Refine
Is part of the Bibliography
- yes (986)
Year of publication
Document Type
- Doctoral Thesis (978)
- Journal article (7)
- Book (1)
Language
- English (763)
- German (222)
- Multiple languages (1)
Keywords
- Maus (38)
- Thrombozyt (32)
- T-Lymphozyt (29)
- Tissue Engineering (29)
- Taufliege (20)
- Dendritische Zelle (19)
- Genexpression (18)
- Angst (16)
- Entzündung (16)
- Myc (15)
- Signaltransduktion (15)
- Angststörung (14)
- Arteriosklerose (13)
- Aufmerksamkeit (13)
- Drosophila (13)
- Staphylococcus aureus (13)
- Biene (12)
- Candida albicans (12)
- G-Protein gekoppelte Rezeptoren (12)
- Immunologie (12)
- Kernspintomografie (12)
- Melanom (12)
- Schlaganfall (12)
- Serotonin (12)
- Fluoreszenzmikroskopie (11)
- Krebs <Medizin> (11)
- Mikroskopie (11)
- Stress (11)
- Epigenetik (10)
- GPCR (10)
- Microscopy (10)
- Regulation (10)
- Therapie (10)
- Thrombose (10)
- Transkriptionsfaktor (10)
- miRNS (10)
- ADHD (9)
- Apoptosis (9)
- DNS-Reparatur (9)
- Depression (9)
- Escherichia coli (9)
- Fluoreszenz-Resonanz-Energie-Transfer (9)
- Furcht (9)
- Megakaryozyt (9)
- Motoneuron (9)
- Plasmozytom (9)
- Stammzelle (9)
- Tagesrhythmus (9)
- Zellskelett (9)
- Zellzyklus (9)
- platelets (9)
- Aspergillus fumigatus (8)
- Elektrophysiologie (8)
- Fuchsbandwurm (8)
- Induzierte pluripotente Stammzelle (8)
- Konditionierung (8)
- Multiples Myelom (8)
- Neisseria meningitidis (8)
- Rezeptor (8)
- Spinale Muskelatrophie (8)
- Synapse (8)
- Alzheimerkrankheit (7)
- Arzneimitteldesign (7)
- Biomarker (7)
- DNS-Schädigung (7)
- EEG (7)
- Genetik (7)
- Herzinfarkt (7)
- Herzinsuffizienz (7)
- Immuntherapie (7)
- Inhibitor (7)
- Kutikula (7)
- Lernen (7)
- Lungenkrebs (7)
- Makrophage (7)
- Regulatorischer T-Lymphozyt (7)
- Small RNA (7)
- Transcription (7)
- Transkription <Genetik> (7)
- Transplantat-Wirt-Reaktion (7)
- Verhalten (7)
- Zellkultur (7)
- dendritic cells (7)
- fMRI (7)
- Antigen CD28 (6)
- Bacteria (6)
- Bildgebendes Verfahren (6)
- Calcium (6)
- DNA damage (6)
- Diabetes mellitus (6)
- Drosophila melanogaster (6)
- Entwicklung (6)
- Funktionelle Kernspintomografie (6)
- Hippocampus (6)
- Inflammation (6)
- Kognition (6)
- MRI (6)
- MYC (6)
- Masernvirus (6)
- Molekularbiologie (6)
- Monoklonaler Antikörper (6)
- Multiple Sklerose (6)
- Oxidativer Stress (6)
- Platelet (6)
- Platelets (6)
- Ratte (6)
- Thrombosis (6)
- Tiermodell (6)
- Transkriptomanalyse (6)
- Trypanosoma brucei (6)
- Ubiquitin (6)
- Virtuelle Realität (6)
- Zellmigration (6)
- Atherosclerosis (5)
- Aufmerksamkeits-Defizit-Syndrom (5)
- Autoantikörper (5)
- B-Lymphozyt (5)
- Bakterien (5)
- Biologie (5)
- Brain-derived neurotrophic factor (5)
- Cataglyphis (5)
- Chronobiologie (5)
- Cytoskeleton (5)
- Dendritic cells (5)
- Dünndarm (5)
- Evolution (5)
- Gefühl (5)
- Genetics (5)
- Gephyrin (5)
- HIV (5)
- Immunsystem (5)
- In-vitro-Kultur (5)
- Inhibition (5)
- MAP-Kinase (5)
- Metabolismus (5)
- Mutagenität (5)
- NFATc1 (5)
- Neurodegeneration (5)
- Neuroethologie (5)
- Polymorphismus (5)
- Resistenz (5)
- Schmerz (5)
- Transkription (5)
- Zelldifferenzierung (5)
- cancer (5)
- platelet (5)
- virtual reality (5)
- 3D-Druck (4)
- Actin (4)
- Anxiety (4)
- Aufmerksamkeitsdefizit-Syndrom (4)
- Aurora-A (4)
- Autoimmunität (4)
- Biofabrication (4)
- Biofilm (4)
- Bioinformatics (4)
- Biomaterial (4)
- Blut-Hirn-Schranke (4)
- Brustkrebs (4)
- CRISPR/Cas-Methode (4)
- CRISPR/Cas9 (4)
- Cadherine (4)
- Cancer (4)
- Chlamydia trachomatis (4)
- Chromatin (4)
- Chronophin (4)
- Darm (4)
- Dendritic cell (4)
- Dendritische Zellen (4)
- Einzelmolekülmikroskopie (4)
- Elektroencephalographie (4)
- Emotion (4)
- FRET (4)
- Fettsäurestoffwechsel (4)
- Fibroblastenwachstumsfaktor (4)
- Furchtgeneralisierung (4)
- Furchtkonditionierung (4)
- G-Protein gekoppelter Rezeptor (4)
- GPVI (4)
- Genotoxizität (4)
- Genregulation (4)
- Glucosetransportproteine (4)
- Graft-versus-host-disease (4)
- Herzmuskelzelle (4)
- Hämostase (4)
- Immunmodulation (4)
- Immunology (4)
- Immunreaktion (4)
- Immunsuppression (4)
- Infektion (4)
- Insulin (4)
- Interleukin 6 (4)
- Ionenkanal (4)
- Jugend (4)
- Kind (4)
- Knockout <Molekulargenetik> (4)
- MMB (4)
- MYCN (4)
- Malaria (4)
- Maschinelles Lernen (4)
- Mausmodell (4)
- Meeresschwämme (4)
- Mitochondrium (4)
- Mitose (4)
- Molekulargenetik (4)
- Myokardinfarkt (4)
- Natürliche Killerzelle (4)
- Neurobiologie (4)
- Neuroblastom (4)
- Neurowissenschaften (4)
- Opiatrezeptor (4)
- Panikstörung (4)
- Parkinson-Krankheit (4)
- Plasmodium falciparum (4)
- Posttranskriptionelle Regulation (4)
- Probiotikum (4)
- Proteasen (4)
- RNS-Bindungsproteine (4)
- Ribosom (4)
- SMN (4)
- Salmonella (4)
- Sepsis (4)
- Soziale Insekten (4)
- Stoffwechsel (4)
- TFIIH (4)
- Treg (4)
- TrkB (4)
- Tumor (4)
- Virulenz (4)
- Zelle (4)
- active zone (4)
- division of labor (4)
- genotoxicity (4)
- honeybee (4)
- inflammation (4)
- magnetic resonance imaging (4)
- measles virus (4)
- miRNA (4)
- microRNA (4)
- myocardial infarction (4)
- neuroblastoma (4)
- olfaction (4)
- regulation (4)
- 3D tissue model (3)
- ADHS (3)
- Adjuvans (3)
- Altern (3)
- Amygdala (3)
- Angiogenese (3)
- Angsterkrankungen (3)
- Animal model (3)
- Antibody (3)
- Antigen CD8 (3)
- Antikörper (3)
- Atherosklerose (3)
- B-MYB (3)
- B-Zell-Lymphom (3)
- Bacillus subtilis (3)
- Biochemie (3)
- Biodiversität (3)
- Bioinformatik (3)
- Biologische Uhr (3)
- Bioreaktor (3)
- Blutstammzelle (3)
- Blutstillung (3)
- CD1d (3)
- CD28 (3)
- CIDP (3)
- Campylobacter jejuni (3)
- Ceramid (3)
- Chemotherapie (3)
- Cyclo-AMP (3)
- Cytokine (3)
- Dimerisierung (3)
- E. coli Nissle 1917 (3)
- Echinococcus (3)
- Echinococcus multilocularis (3)
- Ecology (3)
- Electroencephalographie (3)
- Emotionsregulation (3)
- Endocytose (3)
- Endothel (3)
- Expansion Microscopy (3)
- Expositionstherapie (3)
- Extinktion (3)
- Fettzelle (3)
- Fibromyalgie (3)
- Fluorescence Microscopy (3)
- Gehirn (3)
- Gehirn-Computer-Schnittstelle (3)
- Generalisierung (3)
- Genmutation (3)
- Genom (3)
- Geruch (3)
- Geruchssinn (3)
- Gewebekultur (3)
- Gewebemodell (3)
- Glioblastom (3)
- Glycinrezeptor (3)
- Graft-versus-host disease (3)
- GvHD (3)
- Helicasen (3)
- Helicobacter pylori (3)
- Herzhypertrophie (3)
- Heubacillus (3)
- Histone (3)
- Hyaliner Knorpel (3)
- Hyaluronsäure (3)
- Immuntoleranz (3)
- In vitro (3)
- Informationsverarbeitung (3)
- Knorpel (3)
- Komplement <Immunologie> (3)
- Koronare Herzkrankheit (3)
- Kristallstruktur (3)
- Kutikularwachs (3)
- Latrophilin (3)
- Lunge (3)
- MRSA (3)
- MRT (3)
- Magnetresonanztomographie (3)
- Massenspektrometrie (3)
- Megakaryocyte (3)
- Metagenom (3)
- Metalloproteinasen (3)
- Mitochondria (3)
- Miz1 (3)
- Molekulare Bildgebung (3)
- Multiproteinkomplex (3)
- Muscarinrezeptor (3)
- NIR-Spektroskopie (3)
- NMR-Tomographie (3)
- Nephroblastom (3)
- Nervennetz (3)
- Nervenzelle (3)
- Neuroanatomie (3)
- Neurogenese (3)
- Neuroinflammation (3)
- Neuronale Plastizität (3)
- Neuropathischer Schmerz (3)
- Neuropeptide (3)
- Neutrophiler Granulozyt (3)
- Non-coding RNA (3)
- Obesity (3)
- Opioide (3)
- Orientierung (3)
- PDXP (3)
- PET (3)
- Paniksyndrom (3)
- Parkinson's disease (3)
- Pathogenität (3)
- Pathophysiologie (3)
- Permeabilität (3)
- Phosphatase (3)
- Phosphoglykolatphosphatase (3)
- Phospholipide (3)
- Phosphorylierung (3)
- Physiologie (3)
- Pilzkörper (3)
- Plastizität (3)
- Pneumolysin (3)
- Proteine (3)
- Proteinkinase D (3)
- Präfrontaler Cortex (3)
- Prävention (3)
- Psychische Störung (3)
- RNS (3)
- Raf-Kinasen (3)
- Rho-Proteine (3)
- Rhodopsin (3)
- Salmonella enterica (3)
- Schmerzforschung (3)
- Secretion (3)
- Sphingolipide (3)
- Stressreaktion (3)
- Structural Biology (3)
- T cell activation (3)
- T-Zelle (3)
- T-Zellen (3)
- TNF (3)
- Thrombozytenaggregation (3)
- Transpiration <Pflanzen> (3)
- Transposon (3)
- Tumorzelle (3)
- Typ 1 (3)
- Ubiquitinierung (3)
- Visuelle Aufmerksamkeit (3)
- Wundheilung (3)
- Zellkern (3)
- Zentralnervensystem (3)
- adrenocortical carcinoma (3)
- apoptosis (3)
- atherosclerosis (3)
- attention (3)
- cAMP (3)
- cell wall (3)
- circadian rhythms (3)
- cytoskeleton (3)
- dopamine (3)
- expression (3)
- fear conditioning (3)
- fear generalization (3)
- gp130 (3)
- iPSC (3)
- ischemic stroke (3)
- learning (3)
- learning and memory (3)
- lung cancer (3)
- megakaryocyte (3)
- megakaryopoiesis (3)
- neurobiology (3)
- protease (3)
- regulatorische T-Zellen (3)
- regulatory T cells (3)
- signalling (3)
- super-resolution microscopy (3)
- translation (3)
- virulence (3)
- 3D cell culture (2)
- 3D-Zellkultur (2)
- 53BP1 (2)
- ALS (2)
- APOBEC3G (2)
- ASM (2)
- Acetylation (2)
- Ackerschmalwand (2)
- Actin-bindende Proteine (2)
- Actinomyceten (2)
- Actinomycetes (2)
- Adaptation (2)
- Adaptorproteine (2)
- Adenosinrezeptor (2)
- Adhesion GPCR (2)
- Adipogenesis (2)
- Adjuvant (2)
- Adrenalin (2)
- Adrenerger Rezeptor (2)
- Adventitia (2)
- Affekt (2)
- Analgesie (2)
- Angeborene Immunität (2)
- Angewandte Mikrobiologie (2)
- Antennallobus (2)
- Antiangiogenese (2)
- Antibiotikum (2)
- Antigen CD40 (2)
- Antigen CD95 (2)
- Aorta (2)
- Apis mellifera (2)
- Apoptose (2)
- Arabidopsis thaliana (2)
- Arbeitsgedächtnis (2)
- Arbeitsteilung (2)
- Arrestine (2)
- Aspergillus (2)
- Assoziation (2)
- Assoziatives Lernen (2)
- Astrozyt (2)
- Audiologie (2)
- Augenbewegung (2)
- Autoaggressionskrankheit (2)
- Autophagy (2)
- Aversive Konditionierung (2)
- Axon (2)
- B cells (2)
- B-Zellen (2)
- BAD (2)
- BCI (2)
- BOLD signal (2)
- BRET (2)
- Bacterial infection (2)
- Bakterielle Hirnhautentzündung (2)
- Bakterielle Infektion (2)
- Bakteriengift (2)
- Bauchspeicheldrüsenkrebs (2)
- Beta-1-Rezeptor (2)
- Beta-Rezeptor (2)
- Bildverarbeitung (2)
- Biogenese (2)
- Bioinks (2)
- Biomechanics (2)
- Biomedicine (2)
- Bioprinting (2)
- Bioreactor (2)
- Blickbewegung (2)
- Blut-Nerven-Barriere (2)
- Blutgerinnung (2)
- Bortezomib (2)
- Brain-computer interface (2)
- CD28 Superagonist (2)
- CD84 (2)
- CDH13 (2)
- COVID-19 (2)
- CRPS (2)
- CXCR4 (2)
- Calciumkanal (2)
- Calciumtransport (2)
- Camponotus floridanus (2)
- Cancer immunotherapy (2)
- Cardiomyocyte (2)
- Cartilage Tissue Engineering (2)
- Cell cycle (2)
- Cell migration (2)
- Cement (2)
- Ceramide (2)
- Cholesterinstoffwechsel (2)
- Chronische Niereninsuffizienz (2)
- Ciliary neurotrophic factor (2)
- Circadian Rhythms (2)
- Cochlear-Implantat (2)
- Colonkrebs (2)
- Compressed Sensing (2)
- Contactin-1 (2)
- Corpus amygdaloideum (2)
- Corti-Organ (2)
- Crosstalk (2)
- Cryptochrom (2)
- Cushing-Syndrom (2)
- Cuticle (2)
- Cuticular waxes (2)
- DNA Repair (2)
- DNA repair (2)
- DNA-Reparatur (2)
- DNS (2)
- DNS-Bindung (2)
- DNS-Doppelstrangbruch (2)
- DRG (2)
- DROSHA (2)
- Darmwandnervensystem (2)
- Deep sequencing (2)
- Diabetische Polyneuropathie (2)
- Diagnostik (2)
- Dichtegewichtung (2)
- Differentiation (2)
- Diffusionsgewichtete Magnetresonanztomografie (2)
- Dopamin (2)
- Dosimetrie (2)
- E. coli Nissle (2)
- EHEC (2)
- Echokardiographie (2)
- Einzelzellanalyse (2)
- Elektroencephalogramm (2)
- Elektrospinning (2)
- Emotionen (2)
- Emotionsverarbeitung (2)
- Encephalomyelitis (2)
- Endozytose (2)
- Enterobacteriaceae (2)
- Enzym (2)
- Epigenetic (2)
- Ereigniskorreliertes Potenzial (2)
- Erwachsener (2)
- Extrazelluläre Matrix (2)
- FKBP52 (2)
- Fabry-Krankheit (2)
- Farbensehen (2)
- Fettsucht (2)
- Fettsäurebiosynthese (2)
- Fitness (2)
- Fluconazol (2)
- Fluconazole (2)
- Fluorescence (2)
- Fluoreszenzkorrelationsspektroskopie (2)
- Fluoxetin (2)
- Frühdiagnostik (2)
- Förster Resonanz Energie Transfer (2)
- G-protein-coupled receptors (2)
- GABA-Rezeptor (2)
- Galectine (2)
- Gametogenese (2)
- Gastrointestinaltrakt (2)
- Gedächtnis (2)
- Gelenkknorpel (2)
- Genomics (2)
- Gerinnungsfaktor XII (2)
- Glia (2)
- Glioblastoma (2)
- Gliom (2)
- Glukose (2)
- Glutamat-Decarboxylase (2)
- Glykosylierung (2)
- Golgi-Apparat (2)
- Growth (2)
- Guanylatcyclase (2)
- HUWE1 (2)
- Harnwegsinfektion (2)
- Hautleitfähigkeit (2)
- Hemostasis (2)
- Herz (2)
- Herzfrequenzvariabilität (2)
- Herzmuskel (2)
- Herzmuskelkrankheit (2)
- Herzschrittmacher (2)
- Hfq (2)
- High throughput screening (2)
- Hippo pathway (2)
- Hirnhautentzündung (2)
- Host-pathogen interaction (2)
- Hydrogel (2)
- Hypertrophie (2)
- Hypothalamus (2)
- Hypoxia (2)
- Hypoxie (2)
- Immunisierung (2)
- Immunological synapse (2)
- Immunotherapy (2)
- Impfung (2)
- Induced pluripotent stem cells (2)
- Infektionsmodell (2)
- Inhibitorische Synapse (2)
- Inhibitory synapse (2)
- Innate immunity (2)
- Innere Uhr (2)
- Insekten (2)
- Interaktion (2)
- Interferon (2)
- Interferon <gamma-> (2)
- Interleukin 2 (2)
- Intrazellulärraum (2)
- Invasion (2)
- Ischemic stroke (2)
- Japankärpfling (2)
- K-Ras (2)
- Kardiologie (2)
- Kardiomyozyt (2)
- Keratinozyt (2)
- Kernspintomographie (2)
- Klassische Konditionierung (2)
- Klimaänderung (2)
- Knochen-Morphogenese-Proteine (2)
- Knochenmark (2)
- Knochenmarktransplantation (2)
- Knockout (2)
- Knockout-Maus (2)
- Komplement C5a (2)
- Konfokale Mikroskopie (2)
- Kontextkonditionierung (2)
- Kostimulation (2)
- Kraniosynostose (2)
- Krankheit (2)
- Krebsforschung (2)
- Krebsimmuntherapie (2)
- LC-MS (2)
- Lactatdehydrogenase (2)
- Larve (2)
- Learning (2)
- Leishmania (2)
- Leishmania major (2)
- Leishmaniose (2)
- Lidschlag (2)
- Lipide (2)
- Lung (2)
- Lysosom (2)
- Macrophages (2)
- Magnetische Resonanz (2)
- Malaria tropica (2)
- Manisch-depressive Krankheit (2)
- Marine sponges (2)
- Mechanosensation (2)
- Medizinphysik (2)
- Megakaryopoese (2)
- Melt Electrowriting (2)
- Meningitis (2)
- Mensch (2)
- Merkel cell carcinoma (2)
- Mesenchymzelle (2)
- Metabolomik (2)
- Metastase (2)
- Methylierung (2)
- Mikrotubuli (2)
- Monarchfalter (2)
- Monozyt (2)
- Motoneuron-Krankheit (2)
- Motorisches Lernen (2)
- Multidrug-Resistenz (2)
- Multiple Myeloma (2)
- Mutation (2)
- Myatrophische Lateralsklerose (2)
- N-MYC (2)
- N-Myc (2)
- NFAT (2)
- NNMT (2)
- Na+/K+-ATPase (2)
- NaV1.9 (2)
- Nahrungserwerb (2)
- Nanoparticles (2)
- Natriumkanal (2)
- Navigation (2)
- Neisseria gonorrhoeae (2)
- Nervendegeneration (2)
- Nervensystem (2)
- Nestbau (2)
- Neuralgie (2)
- Neurofascin (2)
- Neuromodulation (2)
- Neuropathie (2)
- Neuroscience (2)
- Neutrophils (2)
- Nicht-kleinzelliges Bronchialkarzinom (NSCLC) (2)
- Nociceptor (2)
- Nozizeptor (2)
- Nuklearmedizin (2)
- Nukleotid-Exzisions-Reparatur (2)
- Olfaction (2)
- Oligomerisation (2)
- Oncostatin M (2)
- Onkogen (2)
- Optogenetik (2)
- Organische Synthese (2)
- Organoid (2)
- Osteogenesis (2)
- Oxidative stress (2)
- P300 (2)
- PVM (2)
- Parathormon (2)
- Peptide (2)
- Peptidsynthese (2)
- Peyer's patch (2)
- Pflanzen (2)
- Pflanzenschutzmittel (2)
- Phosphatasen (2)
- Phospholipase D (2)
- Phylogenie (2)
- Phänologie (2)
- Plant Protection (2)
- Plant cuticle (2)
- Plasmamembran (2)
- Polyneuropathie (2)
- Positronen-Emissions-Tomographie (2)
- Post-transcriptional regulation (2)
- Probiotic (2)
- Progenitor (2)
- Prognose (2)
- Proliferation (2)
- Protein (2)
- Protein-Protein-Wechselwirkung (2)
- Proteinsynthese (2)
- Proteintyrosinphosphatase (2)
- Proteolyse (2)
- Proteom (2)
- Psychologie (2)
- RNA (2)
- RNA helicase (2)
- RNA modifications (2)
- RNA polymerase II (2)
- RNS-Interferenz (2)
- RNS-Viren (2)
- ROS (2)
- RS-Virus (2)
- RS1 (2)
- Radionuklid (2)
- Ranvier-Schnürring (2)
- Regenerative Medizin (2)
- Resilienz (2)
- Rheumatoid arthritis (2)
- RhoA (2)
- Ribosome (2)
- Risikofaktoren (2)
- Röntgenkristallographie (2)
- SARS-CoV-2 (2)
- SGLT1 (2)
- SOCE (2)
- SPECT (2)
- Salmonella Typhimurium (2)
- Salmonella typhimurium (2)
- Schlaf (2)
- Schwertkärpfling (2)
- Schädel-Hirn-Trauma (2)
- Screening (2)
- Sekundärkrankheit (2)
- Sekundärmetabolit (2)
- Sequenzanalyse (2)
- Serotonerge Nervenzelle (2)
- Signalkette (2)
- Sinneszelle (2)
- Sozialangst (2)
- Soziale Wahrnehmung (2)
- Sphingomyelin (2)
- Spinal Muscular Atrophy (2)
- Spred-Proteine (2)
- Staphylococcus (2)
- Stickstoff (2)
- Streptococcus pneumoniae (2)
- Symbiose (2)
- Synthese (2)
- T cells (2)
- T-Lymphozyten-Rezeptor (2)
- TFIIIC (2)
- TLR3 (2)
- TWEAK (2)
- Therapieresistenz (2)
- Thrombo-inflammation (2)
- Thrombopoiesis (2)
- Tissue engineering (2)
- Toleranz (2)
- Toll-like-Rezeptoren (2)
- Transcriptome (2)
- Transforming Growth Factor beta (2)
- Transgener Organismus (2)
- Transkriptom (2)
- Tuberkelbakterium (2)
- Tumor-Nekrose-Faktor (2)
- Turbo Spin Echo (2)
- Ubiquitin-Protein-Ligase (2)
- VSG (2)
- Vaccine (2)
- Wahrnehmung (2)
- Wilms tumor (2)
- Wirkstoffdesign (2)
- X-ray crystallography (2)
- Xiphophorus (2)
- Zebrabärbling (2)
- Zebrafish (2)
- Zellteilung (2)
- Zellwand (2)
- Zement (2)
- Zink-Finger-Proteine (2)
- adipocytes (2)
- adult neurogenesis (2)
- amygdala (2)
- antennal lobe (2)
- anxiety (2)
- anxiety disorders (2)
- autophagy (2)
- bee (2)
- behavior (2)
- behaviour (2)
- biased agonism (2)
- bioinformatics (2)
- biology (2)
- biomedical applications (2)
- biomedicine (2)
- bioreactor (2)
- blood brain barrier (2)
- bone marrow (2)
- brain (2)
- calcium (2)
- cardiac hypertrophy (2)
- cell (2)
- ceramide (2)
- chlamydia (2)
- cytokines (2)
- cytokinesis (2)
- deep brain stimulation (2)
- dendritic cell (2)
- dendritische Zelle (2)
- depression (2)
- development (2)
- developmental differentiation (2)
- diet (2)
- dosimetry (2)
- drug development (2)
- ecology (2)
- electron microscopy (2)
- electrophysiology (2)
- emotion (2)
- emotion processing (2)
- emotion regulation (2)
- endosomal trafficking (2)
- endothelial cells (2)
- epigenetics (2)
- evolution (2)
- experimental autoimmune encephalomyelitis (2)
- extracellular matrix (2)
- fMRI time series (2)
- fear extinction (2)
- foxtapeworm (2)
- funktionelle Kernspintomographie (2)
- gamma-H2AX (2)
- genetics (2)
- genomics (2)
- gephyrin (2)
- glucose (2)
- hippocampus (2)
- host-pathogen interaction (2)
- immunologische Synapse (2)
- in vitro (2)
- infection (2)
- inflammatory pain (2)
- intestinal in vitro model (2)
- kinase (2)
- lipid rafts (2)
- malaria (2)
- mass spectrometry (2)
- medical physics (2)
- melanoma (2)
- mesenchymal stem cells (2)
- metabolism (2)
- metagenomics (2)
- mitosis (2)
- mitotic gene expression (2)
- monoclonal antibodies (2)
- monoklonale Antikörper (2)
- motoneuron (2)
- mouse model (2)
- multi-unit recording (2)
- multiple myeloma (2)
- neuroethology (2)
- neuropathy (2)
- nuclear medicine (2)
- oligomerization (2)
- p53 (2)
- pancreas (2)
- peptide (2)
