up, and cycling (50 W & 120 W) has been standardized for wear testing. Wear testing of each activity was carried out on a knee wear simulator. Additionally, ISO walking was tested for reasons of comparison. Wear was assessed gravimetrically and wear particles were analyzed. In vivo walking produced the highest overall wear rates, which were determined to be three times higher than ISO walking. Moderate wear rates were determined for walking upstairs and downstairs. Low wear rates were determined for standing up/sitting down and cycling at power levels of 50 W and 120 W. The largest wear particles were observed for cycling. Walking based on in vivo data has been shown to be the most wear-relevant activity. Highly demanding activities (stair climbing) produced considerably less wear. Taking into account the expected number of loads, low-impact activities like cycling may have a greater impact on articular wear than highly demanding activities

Label-Free Characterization of Macrophage Polarization Using Raman Spectroscopy

Macrophages are important cells of the innate immune system that play many different roles in host defense, a fact that is reflected by their polarization into many distinct subtypes. Depending on their function and phenotype, macrophages can be grossly classified into classically activated macrophages (pro-inflammatory M1 cells), alternatively activated macrophages (anti-inflammatory M2 cells), and non-activated cells (resting M0 cells). A fast, label-free and non-destructive characterization of macrophage phenotypes could be of importance for studying the contribution of the various subtypes to numerous pathologies. In this work, single cell Raman spectroscopic imaging was applied to visualize the characteristic phenotype as well as to discriminate between different human macrophage phenotypes without any label and in a non-destructive manner. Macrophages were derived by differentiation of peripheral blood monocytes of human healthy donors and differently treated to yield M0, M1 and M2 phenotypes, as confirmed by marker analysis using flow cytometry and fluorescence imaging. Raman images of chemically fixed cells of those three macrophage phenotypes were processed using chemometric methods of unmixing (N-FINDR) and discrimination (PCA-LDA). The discrimination models were validated using leave-one donor-out cross-validation. The results show that Raman imaging is able to discriminate between pro- and anti-inflammatory macrophage phenotypes with high accuracy in a non-invasive, non-destructive and label-free manner. The spectral differences observed can be explained by the biochemical characteristics of the different phenotypes

Machine Learning Uncovers Natural Product Modulators of the 5-Lipoxygenase Pathway and Facilitates the Elucidation of Their Biological Mechanisms.

Publication status: PublishedMachine learning (ML) models have made inroads into chemical sciences, with optimization of chemical reactions and prediction of biologically active molecules being prime examples thereof. These models excel where physical experiments are expensive or time-consuming, for example, due to large scales or the need for materials that are difficult to obtain. Studies of natural products suffer from these issues─this class of small molecules is known for its wealth of structural diversity and wide-ranging biological activities, but their investigation is hindered by poor synthetic accessibility and lack of scalability. To facilitate the evaluation of these molecules, we designed ML models that predict which natural products can interact with a particular target or a relevant pathway. Here, we focused on discovering natural products that are capable of modulating the 5-lipoxygenase (5-LO) pathway that plays key roles in lipid signaling and inflammation. These computational approaches led to the identification of nine natural products that either directly inhibit the activity of the 5-LO enzyme or affect the cellular 5-LO pathway. Further investigation of one of these molecules, deltonin, led us to discover a new cell-type-selective mechanism of action. Our ML approach helped deorphanize natural products as well as shed light on their mechanisms and can be broadly applied to other use cases in chemical biology