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- additive manufacturing (14)
- borylation (14)
- cardiac hypertrophy (14)
- circadian clock (14)
- diversity (14)
- ecology (14)
- evaluation (14)
- exercise (14)
- fNIRS (14)
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- growth (14)
- guidelines (14)
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- in-vitro (14)
- in-vivo (14)
- inorganic chemistry (14)
- mechanisms (14)
- mental health (14)
- neutrophils (14)
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- quantum dot (14)
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- AIDS (13)
- Alter (13)
- Alzheimerkrankheit (13)
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- Aufmerksamkeitsdefizit-Syndrom (13)
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- blood–brain barrier (12)
- carbenes (12)
- case report (12)
- catalysis (12)
- cell cycle (12)
- colorectal carcinoma (12)
- complications (12)
- database (12)
- dementia (12)
- drug delivery (12)
- dynamics (12)
- endothelium (12)
- enzyme replacement therapy (12)
- fear conditioning (12)
- lymphocytes (12)
- malignant hyperthermia (12)
- measles (12)
- metaanalysis (12)
- microarray (12)
- molecular beam epitaxy (12)
- monoclonal antibodies (12)
- neuropathy (12)
- optimization (12)
- osteoarthritis (12)
- performance (12)
- pollen (12)
- polymers (12)
- rats (12)
- recombination (12)
- reconstruction (12)
- reliability (12)
- screening (12)
- super-resolution microscopy (12)
- systematic review (12)
- therapeutic drug monitoring (12)
- tinnitus (12)
- total knee arthroplasty (12)
- trabeculectomy (12)
- translation (12)
- ubiquitin (12)
- ultrasound (12)
- vitamin D (12)
- vorsprachliche Entwicklung (12)
- zebrafish (12)
- Überleben (12)
- ++ (11)
- 3D (11)
- ARDS (11)
- Anorexia nervosa (11)
- Antigen CD8 (11)
- Arabidopsis (11)
- B-Zelle (11)
- Bioisosterie (11)
- Biosynthese (11)
- Bioverfügbarkeit (11)
- Bruchpilot (11)
- Candida (11)
- Chromatin (11)
- Click-Chemie (11)
- Computertomografie (11)
- Diboren (11)
- Dynamik (11)
- EGFR (11)
- Elektronentransfer (11)
- Enzym (11)
- Epidemiologie (11)
- Epidermaler Wachstumsfaktor-Rezeptor (11)
- Flavonoide (11)
- Fließverhalten (11)
- Fotovoltaik (11)
- Genom (11)
- Hirntumor (11)
- Hypoxie (11)
- Immuncytochemie (11)
- Immunotherapy (11)
- Japankärpfling (11)
- Knochen (11)
- Knochenersatz (11)
- Kolorektales Karzinom (11)
- LPS (11)
- Laser (11)
- Längsschnittuntersuchung (11)
- MS (11)
- Mais (11)
- Mathematik (11)
- Medienkompetenz (11)
- Megakaryozyt (11)
- Mensch-Maschine-Kommunikation (11)
- Mesenchymale Stammzellen (11)
- Mesenchymzelle (11)
- Model (11)
- NIRS (11)
- Nanodiamant (11)
- Neuropathie (11)
- Nigeria (11)
- Operation (11)
- Parathormon (11)
- Perylenbisimid (11)
- Phylogenie (11)
- Proteaseinhibitor (11)
- Proteasen (11)
- Psychiatrie (11)
- Pulmonale Hypertonie (11)
- Quality of Experience (11)
- Quality of life (11)
- Quanten-Hall-Effekt (11)
- Religion (11)
- Reperfusion (11)
- Risikofaktoren (11)
- Rotatorenmanschette (11)
- Salmonella (11)
- Schizophrenia (11)
- Schlafapnoe (11)
- Schließzelle (11)
- Schule (11)
- Schädel-Hirn-Trauma (11)
- Silicate (11)
- Skala (11)
- Small RNA (11)
- Squark (11)
- Stammzellen (11)
- Sterblichkeit (11)
- Supraleitung (11)
- Symbiose (11)
- T cell (11)
- Training (11)
- Tuberkelbakterium (11)
- Tumor-Nekrose-Faktor <alpha> (11)
- USA (11)
- Ubiquitin (11)
- Vasodilatator-stimuliertes Phosphoprotein (11)
- Wahrnehmung (11)
- Westafrika (11)
- Zelltod (11)
- adhesion (11)
- age (11)
- allergy (11)
- amygdala (11)
- antibody (11)
- autoantibodies (11)
- biocompatibility (11)
- biofilm (11)
- cancer treatment (11)
- cell adhesion (11)
- cochlear implant (11)
- coping (11)
- dosimetry (11)
- emotions (11)
- fluorescence microscopy (11)
- gender (11)
- genome (11)
- glioblastoma multiforme (11)
- hypoxia (11)
- interaction (11)
- kinetics (11)
- lung (11)
- mechanism (11)
- meiosis (11)
- membrane proteins (11)
- molecular dynamics (11)
- nervous system (11)
- neural networks (11)
- neuroendocrine tumor (11)
- next generation sequencing (11)
- nuclear envelope (11)
- oncolytic virus (11)
- periodontitis (11)
- plasticity (11)
- pollination (11)
- randomized controlled trial (11)
- reactive oxygen species (11)
- relapse (11)
- review (11)
- safety (11)
- synaptic plasticity (11)
- topological insulator (11)
- transport (11)
- university (11)
- Alkohol (10)
- Alzheimer's disease (10)
- Alzheimer’s disease (10)
- Antigen CD28 (10)
- Arzneimittelüberwachung (10)
- Atherosclerosis (10)
- BERA (10)
- Bacteria (10)
- Beschichtung (10)
- Bildverarbeitung (10)
- Bioinformatics (10)
- Biokompatibilität (10)
- Blazar (10)
- Borole (10)
- CRISPR/Cas-Methode (10)
- CRISPR/Cas9 (10)
- Cadherine (10)
- Calciumphosphate (10)
- Children (10)
- Chronische Niereninsuffizienz (10)
- Churritisch (10)
- Cochlear-Implantat (10)
- Coronaviren (10)
- Cyclo-GMP (10)
- Depressivität (10)
- Diamant (10)
- Dimerisierung (10)
- Dotierung (10)
- Durchflusscytometrie (10)
- Echokardiographie (10)
- Eierstockkrebs (10)
- Elektrochemie (10)
- Elektronenkorrelation (10)
- Enzyminhibitor (10)
- Erbkrankheit (10)
- Ereigniskorreliertes Potenzial (10)
- Erwachsener (10)
- Frau (10)
- Funktionalisierung <Chemie> (10)
- Förderung (10)
- Geldpolitik (10)
- Glatter Krallenfrosch (10)
- Globalisierung (10)
- HSM-Satztest (10)
- Herzmuskelzelle (10)
- Honigbiene (10)
- Hypophosphatasie (10)
- Immunology (10)
- Impfstoff (10)
- Induzierte pluripotente Stammzelle (10)
- Interleukin 4 (10)
- JNK (10)
- Klassifikation (10)
- Kniegelenk (10)
- Kohlenstoff (10)
- Komplikation (10)
- Kultur (10)
- LASP1 (10)
- LC-MS (10)
- Ladungstransfer (10)
- Ligand (10)
- Lokalisation (10)
- Mathematikunterricht (10)
- Mathematisches Modell (10)
- Medulloblastom (10)
- Metallocene (10)
- Methylphenidat (10)
- Mikrokerne (10)
- Mittelalter (10)
- Multiple Myeloma (10)
- Mutagenität (10)
- Nahrungserwerb (10)
- Nebennierenrindenkarzinom (10)
- Neuropathischer Schmerz (10)
- Niereninsuffizienz (10)
- Niger (10)
- Osteosynthese (10)
- Pain (10)
- Panikstörung (10)
- Parasit (10)
- Parodontitis (10)
- Pathogenität (10)
- Plattenepithelkarzinom (10)
- Prevalence (10)
- Prostaglandine (10)
- Präfrontaler Cortex (10)
- Pädagogik (10)
- Qualitätskontrolle (10)
- RNA interference (10)
- RNA-seq (10)
- RNS-Spleißen (10)
- Remote Sensing (10)
- Rhodium (10)
- Rituximab (10)
- SAR (10)
- Schilddrüse (10)
- Schultergelenk (10)
- Silaanaloga (10)
- Silber (10)
- Silicon (10)
- South Africa (10)
- Sphingolipide (10)
- Sprachverstehen (10)
- Starke Kopplung (10)
- Stickstoffmonoxid-Synthase (10)
- Stroke (10)
- Struktur (10)
- T-Lymphozyten (10)
- T-Lymphozyten-Rezeptor (10)
- Tagesrhythmus (10)
- Totalsynthese (10)
- Toxizität (10)
- Transplantat-Wirt-Reaktion (10)
- Treg (10)