- perception (2)
- phosphatase (2)
- receptor (2)
- regulatory t cells (2)
- resistance (2)
- serotonergic system (2)
- shRNA (2)
- signal transduction (2)
- single particle tracking (2)
- skin equivalent (2)
- small RNA (2)
- snRNP (2)
- social anxiety (2)
- social attention (2)
- social insects (2)
- sphingolipids (2)
- spontaneous blinks (2)
- stress (2)
- stroke (2)
- symbiosis (2)
- synaptic plasticity (2)
- synaptic proteins (2)
- synaptische Proteine (2)
- t cells (2)
- tDCS (2)
- targeted therapy (2)
- thrombopoiesis (2)
- tissue engineering (2)
- transcription (2)
- transkranielle Gleichstromstimulation (2)
- vascularization (2)
- virtuelle Realität (2)
- wild bees (2)
- working memory (2)
- wound healing (2)
- Ökologie (2)
- Übergewicht (2)
- "Lipid Rafts" (1)
- (hem)ITAM signaling (1)
- 13C-isotopologue profiling (1)
- 177Lu-OPS201 (1)
- 18S rRNA (1)
- 2-point Dixon (1)
- 3 D bioprinting (1)
- 3-D liver model (1)
- 3D Gewebemodelle (1)
- 3D Modell (1)
- 3D Printing (1)
- 3D Tumormodell (1)
- 3D model (1)
- 3D models (1)
- 3D muscle (1)
- 3D spheroid culture (1)
- 3D tumour model (1)
- 3D-Elektrode (1)
- 3D-Gewebemodell (1)
- 3D-Kultur (1)
- 3D-Modell (1)
- 4-HNE (1)
- 5-FU (1)
- 5`-UTR (1)
- 68Ga-OPS202 (1)
- 7 T (1)
- 7SK (1)
- 7T (1)
- A-RAF (1)
- A2B adenosine receptor (1)
- A2BAR (1)
- AAC (1)
- ABC-Transporter (1)
- ACC (1)
- ACL (1)
- ADGRL3 (1)
- ADORA2A (1)
- ADP (1)
- AI (1)
- AMPK (1)
- AMP‐activated protein kinase (1)
- ARF6 GTPase (1)
- ASM-Inhibition (1)
- ATF5 (1)
- ATGL (1)
- ATR-FTIR (1)
- ATRA (1)
- Absorbed Doses (1)
- Accelerated imaging (1)
- Acetylcholinrezeptor (1)
- Acetylierung (1)
- Acid adaptation (1)
- Actin Dynamics (1)
- Actin binding proteins (1)
- Actin cytoskeleton-related protein (1)
- Actinomyces (1)
- Acute myeloid leukemia (AML) (1)
- Acyrthosiphon pisum (1)
- Adapterprotein (1)
- Adenomatous-polyposis-coli-Protein (1)
- Adenosine receptors (1)
- Adhesion and Invasion (1)
- Adhesion and degranulation promoting adapter protein (1)
- Adhesion-GPCR (1)
- Adhesive Hydrogels (1)
- Adhärenz (1)
- Adhäsion (1)
- Adipose (1)
- Adipose Tissue Engineering (1)
- Adipose-Derived Stromal/Stem Cells (1)
- Adipositas (1)
- Administered Activities (1)
- Adrenergic receptor (1)
- Adrenocortical Carcinoma (1)
- Adrenokortikales Karzinom (1)
- Adulte Neurogenese (1)
- Adultschlupfes (1)
- Advanced Therapy Medicinal Products (ATMP) (1)
- Affinity probe (1)
- Agaphelin (1)
- Agarose (1)
- Aggregation (1)
- Agoraphobie (1)
- Agrobacterium vitis (1)
- Aiolos (1)
- Airway epithelia (1)
- Aktive Implantate (1)
- Aktivierung <Physiologie> (1)
- Aktivierungsenergie (1)
- Aktivitätstest (1)
- Akute lymphatische Leukämie (1)
- Akute myeloische Leukämie (1)
- Akzeptanz (1)
- Akzeptanz- und Commitment Therapie (1)
- Algorithmus (1)
- Aliphatics (1)
- Alkaloid (1)
- Allergie (1)
- Allergy (1)
- Allgemeine Zelle (1)
- Alloantigen (1)
- Alloantigen Expression (1)
- Allogene Zelle (1)
- Allogeneic stem cell transplantation (1)
- Allogenic hematopoietic stem cell transplantation (1)
- Allosterie (1)
- Alpaca (1)
- Alpha Neurofeedback (1)
- Alpha-Aktivität (1)
- Alternative polyadenylation (1)
- Alternative zum Tierversuch (1)
- Alveolar Echinococcosis (1)
- Alveoläre Echinokokkose (1)
- Alzheimer's Dementia (1)
- Alzheimer's disease (1)
- Alzheimer`s disease (1)
- AmGr1, AmGr2, AmGr3 (1)
- Ambrosia beetles (1)
- Ambrosiakäfer (1)
- Ameise (1)
- Ameisen (1)
- Ameisenstaat (1)
- Aminobisphosphonat (1)
- Aminobisphosphonate (1)
- Aminobuttersäure <gamma-> (1)
- Ammoniumpermease (1)
- Amphibien (1)
- Amyotrophe Lateralsklerose (1)
- Amyotrophic lateral sclerosis (1)
- Analyse (1)
- Anaphylatoxine (1)
- Anaphylatoxinrezeptoren (1)
- Anastomoseninsuffizienz (1)
- Ancistrocladaceae (1)
- Ancistrocladus (1)
- Androstadienon (1)
- Aneuploidy (1)
- Angewandte Toxikologie (1)
- Angiogenesis (1)
- Angiotensin II (1)
- Angst als Eigenschaft (1)
- Angst als Zustand (1)
- Angsterkrankung (1)
- Angstreaktionen (1)
- Angstsensitivität (1)
- Angstverhalten (1)
- Animal behavior IntelliCage system (1)
- Animales Nervensystem (1)
- Anisotropic structures (1)
- Annotation (1)
- Anoikis (1)
- Anpassung (1)
- Ant (1)
- Anthocyane (1)
- Anthropocene (1)
- Anti-CD52-Antikörper (1)
- Anti-infective (1)
- Antibiotic (1)
- Antibodies (1)
- Antifungal (1)
- Antigen CD3 (1)
- Antigen CD4 (1)
- Antigen CD44 (1)
- Antigen presentation (1)
- Antigen recognition (1)
- Antigenpräsentation (1)
- Antigenrezeptor (1)
- Antikörper-Antwort (1)
- Antimicrobial activities (1)
- Antimicrobial activity (1)
- Antimicrobial proteins (1)
- Antimikrobielle Aktivitäten (1)
- Antimikrobieller Wirkstoff (1)
- Antimykotika (1)
- Antimykotikum (1)
- Antioxidantien (1)
- Antivirale Substanzen (1)
- Antworthemmung (1)
- Antwortverhalten (1)
- Anxiety Disorders (1)
- Anxiety disorder (1)
- Anxiety disorders (1)
- Aortenaneurysma (1)
- Apallisches Syndrom (1)
- Aplysina aerophoba (1)
- Apolipoprotein E (1)
- Approved immunomodulators (1)
- Archaea (1)
- Archaebakterien (1)
- Arginine (1)
- Arid biomes (1)
- ArsZ (1)
- Artefakt (1)
- Artenreichtum (1)
- Arterielles Blut (1)
- Artery Models (1)
- Artesunate (1)
- Arthrose (1)
- Arthrosis deformans (1)
- Arzneimittel (1)
- Arzneimittelzulassung (1)
- Aspartatprotease (1)
- Aspergillose (1)
- Aspergillosis (1)
- Associative learning (1)
- Assoziatives Gedächtnis (1)
- AstA (1)
- Atemwege (1)
- Atemwegsschleimhaut (1)
- Atherogenese (1)
- Atmungsinsuffizienz (1)
- Atomic-force-microscopy (1)
- Atriales natriuretisches Hormon (1)
- Atta vollenweideri (1)
- Attention (1)
- Attention deficit / hyperactivity disorder (1)
- Attentional Avoidance (1)
- Attentional Bias (1)
- Attentional bias (1)
- Attraction (1)
- Attraktion (1)
- AuNPs (1)
- Auditorische Neuropathie (1)
- Auditorischer Kortex (1)
- Aufmerksamkeitsprozesse (1)
- Aurora B (1)
- Aurora-A-MYCN-Komplex (1)
- Ausbreitungsverhalten (1)
- Aussterbedynamik (1)
- Autoantigen (1)
- Autofocus (1)
- Autoimmunity (1)
- Autoinhibition (1)
- Automatische Klassifikation (1)
- Autophagie (1)
- Autophagie <Physiologie> (1)
- Autophagosom (1)
- Autophagozytose (1)
- Autoproteolysis (1)
- Autotransporter (1)
- Autotransporterprotein (1)
- Awareness (1)
- Axon Branching (1)
- B Lymphocytes (1)
- B-Lymphozyten (1)
- B0 (1)
- B0 correction (1)
- B0-Korrektur (1)
- B7-H1 (1)
- BCG (1)
- BCL-2 Familie (1)
- BCL-2 family (1)
- BCMA (1)
- BDNF (1)
- BDNF stimulation (1)
- BH3-only proteins (1)
- BIN2 (1)
- BMP (1)
- BMP signaling pathway (1)
- BMP-2 (1)
- BMP-Signaltransduktionsweg (1)
- BMP2 (1)
- BMS-5 (1)
- BRAF (1)
- BRCA1 (1)
- BTB domain (1)
- BTN3A1 and Phospho-antigen presentation (1)
- Bach (1)
- Bacteria-Host Cell Interaction (1)
- Bacterial Toxins (1)
- Bacterial community analysis (1)
- Bacterial meningitis (1)
- Bacteroides thetaiotaomicron (1)
- Baff (1)
- Baghdadite (1)
- Bahnung (1)
- Bakterielle Nanocellulose (1)
- Bakterienzellwand (1)
- Bakteriophagen (1)
- Bandwürmer (1)
- Bariatric surgery (1)
- Barth Syndrome (1)
- Barth syndrome (1)
- Basal Ganglia (1)
- Basalganglien (1)
- Basalmembran (1)
- Basement membrane (1)
- Batten disease (1)
- Bauchspeicheldrüse (1)
- Baumphysiologie (1)
- Bcl-2-Proteinfamilie (1)
- Bed nucleus of stria terminalis (1)
- Behavior (1)
- Behaviour (1)
- Benzimidazolderivate (1)
- Benzoate (1)
- Benzoate degradation (1)
- Beschleunigte Bildgebung (1)
- Bestimmung des Relaxations-Parameters in der Magnetresonanztomografie (1)
- Bestrahlung (1)
- Bestärkendes Lernen <Künstliche Intelligenz> (1)
- Bestäubung (1)
- Bestäubung Pollination Honigbiene Hummel (1)
- Beta-1-receptor (1)
- Beta-2-Rezeptor (1)
- Beta-Adrenergic Receptor (1)
- Beta-Adrenozeptor (1)
- Bevacizumab (1)
- Bewegungsstörung (1)
- Bienen <Familie> (1)
- Bienen Dressur Lernen (1)
- Bierhefe (1)
- Bildanalyse (1)
- Bildartefakte (1)
- Bildbearbeitung (1)
- Bilderzeugung (1)
- Bildgebung intakten Knochens (1)
- Bimolekulare Lipidschicht (1)
- Bindegewebe (1)
- BioVaSc (1)
- Biochemische Analyse (1)
- Biochemistry (1)
- Biodistribution (1)
- Biodiversity assessment (1)
- Biodiversity conservation (1)
- Biofabrikation (1)
- Biofilm formation (1)
- Biofilmarchitektur (1)
- Biofilms (1)
- Bioink (1)
- Biokinetics (1)
- Biolayerinterferometrie (1)
- Biologie / Zellbiologie (1)
- Biologischer Abbau (1)
- Biologisches Modell (1)
- Biology (1)
- Bioluminescence resonance energy transfer (1)
- Biolumineszenzmessung (1)
- Biomechanik (1)
- Bioprozessmethode (1)
- Bioreaktorplattform (1)
- Biosynthese der aromatischen Aminosäuren (1)
- Biosynthese-Genclustern (1)
- Bipolar (1)
- Bipolar Disorder (1)
- Bisphosphonate (1)
- Bitopic Ligands (1)
- Bitopische Liganden (1)
- Blick (1)
- Blickinteraktion (1)
- Blimp-1 (1)
- Blinatumomab (1)
- Blood nerve barrier (1)
- Blumeria graminis (1)
- Blut-Liquor-Schranke (1)
- Blut-Rückenmarkschranke (1)
- Blutgefäßsystem (1)
- Blutgerinnungsfaktor XII (1)
- Bluthirnschranke (1)
- Bone (1)
- Bone Marrow Dosimetry (1)
- Bone Quantification (1)
- Bone marrow stromal cell (1)
- Bone marrow stromal cell (BMSC) (1)
- Bone marrow transplantation (1)
- Bone morphogenetic protein 2 (1)
- Bone morphogenetic proteins (1)
- Bone regeneration (1)
- Bone-replacement (1)
- Bordetella (1)
- Borkenkäfer (1)
- Borneo (1)
- Borrelia (1)
- Bovine Mastitis (1)
- Brain (1)
- Brain derived neurotorphic factor (1)
- Brain endothelial cells (1)
- Brownsche Bewegung (1)
- Bruchpilot (1)
- Brustdrüsenentzündung (1)
- Burkholderia (1)
- Burkholderia pseudomallei (1)
- Burkitt Lymphom (1)
- Burkitt Lymphoma (1)
- Butyrophilin (1)
- Bäckerhefe (1)
- Bärtierchen (1)
- C. albicans (1)
- C. elegans (1)
- C/EBP (1)
- C1-inibitor (1)
- C5aR1-Antagonist (1)
- CA3 (1)
- CAMP production (1)
- CAR T Zellen (1)
- CAR T cell (1)
- CAR T-Zelltherapie (1)
- CAR T-cells (1)
- CAR-T cell (1)
- CAR-T-Zell-Therapie (1)
- CCR4 (1)
- CD11b+ myeloid cells (1)
- CD150 (1)
- CD28 Molekül (1)
- CD28 Superagonisten (1)
- CD28 antigen (1)
- CD28-SA (1)
- CD28-Superagonist (1)
- CD28-superagonist (1)
- CD4 T-Zellen (1)
- CD4 positiv T cells (1)
- CD4+ (1)
- CD4+ T cell activation (1)
- CD40 gerichteter bifunktioneller scFv-TRAIL Fusionsproteine (1)
- CD40-targeted bifunctional scFv-TRAIL fusion proteins (1)
- CD44 (1)
- CD8 Effektorfunktionen (1)
- CD8 Gedächtnisreaktionen (1)
- CD8 T cell (1)
- CD8 T-Zelle (1)
- CD8 T-Zellen (1)
- CD8+ T cells (1)
- CD8+ T-Zellen (1)
- CD95 (1)
- CD95L (1)
- CDC42 (1)
- CDK4 Inhibitor (1)
- CDK4 inhibitor (1)
- CDR1-Effluxpumpe (1)
- CIP2A (1)
- CLEC-2 (1)
- CLEC16A (1)
- CMT1A (1)
- CMV (1)
- CNBP (1)
- COMT polymorphism (1)
- CRHR1 (1)
- CRISPR Cas9 (1)
- CRISPR Cas9 system (1)
- CTLA-4 (1)
- CVT (1)
- CX-5461 (1)
- CXCL13 (1)
- CXCR5 (1)
- CYP24A1 (1)
- CYP7A1 (1)
- CaCo-2 cell (1)
- Cadherin 13 (1)
- Cadherin-13 (1)
- Caenorhabditis elegans (1)
- Calcium Phosphate (1)
- Calcium phosphate-based biomaterials (1)
- Calcium signalling (1)
- Calcium-bindende Proteine (1)
- Calciumhomöostase (1)
- Calciumphosphat (1)
- Camponotus (1)
- Camponotus rufipes (1)
- Cancer Immune Therapy (1)
- Cancer Metabolism (1)
- Cancer biology (1)
- Cancer models (1)
- Cancer therapeutic resistance (1)
- Carabid beetles (1)
- Carcinogenese (1)
- Cardiac (1)
- Cardiac Efficiency (1)
- Cardiac MRI (1)
- Cardiac hypertrophy (1)
- Cardiac imaging (1)
- Cardiolipin (1)
- Cardiomyocytes (1)
- Cardiomyopathy (1)
- Carotis Intima Media Dicke (1)
- Cartiage Integration (1)
- Cartilage (1)
- Cartilage Regeneration (1)
- Cartilage defect (1)
- Cas9 (1)
- Caspr-1 (1)
- Catechol-0-Methyltransferase (1)
- Catechol-O-Methyltransferase (1)
- Catecholmethyltransferase <Catechol-0-Methyltransferase> (1)
- Cation Homeostasis (1)
- Caveolin-1 (1)
- Ccr4-Not (1)
- Cdh13 (1)
- Celecoxib (1)
- Cell (1)
- Cell Cycle (1)
- Cell Sheet Engineering (1)
- Cell adhesion (1)
- Cell culture (1)
- Cell differentiation (1)
- Cell line (1)
- Cell-Assisted Lipotransfer (1)
- Cells (1)
- Cellular invasion (1)
- Ceramides (1)
- Cerebrospinal fluid (CSF) (1)
- ChIP-Seq (1)
- ChIP-sequencing (1)
- Chaetomium thermophilum (1)
- Chain-length distribution (1)
- Characterization (1)
- Charcot-Marie-Tooth 1A (1)
- Chemische Synapsen (1)
- Chemokin CXCL10 (1)
- Chemokin CXCL12 (1)
- Chemokine (1)
- Chemokine receptors (1)
- Chemoresistenz (1)
- Chemotaxis (1)
- Chemotherapeutic resistance (1)
- Children (1)
- Chimeric Antigen Receptor (1)
- Chimeric antigen receptor (1)
- Chimeric antigen receptor (CAR) (1)
- Chimpanzee (1)
- Chimärer Antigenrezeptor (1)
- Chirurgie (1)
- Chlamydia (1)
- Chlamydienkrankheit (1)
- Cholesterintransporter (1)
- Chondrogenic Differentiation (1)
- Chordontonal organ (1)
- Chromatinremodeling (1)
- Chromatinremodelling (1)
- Chromosom 6 (1)
- Chromosome 6 (1)
- Chronic Mild Stres (1)
- Chronisch-myeloische Leukämie (1)
- Chronische Nierenerkrankung (1)
- Chronischer Schmerz (1)
- Chylomicron (1)
- Chylomicrons (1)
- Circadian (1)
- Circadian Clock (1)
- Circadian clock (1)
- Circadiane Uhr (1)
- Circadianer Rhythmus (1)
- Citalopram (1)
- Claudin (1)
- Click-Chemie (1)
- Climate change (1)
- Clonal competition assay (1)
- Clostridium difficile (1)
- Cluster (1)
- Co-Analgetika (1)
- Co-Kultur (1)
- Co-culture (1)
- Co-culture system (1)
- CoQ10 (1)
- Coactosin-like (1)
- Coffin-Lowry syndrome (1)
- Cofilin (1)
- Cognition (1)
- Cognitive Distortions (1)
- Collagen gels (1)
- Colliculus inferior (1)
- Collybistin (1)
- Colon cancer (1)
- Commensalism (1)
- Comorbidity (1)
- Comoé National Park (1)
- Complement system (1)
- Complex Regional Pain Syndrome (1)
- Complexes (1)
- Complexome (1)
- Compound eyes (1)
- Compressed sensing (1)
- Computational Fluid Dynamics (1)
- Computational drug design (1)
- Computersimulation (1)
- Computerunterstütztes Verfahren (1)
- Conditioning (1)
- Contact-Kinin System (1)
- Contrast Agent Bolus Based Perfusion Magnetic Resonance Imaging (1)
- Convolutional Neural Network (1)
- Coronary heart disease (1)
- Correlative microscopy (1)
- Cortico-striatal projection neurons (1)
- Corticoliberin (1)
- Counting (1)
- Coxiella burnetii (1)
- Craniosynostosis (1)
- Craving (1)
- CsrA (1)
- Cul4b (1)
- Cuticular transpiration (1)
- Cuticular water permeabilities (1)
- Cyanobacteria (1)
- Cyanobakterien (1)
- Cyclic peptides (1)
- Cyclics (1)
- Cystein (1)
- Cysteinproteasen (1)
- Cytologie (1)
- Cytomegalie-Virus (1)
- Cytoplasma (1)
- Cytotoxic T cell (1)
- D-4F (1)
- DC (1)
- DC/T-Zell-Konjugate (1)
- DEQCT (1)
- DExD/H box protein (1)
- DG diacyglycerol (1)
- DGCR8 (1)
- DHX30 (1)
- DNA Barcoding (1)
- DNA Damage (1)
- DNA Doppelstrangbrüche (1)
- DNA Schaden (1)
- DNA Sekundärstruktur (1)
- DNA double strand breaks (1)
- DNA lesion (1)
- DNA methylation (1)
- DNA secondary structure (1)
- DNA-Methylierung (1)
- DNA-Schaden (1)
- DNA-Schäden (1)
- DNS-Bindungsproteine (1)
- DNS-Schaden (1)
- DNS-Sequenz (1)
- DOT1 (1)
- DREAM (1)
- DREAM complex (1)
- DSIF (1)
- DTI (1)
- DUB Mutante (1)
- DYT-TOR1A (1)
- Danaus plexippus (1)
- Danio rerio (1)
- Darmepithel (1)
- Darmflora (1)
- Dasatinib (1)
- Datenbank (1)
- Decapping (1)
- Deep-sequencing (1)
- Deeplearning (1)
- Defensine (1)
- Defäkographie (1)
- Deletion (1)
- Denaturierende Gradienten-Gelelektrophorese (1)
- Denaturing Gradient Gel Electrophoresis (1)
- Density Weighting (1)
- Deoxyribozymes (1)
- Dephosphorylierung (1)
- Depletion (1)
- Dereplicaiton (1)
- Desmoglein 2 (1)
- Desmoglein 3 (1)
- Desmogleine (1)
- Desmosom (1)
- Deubiquitination (1)
- Development (1)
- Dexamethason (1)
- Diabetes (1)
- Diabetes Typ 1 (1)
- Diabetes mellitus Typ 1 (1)
- Diabetic painful neuropathy (1)
- Diabetic polyneuropathy (1)
- Diagnostic Imaging Exams (1)
- Dickdarmkrebs (1)
- Differential scanning calorimetry (1)
- Differentielle Genexpression (1)
- Differentielle Ressourcenallokation (1)
- Differenzierung (1)
- Differenzierungszustand (1)
- Diffusion (1)
- Diffusion Tensor Imaging (1)
- Diffusion coefficient (1)
- Dihydrooxazole (1)
- Dilated cardiomyopathy (1)
- Dimension 3 (1)
- Dimere (1)
- Dimeric Naphthylisoquinoline Alkaloids (1)
- Diphenylether (1)
- Diskriminationslernen (1)
- Diskriminationstraining (1)
- Dispersal (1)
- Disulfidbrücken (1)
- Diversifikation <Biologie> (1)
- Diversity (1)
- Dolutegravir (1)
- Domäne <Biochemie> (1)
- Domänenprotein (1)
- Dopamin-Transporter (1)
- Dopamine transporter (1)
- Dopaminstoffwechsel (1)
- Dopamintransporter (1)
- Dorsal Root Ganglion (1)
- Dorsal root ganglion (1)
- Dosimetry (1)
- Double Negative B cells (1)
- Droseraceae (1)
- Drosophila Larva (1)
- Drosophila Larve (1)
- Drosophila Myc transcription growth PAF1 (1)
- Drug delivery system (DDS) (1)
- Drug resistance (1)
- Drug targets (1)
- Drug testing (1)
- Dual RNA-seq (1)
- Dual RNA-seq data analysis (1)
- Dual-setting (1)
- Dualstere Liganden (1)
- Dualsteric Ligands (1)
- Duodenal Jejunal Bypass (1)
- Duplikation (1)
- Durchflusscytometrie (1)
- Dynabeads (1)
- Dynamic MR imaging (1)
- Dynamics of ribosome assembly (1)
- Dynamik (1)
- Dynamik von Membranrezeptoren (1)
- Dynamische MR-Bildgebung (1)
- Dynein Light Chain (1)
- Dystonie (1)
- Dürrestress (1)
- E.coli Nissle 1917 (1)
- E1 inhibitoren (1)
- E1 inhibitors (1)
- E2 (1)
- EAE (1)
- ECAP (1)
- ECG-gated PET (1)
- ELISA (1)
- EMT (1)
- ER dynamics in axon terminals (1)
- ERK (1)
- Eap (1)
- Early-Life Stress (1)
- Echo Planar Imaging (1)
- Echo planar imaging (1)
- Echocardiography (1)
- Echoplanar-Bildgebung (1)
- Echtzeit (1)
- Echtzeitbildgebung (1)
- Ecological Momentary Assessment (1)
- Ecological Momentary Assessments (1)
- Ecosystem services (1)
- Effect anticipation (1)
- Effektantizipation (1)
- Efflux pump (1)
- Effluxpumpen (1)
- Eierstockkrebs (1)
- Eierstocktumor (1)
- Einfühlung (1)
- Einfühlung <Motiv> (1)
- Eingeweidewürmer (1)
- Einsamkeit (1)
- Einzel-Molekül (1)
- Einzelpartikelverfolgung (1)
- Einzelzellgelelektrophorese (1)
- Electrochemical Impedance Spectroscopy (1)
- Electrode (1)
- Electroencephalography (1)
- Electromyography (1)
- Electrophysiology (1)
- Elektroakupunktur (1)
- Elektrode (1)
- Elektromyographie (1)
- Elektronencephalographie (1)
- Elektrospinnen (1)
- Elektrostimulation (1)
- Elektrotherapie (1)
- Elevated Plus-Maze (1)
- Elongation (Transkription) (1)
- Embryo (1)
- Embryonale Stammzelle (1)
- Emotionale Informationsverarbeitung (1)
- Emotionales Verhalten (1)
- Empfindlichkeit (1)
- Endobrachyösophagus (1)
- Endocytosis (1)
- Endogenous clock (1)
- Endokrinologie (1)
- Endophenotype (1)
- Endophänotyp (1)
- Endophänotypen (1)
