Pre-gelation staining expansion microscopy for visualisation of the Plasmodium liver stage.

Fluorescence and light microscopy are important tools in the history of natural science. However, the resolution of microscopes is limited by the diffraction of light. One possible method to circumvent this physical restriction is the recently developed expansion microscopy (ExM). However, the original ultrastructure ExM (U-ExM) protocol is very time-consuming, and some epitopes are lost during the process. In this study, we developed a shortened pre-gelation staining ExM (PS-ExM) protocol and tested it to investigate the Plasmodium liver stage. The protocol presented in this study allows expanding of pre-stained samples, which results in shorter incubation times, better preservation of some epitopes and the advantage that non-expanded controls can be performed alongside using the same staining protocol. The protocol applicability was accessed throughout the Plasmodium liver stage, showing isotropic five-fold expansion. Furthermore, we used PS-ExM to visualise parasite mitochondria as well as the association of lysosomes to the parasitophorous vacuole membrane (PVM) as an example of visualising host-pathogen interaction. We are convinced that this new tool will be helpful for a deeper understanding of the biology of the Plasmodium liver stage.

Single p197 molecules of the mitochondrial genome segregation system of Trypanosoma brucei determine the distance between basal body and outer membrane.

The tripartite attachment complex (TAC) couples the segregation of the single unit mitochondrial DNA of trypanosomes with the basal body (BB) of the flagellum. Here, we studied the architecture of the exclusion zone filament (EZF) of the TAC, the only known component of which is p197, that connects the BB with the mitochondrial outer membrane (OM). We show that p197 has three domains that are all essential for mitochondrial DNA inheritance. The C terminus of p197 interacts with the mature and probasal body (pro-BB), whereas its N terminus binds to the peripheral OM protein TAC65. The large central region of p197 has a high α-helical content and likely acts as a flexible spacer. Ultrastructure expansion microscopy (U-ExM) of cell lines exclusively expressing p197 versions of different lengths that contain both N- and C-terminal epitope tags demonstrates that full-length p197 alone can bridge the ∼270-nm distance between the BB and the cytosolic face of the OM. Thus U-ExM allows the localization of distinct domains within the same molecules and suggests that p197 is the TAC subunit most proximal to the BB. In addition, U-ExM revealed that p197 acts as a spacer molecule, as two shorter versions of p197, with the repeat domain either removed or replaced by the central domain of the Trypanosoma cruzi p197 ortholog reduced the distance between the BB and the OM in proportion to their predicted molecular weight.

[Bed confinement in old people-A literature review]. / Bettlägerigkeit bei alten Menschen ­ eine Literaturübersicht.

BACKGROUND: Particularly older people are affected by mobility restrictions and can go through the process of gradually becoming bedridden. This can potentially lead to extensive consequences for the individuals involved, which must be considered in their care. AIMS: To bundle nursing implications related to the phenomenon of being bedridden in the field of long-term care and to provide impulses for research in nursing science. METHODS: Research in relevant specialist databases (2003-2023) based on the criteria of a scoping review. RESULTS: In Germany there is a standard for maintaining and promoting mobility that addresses bed and local confinements but German language studies on these phenomena are rare. In the international context, these issues are researched more intensively, focussing on risk factors for the development of immobility and the negative consequences for those affected. The publications focus on the reduction of these factors, while less attention is paid to the organization of life in bed and the participation and involvement of those affected. DISCUSSION: The complexity of the phenomenon is not fully illustrated in current research. In order to develop a nursing perspective research projects that also include aspects of life in bed are a central aspect in order to take greater account of the reality of bedridden people's lives and their potential for participation and involvement.

[Medication supply and polypharmacy in long-term care : An overview of possible interventions and the question of a functioning concept]. / Medikamentenversorgung und Polypharmazie in der Langzeitpflege : Eine Übersicht zu möglichen Interventionen und die Frage nach einem funktionierenden Konzept.

BACKGROUND: Polypharmacy and the resulting problems lead to considerable consequences for those affected. There are also considerable problems with the medication management. OBJECTIVE: Which interventions and programs for optimizing the supply of medication are available for nursing homes and which implementation problems can be expected? MATERIAL AND METHOD: A literature search was carried out for interventional studies in nursing homes in Germany, with a focus on improving medication safety. RESULTS: A total of six programs were identified for which evaluation results are available. Despite a mostly multimodal approach with several pillars of intervention (e.g., medication reviews, further education and training, development of aids), the results are largely disappointing. The effects on the number of prescriptions in general, specific medication groups or outcome parameters such as hospital admissions could only be shown in one study, whereby, selection bias could also be at least partly responsible for this. Interdisciplinary collaboration and the implementation of medication recommendations formulated in reviews by the responsible physicians are the main problem areas. At the same time, too little attention is paid to the central role of nurses in the entire process and they are not actively promoted enough. This could be one of the reasons for the difficulties in implementation in practice. CONCLUSION: There are nearly no significant changes as a result of the interventions implemented in the studies reviewed. In particular, interprofessional cooperation, especially the skills of nurses and the reluctance on the part of physicians, should probably be given more attention.