- Trypanosomen (10)
- Vitamin D (10)
- Vor- und Frühgeschichte (10)
- Vorschulkind (10)
- Wachstum (10)
- Zebrabärbling (10)
- amino acids (10)
- antimicrobial resistance (10)
- bariatric surgery (10)
- biosynthesis (10)
- boranes (10)
- cartilage (10)
- chemokines (10)
- circadian rhythms (10)
- clinical trial (10)
- comparison (10)
- complex (10)
- coronary artery disease (10)
- decision-making (10)
- density functional calculations (10)
- eNOS (10)
- emotion regulation (10)
- fear (10)
- genome-wide association (10)
- head and neck cancer (10)
- heart rate (10)
- hydrogels (10)
- immunomodulation (10)
- injury (10)
- insect (10)
- knockout (10)
- leaf-cutting ants (10)
- leukemia (10)
- lymph nodes (10)
- membrane potential (10)
- microbiome (10)
- microglia (10)
- molecular imaging (10)
- motivation (10)
- oncolytic virotherapy (10)
- oxidativer Stress (10)
- phenotype (10)
- proteomics (10)
- regulatorische T-Zellen (10)
- remodeling (10)
- senescence (10)
- silicon (10)
- social interaction (10)
- stem cell transplantation (10)
- tDCS (10)
- thrombosis (10)
- tight junctions (10)
- tumor microenvironment (10)
- tumors (10)
- vaccination (10)
- vertigo (10)
- water oxidation (10)
- Östrogene (10)
- 3 (9)
- Adenosin (9)
- Adenosine receptors (9)
- Afrika (9)
- Alps (9)
- Angeregter Zustand (9)
- Anxiety (9)
- Aorta (9)
- BMP-2 (9)
- Bakterielle Infektion (9)
- Bauchspeicheldrüsenkrebs (9)
- Beyond Standard Model (9)
- Bordetella pertussis (9)
- Borylierung (9)
- Boğazkale (9)
- Butyrat (9)
- Carbene (9)
- Carcinogenese (9)
- Catalysis (9)
- Chiralität <Chemie> (9)
- Computational chemistry (9)
- Dendritic cells (9)
- Diborane (9)
- Digital Humanities (9)
- Dünndarm (9)
- Einzelphotonenemission (9)
- Elektroencephalographie (9)
- Elektronenmikroskopie (9)
- Elektronenstruktur (9)
- Elementarteilchenphysik (9)
- Englisch (9)
- Epithel (9)
- Ernährung (9)
- Experimentelle Psychologie (9)
- Extremwertstatistik (9)
- Familie (9)
- Fanconi Anämie (9)
- Fettgewebe (9)
- Fibroblastenwachstumsfaktor (9)
- Fibromyalgie (9)
- Franken (9)
- Frühgeborene (9)
- Funktionelle Kernspintomografie (9)
- GIS (9)
- Geistigbehindertenpädagogik (9)
- Gentherapie (9)
- Geoinformationssystem (9)
- Geruchswahrnehmung (9)
- Geschlecht (9)
- Glioblastoma (9)
- Google Earth Engine (9)
- Grammatik (9)
- Grenzfläche (9)
- Handlungsorientierung (9)
- Hethiter (9)
- Hyaluronsäure (9)
- IL-4 (9)
- Immunfluoreszenz (9)
- Immunisierung (9)
- Instrumentelle Analytik (9)
- Integrine (9)
- Iran (9)
- Isolierung <Chemie> (9)
- Jugendliche (9)
- Kapillarelektrophorese (9)
- Knorpel (9)
- Kommunikation (9)
- Kooperation (9)
- Koordinationslehre (9)
- Korpus <Linguistik> (9)
- Landsat (9)
- Leber (9)
- Linguistik (9)
- Löslichkeit (9)
- MAPK (9)
- Macrophage (9)
- Magnesiumphosphate (9)
- Magnetische Resonanz (9)
- Magnetismus (9)
- Mehrfachbindung (9)
- Messenger-RNS (9)
- Metakognition (9)
- Mice (9)
- Molekulare Erkennung (9)
- Monitoring (9)
- Morphologie (9)
- Mukoviszidose (9)
- Musik (9)
- Myokardprotektion (9)
- NAFLD (9)
- NO (9)
- Nebennierenrindenkrebs (9)
- Neuroinflammation (9)
- Nierentransplantation (9)
- Oberfläche (9)
- Oberflächenphysik (9)
- Optoelektronik (9)
- Osteoinduktion (9)
- PD-1 (9)
- Pathogenese (9)
- Pharmakotherapie (9)
- Photolumineszenzspektroskopie (9)
- Physiologie (9)
- Plasmamembran (9)
- Platelets (9)
- Protein (9)
- Pseudomonas syringae (9)
- Psychoonkologie (9)
- Psychotherapie (9)
- Quantenmechanik (9)
- Radiotherapy (9)
- Rechenzentrum (9)
- Rechtsvergleich (9)
- Regeneration (9)
- Regionalentwicklung (9)
- Revision (9)
- Sarkoidose (9)
- Satellit (9)
- Schmalwand <Arabidopsis> (9)
- Schwindel (9)
- Schüttgut (9)
- Sentinel-1 (9)
- Sprache (9)
- Squaraine (9)
- Staphylococcus (9)
- Stathmin (9)
- Stereoselektive Synthese (9)
- Stressreaktion (9)
- Systembiologie (9)
- Südafrika (9)
- TWEAK (9)
- Textverstehen (9)
- Theologie (9)
- Tourismus (9)
- Trabekulektomie (9)
- Transplantat (9)
- Transport (9)
- Trypanosoma (9)
- Tumorantigen (9)
- Vakuole (9)
- Vergleich (9)
- Visuelle Aufmerksamkeit (9)
- Wurzel (9)
- X-ray crystallography (9)
- Zellkern (9)
- Zinkselenid (9)
- aggregation (9)
- agriculture (9)
- aldosterone (9)
- antennal lobe (9)
- anxiety disorders (9)
- aromaticity (9)
- atrial fibrillation (9)
- biomechanics (9)
- blood brain barrier (9)
- body size (9)
- bone cement (9)
- cancer therapy (9)
- cell culture (9)
- cell wall (9)
- central nervous system (9)
- chronic heart failure (9)
- collagen (9)
- communication (9)
- coronary heart disease (9)
- cystic fibrosis (9)
- decision making (9)
- dendritische Zellen (9)
- dialysis (9)
- dispersal (9)
- earth observation (9)
- education (9)
- electron microscopy (9)
- electrophysiology (9)
- energy (9)
- experimental autoimmune encephalomyelitis (9)
- family (9)
- fatigue (9)
- forest (9)
- gene therapy (9)
- genomics (9)
- glioma (9)
- global change (9)
- glucose (9)
- high energy physics (9)
- hip (9)
- hyaluronic acid (9)
- hydrogel (9)
- impact (9)
- incidence (9)
- inhibitor (9)
- interferon (9)
- mapping (9)
- mechanotransduction (9)
- mesenchymale Stammzellen (9)
- metapopulation (9)
- methylation (9)
- mitosis (9)
- monitoring (9)
- monoklonale Antikörper (9)
- myocardium (9)
- near-infrared spectroscopy (9)
- neurology (9)
- organic chemistry (9)
- organic semiconductors (9)
- pathway (9)
- permeability (9)
- platelet activation (9)
- platelet aggregation (9)
- prognostic factors (9)
- quality assurance (9)
- quantification (9)
- rectal cancer (9)
- reveals (9)
- serum (9)
- spinal muscular atrophy (9)
- synapse (9)
- systematic uncertainty (9)
- temperature (9)
- tolerance (9)
- transcription factor (9)
- transient absorption (9)
- transmission (9)
- type 2 diabetes (9)
- vaccine (9)
- walking (9)
- water (9)
- wound healing (9)
- ATLAS (8)
- Adhärenz (8)
- Affekt (8)
- Aggregat <Chemie> (8)
- Akt (8)
- Aktiver galaktischer Kern (8)
- Analyse (8)
- Ancistrocladaceae (8)
- Anthocyane (8)
- Antigen (8)
- Antigen CD40 (8)
- Aortenklappenersatz (8)
- Aortenstenose (8)
- Arthroskopie (8)
- Arzneimittelforschung (8)
- Asymmetrie (8)
- Asymmetrische Synthese (8)
- Auge (8)
- Autoaggressionskrankheit (8)
- Autonomer Roboter (8)
- Autophagie (8)
- Bauchspeicheldrüse (8)
- Biotransformation (8)
- Biradikal (8)
- Blimp-1 (8)
- Blutdruck (8)
- Blutstammzelle (8)
- Botanik (8)
- Brain (8)
- Burkina Faso (8)
- CD28 (8)
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- CD95 (8)
- COMT (8)
- CT (8)
- Caenorhabditis elegans (8)
- Camponotus floridanus (8)
- Chlamydia (8)
- Cloud Computing (8)
- Compressed Sensing (8)
- Cross-Section (8)
- Cysteinproteasen (8)
- Cytomegalie-Virus (8)
- DLBCL (8)
- Darm (8)
- Data Mining (8)
- Datenbank (8)
- Deep learning (8)
- Demenz (8)
- Deutschunterricht (8)
- Diagnose (8)
- Diborene (8)