- Endoplasmatisches Retikulum (1)
- Endorganschaden (1)
- Endosomes (1)
- Endothelium (1)
- Endothelzelle (1)
- Enoyl-Reduktase (1)
- Enoyl-acyl-carrier-protein-Reductase (1)
- Enoyl-acyl-carrier-protein-Reductase <Enoyl-[acyl-carrier-protein]-Reductase> (1)
- Enterische Glia (1)
- Enterisches Nervensystem (1)
- Enterohemorrhagic Escherichia coli (1)
- Entscheidungsverhalten (1)
- Entwicklung chimärer Antigenrezeptor T-Zellen (1)
- Entzündungschmerz (1)
- Entzündungsreaktion (1)
- Entzündungsschmerz (1)
- Enzymaktivierung (1)
- Enzyminhibitor (1)
- Eosinophiler Granulozyt (1)
- EphA2 (1)
- Ephrin ligand (1)
- Ephrine (1)
- Epidemiologie (1)
- Epigenetische Mechanismen (1)
- Epithel (1)
- Epithelial layer (1)
- Epithelial lineage (1)
- Epithelial-mesenchymale Transition (1)
- Epithelzelle (1)
- Erk1/2 (1)
- Erkennung (1)
- Erregbarkeit (1)
- Esophageal adenocarcinoma (1)
- Esophageal disease (1)
- Essgewohnheit (1)
- Essstörung (1)
- Essverhalten (1)
- Estrogens (1)
- Excitatory/inhibitory imbalance (1)
- Excitotoxicity (1)
- Exon Array (1)
- Expansion microscopy (1)
- Expansionsmikroskopie (1)
- Experimental Autoimmune Encephalomyelitis (EAE) (1)
- Experimentelle Autoimmune Enzephalomyelitis (1)
- Experimentelle Autoimmune Enzephalomyelitis (EAE) (1)
- Experimentelle autoimmune Enzephalomyelitis (1)
- Experimentellen Autoimmun-Enzephalomyelitis (1)
- Experimenteller Schlaganfall (1)
- Explainable AI (1)
- Explainable Artificial Intelligence (1)
- Explorationsverhalten (1)
- Exposition (1)
- Expositionslernen (1)
- Exposure treatment (1)
- Expression (1)
- Extracellular Matrix (1)
- Extracellular Vesicles (1)
- Extracellular matrix proteins (1)
- Extrazellulärmatrix (1)
- Exzitatorische Synapse (1)
- Eye irritation (1)
- FAS (1)
- FAS-II (1)
- FAT10ylation (1)
- FBXO6 (1)
- FCS (1)
- FGF (1)
- FGF Signaling (1)
- FLT3 inhibitor (1)
- FMMs (1)
- FMS-like tyrosine kinase 3 (FLT3) (1)
- FOXM1 (1)
- FOXP2 (1)
- FP635 (1)
- FRAP (1)
- FRET sensors (1)
- FT-IR-Spektroskopie (1)
- FabI (1)
- FabV (1)
- Fabry disease (1)
- Factor XII (1)
- Faktor XII (1)
- Fanconi Anemia (1)
- Fanconi-Anämie (1)
- Farnesyl Pyrophosphate Synthase (1)
- Farnesylpyrophosphatsynthase (FPPS) (1)
- Fas-Ligand (1)
- Fat Quantification (1)
- FcgR (1)
- FeS cluster (1)
- Fear (1)
- Fear Conditioning (1)
- Fear Extinction (1)
- Fear Generalization (1)
- Fear Learning (1)
- Fear conditioning (1)
- Fear generalization (1)
- Feedforward loop (1)
- FemX (1)
- Ferroptosis (1)
- Fertilization in angiosperm (1)
- Fettgewebe (1)
- Fettleber (1)
- Fettsäure-Synthase (1)
- Fibroblast (1)
- Fibroblasts (1)
- Fibromyalgia (1)
- Fibromyalgia syndrome (1)
- Fibromyalgiesyndrom (1)
- Fibrose (1)
- Filter (1)
- Flagelle (1)
- Flavonoide (1)
- Flavonoids (1)
- Flippase (1)
- Flotillin (1)
- Flow (1)
- Flugzeitmassenspektrometrie (1)
- FluidFM (1)
- Fluorescence Correlation Spectroscopy (1)
- Fluorescence Resonance Energy Transfer (1)
- Fluorescence correlation spectroscopy (1)
- Fluorescence microscopy (1)
- Fluorescence-resonance-energy-transfer (1)
- Fluorescent probes (1)
- Fluoreszenz (1)
- Fluoreszenzproteine (1)
- Fluoreszenzsonde (1)
- Fluoreszenzspektroskopie (1)
- Fluoxetine (1)
- Fluss (1)
- Foamyviren (1)
- Folliculin (1)
- Forkhead Transcription Factors (1)
- Forkhead-Box-Proteine (1)
- Fox tapeworm (1)
- FoxO transcription factors (1)
- Foxo1 (1)
- Foxp3+CD4+ regulatorische T-Zelle (1)
- Foxp3+CD4+ regulatory T cell (1)
- Fragebogen (1)
- Fragmentscreening (1)
- Französische Feldwespe (1)
- Freezing (1)
- Freies Molekül (1)
- Fremdkörpermodell (1)
- Frizzled 5 (1)
- Frontal asymmetry (1)
- Froschlurche (1)
- Fructosebisphosphat-Aldolase (1)
- Fruit (1)
- Frühe Gene (1)
- Function (1)
- Functional analyses (1)
- Functioning Assessment Short Test (1)
- Funktionelle Annotation (1)
- Funktionelle Bildgebung (1)
- Funktionelle Kernspintomographie (1)
- Funktionsausfälle (1)
- Furagieraktivität (1)
- Furchkonditionierung (1)
- Furchtentwicklung (1)
- Furchtextinktion (1)
- Fusarium (1)
- Fusobacterium nucleatum (1)
- Förster Resonance Energy Transfer (1)
- G Protein-Coupled Receptor (1)
- G Protein-Coupled Receptors (1)
- G Protein-coupled receptor (1)
- G glycoprotein (1)
- G protein coupled receptor (1)
- G protein-coupled receptor kinase (1)
- G protein-coupled receptor kinase 2 (GRK2) (1)
- G protein-coupled receptors (1)
- G protein-gekoppelte Rezeptor Kinase 2 (GRK2) (1)
- G-Protein (1)
- G-Protein gekoppelte Rezeptor (1)
- G-Protein-gekoppelte Rezeptoren (1)
- G-protein (1)
- G-quadruplex (1)
- G2/M genes (1)
- GABA (1)
- GABA A Receptors (1)
- GABA A Rezeptoren (1)
- GABA receptor (1)
- GABA(A) receptor (1)
- GABAA-Rezeptor (1)
- GABAerge Nervenzelle (1)
- GAD1 (1)
- GAD65 (1)
- GAS2L3 (1)
- GC1 cells (1)
- GDF-15 (1)
- GLA KO mouse model (1)
- GLP-1 (1)
- GMP (1)
- GPCR dimerisation (1)
- GPCR nanodomains (1)
- GPCR signaling (1)
- GRM8 (1)
- GWAS (1)
- Ga-68-labelled Peptides (1)
- Galactosidase <alpha-> (1)
- Galectin 1 (1)
- Gambler's Fallacy (1)
- Gametocyt (1)
- Gametozyt (1)
- Ganzkörperbestrahlung (1)
- Gap Junction (1)
- Gas chromatography (1)
- Gaschromatographie (1)
- Gastroesophageal reflux (1)
- Gastrointestinal tract (1)
- Gauss Prozess Klassifikation (1)
- Gaussian Process Classification (1)
- Gaze interaction (1)
- Gb3 accumulation (1)
- Gefrierstrukturierung (1)
- Gefäßalter (1)
- Gefäßwand (1)
- Gefäßwand-residente Stamm- und Vorläuferzellen (1)
- Gehirn-Computer Schnittstelle (1)
- Geißel <Biologie> (1)
- Gelernte Hilflosigkeit (1)
- Gemcitabin (1)
- Gen BRCA 1 (1)
- Gen Umweltinteraktion (1)
- Gen notch (1)
- Gen-Umwelt Interaktion (1)
- Gene by Environment (1)
- Gene expression regulation (1)
- Gene regulation (1)
- Gene therapy (1)
- General Transcription Factor II H (1)
- General amplifier model (1)
- Generalization (1)
- Genetic etiology (1)
- Genome Annotation (1)
- Genome Instability (1)
- Genome editing (1)
- Genomeditierung (1)
- Genomic Selection (1)
- Genomik (1)
- Genotoxicity (1)
- Genotyping (1)
- Gentoxizität (1)
- Geomagnetic Field (1)
- Gerinnungsfaktor (1)
- Germination and differentiation (1)
- Germinative Zellen (1)
- Geruchswahrnehmung (1)
- Geschlechtsunterschiede (1)
- Geschmack (1)
- Gestational diabetes (1)
- Gestationsdiabetes (1)
- Gewebe (1)
- Gewebemodelle (1)
- Glatte Muskulatur (1)
- Glatter Krallenfrosch (1)
- Glioblastomtherapie (1)
- Glioma (1)
- Global change (1)
- Glottoplastik (1)
- Glucocorticoids (1)
- Glucose Starvation (1)
- Glucose metabolism (1)
- Glucosetransport (1)
- Glucosetransporter (1)
- Glukosemetabolismus (1)
- Glukosetransporter -1 (1)
- Glutamin (1)
- Glutathion (1)
- Glycerin-3-phosphat (1)
- Glycerinphosphate (1)
- Glycin (1)
- Glycine Receptors (1)
- Glycocalyx (1)
- Glycolipid (1)
- Glycophyten (1)
- Glycoprotein GPV (1)
- Glycoprotein hormone (1)
- Glykane (1)
- Glykobiologie (1)
- Glykokalyx (1)
- Glykomodifizierung (1)
- Glykosyltransferase (1)
- Glyzin Rezeptoren (1)
- Glyzinrezeptor (1)
- Glücksspiel (1)
- Gold Nanoparticles (1)
- Golgi apparatus (1)
- Gonococcal invasion (1)
- Grad-seq (1)
- Gradient System Transfer Function (1)
- Graft versus Host Disease (1)
- Graft versus Host disease (1)
- Graft versus host disease (1)
- Graft-versus-host disease (GvHD) (1)
- Graft-versus-leukemia (1)
- Gram-positive (1)
- Gram-positive bacteria (1)
- Granulozyten (1)
- Growth-differentiation Factor 15 (1)
- Guanin Nukleotid Austauschfaktor (1)
- Guanine nucleotide exchange factor (GEF) (1)
- Guaninnucleotid-Austauschfaktoren (1)
- Guanylyl cyclase A (1)
- Guillain-Barré-Syndrom (1)
- GvL (1)
- Gvhd (1)
- H2A.Z (1)
- HAD phosphatase (1)
- HASH (1)
- HBD2 (1)
- HCMV (1)
- HD5 (1)
- HDAC (1)
- HECTD1 (1)
- HERV (1)
- HFpEF (1)
- HIF1alpha (1)
- HIV Drug resistance (1)
- HIV South Africa (1)
- HIV-1 (1)
- HIV-1 Vif (1)
- HIV-associated neurocognitive disorder (HAND) (1)
- HIV-assoziierte neurokognitive Störung (HAND) (1)
- HLA-G (1)
- HMBPP (1)
- HNE (1)
- HP1432, Hpn2 (1)
- HPLC-MS (1)
- HRV (1)
- HSF1 (1)
- HST1 (1)
- HSV-1 (1)
- Haarzelle (1)
- Habituationstraining (1)
- Haemostasis (1)
- Haloacid dehalogenase (1)
- Halophyten (1)
- Halophytes (1)
- Harze (1)
- Hautkrebs (1)
- Hautmodell (1)
- Hautverbrennung (1)
- Heart (1)
- Heart Failure (1)
- Heart Period (1)
- Heart development (1)
- Heart failure (1)
- Heat shock protein 90 (1)
- Heat stress (1)
- Heilmittel (1)
- Heilung (1)
- Heißhunger (1)
- Helferzelle (1)
- Helicase (1)
- Helikasen (1)
- Helminths (1)
- Hematopoietic Cell Transplantation (1)
- Hematopoietic Stem Cell (1)
- Hematopoietic cell transplantation (1)
- Hemmung der Proliferation schnell wachsender Krebszellen (1)
- Hemodynamics (1)
- Hemofiltration (1)
- Heparin (1)
- Hereditary spastic paraplegia (1)
- Hereditäre spastsiche Paraplegie (1)
- Herpes (1)
- Herpes simplex Virus DUB C65A (1)
- Herpes simplex virus C65A (1)
- Herpesviren (1)
- Herpesviruses (1)
- Herz-Kreislauf-Erkrankung (1)
- Herzbildgebung (1)
- Herzfrequenz (1)
- Herzfunktion (1)
- Herzruptur (1)
- Heterogenität von Mikroorganismen (1)
- Hey Proteine (1)
- Hey proteins (1)
- Hi-C (1)
- Hic-5 (1)
- Hidden Markov Model (1)
- Hidden-Markov-Modell (1)
- High-thropughput screening (1)
- Himmelskompass (1)
- Hippokampus (1)
- Hirnendothelzellen (1)
- Hirnhaut (1)
- Hirnstimulation (1)
- Histatin 5 (1)
- Histon-Deacetylase (1)
- Histon-Methyltransferase (1)
- Histonacetyltransferase (1)
- Histone Acetylation (1)
- Histone modification (1)
- Histone variant (1)
- Histones (1)
- Hitzeschock Proteine (1)
- Hitzeschock-Proteine (1)
- Hitzeschockprotein 90 (1)
- Hitzeschocktranskriptionsfaktor (1)
- Hitzestress (1)
- Hochauflösende Mikroskopie (1)
- Hochauflösungsmikroskopie (1)
- Hochdurchsatz-Screening (1)
- Hochdurchsatzsequenzierung (1)
- Holobiont (1)
- Homing (1)
- Homöostase (1)
- Honeybee (1)
- Honigbiene (1)
- Honigbienen (1)
- Hormone Receptor Signaling (1)
- Hornhaut (1)
- Hornhautinfektionen (1)
- Hospitalismus <Hygiene> (1)
- Host Colonization (1)
- Host Genome Integrity (1)
- Host defense (1)
- Hot Hand Fallacy (1)
- Huh6 (1)
- Human Herpesvirus 6 (1)
- Human Transcriptome (1)
- Human-pathogenic (1)
- Humanes Herpesvirus 6 (1)
- Humanparasitologie (1)
- Huwe1 (1)
- Hyaluronic Acid (1)
- Hyaluronic acid (1)
- Hybrid-Molecules (1)
- Hydrocortison (1)
- Hydrogels (1)
- Hyperactivity (1)
- Hyperekplexie (1)
- Hypertension (1)
- Hypertrophy (1)
- Hyrogels (1)
- Hämatopoetische Stammzellen (1)
- Hämatopoetische Stammzelltransplantation (1)
- Hämodynamik (1)
- Hämophilie A (1)
- Hämsotase (1)
- Höhenangst (1)
- Hörbahn (1)
- Hörrinde (1)
- ICP22 (1)
- IE3 (1)
- IEG (1)
- IFN-g (1)
- IFNAR (1)
- IKZF1 (1)
- IKZF3 (1)
- IL-12p70 (1)
- IL-2 (1)
- IL-23 (1)
- IL-4 (1)
- IL-4 Rezeptor alpha (1)
- IL-4 receptor alpha chain (1)
- IL-6 Inhibition (1)
- IMP1/ZBP1 (1)
- IP6 (1)
- IPP (1)
- IRAK2 (1)
- ITAM (1)
- ITAM-signalling (1)
- IVIG (1)
- Ibrutinib (1)
- Ideomotor gaze control (1)
- Ideomotorik (1)
- Ideomotorische Blickkontrolle (1)
- IgG-Subklasse (1)
- Ighmbp2 (1)
- Ikaros (1)
- Ileal Trasposition (1)
- Illumina HiSeq (1)
- Image Processing (1)
- Image Quality (1)
- Immunaktivierung (1)
- Immunantwort (1)
- Immune Checkpoint Therapy (1)
- Immune System (1)
- Immune Thrombocytopenia (1)
- Immune activation (1)
- Immune cell assays (1)
- Immune cells (1)
- Immunglobulin G (1)
- Immunität <Medizin> (1)
- Immunkardiologie (1)
- Immunocardiology (1)
- Immunologe (1)
- Immunomodulation (1)
- Immunophilin (1)
- Immunosuppressant (1)
- Immunozyt (1)
- Immunpathogenese (1)
- Immunsuppressiva (1)
- Immunthrombozytopenie (1)
- Immunzellassays (1)
- Immunzellrekrutierung (1)
- Impfstoff (1)
- Implantatentwicklung (1)
- Impulsivität (1)
- In vitro models (1)
- In vivo Imaging (1)
- Individual based model (IBM) (1)
- Individualisierte Medizin (1)
- Infection imaging (1)
- Infection models (1)
- Infectious diseases (1)
- Infektiologie (1)
- Infektionsbildgebung (1)
- Infektionsbiologie (1)
- Infektionsprozess (1)
- Infektionsstudien (1)
- Inferior colliculus (1)
- Inflamamtion (1)
- Inflammatory Pain (1)
- Inflammatory pain (1)
- Informationsnutzung (1)
- Infrared radiation (1)
- Inhibitory-postsynapse (1)
- Injectability (1)
- Insect (1)
- Insekt (1)
- Insektenstaaten (1)
- Instrumental Activities of Daily Living (1)
- Insulin-like Growth (1)
- Insulin-like-Growth-Factor-Binding-Protein-5 (1)
- Insulinresistenz (1)
- Integrase inhibitor (1)
- Integrated Defensive States (1)
- Integrin (1)
- Integrine (1)
- Interaction analysis (1)
- Interaction of 7SK with the Smn complex modulates snRNP production (1)
- Interaktionsrezeptor (1)
- Interferon <Gamma-> (1)
- Interferonrezeptor (1)
- Interleukin (1)
- Interleukin 12 (1)
- Interleukin 17 (1)
- Interleukin 5 (1)
- Interleukin-4 Antagonist (1)
- Interleukine (1)
- Internal Dosimetry (1)
- Internalisierende Störung (1)
- Intestinal metaplasia (1)
- Intestinal stem cell (1)
- Intracellular bacteria (1)
- Intracellular virulence (1)
- Intratumorale Heterogenität (1)
- Intravoxel Incoherent Motion (1)
- Intrazelluläre Bakterien (1)
- Intrinsische Gerinnungskaskade (1)
- Ion channel function (1)
- Ionisierende Strahlung (1)
- Irradiation (1)
- Ischemia (1)
- Ischämischer Schlaganfall (1)
- Isolation (1)
- Isolation and Characterization (1)
- Isomer (1)
- Isopentenyl pyrophosphate (IPP) (1)
- Isopentenyl-pyrophosphat (IPP) (1)
- Isoprenoid Synthesis (1)
- Isoprenoide (1)
- Isoprenoidsynthese (1)
- Ixolaris (1)
- JNCL (1)
- JNK (1)
- JURKAT-Zelle (1)
- Judgment (1)
- Jugendliche (1)
- Jun (1)
- K-RAS (1)
- K5 capsule (1)
- KDELR2 (1)
- Kaliumkanal (1)
- Kallikrein-Kinin-System (1)
- Kardiale MR-Bildgebung (1)
- Kardiovaskuläre Krankheit (1)
- Karotis-Intima-Media-Dicke (1)
- Kation (1)
- Kationen-Homöostase (1)
- Kehlkopfchirurgie (1)
- Keimzentrumsreaktion (1)
- Kern (1)
- Kidney (1)
- Kilimandscharo (1)
- Kinase (1)
- Kinase signaling (1)
- Kinasen (1)
- Kinder (1)
- Kinderpsychiatrie (1)
- Kinderschlaf (1)
- Kindesalter (1)
- Kindheit und Jugend (1)
- Kinetik (1)
- Klassifikations- und Regressionsbaum (1)
- Kleine Proteine (1)
- Kleinkern (1)
- Kleinkind (1)
- Klima (1)
- Klimawandel (1)
- Klinischer Behandlungspfad (1)
- Kniegelenkarthrose (1)
- Knochenbildung (1)
- Knochenersatz (1)
- Knochenmarkzelle (1)
- Knochenmetastase (1)
- Knochenmetastasen (1)
- Kofaktorbindung (1)
- Koffein (1)
- Kognitive Beeinträchtigung (1)
- Kognitive Inhibition (1)
- Kognitive Störung (1)
- Kognitiver Prozess (1)
- Kognitives Lernen (1)
- Kohlendioxid (1)
- Kohlenhydrate (1)
- Kohlenstoff (1)
- Kohlenstofffaser (1)
- Kohlenstoffstoffwechsel (1)
- Kohlenwasserstoffe (1)
- Kollektive Invasion (1)
- Kolloid (1)
- Kolonieerkennung (1)
- Kolonkarzinom (1)
- Kommunikation (1)
- Kommunikationshilfe (1)
- Komorbidität (1)
- Kompass (1)
- Komplement (1)
- Komplement C3a (1)
- Komplement C5a Rezeptor 1 (1)
- Komplementsystem (1)
- Komplexauge (1)
- Komplexes regionales Schmerzsyndrom (1)
- Komprimierte Abtastung (1)
- Kongestive Herzmuskelkrankheit (1)
- Kongo (1)
- Konnektivitätsschätzung (1)
- Konsolidierung (1)
- Kontingenz (1)
- Kontrastmittel (1)
- Kontrastmittel-gestützte MRT-Perfusionsmessung (1)
- Kontrolle (1)
- Kopfschmerz (1)
- Kopienzahlvariation (1)
- Korrelative Mikroskopie (1)
- Krebs (1)
- Kristallographie (1)
- Kryokonservierung (1)
- Kuh (1)
- Körperachsen (1)
- Körperliche Leistungsfähigkeit (1)
- Künstliche Ingelligenz (1)
- Künstliche Intelligenz (1)
- L-typ Calciumkanal Antagonist (1)
- LAD (1)
- LASP1 (1)
- LASP1 (LIM and SH3 Protein 1) (1)
- LASP1 (LIM und SH3 Protein 1) (1)
- LAT2 (1)
- LIMK (1)
- LIS (1)
- LPS (1)
- LPS Biosynthese (1)
- LV Function (1)
- Labeling (1)
- Lactobacillus reuteri (1)
- Lambdoide Prophagen (1)
- Langerhans cells (1)
- Langzeitgedächtnis (1)
- Lasermikrodissektion (1)
- Lasiocarpin (1)
- Lateral root development (1)
- Latrophilin receptor (1)
- Latrophilin-3 (1)
- Ldlr-Knockout (1)
- Leaf (1)
- Learning & Memory (1)
- Learning Walk (1)
- Learning walk (1)
- Lebendzellmikroskopie (1)
- Lebensmittelchemie (1)
- Lebensqualität (1)
- Leber-Metabolismus (1)
- Leberepithelzelle (1)
- Leichte kognitive Beeinträchtigung (1)
- Leitlinien (1)
- Lenalidomid (1)
- Lentiviral transgenesis (1)
- Leptomeningeal cells (1)
- Leptomeningealzellen (1)
- Lernverhalten (1)
- Letalität (1)
- Leukaemia-inhibitory factor (1)
- Lgr5 (1)
- Library Screening (1)
- Library of Phytochemicals (1)
- Library of plant species (1)
- Licht (1)
- Lichtblattmikroskopie (1)
- Lichtscheibenmikroskopie (1)
- Licl (1)
- Ligand <Biochemie> (1)
- Ligand uncaging (1)
- Light Sheet Fluorescence Microscopy (1)
- Light sheet microscopy (1)
- Lipid Raft (1)
- Lipid Rafts (1)
- Lipid Transfer Protein (1)
- Lipidom (1)
- Lipidomics (1)
- Lipidomik (1)
- Lipidumbau (1)
- Lipolysis (1)
- Liquor (1)
- Locomotor activity (1)
- Lokalanästhesie (1)
- Luftfeuchtigkeit (1)
- Lung Cancer metastasis (1)
- Lung cancer (1)
- Lung squamous cancer cells (1)
- Lungenentzündung (1)
- Lungentumor (1)
- Lurche (1)
- Lysin-Oxidase (1)
- Lysosome (1)
- Lysyl oxidases (1)
- Lysyloxidasen (1)
- Ländlicher Raum (1)
- M1 Muscarinic Receptor (1)
- MAP kinase (1)
- MAX (1)
- MCMV (1)
- MDR1 (1)
- MDSC (1)
- MDSCs (1)
- MEK5/ERK5 (1)
- MEK5/ERK5 cascade (1)
- MET-T-box riboswitch (1)
- MGMT (1)
- MIPs (1)
- MIZ1 (1)
- MLN7243 (1)
- MM (1)
- MMB complex (1)
- MMN (1)
- MMP (Matrix-Metalloproteasen) (1)
- MMP (Matrix-Metalloproteases) (1)
- MND (1)
- MRR1 (1)
- MS2-affinity purification and RNA-seq (1)
- MT02 (1)
- MYCN-amplified (1)
- Machine Learning (1)
- Macromolecular machine (1)
- Macrophage (1)
- Macrophage extracellular traps (1)
- Macrophytes (1)
- Magen (1)
- Magenchirurgie (1)
- Magenkrankheit (1)
- Magenkrebs (1)
- Magnetic Resonance Imaging (1)
- Magnetic Resonance Relaxometry (1)
- Magnetic resonance imaging (1)
- Magnetresonanz-Relaxometrie (1)
- Major depression (1)
- Makrokolonie (1)
- Makrophagen (1)
- Makrophyten (1)
- Malignant melanoma (1)
- Mamma carcinoma (1)
- Mammakarzinom (1)
- Mantle cell lymphoma (1)
- Marine natural products (1)
- Marine sponge (1)
- Marine sponge-derived actinomycetes (1)
- Marker (1)
- Marketing authorisation of pharmaceuticals (1)
- Marrow (1)
- Masern (1)
- Masern-Virus (1)
- Masernvirus-Infektion (1)
- Mass Spectrometry (1)
- Mass spectrometry (1)
- Massenspektrometrie/Proteomics (1)
- Masspectrometry (1)
- Mast cells (1)
- Mastzelle (1)
- Mathematical modeling (1)
- Mathematische Modellierung (1)
- Matrix-Metalloprotease (1)
- Matrix-Metalloproteasen (1)
- Matrix-Metalloproteinase (1)
- Mauerbiene (1)
- Mc4r (1)