[Estimation of the risk of drug-related problems in nursing home residents : A retrospective analysis of routine data]. / Einschätzung des Risikos für arzneimittelbezogene Probleme bei Pflegeheimbewohner*innen : Eine retrospektive Analyse von Routinedaten.

BACKGROUND: Polypharmacy and drug-related problems are major challenges in the care and treatment of nursing home residents. Many interventional studies showed disappointing results, which lead to the question if this could also be due to the selection of the target parameters of these studies. MATERIAL AND METHODS: A routine data set from six long-term care facilities was retrospectively analyzed. The question is if the recently validated medication risk score (MERIS) is suitable for carrying out a risk assessment in a population of nursing home residents. Associations between MERIS and the dependent variables hospital admissions and falls over 12 months and a weight loss of ≥ 5% over 3 months were examined. RESULTS: Out of 495 residents 38.6% (n = 191) have a high risk of drug-related problems according to MERIS. A univariate regression analysis showed a significantly increased risk of hospital admissions (OR 2.2; p < 0.001) and weight loss of ≥ 5% (OR 1.95; p = 0.041) with high MERIS, but no significant association with falls. In the multivariate regression the risk of hospitalization was increased by diabetes mellitus (OR 1.88; p = 0.004), falls in the same period (OR 1.91; p = 0.001), positive MERIS (OR 1.75; p = 0.006) and decreased with stable weight (OR 0.88; p = 0.004). CONCLUSION: The results indicate the potential of the score for future research projects and individual risk assessment; however, due to the limitations of retrospective secondary analyses further studies are required.

7-(2-Anilinopyrimidin-4-yl)-1-benzazepin-2-ones Designed by a "Cut and Glue" Strategy Are Dual Aurora A/VEGF-R Kinase Inhibitors.

Although overexpression and hyperactivity of protein kinases are causative for a wide range of human cancers, protein kinase inhibitors currently approved as cancer drugs address only a limited number of these enzymes. To identify new chemotypes addressing alternative protein kinases, the basic structure of a known PLK1/VEGF-R2 inhibitor class was formally dissected and reassembled. The resulting 7-(2-anilinopyrimidin-4-yl)-1-benzazepin-2-ones were synthesized and proved to be dual inhibitors of Aurora A kinase and VEGF receptor kinases. Crystal structures of two representatives of the new chemotype in complex with Aurora A showed the ligand orientation in the ATP binding pocket and provided the basis for rational structural modifications. Congeners with attached sulfamide substituents retained Aurora A inhibitory activity. In vitro screening of two members of the new kinase inhibitor family against the cancer cell line panel of the National Cancer Institute (NCI) showed antiproliferative activity in the single-digit micromolar concentration range in the majority of the cell lines.

Expansion of metabolically labelled endocytic organelles and cytoskeletal cell structures in Giardia lamblia using optimised U-ExM protocols.

Understanding cellular ultrastructure is tightly bound to microscopic resolution and the ability to identify individual components at that resolution. Expansion microscopy has revolutionised this topic. Here we present and compare two protocols of ultrastructure expansion microscopy that allow for 4.5-fold mostly isotropic expansion and the use of antibodies, metabolic labelling, and DNA stains to demarcate individual regions such as the endoplasmic reticulum, the nuclei, the peripheral endocytic compartments as well as the ventral disc and the cytoskeleton in Giardia lamblia. We present an optimised, shortened, and modular protocol that can be swiftly adjusted to the investigators needs in this important protozoan model organism.

Incorporation of ovalbumin into ISCOMs and related colloidal particles prepared by the lipid film hydration method.

The aim of this study was to investigate the incorporation of a model antigen, fluorescently labelled ovalbumin (FITC-OVA), into various colloidal particles including immune stimulating complexes (ISCOMs), liposomes, ring and worm-like micelles, lamellae and lipidic/layered structures that are formed from various combinations of the triterpene saponin Quil A, cholesterol and phosphatidylethanolamine (PE) following hydration of PE/cholesterol lipid films with aqueous solutions of Quil A. Colloidal dispersions of these three components were also prepared by the dialysis method for comparison. FITC-OVA was conjugated with palmitic acid (P) and PE to produce P-FITC-OVA and PE-FITC-OVA, respectively. Both P-FITC-OVA and PE-FITC-OVA could be incorporated in all colloidal structures whereas FITC-OVA was incorporated only into liposomes. The incorporation of PE-FITC-OVA into all colloidal structures was significantly higher than P-FITC-OVA (P < 0.05). The degree of incorporation of protein was in the order: ring and worm-like micelles < liposomes and lipidic/layered structures < ISCOMs and lamellae. The incorporation of protein into the various particles prepared by the lipid film hydration method was similar to those for colloidal particles prepared by the dialysis method (provided both methods lead to the formation of the same colloidal structures). In the case of different colloidal structures arising due to the preparation method, differences in encapsulation efficiency were found (P < 0.05) for formulations with the same polar lipid composition. This study demonstrates that the various colloidal particles formed as a result of hydrating PE/cholesterol lipid films with different amounts of Quil A are capable of incorporating antigen, provided it is amphipathic. Some of these colloidal particles may be used as effective vaccine delivery systems.