- Diskursanalyse (8)
- ERK (8)
- Elektronenspin (8)
- Elektronenspinresonanz (8)
- Emotionsregulation (8)
- Endothelzellen (8)
- Engagement (8)
- Erbrechen (8)
- Europe (8)
- Fachdidaktik (8)
- Femtosekundenbereich (8)
- Fibroblasten (8)
- Finite-Elemente-Methode (8)
- Fitness (8)
- Fluoreszenzspektroskopie (8)
- Forschung (8)
- Fußball (8)
- Gammastrahlung (8)
- Gedächtnisleistung (8)
- Geschlechtsunterschied (8)
- Gewebe (8)
- Gliom (8)
- HPLC-MS (8)
- Harnwegsinfektion (8)
- Haut (8)
- Hernie (8)
- Herzmuskelkrankheit (8)
- Hochbegabung (8)
- Hubbard-Modell (8)
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- Hypertrophie (8)
- Hüftgelenk (8)
- IL-10 (8)
- Immunantwort (8)
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- In vivo (8)
- Inklusion (8)
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- Knockout (8)
- Komposit <Zahnmedizin> (8)
- Kondo-Effekt (8)
- Korrelation (8)
- Kreuzband (8)
- Kristallstruktur (8)
- Landwirtschaft (8)
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- Lehre (8)
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- N-heterocyclic carbenes (8)
- NSCLC (8)
- Nachsorge (8)
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- Neutrino (8)
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- Onkolyse (8)
- Optogenetik (8)
- Organisches Molekül (8)
- Oxylipine (8)
- PD-L1 (8)
- PONV (8)
- Pankreaskarzinom (8)
- Parton distributions (8)
- Pemphigus (8)
- Peptide (8)
- Periphere arterielle Verschlusskrankheit (8)
- Permeabilität (8)
- Persönlichkeit (8)
- Pharmakodynamik (8)
- Photodissoziation (8)
- Photovoltaik (8)
- Platin (8)
- Pollen (8)
- Polycyclische Aromaten (8)
- Protein p53 (8)
- Proteinbindung (8)
- Protonen-NMR-Spektroskopie (8)
- Präkonditionierung (8)
- Psychische Störung (8)
- Pump-Probe-Technik (8)
- Quantendynamik (8)
- Quelle (8)
- Quran (8)
- Radikal <Chemie> (8)
- Raf <Biochemie> (8)
- Raf-Kinasen (8)
- Raman spectroscopy (8)
- Rastertunnelmikroskop (8)
- Rechenzentrum Universität Würzburg (8)
- Regenerative Medizin (8)
- Rekombination (8)
- Retroviren (8)
- Ringöffnungspolymerisation (8)
- Röntgen-Photoelektronenspektroskopie (8)
- Salmonella typhimurium (8)
- Schmerzforschung (8)
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- Semimagnetischer Halbleiter (8)
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- Syntax (8)
- Synthesis (8)
- T cell activation (8)
- T lymphocytes (8)
- T-Zelle (8)
- TDM (8)
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- Titan (8)
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- Transthorakale Echokardiographie (8)
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- UAV (8)
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- Wirkstofffreisetzung (8)
- Zahnmedizin (8)
- Zeitreihenanalyse (8)
- Zentralnervensystem (8)
- actin cytoskeleton (8)
- adherence (8)
- amyotrophic lateral sclerosis (8)
- analysis of variance (8)
- animal models (8)
- antigen (8)
- astrocytes (8)
- bioinformatics (8)
- body mass index (8)
- borylene (8)
- calcium phosphate (8)
- cardiac surgery (8)
- cardiovascular disease (8)
- cell migration (8)
- central complex (8)
- ceramide (8)
- cerebrospinal fluid (8)
- charge transfer (8)
- chromatin (8)
- chronic pain (8)
- cleft lip and palate (8)
- coagulation (8)
- cognitive impairment (8)
- coherence (8)
- computed tomography (8)
- copper (8)
- cortisol (8)
- cultural studies (8)
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- decay (8)
- division of labor (8)
- domain (8)
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- embodiment (8)
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- histology (8)
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- immunology (8)
- implant (8)
- in vivo imaging (8)
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- integrins (8)
- iron (8)
- jet energy scale (8)
- juvenile idiopathic arthritis (8)
- kinematic alignment (8)
- latency (8)
- mHealth (8)
- macrophage (8)
- major depression (8)
- meta-analysis (8)
- methylphenidate (8)
- microenvironment (8)
- microstructure (8)
- monocyte (8)
- mutualism (8)
- neurogenesis (8)
- oncology (8)
- optimal control (8)
- organic solar cells (8)
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- --- (4)
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Schriftenreihe
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- Spezielle Didaktik der Sportarten (2)
- Alter Orient und Altes Testament : Sonderreihe Veröffentlichungen zur Kultur und Geschichte des Alten Orients ; 3 (1)
- Berichte aus der Informatik (1)
- Deuterocanonical and Cognate Literature Studies (1)
- Deuterocanonical and Cognate Literature Yearbook (1)
- European Journal of Clinical Nutrition ; 70 (1)
- Forum Junge Romanistik 18 (1)
- Grundzüge ; 45 (1)
- Journal of Neuroimaging; Volume 28 (1)
Sonstige beteiligte Institutionen
- Johns Hopkins School of Medicine (18)
- Fraunhofer-Institut für Silicatforschung ISC (8)
- IZKF Nachwuchsgruppe Geweberegeneration für muskuloskelettale Erkrankungen (7)
- Akademie der Wissenschaften und der Literatur, Mainz (6)
- DFG Forschungsgruppe 2757 / Lokale Selbstregelungen im Kontext schwacher Staatlichkeit in Antike und Moderne (LoSAM) (6)
- Clinical Trial Center (CTC) / Zentrale für Klinische Studien Würzburg (ZKSW) (5)
- Helmholtz Institute for RNA-based Infection Research (HIRI) (5)
- Johns Hopkins University School of Medicine (5)
- Universität Leipzig (5)
- Universitätsklinikum Würzburg (5)
ResearcherID
- B-1911-2015 (1)
- B-4606-2017 (1)
- C-2593-2016 (1)
- D-1221-2009 (1)
- D-1250-2010 (1)
- D-3057-2014 (1)
- I-5818-2014 (1)
- J-8841-2015 (1)
- M-1240-2017 (1)
- N-2030-2015 (1)
Infection research largely relies on classical cell culture or mouse models. Despite having delivered invaluable insights into host-pathogen interactions, both have limitations in translating mechanistic principles to human pathologies. Alternatives can be derived from modern Tissue Engineering approaches, allowing the reconstruction of functional tissue models in vitro. Here, we combined a biological extracellular matrix with primary tissue-derived enteroids to establish an in vitro model of the human small intestinal epithelium exhibiting in vivo-like characteristics. Using the foodborne pathogen Salmonella enterica serovar Typhimurium, we demonstrated the applicability of our model to enteric infection research in the human context. Infection assays coupled to spatio-temporal readouts recapitulated the established key steps of epithelial infection by this pathogen in our model. Besides, we detected the upregulation of olfactomedin 4 in infected cells, a hitherto unrecognized aspect of the host response to Salmonella infection. Together, this primary human small intestinal tissue model fills the gap between simplistic cell culture and animal models of infection, and shall prove valuable in uncovering human-specific features of host-pathogen interplay.