- Measles virus (1)
- Mechanical deformation (1)
- Mechanisms of Social Attention (1)
- Mechanismus (1)
- Mechanistic model (1)
- Mechanorezeptor (1)
- Medaka (1)
- Mediator Komplex (1)
- Medium spiny neurons (1)
- Medizinprodukt (1)
- Medizintechnik (1)
- Medulloblastom (1)
- Megakaryocytes (1)
- Megakaryozytopoese (1)
- Mek5/Erk5-Signalweg (1)
- Melanin (1)
- Melanin release (1)
- Melanoma (1)
- Melanophor (1)
- Melt electrowriting (1)
- Membrane Receptor Dynamics (1)
- Membrane transporters (1)
- Membranlipide (1)
- Membranrezeptor (1)
- Membrantransporter (1)
- Memory (1)
- Memory B cells (1)
- Menaquinon-BIosynthese (1)
- Menaquinone Biosynthesis (1)
- Meninges (1)
- Meningokokken-Sepsis (1)
- Meniskus (1)
- Meniskusimplantat (1)
- Meniskustransplantation (1)
- Merkel cell polyomavirus (1)
- Merkelzellkarzinom (1)
- Mesenchymal Stem Cell (1)
- Mesenchymal Stromal Cell (1)
- Mesenchymal Stromal Cells (1)
- Mesenchymal stem cell (1)
- Mesenchymal stromal cells (1)
- Mesenchymale Stammzelle (1)
- Mesenchymale Stromazelle (1)
- Messenger-RNS (1)
- Meta-barcoding (1)
- Metabolic Glycoengineering (1)
- Metabolic Labeling (1)
- Metabolism (1)
- Metabolite von Morphin (1)
- Metabolites of morphine (1)
- Metabolomics (1)
- Metagenomics (1)
- Metalloprotease (1)
- Metalloproteasen (1)
- Metalloproteases (1)
- Metalloproteinase (1)
- Metapopulation (1)
- Metastasierung (1)
- Metatranscriptomics (1)
- Methioninbiosynthese (1)
- Method development (1)
- Methodenentwicklung (1)
- Methylphenidat (1)
- Methyltransferase (1)
- Mevalonate Pathway (1)
- MgrB (1)
- MicF (1)
- MicroRNA (1)
- MicroRNAs (1)
- Microarray (1)
- Microbial community (1)
- Microbiota (1)
- Microglia (1)
- Micronucleus (1)
- Microvesicle (1)
- Microwave Assisted Extraction (1)
- Migraine (1)
- Migration (1)
- Migräne (1)
- Mikroarray basierte vergleichende Genomhybridisierung (Array-CGH) (1)
- Mikrobiota (1)
- Mikroglia (1)
- Mikroglomeruli (1)
- Mikrokerne (1)
- Mikroorganismus (1)
- Mikrotubuli-assoziiertes Protein (MAP) (1)
- Mikrotubulus (1)
- Mimik (1)
- Minor histocompatibility antigen mismatch transplantation (1)
- Mitochondrien (1)
- Mittelohrimplantat (1)
- Mobile Crowdsensing (1)
- Mobile Health (1)
- Mobiles Endgerät (1)
- Mobilfunkstrahlung (1)
- Moco biosynthesis (1)
- Model (1)
- Model Based Reconstruction Algorithms in Magnetic Resonance Imaging (1)
- Model Organism (1)
- Model simulation (1)
- Modell (1)
- Modellbasierte-Rekonstruktionsalgorithmen in der Magnetresonanztomografie (1)
- Modellierung (1)
- Modellorganismus (1)
- Modellsystem (1)
- Moderator (1)
- Modification (1)
- Modifizierung (1)
- ModulationTregs (1)
- Molecular Pharmacology (1)
- Molecular Radiotherapy (1)
- Molekulare Zielstrukturen (1)
- Mongolische Rennmaus (1)
- Monitoring (1)
- Monoklonale Gammopathie (1)
- Monoklonaler Antikoerper (1)
- Monoklonaler bispezifischer Antikörper (1)
- Moonlight (1)
- Moosfaserterminalen (1)
- Morbus Alzheimer (1)
- Morbus Fabry (1)
- Morphin (1)
- Morphology (1)
- Morris Water Maze (1)
- Motivation (1)
- Motoneuron diseases (1)
- Motoneuronenerkrankung (1)
- Motoneurons (1)
- Motor Memory (1)
- Motor learning (1)
- Motor neuron disease (1)
- Motorische Endplatte (1)
- Motorisches Gedächtnis (1)
- Mouse Model (1)
- Mrap2 (1)
- Mrr1 (1)
- MscS (1)
- Mucorales (1)
- Mucormycosis (1)
- Mucormykose (1)
- Multi-Unit Aufnahmen (1)
- Multicellular aggregates (1)
- Multidrug-Resistance-Related Proteine (1)
- Multidrugresistant (1)
- Multilayered skin tissue model (1)
- Multiphotonenmikroskopie (1)
- Multiple Sclerosis (1)
- Multiple myeloma (1)
- Multizellulären Bakteriengemeinschaften (1)
- Mundschleimhautzellen (1)
- Murine models of thrombosis and haemostasis (1)
- Muskelin (1)
- Muskelzelle (1)
- Mustererkennungsrezeptoren (1)
- Myb-MuvB complex (1)
- Mycobacterium tuberculosis (1)
- Mycobacterium tuberculosis InhA (1)
- Myeloblast (1)
- Myeloid (1)
- Myeloid-derived suppressor cells (1)
- Myeloide Suppressorzellen (1)
- Myelom (1)
- Myeloma (1)
- Myocardial Work (1)
- Myocardial infarction (1)
- Myokardiale Deformation (1)
- Myokarditis (1)
- Myosin IIA (1)
- Myrmecology (1)
- N-RAS (1)
- N2pc (1)
- N400 (1)
- NA (1)
- NAFLD (1)
- NEDMIAL (1)
- NES (1)
- NF-kB (1)
- NF-kappaB (1)
- NFATc3 (1)
- NHERF (1)
- NK (1)
- NK cell (1)
- NK-DC cross-talk (1)
- NKT (1)
- NLS (1)
- NMJ (neuromuscular junction) (1)
- NMR (1)
- NNMT activity assay (1)
- NO-GC (1)
- NOS-I (1)
- NOS1AP (1)
- NOTES <Chirurgie> (1)
- NRF2 (1)
- NSM2 (1)
- NTRK fusions (1)
- NaV1.9. oxidized phospholipids (1)
- Nachtschattengewächse (1)
- Nahrung (1)
- Nahrungsaufnahme (1)
- Nanofabrication (1)
- Nanofabrikation (1)
- Nanofaser (1)
- Nanomedicine (1)
- Nanomedizin (1)
- Nanopartikel (1)
- Nanotubes (1)
- Naphthalinderivate (1)
- Naphthochinonen (1)
- Naphthoquinones (1)
- Naphthylisochinolinalkaloide (1)
- Naphthylisoquinoline (1)
- Narrow escape problem (1)
- Nasenschleimhaut (1)
- Natrium-Kalium-Pumpe (1)
- Natrium-abhängigen Glukosetransporter-1 (1)
- Natriumoxamat (1)
- Natural pest control (1)
- Natural products (1)
- Nature-Insipired Synthesis (1)
- Naturschutz (1)
- Naturstoff (1)
- Near Miss (1)
- Nebenniere (1)
- Nebennierenrindenkarzinom (1)
- Nebennierenrindenkrebs (1)
- Nebennierentumor (1)
- Nectin4 (1)
- Neogenin-1 (1)
- Nephrogenese (1)
- Nerven (1)
- Nervenstimulation (1)
- Netrin-1 (1)
- Netzhaut (1)
- Netzwerkanalyse (1)
- Neuartige Arzneimittel (1)
- Neural Stem cells (1)
- Neurobiology (1)
- Neuroblastoma (1)
- Neurodegenerative Erkrankung (1)
- Neurodevelopment (1)
- Neurodevelopmental Disorder (1)
- Neurodevelopmental diseases (1)
- Neuroepigenomics (1)
- Neuroethology (1)
- Neurofeedback (1)
- Neurofilament (1)
- Neurogenesis (1)
- Neurogenetik (1)
- Neuroligin 2 (1)
- Neuromuskuläre Endplatte (1)
- Neuronale Stammzellen (1)
- Neuronales visuelles System (1)
- Neuropathic Pain (1)
- Neuropathic pain (1)
- Neuropathologie (1)
- Neuropathy (1)
- Neuropeptid S Rezeptor Gen (1)
- Neuropeptid Y (1)
- Neuropeptidom (1)
- Neurophysiologie (1)
- Neuroplasticity (1)
- Neuroprotection (1)
- Neuroprotektivum (1)
- Neurotransmitter Receptors (1)
- Neurotransmitter Rezeptoren (1)
- Neurotransmitter-Rezeptor (1)
- Neurotrophic factors (1)
- Neutrophil (1)
- Neutrophil granulocyte (1)
- Neutrophil granulocytes interaction (1)
- Newman strain sae (1)
- Next Generation Sequencing (NGS) (1)
- Nicht-kartesische Bildgebung (1)
- Nicht-kleinzelliges Bronchialkarzinom (1)
- Nichtionisierende Strahlung (1)
- Nichtstrukturproteine (1)
- Nicotiana tabacum (1)
- Niere (1)
- Nierenversagen (1)
- Nimodipin (1)
- Nitrogen (1)
- Nodo-Paranodopathie (1)
- Non-Hodgkin-Lymphom (1)
- Non-conventional T cell (1)
- Non-invasive imaging (1)
- Non-ribosomal peptide synthetase (1)
- Non-viral genome engineering (1)
- Noonan-Syndrom (1)
- Noradrenalin (1)
- Noradrenalinstoffwechsel (1)
- Nosocomial Infections (1)
- Nosokomiale Infektionen (1)
- Notch Signalweg (1)
- Notch signalling (1)
- Nozizeption (1)
- Nuclar Medicine (1)
- Nuclear Factor of Activated T cells (NFAT) (1)
- Nuclear Medicine (1)
- Nuclear imaging (1)
- Nuclease (1)
- Nucleasen (1)
- Nucleotide excision repair (1)
- Nucleotide-Excision-Repair (1)
- Nucleus subthalamicus (1)
- Nuklearfaktor Kappa B (1)
- Nukleäre Hormonrezeptoren (1)
- Numerische Fluidmechanik (1)
- O(6)-Methylguanine-DNA Methyltransferase (1)
- OCT1 (1)
- OIS (1)
- OSM (1)
- OSMR (1)
- Oberflächenproteine der Sexualstadien (1)
- Olfaktion (1)
- Olfaktorik (1)
- Oligomerization (1)
- OmoMYC (1)
- Onchocerca volvulus (1)
- Onchozerkose (1)
- Oncogenes (1)
- Oncolytic Virotherapy (1)
- Onkologie (1)
- Onkolyse (1)
- Oozyte (1)
- Open chromatin (1)
- Opioidpeptide (1)
- Optimal foraging (1)
- Optogenetics (1)
- Orai1 (1)
- Organ of Corti (1)
- Organoids (1)
- Orientia tsutsugamushi (1)
- Osmr-Knockout (1)
- Osteoporose (1)
- OxPL (1)
- Oxidative Stress (1)
- Oxidized Phospholipids (1)
- Oxidized phospholipids (1)
- Oxygen partial pressure (1)
- Oxytocin (1)
- Oxytosis (1)
- P4-ATPase (1)
- PAF1c (1)
- PAR-CLIP (1)
- PCDHGC3 (1)
- PD-L1 (1)
- PDE (1)
- PDI (1)
- PE Phosphoethanolamine (1)
- PEG (1)
- PER (1)
- PG neurons (1)
- PGE2 (1)
- PHMB (1)
- PI3K (1)
- PKA signaling (1)
- PLEKHG5 (1)
- PLP phosphatase (1)
- PP2A (1)
- PRKACA (1)
- PROTAC (1)
- PRR (1)
- PTH1R (1)
- PTMs (1)
- PTPN22 (1)
- PacBio sequencing (1)
- Palbociclib (1)
- Panic Disorder (1)
- Panik (1)
- Pankreas (1)
- Paranodopathie (1)
- Parasitology (1)
- Parc National de la Comoé (1)
- Parkinson Krankheit (1)
- Partial Agonists (1)
- Partialagonismus (1)
- Paternal age and BMI effects (1)
- Pathogene Bakterien (1)
- Pathogenese (1)
- Pathogenesis (1)
- Pathogenicity (1)
- Pathogens (1)
- Pathologie (1)
- Pathology (1)
- Pathophysiologische Mechanismen (1)
- Patienten-orientierte Versorgung (1)
- Patientenklassifikation (1)
- Patientenorientierte Medizin (1)
- Pattern recognition receptors (1)
- Pattern-Recognition-Receptors (1)
- Patulin (1)
- Pediatric Nuclear Medicine (1)
- Pediatric Patients (1)
- Pemphigus (1)
- Peptides (1)
- Perforation <Medizin> (1)
- Perforine (1)
- Perfusion Bioreactor (1)
- Perianova, Irina (1)
- Periaqueductal gray (1)
- Period2 (1)
- Peripheral eosinophils (1)
- Periphere Analgesie (1)
- Peripheres Nervensystem (1)
- Permeability (1)
- Permeation (1)
- Peroxisomen-Proliferator-aktivierter Rezeptor (1)
- Persistence (1)
- Pertussis (1)
- Pesticide (1)
- Peyersche Plaques (1)
- Pflanzen-Bienen-Netzwerke (1)
- Pflanzenaussterben (1)
- Pflanzenhydraulik (1)
- Pflanzenzelle (1)
- Pflanzenzellkulturen (1)
- Pflanzenökologie (1)
- PhD thesis pharmacology (1)
- Pharmaceutische Biologie (1)
- Pharmacology (1)
- Pharmakodynamik (1)
- Phasenvariation (1)
- Phenotypic switch (1)
- Pheromon (1)
- Phonochirurgie (1)
- Phosphatidylinositolkinase <Phosphatidylinositol-3-Kinase> (1)
- Phosphoantigen (1)
- Phosphodiesterase (1)
- Phosphoglykolat (1)
- Phosphoglykolat-Phosphatase (1)
- Phospholamban (1)
- Phosphorylation (1)
- Photoreceptor (1)
- Photoswitch (1)
- Phyllosphere (1)
- Phyllosphäre (1)
- Phylogeny (1)
- Pilze (1)
- PinT (1)
- Plant Biology (1)
- Plant Ecology (1)
- Plant cell cultures (1)
- Plant extracts (1)
- Plant hydraulic (1)
- Plants (1)
- Plasmonic (1)
- Plastin 3 (1)
- Platelet activating Factor (1)
- Platelet count (1)
- Platelet granules (1)
- Platelet-Membranglykoprotein p62 (1)
- Platy (1)
- Pluripotenz (1)
- Pmn mutant mouse (1)
- Pneumonievirus der Maus (1)
- Pneumoviruses (1)
- Point Spread Function (1)
- Polistes (1)
- Pollen (1)
- Polo-like kinase 1 (1)
- Poly(2-oxazoline) (1)
- Poly(2-oxazoline)s (1)
- Poly(glycidol) (1)
- Polyadenylierung (1)
- Polyethism (1)
- Polymer Science (1)
- Polymere (1)
- Polymers (1)
- Polyphosphate (1)
- Polysaccharide (1)
- Polysaccharide intercellular adhesin (PIA) (1)
- Pore (1)
- Pore formation (1)
- Pore-formation (1)
- Porenbildung (1)
- Porifera (1)
- Porins (1)
- Positron emission tomography (1)
- Positronen-Emissions-Tomografie (1)
- Posttranslationale Änderung (1)
- Powdery mildew fungus (1)
- Pra1 (1)
- Preclinical studies (1)
- Prefrontal cortex (1)
- Prefrontalt Cortex (1)
- Primaten (1)
- Primär-basierte immortalisierte Zelllinie (1)
- Primäre Ziliendyskinesie (1)
- Primärelement (1)
- Primärprevention (1)
- Primärtumor (1)
- Primärzellen (1)
- ProQ (1)
- Problem Gambling (1)
- Processing fluency (1)
- Profilin (1)
- Programmed Cell Death 1 (1)
- Programmed Cell Death Ligand 1 (1)
- Promotor (1)
- Prosoziales Verhalten (1)
- Prostaglandin E2 (1)
- Protease (1)
- Proteaseaktivität (1)
- Proteaseinhibitor (1)
- Proteases (1)
- Proteasom (1)
- Proteasomaler Abbau (1)
- Proteasome (1)
- Protein Disulfid Isomerase (1)
- Protein Kinase D2 (1)
- Protein TRPC6 (1)
- Protein chemistry (1)
- Protein crosslinking (1)
- Protein folding (1)
- Protein interactions (1)
- Protein interactor (1)
- Protein purification (1)
- Protein-Protein Interaktion (1)
- Protein-Protein-Interaktion (1)
- Protein-Tyrosin-Kinasen (1)
- Proteinen mit antimikrobieller Wirkung (1)
- Proteinfaltung (1)
- Proteininteraktion (1)
- Proteininteraktionen (1)
- Proteinkinase A (1)
- Proteinkinase C (1)
- Proteinmodifizierung (1)
- Proteinquervernetzungen (1)
- Proteolysis (1)
- Proteolysis-Targeting-Chimera (1)
- Proteomics (1)
- Prozessierung (1)
- Prädiktoren und Korrelate (1)
- Präklinische Bildgebung (1)
- Präklinische Studien (1)
- Pränatale Entwicklung (1)
- Präzisionsmedizin (1)
- Pseudouridin (1)
- Psoriasis (1)
- Psychische Belastung (1)
- Psychische Gesundheit (1)
- Psychobiologie (1)
- Psychotherapie (1)
- Ptpn22 (1)
- Puberty (1)
- Pubertät (1)
- Pulswelle (1)
- Punktmutation (1)
- Purinozeptor (1)
- Pyridoxal 5'-phosphate phosphatase (1)
- Pyridoxalphosphat (1)
- Pyrrolidinderivate (1)
- Pyrrolidine carboxamides (1)
- Pyrrolizidinalkaloide (1)
- QPCR (1)
- QTOF (1)
- Quadruplex-DNS (1)
- Qualität (1)
- Qualitätsindikator (1)
- Qualitätssicherung (1)
- Quantifizierung (1)
- Quantitation (1)
- Quantitative Genetics (1)
- Quartäre Bisnaphthalimide (1)
- Quaternary Bisnaphthalimide (1)
- RAF (1)
- RAF Kinasen (1)
- RAMP (1)
- REDD1 (1)
- REST-Complex (1)
- RESTORE protocol (1)
- RIL-seq (1)
- RIM1α (1)
- RNA Abbau (1)
- RNA G-quadruplex (1)
- RNA Sequencing (1)
- RNA binding potein CNBP (1)
- RNA binding proteins (1)
- RNA decay (1)
- RNA metabolism (1)
- RNA protein interactions (1)
- RNA secondary structures (1)
- RNA sequencing (1)
- RNA structures (1)
- RNA virus (1)
- RNA-Protein Interaktom (1)
- RNA-RNA interactions (1)
- RNA-Seq (1)
- RNA-Sequenzierung (1)
- RNA-bindendes Protein (1)
- RNA-binding protein (1)
- RNA-binding proteins (1)
- RNA-protein interactome (1)
- RNAi (1)
- RNS Polymerase I (1)
- RNS-Polymerase II (1)
- RNS-Spleißen (1)
- ROR2 (1)
- RS1 Peptides (1)
- RS1 derived peptides (1)
- RSK2 (1)
- RSV (1)
- RYGB (1)
- Rac1 (1)
- Radiale MR-Bildgebung (1)
- Radiation Protection (1)
- Radiation-associated Cancer Risk (1)
- Radiologische Diagnostik (1)
- Radionuklidtherapie (1)
- Raf <Biochemie> (1)
- Raf Kinase Inhibitor Protein (RKIP) (1)
- Raf kinase inhibitor protein (1)
- Raf1 (1)
- Random Forest (1)
- Random-walk simulations (1)
- Raphe (1)
- Raphe Kerne (1)
- Rasterionenmikroskop (1)
- Rasterkraft-Mikroskopie (1)
- Rauch (1)
- Rbm8a (1)
- Read-through transcription (1)
- Reaktive Sauerstoffspezies (1)
- Real-time imaging (1)
- RecQ helicase (1)
- Receptor (1)
- Receptor Anchoring (1)
- Receptor dynamics (1)
- Receptor internalization (1)
- Receptor signaling (1)
- Recombinant defensins (1)
- Recycling- Endosomen (1)
- Red Sea (1)
- Regenerative Medicine (1)
- Regulation of Gene Expression (1)
- Regulator of G protein signaling 2 (1)
- Regulatorische T-Zellen (1)
- Regulatory T Cells (1)
- Regulatory T-cell (1)
- Rekombinante DNS (1)
- Rekonsolidierung (1)
- Rekonstruktion (1)
- Relaxation Parameter Mapping in Magnetic Resonance Imaging (1)
- Remission (1)
- Remodeling (1)
- Renaturierung <Ökologie> (1)
- Repeats (1)
- Reperfusion (1)
- Reporter Cells (1)
- Reporterzellen (1)
- Repression (1)
- Repression <Genetik> (1)
- Reproducibility challenges (1)
- Reprogrammming (1)
- Rescue behaviour (1)
- Resistance (1)
- Resistenzmechanismen (1)
- Respiratorisches System (1)
- Response Inhibition (1)
- Ressourcenallokation (1)
- Restriktionsenzym (1)
- Resveratrol (1)
- Retinales S-Antigen (1)
- Retinoesäure (1)
- Retinoic acid (1)
- Reverse Transkriptase (1)
- Rezeptor Verankerung (1)
- Rezeptor-Tyrosinkinasen (1)
- Rezeptorblocker (1)
- Rezeptorpharmakologie (1)
- Rezidiv (1)
- Rgs2 (1)
- Rheologie (1)
- Rheology (1)
- Rheumatoider arthritis (1)
- Rhizopus arrhizus (1)
- Rho GTPase (1)
- Rho GTPasen (1)
- Rho GTPasw (1)
- Rho-GTPasen (1)
- Rho-Kinasen (1)
- RhoGTPase (1)
- Rhodopseudomonas palustris (1)
- Rhodopsin 7 (1)
- Ribonuclease H2 (1)
- Riboregulation (1)
- Ribosomale RNA (1)
- Ribosome biogenesis (1)
- Ribosome profiling (1)
- Ribozymes (1)
- Riesensynapsen (1)
- Risk Assessment (1)
- Rituximab (1)
- Rodents (1)
- Rodung (1)
- Roscovitine (1)
- Rossameise (1)
- Rotes Meer (1)
- Rothmund-Thomson-Syndrome (1)
- Rupturen (1)
- Räumliches Gedächtnis (1)
- SAP2 (1)
- SCV maturation (1)
- SEPHCHC Synthase (1)
- SERCA2a (1)
- SET7 (1)
- SGLT (1)
- SIS-muc (1)
- SIX1 (1)
- SLC2A3 (1)
- SLFN5 (1)
- SMARD1 (1)
- SMN complex (1)
- SNAP-Rezeptor (1)
- SNPs (1)
- SOC (1)
- SOD1 (1)
- SPECT/CT (1)
- SR proteins (1)
- SREBP (1)
- STAAB Cohort Study (1)
- STAAB study (1)
- STAAB-Studie (1)
- STD patients (1)
- STEC (1)
- STED (1)
- STIM2 (1)
- STP1 (1)
- SUMOylation (1)
- SWI/SNF (1)
- Saatgutbeizung (1)
- Saccharomyces cerevisiae (1)
- Salmonellose (1)
- Samenschale (1)
- Sandminen (1)
- Sap47 (1)
- Satellite glial cell (1)
- Sauerstoffkonzentration (1)
- Saure Sphingomyelinase (1)
- Scaffold bone implant (1)
- Schaumzelle (1)
- Scherstress (1)
- Schilddrüse (1)
- Schimmelpilze (1)
- Schimpanse (1)
- Schizosaccharomyces pombe (1)
- Schlafstörung (1)
- Schlaganfallversorgung (1)
- Schließzellfunktion (1)
- Schmerztherapie (1)
- Schmetterlinge (1)
- Schnelle MR-Bildgebung (1)
- Schreckreaktion (1)
- Schuppenflechte (1)
- Schwamm (1)
- Schwämme (1)
- Sehen (1)
- Sekretion (1)
- Sekretion von Aspartatproteasen (1)
- Sekundäre Inflammation (1)
- Sekundärprävention (1)
- Sekundärstruktur (1)
- Selbstregulation (1)
- Selective extraction (1)
- Selektiver Serotonin Wiederaufnahmehemmer / SSRI (1)
- Senecionin (1)
- Seneciphyllin (1)
- Seneszenz (1)
- Sensory Gating (1)
- Sequence Analysis (1)
- Sequence-Structure (1)
- Sequenz (1)
- Sequenzanalyse <Chemie> (1)
- Sequenzdaten (1)
- Sequenzwiederholung (1)
- Serinprotease (1)
- Serotonin Transporter (1)
- Serotonin transporter (1)
- Serotonin-Reuptake-Hemmer (1)
- Serotonin-Wiederaufnahmehemmer (1)
- Serotonindefizienz (1)
- Serotonintransporter (1)
- Sertralin (1)
- Setting Control (1)
- Sexuelle Entwicklung (1)
- Sezernierte Faktoren (1)
- Sglt1 (1)
- Shear Stress (1)
- Shigella (1)
- Shigella flexneri (1)
- Shimming (1)
- Sialoadhesin (1)
- Signal-Übertragung (1)
- Signaling (1)
- Signalregenerierung (1)
- Silica precursor (1)
- Simulation (1)
- Single Particle Tracking (1)
- Single-cell RNA Sequencing (1)
- Single-cell RNA-sequencing (1)
- Single-molecule (1)
- Sinusthrombose (1)
- Skin (1)
- Skin Tissue Engineering (1)
- Skin conductance (1)
- Sleep (1)
- Sleep in children (1)
- Sleeping Beauty Transposon System (1)
- Sleeping Beauty transposon (1)
- Smad (1)
- Small angle X-ray scattering (1)
- Small intestine (1)
- Small non-messenger RNS (1)
- Small nuclear RNP (1)
- Small-angle X-ray Scattering (1)