In the fast-evolving landscape of biomedical research, the emergence of big data has presented researchers with extraordinary opportunities to explore biological complexities. In biomedical research, big data imply also a big responsibility. This is not only due to genomics data being sensitive information but also due to genomics data being shared and re-analysed among the scientific community. This saves valuable resources and can even help to find new insights in silico. To fully use these opportunities, detailed and correct metadata are imperative. This includes not only the availability of metadata but also their correctness. Metadata integrity serves as a fundamental determinant of research credibility, supporting the reliability and reproducibility of data-driven findings. Ensuring metadata availability, curation, and accuracy are therefore essential for bioinformatic research. Not only must metadata be readily available, but they must also be meticulously curated and ideally error-free. Motivated by an accidental discovery of a critical metadata error in patient data published in two high-impact journals, we aim to raise awareness for the need of correct, complete, and curated metadata. We describe how the metadata error was found, addressed, and present examples for metadata-related challenges in omics research, along with supporting measures, including tools for checking metadata and software to facilitate various steps from data analysis to published research.
Highlights
• Data awareness and data integrity underpins the trustworthiness of results and subsequent further analysis.
• Big data and bioinformatics enable efficient resource use by repurposing publicly available RNA-Sequencing data.
• Manual checks of data quality and integrity are insufficient due to the overwhelming volume and rapidly growing data.
• Automation and artificial intelligence provide cost-effective and efficient solutions for data integrity and quality checks.
• FAIR data management, various software solutions and analysis tools assist metadata maintenance.
Machine learning techniques are excellent to analyze expression data from single cells. These techniques impact all fields ranging from cell annotation and clustering to signature identification. The presented framework evaluates gene selection sets how far they optimally separate defined phenotypes or cell groups. This innovation overcomes the present limitation to objectively and correctly identify a small gene set of high information content regarding separating phenotypes for which corresponding code scripts are provided. The small but meaningful subset of the original genes (or feature space) facilitates human interpretability of the differences of the phenotypes including those found by machine learning results and may even turn correlations between genes and phenotypes into a causal explanation. For the feature selection task, the principal feature analysis is utilized which reduces redundant information while selecting genes that carry the information for separating the phenotypes. In this context, the presented framework shows explainability of unsupervised learning as it reveals cell-type specific signatures. Apart from a Seurat preprocessing tool and the PFA script, the pipeline uses mutual information to balance accuracy and size of the gene set if desired. A validation part to evaluate the gene selection for their information content regarding the separation of the phenotypes is provided as well, binary and multiclass classification of 3 or 4 groups are studied. Results from different single-cell data are presented. In each, only about ten out of more than 30000 genes are identified as carrying the relevant information. The code is provided in a GitHub repository at https://github.com/AC-PHD/Seurat_PFA_pipeline.
CRISPR/Cas9 gene editing has revolutionised loss-of-function experiments in Leishmania, the causative agent of leishmaniasis. As Leishmania lack a functional non-homologous DNA end joining pathway however, obtaining null mutants typically requires additional donor DNA, selection of drug resistance-associated edits or time-consuming isolation of clones. Genome-wide loss-of-function screens across different conditions and across multiple Leishmania species are therefore unfeasible at present. Here, we report a CRISPR/Cas9 cytosine base editor (CBE) toolbox that overcomes these limitations. We employed CBEs in Leishmania to introduce STOP codons by converting cytosine into thymine and created http://www.leishbaseedit.net/ for CBE primer design in kinetoplastids. Through reporter assays and by targeting single- and multi-copy genes in L. mexicana, L. major, L. donovani, and L. infantum, we demonstrate how this tool can efficiently generate functional null mutants by expressing just one single-guide RNA, reaching up to 100% editing rate in non-clonal populations. We then generated a Leishmania-optimised CBE and successfully targeted an essential gene in a plasmid library delivered loss-of-function screen in L. mexicana. Since our method does not require DNA double-strand breaks, homologous recombination, donor DNA, or isolation of clones, we believe that this enables for the first time functional genetic screens in Leishmania via delivery of plasmid libraries.
Increased intestinal permeability and inflammation, both fueled by dysbiosis, appear to contribute to rheumatoid arthritis (RA) pathogenesis. This single-center pilot study aimed to investigate zonulin, a marker of intestinal permeability, and calprotectin, a marker of intestinal inflammation, measured in serum and fecal samples of RA patients using commercially available kits. We also analyzed plasma lipopolysaccharide (LPS) levels, a marker of intestinal permeability and inflammation. Furthermore, univariate, and multivariate regression analyses were carried out to determine whether or not there were associations of zonulin and calprotectin with LPS, BMI, gender, age, RA-specific parameters, fiber intake, and short-chain fatty acids in the gut. Serum zonulin levels were more likely to be abnormal with a longer disease duration and fecal zonulin levels were inversely associated with age. A strong association between fecal and serum calprotectin and between fecal calprotectin and LPS were found in males, but not in females, independent of other biomarkers, suggesting that fecal calprotectin may be a more specific biomarker than serum calprotectin is of intestinal inflammation in RA. Since this was a proof-of-principle study without a healthy control group, further research is needed to validate fecal and serum zonulin as valid biomarkers of RA in comparison with other promising biomarkers.