- Smoke (1)
- SnRK1-bZIP complex (1)
- Snap25 (1)
- Social Buffering (1)
- Social Distancing (1)
- Social action effects (1)
- Social buffering (1)
- Sociomotor gaze control (1)
- Sodium dependent glucose transporter-1 (1)
- Sol-gel (1)
- Solanaceae (1)
- Solanum species (1)
- Solid-phase peptide synthesis (1)
- South Africa (1)
- Soziale Handlungseffekte (1)
- Soziale Phobie (1)
- Sozialer Kontakt (1)
- Soziobiologie (1)
- Soziomotorische Blickkontrolle (1)
- Spatial heterogeneity (1)
- Spatiotemporal analysis (1)
- Specialized pro resolving mediators (1)
- Species richness (1)
- Speckle Tracking (1)
- Speicheldrüse (1)
- Sphingolipidstoffwechsel (1)
- Sphingomyelinase (1)
- Sphingomyelinphosphodiesterase (1)
- Sphingosine-1-phosphate (1)
- Sphingosine-1-phosphats (1)
- Spielsucht (1)
- Spinal muscular Atrophy (1)
- Spinal muscular atrophy (1)
- Spinal muscular atrophy (DLC) (1)
- Spinalganglion (1)
- Spirale (1)
- Spleen tyrosine kinase (1)
- Spliceosome (1)
- Splicing (1)
- Spo0A (1)
- Sponge diseases (1)
- Sponges (1)
- Sporenbildung (1)
- Sporulation (1)
- Spumaviren (1)
- Squamous cell carcinoma (1)
- Ssl1 (1)
- Stack-of-Stars Sequenz (1)
- Stack-of-Stars sequence (1)
- Stammzellen (1)
- Stammzelltransplantation (1)
- Staphylococci (1)
- Staphylococcus epidermidis (1)
- Stat3 (1)
- Stat5 (1)
- Stathmin (1)
- Staupevirus (1)
- SteC (1)
- Stechameisen (1)
- Stem Cells (1)
- Stem cells (1)
- Sterblichkeit (1)
- Steroide (1)
- Steroidhormon (1)
- Stickstoffkontrolle (1)
- Stickstoffmetabolismus (1)
- Stickstoffmonoxid-Synthase (1)
- Stickstoffwechsel (1)
- Stiff person syndrome (1)
- Stiff-Person Syndrom (1)
- Stiff-person syndrome (1)
- Stimmbandchirurgie (1)
- Stimmerhöhung (1)
- Stimulation (1)
- Stimulationsversuche (1)
- Stk (1)
- Stofftransport <Biologie> (1)
- Stoffwechselinhibitoren (1)
- Store-Operated (1)
- Stp (1)
- Strahlenpilze (1)
- Streptomyces (1)
- Stressresistenz (1)
- Striatum (1)
- Stroke (1)
- Stroke Unit (1)
- Stroma (1)
- Stroop-Verfahren (1)
- Structural elucidation (1)
- Structural organisation (1)
- Structure-based (1)
- Structured Illumination Microscopy (1)
- Struktur (1)
- Struktur-basiertes Wirkstoff Design (1)
- Strukturanalyse (1)
- Strukturbiologie (1)
- Strumpellin (1)
- Störabstand (1)
- Störung des Sozialverhaltens (1)
- Subitizing (1)
- Subtyp C (1)
- Sumo (1)
- Sumoylierung (1)
- Super-Resolution Microscopy (1)
- Super-resolution microscopy (1)
- Super-resolution microsopy (1)
- Superhochauflösende Mikroskopie (1)
- Superparamagnetische Eisenoxid Kontrastmittel (1)
- Superresolution microscopy (1)
- Suppressorzelle (1)
- Survival Motor Neuron (1)
- Suspensionskultur (1)
- Syap1 (1)
- Symbionts (1)
- Symbiosis (1)
- Synapse-associated protein (1)
- Synapsen assoziiert (1)
- Synapses (1)
- Synapsine (1)
- Synaptic plasticity (1)
- Synaptische Plastizität (1)
- Synaptische Transmission (1)
- Synaptische Vesikel (1)
- Synchronitätsmessung (1)
- Syncytin (1)
- Synthesis (1)
- Systematik (1)
- Systembiologie (1)
- Säugerzellen (1)
- Säugetiere (1)
- Südafrika (1)
- Südostasien (1)
- T Helper Cell (1)
- T Lymphocyte (1)
- T Zell Selektion (1)
- T Zellen (1)
- T cell (1)
- T cell cytotoxicity (1)
- T cell engaging Antibody (1)
- T cell homing (1)
- T cell receptor (1)
- T cell selection (1)
- T cell specificity (1)
- T zellen (1)
- T-Zell-Aktivierung (1)
- T-Zellaktivierung (1)
- T-Zellhoming (1)
- T-Zellmigration (1)
- T-cell (1)
- T-cell engineering (1)
- T-cell therapy (1)
- TAC (1)
- TCTP (1)
- TGF-ß (1)
- TGF-β (1)
- TGN1412 (1)
- TH2 Immunantwort (1)
- TLR signaling (1)
- TMEM16F (1)
- TNBC (1)
- TNFR2 (1)
- TP53 (1)
- TP53 lesions (1)
- TRAIL mutants (1)
- TRPA1 (1)
- TRPA1 channel (1)
- TRPC (1)
- TRPC-Ionenkanäle (1)
- TRPC3 (1)
- TRPC6 (1)
- TRPM7 kinase (1)
- TRPV1 (1)
- TRRAP (1)
- TT-Fields (1)
- TTF (1)
- Tabakkonsum (1)
- Tageslänge (1)
- Tagesrhythmik (1)
- Tamoxifen (1)
- Tansania (1)
- Tanzania (1)
- Tapeworm (1)
- Targeted therapies (1)
- Targeted therapy (1)
- Tc-99m-MAG3 Scans (1)
- Telomer <Molekulargenetik> (1)
- Telomerase (1)
- Temperatur (1)
- Temperaturabhängigkeit (1)
- Temporal heterogeneity (1)
- Temporal predictability (1)
- Terahertzbereich (1)
- Terahertzstrahlung (1)
- Terpene (1)
- Test system (1)
- Tfb4 (1)
- Thebain (1)
- Theoretical Ecology (1)
- Theory of Mind (1)
- Therapeutical application (1)
- Therapieoutcome (1)
- Therapiesimulation (1)
- Therapy (1)
- Thermoregulation (1)
- Thermoresponsive Polymere (1)
- Thermotoleranz (1)
- Theta Burst Stimulation (1)
- Thigmotaxis (1)
- Think/No-Think (1)
- Thiolase (1)
- Thiostrepton (1)
- Thrombin (1)
- Thromboinflammation (1)
- Thrombopoese (1)
- Thrombozyten (1)
- Thrombozytenfunktion (1)
- Thrombozytenfunktionsanalyse (1)
- Thrombozytopathie (1)
- Thrombozytopenie (1)
- Thrombozytopoese (1)
- Thrombus (1)
- Thymocytes (1)
- Thymosin b4 (1)
- Thymus (1)
- Tierphysiologie (1)
- Tight Junction Proteins (1)
- Tight junction (1)
- Tight-Junction-Protein (1)
- Time-of-flight (1)
- Timing (1)
- Tissue staining (1)
- Tobacco smoking (1)
- Tolerance (1)
- Toleranz <Biologie> (1)
- Toxin (1)
- TraDIS (1)
- Tracer (1)
- Trailmaking Test (1)
- Transcranial Magnetic Stimulation (1)
- Transcription Factor (1)
- Transcription factor (1)
- Transgenic mice (1)
- Transkriptomik anlyze (1)
- Translation <Genetik> (1)
- Transmission (1)
- Transmissionsblockierende Impfstoffe (1)
- Transpiration barrier (1)
- Transplantatabstoßung (1)
- Transportbarriere (1)
- Transporters (1)
- Transsexualismus (1)
- Transthorakale Echokardiographie (1)
- Trauma (1)
- Traumatic neuropathy (1)
- Tree physiology (1)
- Trem2 (1)
- Trend (1)
- Trennungsangst (1)
- Triclosan (1)
- Triglyceride (1)
- Tripartite Model (1)
- Trockenstress (1)
- Trophic Factors (1)
- Tropomyosin receptor kinase B (1)
- TruD (1)
- Trypanosoma (1)
- Trypanosoma Brucei (1)
- Trypanosoma brucei brucei (1)
- Trypanosomiasis (1)
- Tryptophan hydroxylase (1)
- Tryptophan hydroxylase 2 (1)
- Tuberkulose (1)
- Tumor Treating Fields (1)
- Tumor microenvironment (1)
- Tumor-Nekrose-Faktor <alpha> (1)
- Tumorangiogenese (1)
- Tumorgefäßmorphologie (1)
- Tumorhypoxie (1)
- Tumorimmunologie (1)
- Tumormetabolismus (1)
- Tumormikroumgebung (1)
- Tumortherapiefeld (1)
- Tumorwachstum (1)
- Tumorzellen (1)
- Tumour (1)
- Tumour angiogenesis (1)
- Twilight (1)
- Typ 2 (1)
- Type 1 Diabetes (1)
- Type 1 diabetes (1)
- Type II-C CRISPR/Cas (1)
- Type VIIb secretion system (1)
- Tyrosinkinaseinhibitor (1)
- U snRNPs (1)
- U1 snRNA (1)
- UBA6 (1)
- UBE2S (1)
- UBE2Z (1)
- UNC5B (1)
- UPEC (1)
- USA300 (1)
- USP (1)
- USP28 (1)
- Ubiquitin activating eznyme 1 (1)
- Ubiquitin system (1)
- Ubiquitin-PA (1)
- Ubiquitin-aktivierende Enzym 1 (1)
- Ubiquitin-conjugating enzyme (1)
- Ubiquitylation (1)
- Ultrahigh field (1)
- Ultraschall (1)
- Ultrashort echo time - UTE (1)
- Unidirectional Freezing (1)
- Untertyp (1)
- Uremic cardiomyopathy (1)
- Urinary tract infection (1)
- Urteilen (1)
- Urämie (1)
- Urämische Kardiomyopathie (1)
- VCAM (1)
- VE-Cadherin (1)
- VE-PTP (1)
- VLA-1 (1)
- VSMC (1)
- VW-SCs (1)
- Vaccinia-Virus (1)
- Vagus (1)
- Vagusnervstimulation (1)
- Vakuole (1)
- Vakzinen (1)
- Validierung (1)
- Validierungsstudie (1)
- Validität (1)
- Valscularization (1)
- Variant Surface Glycoprotein (1)
- Variants (1)
- Vascular system (1)
- Vaskularisation (1)
- Vaskularisierung (1)
- Vaskuläre Integrität (1)
- Vasopressin (1)
- Venusfliegenfalle (1)
- Verbreitungsökologie (1)
- Verhaltenplastizität (1)
- Verhaltenskontrolle (1)
- Verhaltensplastizität (1)
- Verhaltensstörung (1)
- Vermehrung (1)
- Vermeidungsreaktion (1)
- Vermeidungsverhalten (1)
- Vernetzung <Chemie> (1)
- Versorgungsqualität (1)
- Verweildauer (1)
- Verwundbarkeit (1)
- Very-long-chain aliphatic (1)
- Vesikel (1)
- Vesikelbildung (1)
- Vessel wall (1)
- Vg9Vd2 T Zellaktivierung (1)
- Vg9Vd2 T cell (1)
- Vgamma9Vdelta2 T cell (1)
- Vgamma9Vdelta2 T cells (1)
- Vibrationstraining (1)
- Vielfalt (1)
- Virologische Synapse (1)
- Virology (1)
- Virulenzfaktor (1)
- Virus infection (1)
- Virus-Transmission (1)
- Virusreplikation (1)
- Visual attention (1)
- Visuelle Orientierung (1)
- Visuelle Wahrnehmung (1)
- Visuelles Gedächtnis (1)
- Visuelles System (1)
- Vitamin D (1)
- Vitamin D-Rezeptor (1)
- Vitamin D3 (1)
- Voltage-Clamp-Methode (1)
- Vorhersage (1)
- Vorläufer (1)
- Vulnerable plaque (1)
- Vγ9Vδ2 T cells (1)
- WASH complex (1)
- WNT (1)
- WNT signaling (1)
- Wachstum (1)
- Wachstumsfaktor (1)
- Waiting Impulsivity (1)
- Warburg, Otto (1)
- Warburg-Effekt (1)
- Wasser Fett Trennung (1)
- Wasted Work (1)
- Water stress (1)
- Web services (1)
- Wechselwirkungen (1)
- Weinrebe (1)
- Wildbienen (1)
- Wilms Tumor (1)
- Wilms tumor protein 1 (1)
- Wilms-Tumor (1)
- Wirbelströme (1)
- Wirkstoff (1)
- Wirkstofftestung (1)
- Wirt-Erreger Interaktion (1)
- Wirtszelle (1)
- Wnt-Proteine (1)
- Wnt-Signalweg (1)
- Wnt-pathway (1)
- Wurzelhalsgalle (1)
- Würmer (1)
- Wüstenpflanze (1)
- X-ray Crystallography (1)
- X-ray diffraction (1)
- XPD (1)
- Xenograft (1)
- Xeroderma pigmentosum (1)
- Xerostomie (1)
- Xiphophorus Melanom (1)
- Xmrk (1)
- YAP (1)
- YB-1 (1)
- Yb1 (1)
- Yeast (1)
- Yersinia (1)
- Yersinia pestis (1)
- YnaI (1)
- ZFAND1 (1)
- Zebrafisch (1)
- Zeitgeber (1)
- Zell Migration (1)
- Zell-Migration (1)
- Zelladhäsion (1)
- Zellfilamente (1)
- Zellkernarchitektur (1)
- Zellkontakt (1)
- Zelllinie (1)
- Zellteilung (Zytokinese) (1)
- Zelltod (1)
- Zelltransport (1)
- Zielstruktur (1)
- Zink-Cluster-Transkriptionsfaktoren (1)
- Zytokin (1)
- Zytokingenpolymorphismus (1)
- Zytoskelett (1)
- Zytoskelettreorganisation (1)
- Zählen (1)
- aGPCR (1)
- acceptance-based strategies (1)
- acid sphingomyelinase (1)
- actin cytoskeleton (1)
- actin-binding proteins (1)
- actinomycetes (1)
- acute brain slices (1)
- acute graft-versus host disease (1)
- acute lymphoblastic leukaemia (1)
- adhesion (1)
- adhesion-GPCR (1)
- adipogenic differentiation (1)
- adipose (1)
- adipose-derived (1)
- adolescents (1)
- adrenal gland (1)
- adrenal incidentaloma (1)
- adrenerge Rezeptoren (1)
- adrenergic receptor (1)
- adrenergic receptors (1)
- adrenocortical adenoma (1)
- adult ADHD (1)
- adult attention deficit hyperactivity disorder (aADHD) (1)
- adulte Neurogenese (1)
- aggressive behavior (1)
- aging (1)
- agonist (1)
- airflow (1)
- airways (1)
- akute lymphatische Leukämie bei Kindern (1)
- alfa-cyano-4-hydroxycinnamat (1)
- alkaloids (1)
- allogen (1)
- allogeneic stem cell transplantation (1)
- allogenic (1)
- allogenic stem cell transplantation (1)
- allografts (1)
- alloreactive T cells (1)
- alltägliche Funktionsfähigkeit (1)
- alpha-IIb beta-3 (1)
- alternative intronic polyadenylation (1)
- ammonium (1)
- ammonium permease (1)
- amphibia (1)
- amphiphysin (1)
- amyotrophic lateral sclerosis (1)
- anaerobe (1)
- analysis (1)
- anaphylatoxin receptors (1)
- androstadienone (1)
- anoikis (1)
- anterior insula (1)
- anthocyanins (1)
- anti-hDEC205-WT1 antibody fusion protein (1)
- anti-infective (1)
- antigen (1)
- antigenic variation (1)
- antileukemia vaccine (1)
- antimicrobial (1)
- antithrombotic (1)
- antivirale Gene (1)
- anxiety conditioning (1)
- arf (1)
- aromatic amino acid biosynthesis (1)
- articular cartilage progenitor cells (1)
- artificial diet (1)
- artificial human skin (1)
- artifizielle Aktivierung (1)
- artifizielle Hautmodelle (1)
- asialoGM1 (1)
- aspartic protease (1)
- aspergillus fumigatus (1)
- assembly (1)
- association studies (1)
- astrocytoma (1)
- asymptomatic bacteriuria (1)
- atomic force microscopy (1)
- atopic diseases (1)
- atrial natriuretic peptide (1)
- attributable fraction (1)
- attributable risk (1)
- auditorisch (1)
- auditory (1)
- auditory cortex (1)
- auditory pathway (1)
- autoantibodies (1)
- autoantibody (1)
- autoimmunity (1)
- autophagocytose (1)
- autophagocytosis (1)
- autophagosome (1)
- autotransporter (1)
- bSSFP (1)
- bac-genomics-scripts (1)
- bacterial cancer therapy (1)
- bacterial fatty-acid biosynthesis (1)
- bacterial lipid rafts (1)
- bacterial nanocellulose (1)
- bacterial tumor targeting (1)
- bakterielle Flora (1)
- balanced steady state free precession (1)
- bark beetles (1)
- bee microbiota (1)
- bee-associated bacteria (1)
- bee-lining (1)
- bees (1)
- behavioral maturation (1)
- behavioral rhythms (1)
- beige adipocytes (1)
- benzimidazole (1)
- bestäuberfreundliche Pflanzen (1)
- beta actin (1)
- beta cell (1)
- beta-Arrestin2 (1)
- beta-Catenin (1)
- beta-adrenerge Signalwege (1)
- bicuculline (1)
- bilateral BAS model (1)
- binding mode (1)
- biochemistry (1)
- biodistribution (1)
- biodiversity (1)
- biofabrication (1)
- biofilm architecture (1)
- bioimage analysis (1)
- bioink (1)
- biokinetics (1)
- biolayerinterferometry (1)
- biological scaffolds (1)
- bioluminescence imaging (1)
- biomaterials (1)
- bioprocessing (1)
- bioreactor plattform (1)
- biosynthetic gene clusters (1)
- bipolar disorder (1)
- bipolare Störung (1)
- bispecific (1)
- bispezifisch (1)
- bitter taste (1)
- blood (1)
- blood cerebrospinal fluid barrier (1)
- blood glucose regulation (1)
- blood nerve barrier (1)
- blood-brain barrier (1)
- body axis (1)
- bone marrow niche (1)
- bone marrow transplantation (1)
- bone metastases (1)
- bone regeneration (1)
- brood rearing (1)
- building behavior (1)
- bumblebee*s (1)
- burn wound (1)
- butyrophilin 3A (1)
- c-Jun Phosphorylierung (1)
- c-myc (1)
- cAMP signaling (1)
- cadherin-13 (1)
- caffeine (1)
- calcium activity (1)
- calcium homeostasis (1)
- calcium imaging (1)
- cancer therapy (1)
- candida (1)
- candida albicans (1)
- canine (1)
- carbon dioxide (1)
- cardiac imaging (1)
- cardiac magnetic resonance imaging (1)
- cardiac tissue (1)
- cardiac tissue engineering (1)
- cardiovascular (1)
- carnivorous plants (1)
- cartilage (1)
- cartilage regeneration (1)
- cd28 superagonists (1)
- cell adhesion (1)
- cell cycle (1)
- cell filaments (1)
- cell migration (1)
- cell therapy (1)
- cellular model (1)
- cellular-trafficking (1)
- central complex (1)
- chaperone (1)
- chemotherapy (1)
- children (1)
- chimeric antigen receptor (1)
- chlamydia trachomatis (1)
- chondrogene Differenzierung (1)
- chromatin accessibility (1)
- chromosome conformation capture (1)
- chronic kidney disease (1)
- chronic pain (1)
- chronophin (1)
- cine loop (1)
- circadian clock (1)
- circadian clocks (1)
- circuitopathies (1)
- classical conditioning (1)
- claudin-12 (1)
- climate change (1)
- climate control (1)
- clock genes (1)
- closing of chromatin (1)
- co-culture (1)
- co-dependent expression (1)
- co-infection (1)
- coagulation factor XII (1)
- cochlea implant (1)
- cofactorbinding (1)
- cognitive decline (1)
- cognitive deficits (1)
- cognitive inhibition (1)
- cognitive remediation (1)
- cohesin (1)
- collective invasion (1)
- collybistin (1)
- colony recognition (1)
- color vision (1)
- comet assay (1)
- compartments (1)
- compound eyes (1)
- compressed sensing (1)
- conditional Knockout (1)
- confocal microscopy (1)
- conformational activation (1)
- context conditioning (1)
- contextual conditioning (1)
- contingency awareness (1)
- continuous wavelet analysis (1)
- control (1)
- convolutional neural network (1)
- corneal confocal microscopy (1)
- coronary heart disease (1)
- correlation (1)
- corticosteroids and cyclophosphamide (1)
- cryokonservation (1)
- crystal structure (1)
- ctr (1)
- current source density (1)
- cushing's syndrome (1)
- cuticular hydrocarbons (1)
- cuticular leaf wax (1)
- cuticular transpiration barrier (1)
- cuticular water permeability (1)
- cuticular waxes (1)
- cyclase-associated protein (1)
- cyclase-associated protein 2 (1)
- cyclo-AMP (1)
- cytokine release syndrome (1)
- cytoskeletal reorganisation (1)
- dCIRL (1)
- dSTORM (1)
- decellularization (1)
- decision-making (1)
- defense (1)
- degeneratives Nervengewebe (1)
- delipidation (1)
- dendritic cells (1)
- dendritic cell-targeting (1)
- density weighting (1)
- desert plant (1)
- deubiquitinase (1)
- dexamethasone suppression test (1)
- diabetic cardiomyopathy (1)
- diagnostic Microarray (1)
- diagnostic accuracy (1)
- diagnostics (1)
- diagnostischer Microarray (1)
- diastolic dysfunction (1)
- differential RNA-seq (1)
- differential coverage binning (1)
- differential geneexpression (1)
- differentiation status (1)
- differenzielle Genexpression (1)
- dilated cardiomyopathy with ataxia (DCMA) (1)
- dimerer Naphthylisochinolin-Alkaloide (1)
- discrimination training (1)
- disease model (1)
- dispersal (1)
- distance (1)
- distribution (1)
- disulfide bonds (1)
- diversity (1)
- double-strand break repair (1)
- drift-diffusion model (1)
- drug (1)
- drug repurposing (1)
- dual RNA-seq (1)
- early detection (1)
- early neural precursors (1)
- early-life stress (1)
- early-onset isolated dystonia (1)
- eating behavior (1)
- eating disorders (1)
- echocardiography (1)
- ecoli_VF_collection (1)
- ecological validity (1)
- ectopic bone formation (1)
- ectopic release (1)
- effector protein (1)
- efflux pump (1)
- electroacupuncture (1)
- electrode scaffold (1)
- electroencephalogram (1)
- electron cryomicroscopy (1)
- electron tomography (1)
- electrospinning (1)
- embryonale Maus (1)
- emojis (1)
- emotional (1)
- emotional feedback (1)
- emotional information processing (1)
- emotional interference (1)
- emotional processing (1)
- emotionale Anspannung (1)
- emotionale Interferenz (1)
- emotions (1)
- endocytic recycling (1)
- endocytosis (1)
- endophyte (1)
- enhancers (1)
- enoyl ACP reductase (1)
- enoyl reductase (1)
- enoyl-ACP reductase (1)
- entero-aggregative-haemorrhagic Escherichia coli (EAHEC) (1)
- enteroinvasive (1)
- eosinophil (1)
- epidemiology (1)
- epigenetic (1)
- epithelial to mesenchymal transition (1)
- epithelial-mesenchymal transition (1)
- ereigniskorreliertes Potential (1)
- etiology (1)
- evozierte Potentiale (1)
- exotische Pflanzen (1)
- experimentelle autoimmune Enzephalomyelitis (1)
- exposition training (1)
- exposure therapy (1)
- extinction (1)
- extinction dynamics (1)
- extracellular signal–regulated kinases 1/2 (1)
- extrazelluläre Matrix (1)
- eye contact (1)
- eye tracking (1)
- eye-tracking (1)
- eyetracking (1)
- fabry disease (1)
- facial expressions (1)
- fast MR imaging (1)
- fatty acid biosynthesis (1)
- fatty acid synthesis (1)
- fear potentiated startle response (1)
- feasibility (1)
- feral bees (1)
- fgf (1)
- fibroblast growth factors (1)
- fibromyalgia sydrome (1)
- filtering (1)
- fitness (1)
- flagellum (1)
- fluorescence imaging (1)
- fluorescence microscopy (1)
- fluorescence resonance energy transfer (1)
- fluorescence resonance energy transfer (FRET) (1)
- flytrap (1)
- fmri activity (1)
- foamy viruses (1)
- follicular regulatory T cell (1)
- follikuläre regulatorische T-Zelle (1)
- food craving (1)
- forager (1)
- foraging (1)
- foraging activity (1)
- forest landscape (1)
- fragment screening (1)
- frameshifting (1)
- freezing of gait (1)
- frequency modulation (1)
- frogs (1)
- fruit cuticle (1)
- frühe neurale Vorläufer (1)
- functional connectivity (1)
- functional imaging (1)
- functional membrane microdomains (1)
- functional modules (1)
- functional neuroimaging (1)
- functional resting-state connectivity (1)
- functional selectivity (1)
- functional studies (1)
- fungal endophytes of grasses (1)
- fungi (1)
- fungus (1)
- funktionale Bildgebung (1)
- funktionelle Magnetresonanztomographie (1)
- funktionelle Module (1)
- funktionelle Resting-State Konnektivität (1)
- g-factor (1)
- gait analysis (1)
- gait initiation (1)
- gametocyt (1)
- gametocyte (1)
- gametogenesis (1)
- gamma delta T cells (1)
- gastrointestinal infection (1)
- gastronintestinal microbiota (1)
- gene environment interaction (1)
- gene-environmental interaction (1)
- genetic cytokine polymorphism (1)
- genetic modification (1)
- genetic screen (1)
- genome stability (1)
- genomic damage (1)
- genomic imaging (1)
- genomische Schäden (1)
- genotoxic agents (1)
- genotoxische Agenzien (1)
- gepaarte Vagusnerv-Stimulation (1)
- germinal center (1)
- germinative cell (1)
- gezielte Therapie (1)
- glioblastoma multiforme (1)
- glioma (1)
- glutamate decarboxylase 65 (1)
- glycine receptor (1)
- glycine receptor autoantibodies (1)
- glycophytes (1)
- glycoprotein GPV (1)
- gonococcal (1)
- gonococcal infection (1)
- grass (1)
- guanine nucleotide exchange factor (1)
- guanylyl cyclase (GC) (1)
- guard cells (1)
- gustation (1)
- habitat (1)
- haloacid dehalogenase phosphatase (1)
- hearing (1)
- heart failure (1)
- heat shock proteins (1)
- hematopoiesis (1)
- hereditary spastic paraplegia (1)
- hiPSC aggregation (1)
- hibernation (1)
- high-pressure freezing/freeze substitution (1)
- high-throughput sequencing (1)
- histone variants (1)
- hnRNP (1)
- hnRNP R (1)
- homeostasis (1)
- homoFRET (1)
- honey bee density (1)
- honey bees (1)
- honeybee*s (1)
- honeybees (1)
- host colonization (1)
- huh6 (1)
- human (1)
- human adipose tissue (1)
- human chromosome 6 (1)
- human factor H (1)
- human induced pluripotent stem cells (1)
- human intestinal epithelium (1)
- human parthenogenetic neural stem cells (1)
- human parthenogenetic stem cells (1)
- human plasma (1)
- human primary cells (1)
- humanen induzierte pluripotente Stammzellen (1)
- humaner Faktor H (1)
- humans (1)
- hyaline cartilage (1)
- hyaluronic acid (1)
- hybrid (1)
- hybrid assembly (1)
- hydrogel (1)
- hyperekplexia (1)
- iNKT cell (1)
- iPSC-derived CMs (iPSC-CMs) (1)
- iPSCs (1)
- icaADBC (1)
- imaging (1)
- immediate early genes (1)
- immune cell recruitment (1)
- immune escape (1)
- immune evasion (1)
- immune response (1)
- immune system (1)
- immunologic tolerance (1)
- immunological synapse (1)
- immunotherapy (1)
- imprinting. (1)
- imunology (1)
- in situ microscopy (1)
- in vitro Kulturmodelle (1)
- in vitro Modelle (1)
- in vitro Testmodell (1)
- in vitro Testsystem (1)
- in vitro model (1)
- in vitro model system of inherited cardiomyopathies (1)
- in vitro neural differentiation (1)
- in vitro-Testsystem (1)
- in vivo study (1)
- induced pluripotent stem cells (1)
- induced pluripotent stem cells (iPSCs) (1)
- induzierte Phosphatasen MKP-1 und MKP-2 (1)
- infection biology (1)
- infectionmodel (1)
- infectionprocess (1)
- inferior frontal gyrus (1)
- inferiore frontale Gyrus (1)
- inflammatorische Gewebeschäden (1)
- influenza A virus (1)
- information processing (1)
- information use (1)
- inherited macrothrombocytopenia (1)
- inhibitor residence time (1)
- inhibitory postsynapse (1)
- innate immunity (1)
- insect visual learning (1)
- insects (1)
- insertion-site deep sequencing (1)
- instrumentelle Aktivitäten des täglichen Lebens (1)
- insulin (1)
- insulin resistance (1)
- intact bone imaging (1)
- integrins (1)
- interaction stress and circadian system (1)
- interactions (1)
- interferon (1)
- interleukin-5 signaling (1)
- internal dosimetry (1)
- interphase gap (1)
- interpulse interval (1)
- intestinal mucus (1)
- intestine (1)
- intracellular calcium release (1)
- intracellular domain (1)
- intraoperative Ankopplungseffizienz (1)
- intravenöse Immunglobuline (1)
- intrazelluläre Domäne (1)
- invariant NKT cells (1)
- invariante NKT Zellen (1)
- invasive meningococcal diseases (1)
- invasive pulmonary aspergillosis (1)
- ion channel (1)
- iron oxide contrast agent (1)
- ischemia reperfusion injury (1)
- ischämischer Schlaganfall (1)
- islets of Langerhans (1)
- jasmonate (1)
- juvenile hormone (1)
- kardiales Gewebe (1)
- kardiales Tissue Engineering (1)
- kardiovaskuläre Risikofaktoren (1)
- ketamine anaesthesia (1)
- kidney development (1)
- kinesin (1)
- knockout (1)
- knockout-mice (1)
- kognitive Defizite (1)
- kognitive Remediation (1)
- kolorektale Karzinomzelllinien (1)
- konventionelle CD4 T Zellen (1)
- koronare Herzerkrankung (1)
- kutikuläre Kohlenwasserstoffe (1)
- kutikuläre Wasserpermeabilität (1)
- kutikuläres Blattwachs (1)
- l-type calcium channel antagonist (1)
- labeling techniques (1)
- lambdoid phage resistance (1)
- lambdoid prophage (1)
- laminar recording (1)
- lasiocarpine (1)
- lasp (1)
- late enhancement (1)
- late gadolinium-enhancement (1)
- leaf cuticle (1)
- leaf-cutting ants (1)
- learned helplessness (1)
- learning and behaviour (1)
- leishmaniasis (1)
- lightsheet microscopy (1)
- lipid remodeling (1)
- liver (1)
- local point-spread function (1)
- local translation (1)
- locomotor network (1)
- logging (1)
- lokale Proteinsynthese (1)
- long-term memory formation (1)
- luciferase assay (1)
- lymph node stromal cells (1)
- lymph node transplantation (1)
- lysosome (1)
- lytic infection (1)
- lytic phage resistance (1)
- mRNA (1)
- mRNA processing (1)
- mRNA translation (1)
- mRNA-Translation (1)
- mTOR (1)
- machine learning (1)
- macrocolony (1)
- macrophage (1)
- macrophages (1)
- malnourishment (1)
- mammalian cells (1)
- marine sponges (1)
- mastitis (1)
- mastitis-associated Escherichia coli (MAEC) (1)
- mean first passage time (1)
- measles virus infection (1)
- mechanistische Modellierung (1)
- mechanosensing (1)
- mechanosensitive channels (1)
- medaka (1)
- medical device (1)
- megachilid bees (1)
- megakaryocytes (1)
- melanocytic nevi (1)
- melanoma cancer (1)
- melanoma dedifferentiation (1)
- melt electrowriting (1)
- membrane dynamics (1)
- membrane trafficking (1)
- memory (1)
- memory B cells (1)
- meniscus implant (1)
- menschliches Darmepithel (1)
- mental health (1)
- mesenteric lymph node (1)
- mesoscopic (1)
- metabolic modelling (1)
- metabolic theory of ecology (1)
- metabolite repair (1)
- metabolomics (1)
- metallo protease (1)
- metapopulation (1)
- metatranscriptome (1)
- methionine biosynthesis (1)
- methylphenidate (1)
- miR-17~92 (1)
- miR-21 (1)
- miR-223-5p (1)
- miR-23a cluster (1)
- miR-26 (1)
- miRNA Biogenesis (1)
- miRNA target (1)
- mice (1)
- micro processor complex (1)
- microRNA biogenesis (1)
- microbial transmission (1)
- microbiology (1)
- microbiota (1)
- microcircuitry (1)
- microelectrode array (1)
- microglomeruli (1)
- microinjection (1)
- micronuclei (1)
- microphysiologic 3D tumour model (1)
- microprocessor complex (1)
- microrna (1)
- mild cognitive impairment (1)
- minimum leaf conductance (1)
- mitochondria (1)
- mitogen cascade (1)
- mobil phone radiation (1)
- molecular biology of cytokines (1)
- molecular mechanism (1)
- monarch butterfly (1)
- monoclonal antibody (1)
- monoclonal antibody 103.2 (1)
- monoclonal antibody 20.1 (1)
- monocyte-derived dendritic cells (1)
- monozytenderivierte dendritische Zellen (1)
- mossy fiber synapse (1)
- mossy fiber terminal (1)
- motoneurons (1)
- motor neuron (1)
- mouse models (1)
- mouse platelets (1)
- movement disorders (1)
- movement interaction (1)
- mucormycosis (1)
- mukosale Immunantwort (1)
- multi-drug-resistance (1)
- multi-modal stimuli (1)
- multi-photon microscopy (1)
- multi-pinhole collimation (1)
- multiphoton microscopy (1)
- multiple sclerosis (1)
- murine (1)
- murine leishmaniasis (1)
- murines Modell der aufsteigenden Harnwegsinfektion (1)
- muscarinic m ACh receptors (1)
- mushroom body (1)
- mushroom body calyx microglomeruli (1)
- muskarinische Rezeptoren (1)
- mutualism (1)
- mutually exclusive expression (1)
- myelin barrier (1)
- myeloablation (1)
- myeloid derived suppressor cells (1)
- myocardial deformation (1)
- myocarditis (1)
- myokardiale Arbeit (1)
- nanotubes (1)
- ncRNA (1)
- near-infrared spectroscopy (1)
- nest climate (1)
- nesting behaviour (1)
- networkanalysis (1)
- neuer anti-infektiver Substanzen (1)
- neural biomarkers (1)
- neurodevelopment (1)
- neurodevelopmental disorders (1)
- neuroepithelial progenitors (1)
- neuroepitheliale Vorläufer (1)
- neurogenesis (1)
- neuroinflammation (1)
- neurologin-2 (1)
- neuromodulation (1)
- neuron (1)
- neuronal activation (1)
- neuronal cell death (1)
- neuronal excitability (1)
- neuronal network (1)
- neuronal nitric oxide synthase (1)
- neuronal visual system (1)
- neuronale Stickstoffmonoxidsynthase (1)
- neuropathic pain (1)
- neuropaticher Schmerz (1)
- neuropeptide S receptor gene (1)
- neuropeptide Y (1)
- neuropeptides (1)
- neuroprotection (1)
- neuropsychiatric disorders (1)
- neuropsychiatrische Störungen (1)
- neuroscience (1)
- neutrophil (1)
- neutrophils (1)
- next generation sequencing (1)
- nicht-viraler Gentransfer (1)
- nicht-visuell (1)
- nichtinvasive Bildgebung (1)
- nitric oxide (1)
- nitrogen regulation (1)
- nociception (1)
- nociceptors (1)
- non-coding RNA (1)
- non-ionizing radiation (1)
- non-visual (1)
- nuclear architecture (1)
- nuclear export signal (1)
- nuclear localization signal (1)
- nuclesosome positioning (1)
- nucleus (1)
- nurse bee (1)
- nutrients (1)
- nutrition (1)
- object segmentation (1)
- oilpalm plantation (1)
- oligopeptide transport (1)
- oncogenes (1)
- oncogenic signalling network (1)
- onkogenes Signalnetzwerk (1)
- opening of chromatin (1)
- opioid peptide (1)
- opioid receptor (1)
- opioid receptors (1)
- optogenetics (1)
- orientation (1)
- oscillations (1)
- ovarian cancer (1)
- ovarian carcinoma (1)
- oxidative stress (1)
- oxidativer Stress (1)
- oxidierte Phospholipide (1)
- p34 (1)
- p38MAPK (1)
- p44 (1)
- p97 (1)
- pICln (1)
- pain (1)
- pain regulation (1)
- pancreatic cancer (1)
- pancreatic differentiation (1)
- panic disorder (1)
- parasitology (1)
- parathyroid hormone (1)
- parkinson's disease (1)
- pathophysiological mechanisms (1)
- pathotypes (1)
- pdxp (1)
- pea aphid (1)
- pediatric hematology oncology (1)
- peptide engineering (1)
- peptide-based interleukin-5 inhibitor (1)
- performance evaluation (1)
- perfused hydrogel (1)
- peripheral analgesia (1)
- peripheral nerve trauma (1)
- peripheral nervous system (1)
- permeability (1)
- permeance (1)
- permeation (1)
- personality traits (1)
- pflegende Angehörige (1)
- phage resistance (1)
- pharmacology (1)
- pheromone (1)
- phosphoantigen (1)
- phosphoglycolate phosphatase (1)
- phospholipase D (1)
- phosphorylation (1)
- phosphorylation sites (1)
- photoinduced electron transfer (1)
- photoinduzierter Elektronentransfer (1)
- phylogeny (1)
- plant defense (1)
- plant-bee visitation networks (1)
- platelet aggregation (1)
- platelet biogenesis (1)
- pluripotency (1)
- podosome formation (1)
- point-of-care (1)
- polarization (1)
- polarized epithelium (1)
- pollen (1)
- pollen analysis (1)
- pollen foraging (1)
- pollen metabarcoding (1)
- pollinator friendly plants (1)
- polyethism (1)
- polymer (1)
- polymorphism (1)
- polyradiculoneuropathy (1)
- population attributable fraction (1)
- population dynamics (1)
- post-transcriptional regulation (1)
- postradiogene Xerostomie (1)
- posttranscriptional regulation (1)
- potenzielles therapeutisches Target (1)
- precision medicine (1)
- preclinical PET (1)
- predictors and correlates (1)
- prefrontal cortex (1)
- prenatal stress (1)
- presynaptic (1)
- primary ciliary dyskinesia (1)
- primary-cell-derived immortalized cell line (1)
- primate (1)
- probiotica (1)
- proboscis extension response (1)
- profiles (1)
- progenitors (1)
- programmed ribosomal frameshifting (1)
- promoter invasion (1)
- proplatelets (1)
- protease activity (1)
- protease inhibitor (1)
- proteasome (1)
- protein (1)
- protein disulfide isomerase (1)
- protein folding (1)
- protein immobilization (1)
- protein kinase D1 (1)
- protein kinase a (1)
- protein regulation and expression (1)
- protein-DNA interactions (1)
- protein-lipid interactions (1)
- protein-protein-interaction (1)
- pränataler Stress (1)
- psychophysiology (1)
- psychosocial resilience (1)
- psychosocial stress (1)
- psychotherapeutic intervention (1)
- psychotherapeutische Intervention (1)
- pyrrolidine carboxamides (1)
- quiescence (1)
- rIFG (1)
- rRNA processing (1)
- rac1 inhibitors (1)
- radial MR imaging (1)
- radionuclide (1)
- radionuclide therapy (1)
- ranskranielle magnetische Stimulation (1)
- rapid evolution (1)
- rat (1)
- reactive oxygen species (1)
- real-time imaging (1)
- reception (1)
- receptor signaling (1)
- reciprocity (1)
- reconstruction (1)
- recurrence (1)
- reduction of ERK1/2 phosphorylation (1)
- reduction of cells proliferation (1)
- regenerative medicine (1)
- regional analgesia (1)
- regulatorische T Zellen (1)
- regulatory RNA (1)
- reinforcement sensitivity theory (1)
- relapse (1)
- replication (1)
- reporter screen (1)
- research software (1)
- resilience (1)
- resin (1)
- resolution (1)
- responsiveness (1)
- resting tremor (1)
- restriction factors (1)
- resveratrol (1)
- retinal development (1)
- reverse Transkription (1)
- reverse genetics (1)
- reverse transcription (1)
- reversible oxidation (1)
- rheumatoide Arthritis (1)
- rho gtpases (1)
- ribosomal RNA (1)
- ribosomale RNS (1)
- ribosome biogenesis (1)
- ribosome profiling (1)
- riboswitch (1)
- risk factors (1)
- rodent model (1)
- rtPA (1)
- salinen Wachstumsbedingungen (1)
- salt stress (1)
- sand mine (1)
- saure Sphingomyelinase (1)
- scale-up (1)
- schnelle Evolution (1)
- screening (1)
- second messenger (1)
- secondary metabolites (1)
- secondary prevention (1)
- secreted aspartic protease (1)
- secreted factors (1)
- secretion (1)
- secretory properties (1)
- seed coat (1)
- seed treatment (1)
- self-activation (1)
- self-regulation (1)
- self-targeting CRISPR-Cas (1)
- senecionine (1)
- seneciphylline (1)
- senescence (1)
- sensitivity (1)
- sensorische Neurone (1)
- sequence-structure (1)
- serine protease (1)
- serotonerges System (1)
- serotonin (1)
- serotonin deficiency (1)
- serotonin transporter (1)
- serum retention (1)
- sexual stage surface proteins (1)
- short neuropeptide F (1)
- sifA (1)
- sigma factor (1)
- signaling pathway (1)
- signaltransduction (1)
- similarity (1)
- single cell anatomy (1)
- single molecule microscopy (1)
- single-cell RNA sequencing (1)
- single-cell genomics (1)
- single-molecule imaging (1)
- single-molecule localization microscopy (1)
- single-molecule microscopy (1)
- site directed immobilization (1)
- site-specific immobilization (1)
- skin (1)
- skin biopsy (1)
- skin cancer (1)
- skin model (1)
- small RNA expression (1)
- small fiber pathology (1)
- small intestinal submucosa (1)
- small proteins (1)
- small-animal SPECT (1)
- smell (1)
- smells (1)
- snoRNA (1)
- social cognition (1)
- social interaction (1)
- social stimuli (1)
- social understanding (1)
- solid tumour (1)
- solitary bee nests (1)
- somatic resilience (1)
- somatostatin receptor antagonists (1)
- soziale Insekten (1)
- spatial organization (1)
- specific phobia (1)
- speckle tracking (1)
- spezifische Phobie (1)
- sphingolipid (1)
- spider phobia (1)
- spiral trajectory (1)
- splicing (1)
- sponge microbiome (1)
- spontaneous neuronal activity (1)
- ssVEP (1)
- stachellose Biene (1)
- stachellose Bienen (1)
- steady-state visually evoked potentials (1)
- stem (1)
- stem cells (1)
- stiff person syndrome (1)
- stingless bees (1)
- store-operated calcium entry (1)
- strain (1)
- strain rate (1)
- stream (1)
- streptozotocin (1)
- stress response (1)
- striatum (1)
- stroop task (1)
- structural mechanism (1)
- structure analysis (1)
- structure based drug design (1)
- structure-based drug design (1)
- structure-function relationships (1)
- stx-Phagen (1)
- stx-phages (1)
- subcutaneous implanation (1)
- subthalamic nucleus (1)
- sugar perception (fructose, sucrose) (1)
- sugar receptor (1)
- sumo (1)
- sumoylation (1)
- superagonistische Funktion (1)
- superparamagnetische Eisenoxid Kontrastmittel (1)
- suspension culture (1)
- sustained fear (1)
- synaptische Plastizität (1)
- synthetic lethal interaction (1)
- synthetisch lethale Interaktion (1)
- systemic inflammation (1)
- systems biology (1)
- t cell (1)
- tactil (1)
- tactile (1)
- tapeworm (1)
- tdcs (1)
- telomere-associated protein (1)
- temperate zones (1)
- temporal information transfer (1)
- temporal organization (1)
- temporo-parietal junction (1)
- terahertz radiation (1)
- terpenes (1)
- test system (1)
- th1/th2 polarization (1)
- therapeutic strategy (1)
- therapeutisches Target (1)
- therapy (1)
- therapy of glioblastoma (1)
- therapy outcome (1)
- therapy simulation (1)
- thermal orientation (1)
- thermotolerance (1)
- theta burst stimulation (1)
- think/no-think (1)
- thiostrepton (1)
- threat conditioning (1)
- thrombin (1)
- thrombo-inflammation (1)
- thrombosis (1)
- thyroid stimulating hormone receptor (1)
- time-correlated single photon counting (TCSPC) (1)
- time-resolved anisotropy (1)
- timing (1)
- tissue model (1)
- tolerance (1)
- tolerogen (1)
- tolerogenic (1)
- tool (1)
- tool-use (1)
- torque meter (1)
- tracheal cytotoxin (1)
- trachomatis (1)
- trade-offs (1)
- trans-Golgi network (1)
- transcranial Direct Current Stimulation (tDCS) (1)
- transcranial direct current stimulation (1)
- transcription factor (1)
- transcription regulator (1)
- transcription/replication conflicts (1)
- transcriptional termination site (1)
- transcriptome (1)
- transcriptome analysis (1)
- transcriptome data analysis (1)
- transcriptome profiling (1)
- transcriptomic analysis (1)