Making judgments of learning (JOLs) after studying can directly improve learning. This JOL reactivity has been shown for simple materials but has scarcely been investigated with educationally relevant materials such as expository texts. The few existing studies have not yet reported any consistent gains in text comprehension due to providing JOLs. In the present study, we hypothesized that increasing the chances of covert retrieval attempts when making JOLs after each of five to-be-studied text passages would produce comprehension benefits at 1 week compared to restudy. In a between-subjects design, we manipulated both whether participants (N = 210) were instructed to covertly retrieve the texts, and whether they made delayed target-absent JOLs. The results indicated that delayed, target-absent JOLs did not improve text comprehension after 1 week, regardless of whether prior instructions to engage in covert retrieval were provided. Based on the two-stage model of JOLs, we reasoned that participants’ retrieval attempts during metacomprehension judgments were either insufficient (i.e., due to a quick familiarity assessment) or were ineffective (e.g., due to low retrieval success).
Background: The COVID-19 pandemic has led to a flood of — often contradictory — evidence. HCWs had to develop strategies to locate information that supported their work. We investigated the information-seeking of different HCW groups in Germany. Methods: In December 2020, we conducted online surveys on COVID-19 information sources, strategies, assigned trustworthiness, and barriers — and in February 2021, on COVID-19 vaccination information sources. Results were analyzed descriptively; group comparisons were performed using χ\(^2\)-tests. Results: For general COVID-19-related medical information (413 participants), non-physicians most often selected official websites (57%), TV (57%), and e-mail/newsletters (46%) as preferred information sources — physicians chose official websites (63%), e-mail/newsletters (56%), and professional journals (55%). Non-physician HCWs used Facebook/YouTube more frequently. The main barriers were insufficient time and access issues. Non-physicians chose abstracts (66%), videos (45%), and webinars (40%) as preferred information strategy; physicians: overviews with algorithms (66%), abstracts (62%), webinars (48%). Information seeking on COVID-19 vaccination (2700 participants) was quite similar, however, with newspapers being more often used by non-physicians (63%) vs. physician HCWs (70%). Conclusion: Non-physician HCWs more often consulted public information sources. Employers/institutions should ensure the supply of professional, targeted COVID-19 information for different HCW groups.
PRO-Simat is a simulation tool for analysing protein interaction networks, their dynamic change and pathway engineering. It provides GO enrichment, KEGG pathway analyses, and network visualisation from an integrated database of more than 8 million protein-protein interactions across 32 model organisms and the human proteome. We integrated dynamical network simulation using the Jimena framework, which quickly and efficiently simulates Boolean genetic regulatory networks. It enables simulation outputs with in-depth analysis of the type, strength, duration and pathway of the protein interactions on the website. Furthermore, the user can efficiently edit and analyse the effect of network modifications and engineering experiments. In case studies, applications of PRO-Simat are demonstrated: (i) understanding mutually exclusive differentiation pathways in Bacillus subtilis, (ii) making Vaccinia virus oncolytic by switching on its viral replication mainly in cancer cells and triggering cancer cell apoptosis and (iii) optogenetic control of nucleotide processing protein networks to operate DNA storage. Multilevel communication between components is critical for efficient network switching, as demonstrated by a general census on prokaryotic and eukaryotic networks and comparing design with synthetic networks using PRO-Simat. The tool is available at https://prosimat.heinzelab.de/ as a web-based query server.
In recent years, various forms of caloric restriction (CR) and amino acid or protein restriction (AAR or PR) have shown not only success in preventing age-associated diseases, such as type II diabetes and cardiovascular diseases, but also potential for cancer therapy. These strategies not only reprogram metabolism to low-energy metabolism (LEM), which is disadvantageous for neoplastic cells, but also significantly inhibit proliferation. Head and neck squamous cell carcinoma (HNSCC) is one of the most common tumour types, with over 600,000 new cases diagnosed annually worldwide. With a 5-year survival rate of approximately 55%, the poor prognosis has not improved despite extensive research and new adjuvant therapies. Therefore, for the first time, we analysed the potential of methionine restriction (MetR) in selected HNSCC cell lines. We investigated the influence of MetR on cell proliferation and vitality, the compensation for MetR by homocysteine, the gene regulation of different amino acid transporters, and the influence of cisplatin on cell proliferation in different HNSCC cell lines.
Alveolar (AE) and cystic (CE) echinococcosis are two parasitic diseases caused by the tapeworms Echinococcus multilocularis and E. granulosus sensu lato (s. l.), respectively. Currently, AE and CE are mainly diagnosed by means of imaging techniques, serology, and clinical and epidemiological data. However, no viability markers that indicate parasite state during infection are available. Extracellular small RNAs (sRNAs) are short non-coding RNAs that can be secreted by cells through association with extracellular vesicles, proteins, or lipoproteins. Circulating sRNAs can show altered expression in pathological states; hence, they are intensively studied as biomarkers for several diseases. Here, we profiled the sRNA transcriptomes of AE and CE patients to identify novel biomarkers to aid in medical decisions when current diagnostic procedures are inconclusive. For this, endogenous and parasitic sRNAs were analyzed by sRNA sequencing in serum from disease negative, positive, and treated patients and patients harboring a non-parasitic lesion. Consequently, 20 differentially expressed sRNAs associated with AE, CE, and/or non-parasitic lesion were identified. Our results represent an in-depth characterization of the effect E. multilocularis and E. granulosus s. l. exert on the extracellular sRNA landscape in human infections and provide a set of novel candidate biomarkers for both AE and CE detection.
Grading, immunohistochemistry and c-kit mutation status are criteria for assessing the prognosis and therapeutic options of canine cutaneous mast cell tumours (MCTs). As a subset, canine digital MCTs have rarely been explored in this context. Therefore, in this retrospective study, 68 paraffin-embedded canine digital MCTs were analysed, and histological grading was assessed according to Patnaik and Kiupel. The immunohistochemical markers KIT and Ki67 were used, as well as polymerase chain reaction (PCR) for mutational screening in c-kit exons 8, 9, 11 and 14. Patnaik grading resulted in 22.1% grade I, 67.6% grade II and 10.3% grade III tumours. Some 86.8% of the digital MCTs were Kiupel low-grade. Aberrant KIT staining patterns II and III were found in 58.8%, and a count of more than 23 Ki67-positive cells in 52.3% of the cases. Both parameters were significantly associated with an internal tandem duplication (ITD) in c-kit exon 11 (12.7%). French Bulldogs, which tend to form well-differentiated cutaneous MCTs, had a higher proportion of digital high-grade MCTs and ITD in c-kit exon 11 compared with mongrels. Due to its retrospective nature, this study did not allow for an analysis of survival data. Nevertheless, it may contribute to the targeted characterisation of digital MCTs.
Improved wall temperature prediction for the LUMEN rocket combustion chamber with neural networks
(2023)
Accurate calculations of the heat transfer and the resulting maximum wall temperature are essential for the optimal design of reliable and efficient regenerative cooling systems. However, predicting the heat transfer of supercritical methane flowing in cooling channels of a regeneratively cooled rocket combustor presents a significant challenge. High-fidelity CFD calculations provide sufficient accuracy but are computationally too expensive to be used within elaborate design optimization routines. In a previous work it has been shown that a surrogate model based on neural networks is able to predict the maximum wall temperature along straight cooling channels with convincing precision when trained with data from CFD simulations for simple cooling channel segments. In this paper, the methodology is extended to cooling channels with curvature. The predictions of the extended model are tested against CFD simulations with different boundary conditions for the representative LUMEN combustor contour with varying geometries and heat flux densities. The high accuracy of the extended model’s predictions, suggests that it will be a valuable tool for designing and analyzing regenerative cooling systems with greater efficiency and effectiveness.
After priming in Peyer's patches (PPs) and mesenteric lymph nodes (mLN) T- cells infiltrate the intestine through lymphatic draining and homing through the bloodstream. However, we found that in mouse models of acute graft-versus-host disease (GvHD), a subset of alloreactive T-cells directly migrates from PPs to the adjacent intestinal lamina propria (LP), bypassing the normal lymphatic drainage and vascular trafficking routes. Notably, this direct migration occurred in irradiated and unirradiated GvHD models, indicating that irradiation is not a prerequisite for this observed behavior.
Next, we established a method termed serial intravascular staining (SIVS) in mouse models to systematically investigate the trafficking and migration of donor T- cells in the early stages of acute GvHD initiation. We found that the direct migration of T-cells from PPs to LP resulted in faster recruitment of cells after allogeneic hematopoietic cell transplantation (allo-HCT). These directly migrating T-cells were found to be in an activated and proliferative state, exhibiting a TH1/TH17-like phenotype and producing cytokines such as IFN-γ and TNF-α. Furthermore, we observed that the directly migrating alloreactive T-cells expressed specific integrins (α4+, αE+) and chemokine receptors (CxCR3+, CCR5+, and CCR9+). Surprisingly, blocking these integrins and chemokine-coupled receptors did not hinder the direct migration of T- cells from PPs to LP, suggesting the involvement of alternative mechanisms. Previous experiments ruled out the involvement of S1PR1 and topographical features of macrophages, leading us to hypothesize that mediators of cytoskeleton reorganization, such as Coro1a, Dock2, or Cdc42, may play a role in this unique migration process.
Additionally, we observed that directly migrating T-cells created a local inflammatory microenvironment, which attracts circulating T-cells. Histological analysis confirmed that alloreactive PPs-derived T-cells and bloodborne T-cells colocalized. We employed two experimental approaches, including either photoconversion of T-cells in PPs or direct transfer of activated T-cells into the vasculature, to demonstrate this colocalization. We hypothesize that cytokines released by migrating T-cells, such as IFN-γ and TNF-α, may play a role in recruiting T-cells from the vasculature, as inhibiting chemokine-coupled receptors did not impair recruitment.
Gastroesophageal junction (GEJ), demarcating the region where the distal esophagus meets with the proximal stomach region, is known for developing pathological conditions, including metaplasia and esophageal adenocarcinoma (EAC). It is essential to understand the mechanisms of developmental stages which lead to EAC since the incidence rate of EAC increased over 7-fold during the past four decades, and the overall five years survival rate is 18.4%. In most cases, patients are diagnosed in the advanced stage without prior symptoms. The main precursor for the development of EAC is a pre-malignant condition called Barrett's esophagus (BE). BE is the metaplastic condition where the multilayered squamous epithelium of the native esophagus is replaced by specialized single-layered columnar epithelium, which shows the molecular characteristics of the gastric as well as intestinal epithelium. The main risk factors for BE development include chronic gastro-esophageal acid reflux disease (GERD), altered microbiota, and altered retinoic acid signaling (RA). The cell of origin of BE is under debate due to a lack of clear evidence demonstrating the process of BE initiation. Here, I investigated how GEJ homeostasis is maintained in healthy tissue by stem cell regulatory morphogens, the role of vitamin A (RA signaling), and how its alteration contributes to BE development.
In the first part of my thesis, I showed the presence of two types of epithelial cells, the squamous type in the esophagus and the columnar type in the stomach region in the GEJ, using single-molecule RNA in situ hybridization (smRNA-ISH) and immunohistochemistry. Employing lineage tracing in the mouse model, I have demonstrated that the esophageal epithelial and stomach epithelial cells derived from two distinct epithelial stem cell lineages in the GEJ. The border between squamous and columnar epithelial cells in the Squamo-columnar junction (SCJ) of GEJ is regulated by opposing Wnt microenvironments. The regeneration of stomach columnar epithelial stem cells is maintained by Wnt activating signal from the stromal compartment while squamous epithelial stem cells of the esophagus are maintained by the Wnt inhibitory signals. I recapitulated the in vivo GEJ epithelial stem cell maintenance by using in vitro epithelial 3D organoid culture model. The growth and propagation of stomach columnar epithelial organoids depend on Wnt growth factors, while squamous epithelial organoids' development needs Wnt-deficient culture conditions.
Further, single-cell RNA sequence (scRNA-seq) analysis of organoid-derived epithelial cells revealed the non-canonical Wnt/ planar cell polarity (PCP) pathway involvement in regulating the squamous epithelial cells. In contrast, columnar stomach epithelial cells are regulated by the canonical Wnt/ beta-catenin and non-canonical Wnt/Ca2+ pathways. My data indicate that the SCJ epithelial cells that merge at the GEJ are regulated by opposing stromal Wnt factors and distinct Wnt pathway signaling in the epithelial cells.
In the second part of the thesis, I investigated the role of Vitamin A-derived bioactive compound RA on esophageal and stomach epithelial stem cells. In vitro treatment of esophageal and stomach, epithelial organoids with RA or its pharmacological inhibitor BMS 493 revealed that each cell type was regulated distinctly. I observed that enhanced RA promoted esophageal stem cell differentiation and loss of stratification, while RA inhibition led to enhanced stemness and regeneration of the esophagus stratified epithelium. As opposed to the esophagus, RA signaling is active in the stomach organoids, and inhibition of RA reduces the growth of stomach organoids. Global transcriptomic data and scRNA-seq data revealed that RA signaling induces dormancy phenotype in the esophageal cells. In contrast, the absence of RA in stomach epithelial cells induces the expression of genes associated with BE. Thus, spatially defined regulation of Wnt and RA signaling at GEJ is critical for healthy homeostasis, and its perturbation leads to disease development.