- transcutaneous vagus nerve stimulation (1)
- transfer (1)
- transgenes Modell (1)
- transient dynamics (1)
- transmission blocking vaccine (1)
- transpiration barrier (1)
- transport (1)
- transporter regulator (1)
- tuberculosis (1)
- tumor angiogenesis (1)
- tumor metabolism (1)
- tumor vascular morphologie (1)
- tumorspezifische Therapie (1)
- tumour (1)
- tumour microenvironment (1)
- tumour stroma (1)
- two-component (1)
- type 1 diabetes (1)
- tyrosine kinase (1)
- ubiquitin (1)
- ubiquitin chain formation (1)
- ubiquitin linkage specificity (1)
- ubiquitin recognition (1)
- ultimatum game (1)
- unconventional T cells (1)
- unklarer Signifikanz (1)
- vaccine (1)
- vacuole (1)
- validation study (1)
- variant surface glycoprotein (1)
- variational network (1)
- varroa (1)
- vascular cell adhesion molecules (1)
- vascular smooth muscle cells (1)
- vascular system (1)
- vaskuläre Adhäsionsmoleküle (1)
- vaskuläre glatte Muskelzelle (1)
- vaskuläre glatte Muskelzellen (1)
- vasp (1)
- vav2 (1)
- vdr (1)
- ve-cadherin (1)
- venus (1)
- viral genome packaging (1)
- viral miRNAs (1)
- virological synapse (1)
- virtual reality T-maze (1)
- virulence factors (1)
- virus transmission (1)
- vision (1)
- visual attention (1)
- visual cue (1)
- visual perception (1)
- visuelles Langzeitgedächtnis (1)
- vitamin d (1)
- vitamin d receptor (1)
- vmPFC (1)
- waggle dance (1)
- waggle dance decoding (1)
- water fat separation (1)
- water loss (1)
- wax (1)
- wild-living honey bees (1)
- workbench (1)
- x-ray micro computed tomography (1)
- xiphophorus (1)
- yeast (1)
- zeitgeber (1)
- zeitlich Informationsübertragung (1)
- zeitliche Organisation (1)
- zeitliche Trends (1)
- zentrale Spindel und Mittelkörper (1)
- zielgerichtete Therapien (1)
- zonal Hydrogels (1)
- zyklische Peptide (1)
- µ-Opioid Rezeptor (1)
- Ängstliche Depression (1)
- Ätiologie (1)
- Ölpalmenanbau (1)
- Überexpression (1)
- Übertragungsfunktion (1)
- Übung (1)
- ß-adrenerge Rezeptoren (1)
- ß-adrenergic Receptors (1)
- ΔNp63 (1)
- α-synuclein (1)
- β3 adrenergic receptor (1)
- γδ T cells (1)
Institute
- Graduate School of Life Sciences (986) (remove)
Sonstige beteiligte Institutionen
- Helmholtz Institute for RNA-based Infection Research (HIRI) (5)
- Universitätsklinikum Münster (3)
- Rudolf Virchow Center for Integrative and Translational Bioimaging, University of Würzburg (2)
- Zentrum für Infektionsforschung (ZINF) Würzburg (2)
- Bio-Imaging Center Würzburg (1)
- Biomedical Center Munich, Department of Physiological Chemistry, Ludwig-Maximilians-Universität München (1)
- CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - the development agency of the Brazilian Federal Government (1)
- CBIO, University of Cape Town, South Africa (1)
- Carl-Ludwig-Institut für Physiologie, Universität Leipzig (1)
- Chair of Experimental Biomedicine I (1)
Die asexuellen Sporen von Aspergillus fumigatus sind ubiquitär verbreitete Luftkeime. Als Saprophyt ist dieser opportunistisch humanpathogene Pilz darauf spezialisiert, polymere Substanzen aus dem umgebenden Milieu zu zersetzen, um daraus die von ihm benötigten Nährstoffe zu generieren und aufzunehmen. Die Fähigkeit, verschiedene Stickstoff- und Kohlenstoffquellen zu verwerten, trägt dabei zu seiner Virulenz bei und hierbei scheint die extrazelluläre Proteolyse eine wichtige Rolle zu spielen. Sekretierte Proteasen, die das umgebende Gewebe während einer Infektion mit A. fumigatus erschließen, könnten somit zu dessen Pathogenität beitragen. Dementsprechend sollte im Rahmen dieser Arbeit die Bedeutung einer Regulation der extrazellulären proteolytischen Aktivität von A. fumigatus für dessen Virulenz untersucht werden. Dies geschah durch Untersuchungen eines konservierten Transkriptionsfaktors, PrtT. Dabei stellte sich heraus, dass PrtT die Expression der drei Hauptproteasen von A. fumigatus, Alp, Mep und Pep stark beeinflusst, in einem murinen Tiermodell der pulmonaren Aspergillose scheint dieser Regulator jedoch keine Rolle für die Pathogenität von A. fumigatus zu spielen. Um einen weiteren Aspekt des pilzlichen Aminosäurestoffwechsels zu beleuchten, wurde die Biosynthese der aromatischen Aminosäuren als mögliche Virulenzdeterminate untersucht. Für den Menschen sind diese Aminosäuren essentiell, weshalb dieser Syntheseweg ein mögliches Ziel für antimykotische Substanzen darstellen könnte. Es konnten mehrere für A. fumigatus essentielle Komponenten des Shikimatweges identifiziert werden, des Weiteren wurden Deletionsmutanten in den Genen aroC und trpA, die für die Chorismatmutase bzw. Anthranilatsynthase der Biosynthese von Phenylalanin und Tyrosin bzw. Tryptophan kodieren, erzeugt und phänotypisch charakterisiert. Deren Untersuchung in einem alternativen Tiermodell der Aspergillose zeigte eine deutlich attenuierte Virulenz. Diese Ergebnisse verdeutlichen, wie wichtig die Biosynthese der aromatischen Aminosäuren für das Wachstum von A. fumigatus ist, und dass ein Eingriff in diesen Syntheseweg eine lohnende Strategie zur Entwicklung neuer Antimykotika sein könnte. Die hier präsentierten Ergebnisse unterstreichen die für den Schimmelpilz A. fumigatus typische Redundanz bezüglich extrazellulärer proteolytischer Enzyme und dass diese nur bedingt hinsichtlich ihres Virulenzbeitrags untersucht werden können. Im Gegensatz hierzu lassen sich bestimmte Stoffwechselwege, die oftmals durch einzigartige Genprodukte katalysiert werden, unter Umständen besser als unspezifische aber vielversprechende Virulenzdeterminanten identifizieren.
While beneficial sponge-microbe associations have received much attention in recent years, less effort has been undertaken to investigate the interactions of sponges with potentially pathogenic microorganisms. Thus, the aim of this study was to examine two selected Caribbean disease conditions, termed “Sponge Orange Band” and “Sponge White Patch”, via ecological and molecular methods. Sponge Orange Band (SOB) disease affects the prominent Caribbean barrel sponge Xestospongia muta that is counted among the high-microbial-abundance (HMA) sponges, whereas Sponge White Patch (SWP) disease affects the abundant rope sponge Amphimedon compressa that belongs to the low-microbial-abundance (LMA) sponges. I have documented for both Caribbean sponge diseases a disease progression going along with massive tissue destruction as well as loss of the characteristic microbial signatures. Even though new bacteria were shown to colonize the bleached areas, the infection trials revealed in both cases no indication for the involvement of a microbial pathogen as an etiologic agent of disease leaving us still in the dark about the cause of Sponge Orange Band as well as Sponge White Patch disease.
Background: There is extensive evidence that explicit memory, which involves conscious recall of encoded information, can be modulated by emotions; emotions may influence encoding, consolidation or retrieval of information. However, less is known about the modulatory effects of emotions on procedural processes like motor memory, which do not depend upon conscious recall and are instead demonstrated through changes in behaviour. Experiment 1: The goal of the first experiment was to examine the influence of emotions on motor learning. Four groups of subjects completed a motor learning task performing brisk isometric abductions with their thumb. While performing the motor task, the subjects heard emotional sounds varying in arousal and valence: (1) valence negative / arousal low (V-/A-), (2) valence negative / arousal high (V-/A+), (3) valence positive / arousal low (V+/A-), and (4) valence positive / arousal high (V+/A+). Descriptive analysis of the complete data set showed best performances for motor learning in the V-/A- condition, but the differences between the conditions did not reach significance. Results suggest that the interaction between valence and arousal may modulate motor encoding processes. Since limitations of the study cannot be ruled out, future studies with different emotional stimuli have to test the assumption that exposure to low arousing negative stimuli during encoding has a facilitating effect on short term motor memory. Experiment 2: The purpose of the second experiment was to investigate the effects of emotional interference on consolidation of sequential learning. In different sessions, 6 groups of subjects were initially trained on a serial reaction time task (SRTT). To modulate consolidation of the newly learned skill, subjects were exposed, after the training, to 1 of 3 (positive, negative or neutral) different classes of emotional stimuli which consisted of a set of emotional pictures combined with congruent emotional musical pieces or neutral sound. Emotional intervention for each subject group was done in 2 different time intervals (either directly after the training session, or 6 h later). After a 72 h post-training interval, each group was retested on the SRTT. Re-test performance was evaluated in terms of response times and accuracy during performance of the target sequence. Emotional intervention did not influence either response times or accuracy of re-testing SRTT task performance. However, explicit awareness of sequence knowledge was enhanced by arousing negative stimuli applied at 0 h after training. These findings suggest that consolidation of explicit aspects of procedural learning may be more responsive toward emotional interference than are implicit aspects. Consolidation of different domains of skill acquisition may be governed by different mechanisms. Since skill performance did not correlate with explicit awareness we suggest that implicit and explicit modes of SRTT performance are not complementary. Experiment 3: The aim of the third experiment was to analyze if the left hemisphere preferentially controls flexion responses towards positive stimuli, while the right hemisphere is specialized towards extensor responses to negative pictures. To this end, right-handed subjects had to pull or push a joystick subsequent to seeing a positive or a negative stimulus in their left or right hemifield. Flexion responses were faster for positive stimuli, while negative stimuli were associated with faster extensions responses. Overall, performance was fastest when emotional stimuli were presented to the left visual hemifield. This right hemisphere superiority was especially clear for negative stimuli, while reaction times towards positive pictures showed no hemispheric difference. We did not find any interaction between hemifield and response type. Neither was there a triple interaction between valence, hemifield and response type. In our experimental context the interaction between valence and hemifield seems to be stronger than the interaction between valence and motor behaviour. From these results we suppose that under certain conditions a hierarchy scaling of the asymmetry patterns prevails, which might mask any other existing asymmetries.
Ceramide sind biologisch aktive Sphingolipide, die verschiedene zelluläre Signalwege regulieren, meist im Zusammenhang mit der Induktion von Apoptose oder der Regulation des Zellzyklus. Darüber hinaus wurde in der Literatur beschrieben, dass Ceramide die Zytoskelettdynamik unterschiedlicher Zelltypen beeinflussen, die Bedeutung von Ceramiden für die Funktion von T-Zellen wurde allerdings bisher wenig untersucht. In der vorliegenden Arbeit konnte gezeigt werden, dass die exogene Akkumulation von Ceramiden ebenso wie die Generierung von Ceramiden durch bSMase die Adhärenz von T-Zellen an FN bzw. ICAM-1 beeinträchtigt. Des Weiteren konnte eine verminderte T-Zell-Polarisierung auf FN sowie eine reduzierte Chemotaxis und Motilität ceramidmodifizierter T-Zellen in Antwort auf SDF-1 nachgewiesen werden. In Übereinstimmung mit der Unfähigkeit ceramidmodifizierter Zellen morphologisch zu polarisieren wird ferner die Relokalisation von Oberflächenmolekülen und intrazellulärer Proteine durch die Akkumulation von Ceramiden gestört. Überdies konnte in dieser Arbeit gezeigt werden, dass Ceramide mit dem Aktivierungsstatus von Akt und ERM-Proteinen interferieren, da eine verminderte stimulationsabhängige Phosphorylierung von Akt und ERM-Proteinen in ceramidmodifizierten Zellen nachgewiesen wurde. Ein wesentlicher Schritt im Verlauf der T-Zell-Aktivierung ist die Ausbildung einer immunologischen Synapse mit dendritischen Zellen. In dieser Arbeit konnte gezeigt werden, dass, obwohl ceramidreiche Membrandomänen von der Kontaktstelle ausgeschlossen werden, Konjugatfrequenz und Architektur der IS durch die Induktion von Ceramiden nicht beeinflusst werden, da eine normale Verteilung von CD3 und des MTOC beobachtet wurde. Allerdings wird die Funktionalität der Konjugate durch die Induktion von Ceramiden beeinträchtigt. Ceramidmodifizierte Zellen waren nur eingeschränkt in der Lage Orai1 und Stim1 zur Kontaktfläche mit DCs zu translozieren. In Übereinstimmung mit diesen Befunden wurde auch ein verminderter Calcium-Einstrom sowie eine verminderte Proliferation infolge der Akkumulation von Ceramiden detektiert. Zusammenfassend konnte in dieser Arbeit gezeigt werden, dass Ceramide wesentliche Prozesse im Verlauf der T-Zell-Aktivierung beeinflussen, so dass die pathogeninduzierte Generierung von Ceramiden einen möglichen Mechanismus darstellt, die Funktion von T-Zellen zu beeinträchtigen.
Single-molecule microscopy is one of the decisive methodologies that allows one to clarify cellular signaling in both spatial and temporal dimentions by tracking with nanometer precision the diffusion of individual microscopic particles coupled to relevant biological molecules. Trajectory analysis not only enables determination of the mechanisms that drive and constrain the particles motion but also to reveal crucial information about the molecule interaction, mobility, stoichiometry, all existing subpopulations and unique functions of particular molecules. Efficacy of this technique depends on two problematic issues the usage of the proper fluorophore and the type of biochemical attachment of the fluorophore to a biomolecule. The goal of this study was to evolve a highly specific labeling method suitable for single molecule tracking, internalization and trafficking studies that would attain a calculable 1:1 fluorophore-to-receptor stoichiometry. A covalent attachment of quantum dots to transmembrane receptors was successfully achieved with a techinque that amalgamates acyl carrier protein (ACP) system as a comparatively small linker and coenzyme A (CoA)-functionalized quantum dots. The necessity of optimization of the quantum dot usage for more precise calculation of the membrane protein stoichiometries in larger assemblies led to the further study in which methods maximizing the number of signals and the tracking times of diverse QD types were examined. Next, the optimized techniques were applied to analyze behavior of interleukin-5 β-common chain receptor (IL-5Rβc) receptors that are endogenously expressed at low level on living differentiated eosinophil-like HL-60 cells. Obtained data disclosed that perused receptors form stable and higher order oligomers. Additionally, the mobility analysis based on increased in number (>10%) uninterrupted 1000-step trajectories revealed two patterns of confined motion. Thereupon methods were developed that allow both, determination of stoichiometries of cell surface protein complexes and the acquisition of long trajectories for mobility analysis. Sequentially, the aforementioned methods were used to scrutinize on the mobility, internalization and recycling dynamics characterization of a G protein-coupled receptor (GPCRs), the parathyroid hormone receptor (PTHR1) and several bone morphogenetic proteins (BMPs), a member of the TGF-beta superfamily of receptors. These receptors are two important representatives of two varied membrane receptor classes. BMPs activate SMAD- and non-SMAD pathways and as members of the transforming growth factor β (TGF-β) superfamily are entailed in the regulation of proliferation, differentiation, chemotaxis, and apoptosis. For effective ligand induced and ligand independent signaling, two types of transmembrane serine/threonine kinases, BMP type I and type II receptors (BMPRI and BMPRII, respectively) are engaged. Apparently, the lateral mobility profiles of BMPRI and BMPRII receptors differ markedly, which determinate specificity of the signal. Non-SMAD signaling and subsequent osteoblastic differentiation of precursor cells particularly necessitate the confinement of the BMP type I receptor, resulting in the conclusion that receptor lateral mobility is a dominative mechanism to modulate SMAD versus non-SMAD signaling during differentiation. Confined motion was also predominantly observed in the studies devoted to, entailed in the regulation of calcium homeostasis and in bone remodeling, the parathyroid hormone receptor (PTHR1), in which stimulation with five peptide ligands, specific fragments of PTH: hPTH(1–34), hPTHrP(107–111)NH2; PTH(1–14); PTH(1–28) G1R19, bPTH(3–34), first four belonging to PTH agonist group and the last to the antagonist one, were tested in the wide concentration range on living COS-1 and AD293 cells. Next to the mobility, defining the internalization and recycling rates of the PTHR1 receptor maintained in this investigation one of the crucial questions. Internalization, in general, allows to diminish the magnitude of the receptor-mediated G protein signals (desensitization), receptor resensitization via recycling, degradation (down-regulation), and coupling to other signaling pathways (e.g. MAP kinases). Determinants of the internalization process are one of the most addressed in recent studies as key factors for clearer understanding of the process and linking it with biological responses evoked by the signal transduction. The internalization of the PTH-receptor complex occurs via the clathrin-coated pit pathway involving β-arrestin2 and is initiated through the agonist occupancy of the PTHR1 leading to activation of adenylyl cyclase (via Gs), and phosphatidylinositol-specific phospholipase Cβ (via Gq). Taken together, this work embodies complex study of the interleukin-5 β-common chain receptor (IL-5Rβc) receptors, bone morphogenetic proteins (BMPs) and the parathyroid hormone receptor with the application of single-molecule microscopy with the newly attained ACP-quantum dot labeling method and standard techniques.