The unicellular pathogen Trypanosoma brucei is the causative agent of African
trypanosomiasis, an endemic disease prevalent in sub-Saharan Africa. Trypanosoma brucei alternates between a mammalian host and the tsetse fly vector. The extracellular parasite survives in the mammalian bloodstream by periodically exchanging their ˈvariant surface glycoproteinˈ (VSG) coat to evade the host immune response. This antigenic variation is achieved through monoallelic expression of one VSG variant from subtelomeric ˈbloodstream
form expression sitesˈ (BES) at a given timepoint. During the differentiation from the bloodstream form (BSF) to the procyclic form (PCF) in the tsetse fly midgut, the stage specific surface protein is transcriptionally silenced and replaced by procyclins. Due to their subtelomeric localization on the chromosomes, VSG transcription and silencing is partly regulated by homologues of the mammalian telomere complex such as TbTRF, TbTIF2 and TbRAP1 as well as by ˈtelomere-associated proteinsˈ (TelAPs) like TelAP1. To gain more insights into transcription regulation of VSG genes, the identification and characterization of other TelAPs is critical and has not yet been achieved. In a previous study, two biochemical approaches were used to identify other novel TelAPs. By using ˈco-immunoprecipitationˈ (co-IP) to enrich possible interaction partners of TbTRF and by affinity chromatography using telomeric repeat oligonucleotides, a listing of TelAP candidates has been conducted. With this approach TelAP1 was identified as a novel component of the telomere complex, involved in the kinetics of transcriptional BES silencing during BSF to PCF differentiation. To gain further insights into the telomere complex composition, other previously enriched proteins were characterized through a screening process using RNA interference to deplete potential candidates. VSG expression profile changes and overall proteomic changes after depletion were analyzed by mass spectrometry. With this method, one can gain insights into the functions of the proteins and their involvement in VSG expression site regulation. To validate the interaction of proteins enriched by co-IP with TbTRF and TelAP1 and to identify novel interaction proteins, I performed reciprocal affinity purifications of the four most promising candidates (TelAP2, TelAP3, PPL2 and PolIE) and additionally confirmed colocalization of two candidates with TbTRF via immunofluorescence (TelAP2, TelAP3). TelAP3 colocalizes with TbTRF and potentially interacts with TbTRF, TbTIF2, TelAP1 and TelAP2, as well as with two translesion polymerases PPL2 and PolIE in BSF. PPL2 and PolIE seem to be in close contact to each other at the telomeric ends and fulfill different roles as only PolIE is involved in VSG regulation while PPL2 is not. TelAP2 was previously characterized to be associated with telomeres by partially colocalizing with TbTRF and cells show a VSG derepression phenotype when the protein was depleted. Here I show that TelAP2 interacts with the telomere-binding proteins TbTRF and TbTIF2 as well as with the telomere-associated protein TelAP1 in BSF and that TelAP2 depletion results in a loss of TelAP1 colocalization with TbTRF in BSF.
In conclusion, this study demonstrates that characterizing potential TelAPs is effective in gaining insights into the telomeric complex's composition and its role in VSG regulation in Trypanosoma brucei. Understanding these interactions could potentially lead to new therapeutic targets for combatting African trypanosomiasis.
Obwohl es in den letzten 10-15 Jahren gelang, multiple MM-Genome mittels NGS auf eine kosteneffiziente Art und mit geringem Zeit- und Materialaufwand zu sequenzieren und hierdurch zum Teil bahnbrechende Erkenntnisse gewonnen werden konnten, sind molekulargenetische Untersuchungen im diagnostischen Workflow des MMs bisher nicht ausreichend implementiert, um eine personalisierte Therapieentscheidung zu ermöglichen.
Vor diesem Hintergrund wurde in der vorliegenden Arbeit eine Gruppe an Patienten mit NDMM und RRMM anhand klinischer Parameter charakterisiert und durch Verwendung des M³P-Panels auf das Vorliegen bestimmter molekulargenetischer Veränderungen untersucht. Zusammenfassend lässt sich sagen, dass unsere Analyse die bisher veröffentliche M³P-Prävalenz in MM-Tumorproben bestätigt. Zu den am häufigsten mutierten Genen gehörten KRAS, NRAS, DIS3, ATM und BRAF. In der Gruppe der Patienten mit NRAS-Mutation oder del17p war die Zahl der relevanten Mutationen deutlich höher als ohne Vorliegen der entsprechenden Veränderung. Der Nachweis eines Double-Hit-Myeloms war erwartungsgemäß der stärkste ungünstige Faktor in unserer Kohorte. Unter den Patienten mit CRBN-Mutation waren alle IMiD-vorbehandelt und zeigten im Verlauf eine Refraktärität gegenüber dieser Substanzgruppe auf. Bezüglich der Überlebensanalysen bestätigten unsere Ergebnisse bereits bekannte prognostische Risikofaktoren wie Hochrisikozytogenetik, insbesondere del17p und gain1q, eine TP53-Mutation sowie ISS- und R-ISS-Stadium III.
Die Ergebnisse der Mutationsanalysen dieser Arbeit verdeutlichen den großen wissenschaftlichen und therapeutischen Nutzen, der von molekulargenetischen Untersuchungen ausgeht. Zukünftig werden auch beim MM Therapieentscheidungen auf Grundlage genetischer Diagnostik getroffen werden, mit dem Ziel die Behandlung für MM-Patienten weiter zu verbessern.
Magnetic Particle Imaging (MPI) ist ein innovatives tomographisches Bildgebungsverfahren, mit dem Tracerpartikel äußerst sensitiv und schnell mehrdimensional abgebildet werden können. Die Methode basiert auf der nichtlinearen Magnetisierungsantwort superparamagnetischer Eisenoxidnanopartikel (SPION) in einem Messpunkt, welcher ein Messvolumen rastert. In vorliegender Arbeit wurde das sog. Traveling Wave MPI (TWMPI) Verfahren eingesetzt, wodurch im Vergleich zu konventionellen MPI-Scannern ein größeres Field of View (FOV) und eine geringere Latenz bis zur Bildanzeige erreicht werden konnte. TWMPI weist einige für medizinische Zwecke vielversprechende Eigenschaften auf: Es liefert zwei- und dreidimensionale Bildrekonstruktionen in Echtzeit mit hoher zeitlicher und räumlicher Auflösung. Dabei ist die Bildgebung von Grund auf hintergrundfrei und erfordert keinerlei ionisierende Strahlung. Zudem ist die Technik äußerst sensitiv und kann SPION-Tracer noch in mikromolaren Konzentrationen detektieren.
Ziel dieser Arbeit war es daher zu untersuchen, inwiefern es mittels TWMPI möglich ist, künstliche Stenosen im Gefäßmodell visuell in Echtzeit darzustellen und quantitativ zu beurteilen sowie überdies eine perkutane transluminale Angioplastie (PTA) im Gefäßmodell unter TWMPI-Echtzeit-Bildgebung durchzuführen.
Alle Experimente wurden in einem speziell angefertigten TWMPI-Scanner durchgeführt (JMU Würzburg, Experimentelle Physik V (Biophysik), FOV: 65 x 29 x 29 mm³, Auflösung: ca. 1.5 - 2 mm). Die Lumen-Darstellungen erfolgten mittels des SPION-Tracers Ferucarbotran in einer Verdünnung von 1 : 50 (entspr. 10 mmol [Fe]/l). Das PTA-Instrumentarium wurde mit eigens hergestelltem ferucarbotranhaltigem Lack (100 mmol [Fe]/l) markiert. Für die verschiedenen Teilexperimente wurden den jeweiligen speziellen Anforderungen entsprechend mehrere Gefäßmodelle handgefertigt.
Für die visuelle Stenosequantifizierung wurden fünf starre Stenosephantome unterschiedlicher Stenosierung (0%, 25%, 50%, 75%, 100%) aus Polyoxymethylen hergestellt (l: 40 mm, ID: 8 mm). Die Gefäßmodelle wurden mehrfach zentral im FOV platziert und das stenosierte Lumen mittels sog. Slice-Scanning Modus (SSM, Einzelaufnahme inkl. 10 Mittelungen: 200 ms, Bildfrequenz: 5 Bilder pro Sekunde, Latenz: ca. 100 ms) als zweidimensionale Quasi-Projektionen abgebildet. Diese Aufnahmen (n = 80, 16 je Phantom) wurden mit einer einheitlichen Grauskalierung versehen und anschließend entsprechend den NASCET-Kriterien visuell ausgewertet.
Alle achtzig Aufnahmen waren unabhängig vom Stenosegrad aufgrund einheitlicher Fensterung sowie konstanter Scannerparameter untereinander gut vergleichbar. Niedriggradige Stenosen konnten insgesamt genauer abgebildet werden als höhergradige, was sich neben der subjektiven Bildqualität auch in geringeren Standardabweichungen zeigte (0%: 3.70 % ± 2.71, 25%: 18.64 % ± 1.84, 50%: 52.82 % ± 3.66, 75%: 77.84 % ± 14.77, 100%: 100 % ± 0). Mit zunehmendem Stenosegrad kam es vermehrt zu geometrischen Verzerrungen im Zentrum, sodass bei den 75%-Stenosen eine breitere Streuung der Messwerte mit einer höheren Standardabweichung von 14.77% einherging. Leichte, randständige Artefakte konnten bei allen Datensätzen beobachtet werden.
Für die PTA wurden drei interaktive Gefäßmodelle aus Polyvinylchlorid (l: 100 mm, ID: 8 mm) mit zu- und abführendem Schlauchsystem entwickelt, welche mittels Kabelband von außen hochgradig eingeengt werden konnten. Analog zu einer konventionellen PTA mittels röntgenbasierter digitaler Subtraktionsangiographie (DSA), wurden alle erforderlichen Arbeitsschritte (Gefäßdarstellung, Drahtpassage, Ballonplatzierung, Angioplastie, Erfolgskontrolle) unter (TW)MPI-Echtzeit-Bildgebung (Framerate: 2 - 4 FPS, Latenz: ca. 100 ms) abgebildet bzw. durchgeführt.
Im Rahmen der PTA war eine Echtzeit-Visualisierung der Stenose im Gefäßmodell durch Tracer-Bolusgabe sowie die Führung des markierten Instrumentariums zum Zielort möglich. Die Markierung der Instrumente hielt der Beanspruchung während der Prozedur stand und ermöglichte eine genaue Platzierung des Ballonkatheters. Die Stenose konnte mittels Angioplastie-Ballons unter Echtzeit-Darstellung gesprengt werden und der Interventionserfolg im Anschluss durch erneute Visualisierung des Lumens validiert werden.
Insgesamt zeigt sich MPI somit als adäquate Bildgebungstechnik für die beiden in der Fragestellung bzw. Zielsetzung definierten experimentellen Anwendungen. Stenosen im Gefäßmodell konnten erfolgreich in Echtzeit visualisiert und bildmorphologisch nach NASCET-Kriterien quantifiziert werden. Ebenso war eine PTA im Gefäßmodell unter TWMPI-Echtzeit-Bildgebung machbar. Diese Ergebnisse unterstreichen das grundlegende Potenzial von MPI für medizinische Zwecke. Um zu den bereits etablierten Bildgebungsmethoden aufzuschließen, ist jedoch weitere Forschung im Bereich der Scanner-Hard- und -Software sowie bezüglich SPION-Tracern nötig.
The current study presents a new a group of Demotic ostraca in the belongings of the Cairo Museum. A large part of this group stem from Medinet Habu in the western bank of modern Luxor in Upper Egypt and was discovered in the beginning of the thirties of the last century by the Chicago Oriental Institute (recently renamed as Institute for the Study of Ancient Cultures ‘ISAC’). A small portion of the collection under consideration come from other Upper Egyptian provenances including Gebelein, Edfu, Kom Ombo, and possibly elsewhere in Thebes. The main goal of the present dissertation is to decipher, translate, and provide a philological, paleographical, and cultural analysis of the group of texts in question. The results of this study are spread over two main parts, the first of which is dedicated to the main and largest part of the collection, i.e. ostraca from Medinet Habu, while the second is concerned with ostraca from other places. The first part comprises of five sections beginning with receipts of money and in-kind payments including some receipts for the payments of the different capitation charges in the Ptolemaic and Roman Periods, a few for land-related payments, as well as others related to different Ptolemaic monopolies or trades such as a receipt for the price of oil, one for the linen tax, in addition to a unique receipt for the rarely attested fish tax. The second section includes accounts and lists of different kinds be it monetary, in-kind, agriculture, or any other type of lists or accounts that record different everyday transactions. The following section presents a relatively different type of lists, namely lists of personal names. The fourth section incorporates a variety of texts of different concerns, e.g. texts of religious nature, letters, temples oaths, or other private documents. Unidentified texts occupy the fifth and final section of the first part. The second part of the study, which comprises texts that originate from different Upper Egyptian localities, includes three sections, i.e. receipts, accounts, and lists of names.
The mold Aspergillus fumigatus (A. fumigatus) is known as human pathogen and can cause life-threatening infections in humans with a weakened immune system. This is a known complication in patients receiving glucocorticoids, e.g. after hematopoietic stem cell transplantation or solid organ transplantation. Although research in the field of immune cell/fungus interaction has discovered key strategies how immune cells fight against infectious fungi, our knowledge is still incomplete. In order to develop effective treatment options against fungal infections, a detailed understanding of their interactions is crucial. Thus, visualization of immune cell and fungus is an excellent approach to gain further knowledge. For a detailed view of such interaction processes, a high optical resolution on nanometer scale is required. There is a variety of super resolution microscopy techniques, enabling fluorescence imaging beyond the diffraction limit. This work combines the use of three complementary super resolution microscopy techniques, in order to study immune cell/fungus interaction from different points of view.
Aim of this work is the introduction of the recently invented imaging technique named expansion microscopy (ExM) for the study of immune cell/fungus interactions. The core aspect of this method is the physical magnification of the specimen, which increases the distance between protein structures that are close to each other and which can therefore be imaged separately.
The simultaneous magnification of primary human natural killer (NK) cells and A. fumigatus hyphae was established in this work using ExM. Reorganization of cytoskeletal components of interacting NK cells was demonstrated here, by expansion of the immunological synapse (IS), formed between NK cells and A. fumigatus. In addition, reorganization of the microtubule-organizing center (MTOC) towards fungal hyphae and an accumulation of actin at the IS has been observed. Furthermore, ExM has been used to visualize lytic granules of NK cells after degranulation. After magnification of the specimen, lysosome associated protein 1 (LAMP1) was shown to surround perforin. In absence of the plasma membrane-exposed degranulation marker LAMP1, a “ring-shaped” structure was often observed for fluorescently labeled perforin. Volume calculation of lytic granules demonstrated the benefit of ExM. Compared to pre-expansion images, analyses of post-expansion images showed two volume distributions for degranulated and non-degranulated NK cells. In addition, this work emphasizes the importance of determining the expansion factor for a structure in each species, as variations of expansion factors have been observed. This factor, as well as possible sample distortions should be considered, when ExM is used in order to analyze the interaction between two species.
A second focus of this work is the visualization of a chimeric antigen receptor (CAR), targeting an epitope on the cell wall of A. fumigatus. Structured illumination microscopy (SIM) revealed that the CAR is part of the immunological synapse of primary human CAR T cells and CAR-NK-92 cells. At the interaction site, an accumulation of the CAR was observed, as well as the presence of perforin. CAR accumulation at fungal hyphae was further demonstrated by automated live cell imaging of interacting CAR-NK-92 cells, expressing a fluorescent fusion protein.
Additionally, the use of direct stochastic optical reconstruction microscopy (dSTORM) gave first insights in CAR expression levels on the basal membrane of CAR-NK-92 cells, with single molecule sensitivity. CAR cluster analyses displayed a heterogeneous CAR density on the basal membrane of transfected NK 92 cells.
In summary, this work provides insights into the application of ExM for studying the interaction of primary human NK cells and A. fumigatus for the first time. Furthermore, this thesis presents first insights regarding the characterization of an A. fumigatus-targeting CAR, by applying super-resolution fluorescence microscopy, like SIM and dSTORM.