Die Therapie von bakteriellen Infektionen beruht heutzutage zum Großteil auf dem Einsatz von Antibiotika. Die schnelle Entwicklung und rasche Verbreitung von resistenten Stämmen mancher Erreger gegen diese Antibiotika stellt ein enormes Problem für das Gesundheitswesen dar. Da momentan zur Antibiotikatherapie keine Alternativen bestehen, kommt der Erforschung neuer potenzieller Wirkstoffe eine sehr große Bedeutung zu. In einem Screening-Verfahren lagen die minimalen Hemmkonzentrationen einiger bisquartärer Bisnaphthalimide gegen Staphylococcus aureus und S. epidermidis im Bereich von 0,6 bis 2,5 µg/ml. Die Substanz mit den geringsten minimalen Hemmkonzentrationen war MT02. Daraufhin wurde das Wirkungsspektrum von MT02 gegen Bakterien detaillierter untersucht und gefunden, dass die Substanz vorwiegend gegen Gram-positive Erreger und nicht gegen Gram-negative Bakterien wirksam ist. Zytotoxizitätstests ergaben eine geringe bis nicht nachweisbare Toxizität gegen verschiedene Zelllinien im Bereich von 73 bis mehr als 150 µg/ml. Um die Wirkungsweise von MT02 genauer zu untersuchen wurden zunächst DNA-Microarray-Untersuchungen an S. aureus durchgeführt. Deren Ergebnisse ließen einen Einfluss der Substanz auf viele Gene des DNA-Metabolismus erkennen. Inkorporationsstudien mittels radioaktiver Ganzzellmarkierung bestätigten die Auswirkung von MT02 auf den DNA-Stoffwechsel. Durch kompetitive Inkubation wurde festgestellt, dass MT02 in der Lage ist Ethidiumbromid von DNA zu verdrängen bzw. dessen Bindung zu verhindern. Genauere Untersuchungen mittels Oberflächen-Plasmon-Resonanz ergaben, dass MT02 konzentrationsabhängig, reversibel und sequenzunspezifisch an DNA bindet. Die thermodynamischen Dissoziationskonstanten lagen im Mittel bei ca. 4 x 10-8 mol/l und beschrieben somit eine relativ starke Bindung von MT02 an DNA. Neben diesem primären Wirkungsmechanismus der DNA-Bindung gaben mehrere Befunde Hinweise auf einen sekundären Wirkmechanismus, der die Zellwand-Struktur bzw. Zellwand-Biosynthese beinhaltet. Eine MT02-resistente Mutante von S. aureus HG001 konnte durch vielfaches Passagieren in MT02-haltigem Medium generiert werden. Diese erzeugte bei Wachstum mit hohen Konzentrationen an MT02 einen roten Phänotyp. Die Natur dieses roten Farbstoffes konnte bislang nicht aufgeklärt werden, jedoch gibt es Hinweise, dass dieser auf Abbauprodukte von MT02 zurückzuführen ist. In einem weiteren Projekt wurde mittels Transkriptionsstudien die Auswirkung von verschiedenen bekannten Antibiotika sowie von neuen Wirkstoffen auf das Transkriptom von S. epidermidis untersucht. Die Ergebnisse dieser Studien können durch vergleichende Analysen als Grundlage für die Einordnung des Wirkmechanismus neuer Substanzen dienen.
Growth factor induced signaling cascades are key regulatory elements in tissue development, maintenance and regeneration. Deregulation of the cascades has severe consequences, leading to developmental disorders and neoplastic diseases. As a major function in signal transduction, activating mutations in RAF family kinases are the cause of many human cancers. In the first project described in this thesis we focused on B-RAF V600E that has been identified as the most prevalent B-RAF mutant in human cancer. In order to address the oncogenic function of B-RAF V600E, we have generated transgenic mice expressing the activated oncogene specifically in lung alveolar epithelial type II cells. Constitutive expression of B-RAF V600E caused abnormalities in alveolar epithelium formation that led to airspace enlargements. These lung lesions showed signs of tissue remodeling and were often associated with chronic inflammation and low incidence of lung tumors. Inflammatory cell infiltration did not precede the formation of emphysema-like lesions but was rather accompanied with late tumor development. These data support a model where the continuous regenerative process initiated by oncogenic B-RAF-driven alveolar disruption provides a tumor-promoting environment associated with chronic inflammation. In the second project we focused on wild type B-RAF and its role in an oncogenic-C-RAF driven mouse lung tumor model. Toward this aim we have generated compound mice in which we could conditionally deplete B-RAF in oncogenic-C-RAF driven lung tumors. Conditional elimination of B-RAF did not block lung tumor formation however led to reduced tumor growth. The diminished tumor growth was not caused by increased cell death instead was a consequence of reduced cell proliferation. Moreover, B-RAF ablation caused a reduction in the amplitude of the mitogenic signalling cascade. These data indicate that in vivo B-RAF is dispensable for the oncogenic potential of active C-RAF; however it cooperates with oncogenic C-RAF in the activation of the mitogenic cascade.
When there is an imbalance between reactive oxygen species (ROS) and endogenous antioxidants (glutathione (GSH), superoxide dismutase (SOD), catalase etc.) the oxidative stress is increased and results in the oxidation of lipids, proteins and DNA. Although oxidation of lipids and proteins may also accumulates with age, only DNA oxidation leads to altered genomic information. As one pathway for increased ROS production, many endogenous and exogenous substances activate NADPH oxidase (NOX) enzyme and produce ROS. p47phox is a cytosolic organizer protein which plays an important role in NOX activation. Angiotensin II (Ang II) is an example for an endogenous compound which causes ROS through NOX activation. Rosuvastatin is an example for a drug with antioxidative capacity (upregulation of endogenous antioxidants). It is a lipid lowering drug which also reduces an elevated level of angiotensin II type 1 receptor (AT1R). Commonly, oxidative stress is elevated in ageing and age related diseases (eg. Parkinson’s disease (PD)). The aim of the present study was to investigate the role of NOX derived ROS induced oxidative DNA damage and the influence of ROS in ageing and age related diseases, using different in vitro and in vivo models.
There is evidence that pheromones are communicative signals in animals. However, the existence and function of human pheromones are still under discussion. During the last years several substances have been labeled as putative human pheromones and especially 4,16–androstadien-3-one (androstadienone), found in male and female sweat, became subject of intense investigation. In contrast to common odors androstadienone presumably modulates human physiological and psychological reactions. Data suggest that androstadienone might influence the processing of visual cues, specifically faces or affective stimuli, via projections from the fusiform gyrus and the amygdala. Moreover, attentional processes may be modulated, which is supported by explicit and implicit behavioral data. This thesis includes three experimental studies examining effects of androstadienone exposure on behavioral and cortical reactions to visual and emotional stimuli. The main hypotheses were that androstadienone might influence human behavior to and perception of visual cues. The first study sought to clarify androstadienone effects on attention-related reactions as well as on behavioral tendencies. Motoric approach-avoidance reactions in response to happy and angry facial expressions were investigated in 30 women and 32 men. Participants either inhaled androstadienone or a control solution, without knowing the real content, while performing the following task: they had to push away or to pull towards them a joystick as fast as possible in reaction to either an angry or a happy cartoon face, which was presented on a computer screen. Results showed that androstadienone modulated the participant´s task performance by accelerating the reaction speed compared to the control compound. Faster reactions were observed particularly when reacting to angry faces but not when reacting to happy faces. This might be explained by the finding that human body odors, the source of androstadienone, were found to activate the human fear system, i.e. modulating fear-related attentional processes. Therefore, the quicker reaction towards angry faces with exposure to androstadienone could be due to an enhanced allocation of attentional resources towards fear-related cues like angry faces. Results also showed that androstadienone enhanced men´s approach tendency towards faces independent of emotional expressions. This observation might be explained by androstadienone´s former shown ability to improve attractiveness ratings of other persons. In this regard, the endogenous odor might enhance evaluations of faces in men and, thus, might improve their willingness to approach social stimuli. In contrast to men, women already showed in the control condition higher approach tendency towards faces. Therefore, androstadienone might rather maintain than enhance the approach score in women. In the second study event-related brain potentials (ERPs) triggered by social and non-social visual stimuli were investigated by means of electroencephalography. In a double-blind between-subjects design 51 women participated. Twenty-eight women inhaled androstadienone, whereas 23 women inhaled a control solution. Four different picture categories, i.e. real faces, pictures with couples, pictures with social and non-social scenes, each including three different valence categories, i.e. positive, negative and neutral, should clarify the stimulus type or context androstadienone is acting on. The androstadienone compared to the control odor did not influence brain responses significantly. Explorative analyses, however, suggested that androstadienone influences the processing of faces. While in the control group angry faces elicited larger P300 amplitudes than happy faces, the androstadienone group showed similar P300 amplitudes concerning all emotional expressions. This observation tentatively indicates that the endogenous odor might indeed affect the neuronal responses to emotional facial stimuli, especially late components reflecting evaluative processes. However, this observation has to be verified and further investigated, in particular whether androstadienone caused reduced responses to angry faces or enhanced responses to happy faces. The third study investigated androstadienone effects on face processing especially in men. ERPs elicited by happy, angry and neutral cartoon faces, which were presented on a computer screen, were measured while 16 men, not knowing the applicated odor, inhaled either androstadienone or a control solution. Exposure to androstadienone significantly increased later neuronal responses, the P300 amplitude. This belated component of the ERP reflects attention allocation and evaluative processes towards important stimuli. Therefore, androstadienone might facilitate central nervous face processing by enhancing attention towards these stimuli. In sum, the current results corroborate the notion of androstadienone as an active social chemosignal. In minute amounts and not detectable as an odor it influenced cortical and motoric reactions. Therefore, it might be concluded that androstadienone indeed affects cognitive functions like attentional processes and in turn affects our behavior. The current results further support the notion that androstadienone acts like a human modulator pheromone, namely modulating ongoing behavior or a psychological reaction to a particular context, changing stimulus sensitivity, salience and sensory-motor integration. However, these conclusions remain tentative until further replication takes place, best in ecologically valid environments. Furthermore, one has to keep in mind that the current studies could not replicate several previous findings and could not verify some hypotheses assuming communicative effects of androstadienone. Thus, the main assumption of this thesis that androstadienone is an active chemosignal is still challenged. Also, whether the term “pheromone” is indeed suitable to label androstadienone remains open.
The Nuclear Factors of Activated T cells (NFATs) are critical transcription factors playing major roles in the control of the cell cycle, apoptosis and, probably, also cancerogenesis. Of all the four genuine NFATc family members, NFATc1 has the unique induction property which appears to be essential for T and B cell development, along with its considerable role in cytokine gene expression and function in non-lymphoid tissues and during organ development (such as in the development of muscle and heart cells). A number of studies have proved the potential role of NFATc1 protein in development of lymphomas and leukemias and provided evidence of differential expression of the same gene in different tumours (Suppression in classical Hodgkin lymphomas but overexpression in T-ALLs). Although the most commonly accepted pathway is the dephosphorylation of NFAT by calcineurin upon a rise in intracellular Ca++ leading to nuclear translocation followed by transcription of Il2 gene and related cytokines, it is quite possible that signaling mechanisms other than (or in addition to) calcineurin activation lead to NFATc1 induction as well. One of the major isoforms of NFATc1, NFATc1/αA, is the short inducible factor, produced upon full T and B cell activation. Here we used two different conditional knock-out mice as our study model. Inactivation of the murine Nfatc1 gene in bone marrow (of Cd79a/mb-1-cre x Nfatc1flx/flx mice) and spleen (of Cd23-cre x Nfatc1flx/flx mice) resulted in complete ablation of NFATc1 expression in splenic B cells. Although no severe developmental defects were found for the generation of ‘conventional’ B2 cells, NFATc1 inactivation in bone marrow B-cells led to a strong decrease in the peritoneal B1a cell population. In-vitro studies showed a clear-cut decrease in proliferation and an increase in Activation Induced Cell Death (AICD) of NFATc1-/- splenic B cells upon BCR stimulation. While NFATc1 appears to control directly the AICD of peripheral B cells, further studies revealed an effect of NFATc1 on proliferation by a sustained differentiation program controlling Ca++ flux and calcineurin activity which are needed to maintain transcription and proliferation of primary B cells. Re-expression of NFATc1 at a low dose could protect cells against AICD, whereas at a higher dose it initiated AICD. These data suggest an important dual role of NFATc1 in controlling proliferation and apoptosis of peripheral B lymphocytes. NFATc1 ablation also impaired the Ig class switch to IgG3 by T cell-independent (TI) type II antigens and impaired IgG3+ plasmablast formation when studied in-vivo by NP-Ficoll immunization or in-vitro using an in-vitro class-switch model. Contrary to the immunizations with TI-type II antigen, no significant differences were documented in Ig class switch upon immunization with NP-KLH, a T-cell dependent (TD) antigen. Taken together, the data indicate NFATc1/αA as a crucial player in the activation and function of splenic B cells upon BCR stimulation. Missing or incomplete NFATc1/αA induction appears to be one reason for the generation of B cell unresponsiveness, whereas uncontrolled NFATc1/αA expression could lead to unbalanced immune reactions and autoimmune diseases.
BAD (Bcl-2 antagonist of cell death, Bcl-2 associated death promoter) is a pro-apoptotic member of the Bcl-2 protein family that is regulated by phosphorylation in response to survival factors. Although much attention has been devoted to the identification of phosphorylation sites in murine BAD (mBAD), little data are available with respect to phosphorylation of human BAD (hBAD) protein. In this work, we investigated the quantitative contribution of BAD targeting kinases in phosphorylating serines 75, 99 and 118 of hBAD (Chapter 3.1). Our results indicate that RAF kinases phosphorylate hBAD in vivo at these established serine residues. RAF-induced phosphorylation of hBAD was not prevented by MEK inhibitors but could be reduced to control levels by use of the RAF inhibitor Sorafenib (BAY 43-9006). Consistently, expression of active RAF suppressed apoptosis induced by hBAD and the inhibition of colony formation caused by hBAD could be prevented by RAF. In addition, using surface plasmon resonance technique we analyzed the direct consequences of hBAD phosphorylation by RAF with respect to complex formation of BAD with 14-3-3 proteins and Bcl-XL. Phosphorylation of hBAD by active RAF promotes 14-3-3 protein association, whereby the phosphoserine 99 represents the major binding site. Furthermore, we demonstrate in this work that hBAD forms channels in planar bilayer membranes in vitro. This pore-forming capacity is dependent on phosphorylation status and interaction with 14-3-3 proteins. Additionally, we show that hBAD pores possess a funnel-shaped geometry that can be entered by ions and non-charged molecules up to 200 Da (Chapter 3.2). Since both lipid binding domains of hBAD (LBD1 and LBD2) are located within the C-terminal region, we investigated this part of the protein with respect to its structural properties (Chapter 3.3). Our results demonstrate that the C-terminus of hBAD possesses an ordered β-sheet structure in aqueous solution that adopts helical disposition upon interaction with lipid membranes. Additionally, we show that the interaction of the C-terminal segment of hBAD with the BH3 domain results in the formation of permanently open pores, whereby the phosphorylation of serine 118 proved to be necessary for effective pore-formation. In contrast, phosphorylation of serine 99 in combination with 14-3-3 association suppresses formation of channels. These results indicate that the C-terminal part of hBAD controls hBAD function by structural transitions, lipid binding and phosphorylation. Using mass spectrometry we identified in this work, besides the established in vivo phosphorylation sites at serines 75, 99 and 118, several novel hBAD phosphorylation sites (serines 25, 32/34, 97, 124 and 134, Chapter 3.1). To further analyze the regulation of hBAD function, we investigated the role of these newly identified phosphorylation sites on BAD-mediated apoptosis. We found that in contrast to the N-terminal phosphorylation sites, the C-terminal serines 124 and 134 act in an anti-apoptotic manner (Chapter 3.4). Our results further indicate that RAF kinases and PAK1 effectively phosphorylate BAD at serine 134. Notably, in the presence of wild type hBAD, co-expression of survival kinases, such as RAF and PAK1, leads to a strongly increased proliferation, whereas substitution of serine 134 by alanine abolishes this process. Furthermore, we identified hBAD serine 134 to be strongly involved in survival signaling in B-RAF-V600E containing tumor cells and found phosphorylation of this residue to be crucial for efficient proliferation in these cells. Collectively, our findings provide new insights into the regulation of hBAD function by phosphorylation and its role in cancer signaling.
Das Ziel der vorliegenden Arbeit war die Untersuchung der Reaktionen von Migränepatientinnen mit episodischer (EM) und häufiger Migräne (HM) auf verschiedene Aspekte des Triggerfaktors „Negativer Affekt“ wie Stress und negative Emotionen. Die Ergebnisse der beiden Gruppen wurden mit denen gesunder Kontrollpersonen verglichen (KG). Zur Ermittlung des Aufmerksamkeitsverhaltens gegenüber emotionalen Reizen wurden zwei Emotionale Stroop Tests (EST) durchgeführt. Erwartet wurde ein Aufmerksamkeitsbias der Patientinnen hinsichtlich negativer emotionaler Reize. Im EST 1 wurden allgemeine affektive Wörter der Valenzen positiv, neutral und negativ verwendet. Die Probandinnen sollten auf die Wortfarbe mit Tastendruck reagieren und den Wortinhalt ignorieren. Im EST 2 wurden emotionale Gesichtsausdrücke (ärgerlich, freundlich, neutral) als Reize verwendet. Dabei sollte die Rahmenfarbe der Bilder per Tastendruck bestimmt werden und der Inhalt ignoriert werden. Zur Auswertung wurden Emotionale Stroop Interferenzen (ESI) zum Vergleich Reaktionszeitdifferenzen negativ-neutral und negativ-positiv berechnet. Der erwartete Aufmerksamkeitsbias der HM für negative emotionale Reize wurde dabei nicht gefunden. Dafür zeigten im EST 2 die KG einen Aufmerksamkeitsbias für ärgerliche Gesichter. Ein signifikanter Gruppenunterschied in EST 2 mit sehr niedrigen, im Vergleich negativ-positiv sogar negativen ESI der HM ließ auf ein Vermeidungsverhalten dieser Gruppe ärgerlichen Gesichtern gegenüber schließen. Dieses wurde als Vermeidung negativer sozialer Reize interpretiert und zum gelernten, möglicherweise dysfunktionalen Vermeidungsverhalten von Migränepatienten potentiellen Triggersituationen gegenüber in Bezug gesetzt. Weiterhin wurden die Probandinnen mit dem „Paradigma der Öffentlichen Rede“ psychosozialem Stress ausgesetzt, indem sie vor einer Videokamera unter Beobachtung eine Rede halten sowie eine Kopfrechenaufgabe lösen sollten. Vorher und nachher wurden insgesamt vier Speichelproben zur Bestimmung des Stresshormons Kortisol genommen. Zudem wurden die Druckschmerzschwellen vor und nach dem Experimentalteil gemessen. Die erwartete Kortisolreaktion als Antwort auf die psychosoziale Stressaufgabe blieb aus. Ursache dafür kann die Stichprobenzusammensetzung mit 98% Frauen sein, deren Kortisolreaktion auf Stress durch hormonelle Schwankungen im Experiment nur unzuverlässig stimulierbar ist. Bei der Berechnung der Gesamtkortisolausschüttung über die Zeit zeigte sich im Gegensatz zu dem erwarteten erhöhten Kortisolspiegel der Migränepatientinnen ein linearer Abfall des Spiegels von KG, über EM zu HM, mit den niedrigsten Werten der HM. Diese Ergebnisse könnten auf Veränderungen der Hypophysen-Nebennieren (HHN)-Achse im Sinne eines Hypokortisolismus bei Migränepatientinnen widerspiegeln, der weiterer Klärung bedarf, z.B. durch die Bestimmung eines Kortisoltagesprofils bei Patientinnen. Eine veränderte Funktion der HHN-Achse könnte außerdem zu einer inadäquaten Reaktion auf Stresssituationen beitragen. Die bei Patientinnen ausbleibende Veränderung der Druckschmerzschwelle in Reaktion auf Stress lässt ebenfalls auf eine ungenügende Stressreaktion der Patientinnen schließen. Am Ende der Untersuchung, nach einer Entspannungsphase von 50 Minuten, wurde den Probandinnen Blut abgenommen, in dem die mRNA- und Proteinkonzentrationen ausgewählter pro- und antiinflammatorischer Zytokine bestimmt wurden. Die Analyse der Zytokinkonzentrationen mit Luminex ergab für die Proteindaten aufgrund zu geringer verwertbarer Daten kein interpretierbares Bild. Die mittels Real Time Quantitativer PCR erhaltenen mRNA-Konzentrationen spiegelten die Schmerzfreiheit der Patienten wieder, mit im Vergleich zu KG verringerten proinflammatorischen Zytokinen (TNF-alpha, IL-1beta, IL-2, IL-6) und dem ebenfalls verringerten antiinflammatorischen Zytokin IL-10, sowie dem deutlich erhöhten antiinflammatorischen IL-4. Die im Vergleich zur KG überregulierten Zytokine im schmerzfreien Intervall weisen auf veränderte Regulierungsmechanismen des Immunsystems für die Schmerzmediatoren Zytokine hin. Weitere Schmerzmediatoren könnten ebenfalls verändert sein, was weiterer Klärung in nachfolgenden Studien bedarf. Alles in allem konnten verschiedene Veränderungen in den psychologischen und endokrinen Reaktionen der Migränepatientinnen auf Bestandteile des Triggers „Negativer Affekt“ sowie in der Schmerzregulierung gefunden werden, wobei die Veränderungen bei Patientinnen mit Häufiger Migräne stärker auftraten. Dies weist auf eine mögliche Rolle der einzelnen untersuchten Komponenten bei der Migränechronifizierung hin, was in weiteren Studien vertiefend untersucht werden sollte.
Die chronische Herzinsuffizienz stellt nach wie vor eine der häufigsten Todesursachen weltweit dar. Trotz intensiver Forschung ist es bisher nicht möglich die pathophysiologischen Prozesse aufzuhalten. Es wird nach neuen Strategien gesucht, hier therapeutisch eingreifen zu können. Kleine nicht-kodierende RNAs, sogenannte microRNAs (miRNAs), wurden als wichtige Faktoren bei verschiedenen Herzkrankheiten beschrieben. Die Mehrzahl der bisherigen Studien fokussierte sich dabei auf die am stärksten deregulierten miRNAs im erkrankten Herz. In einer automatisierten Analyse im 96 Well-Format untersuchten wir 230 miRNAs auf ihr Potential, in das Größenwachstum von primären Kardiomyozyten einzugreifen. Aus den miRNAs mit den größten Effekten selektierten wir diejenigen, die eine hohe endogene Expression aufwiesen, und unterzogen sie einem Validierungsprozess. Hier konnten wir die Effekte aller pro- (miR-22, miR-30c, miR-30d, miR-212, miR-365) und anti-hypertrophen (miR-27a, miR-27b, miR-133a) miRNAs bestätigen. Die Mehrzahl dieser miRNAs wurde hiermit erstmalig beschrieben, dass sie eine wichtige Rolle beim Größenwachstum von Kardiomyozyten spielen. Sie wären daher interessante Kandidaten für detaillierte funktionelle Studien mit dem Ziel ihr therapeutisches Potential zu evaluieren. In einem früheren genetischen Screen zur Identifizierung von kardialen, sezernierten Faktoren wurde der Protease Inhibitor 16 (PI16) entdeckt, der sich im insuffizienten Herz durch eine starke Akkumulation auszeichnet. Gegenstand des zweiten Teils dieser Arbeit war es, eine Mauslinie zu generieren, in der PI16 global oder konditionell mit Hilfe des Cre/LoxP-Systems ausgeschaltet werden kann. Nach Elektroporation des Pi16floxneo Targeting Vektors in embryonale Stammzellen und Blastozysteninjektion erhielten wir eine Mauslinie, die Träger der zielgerichteten Modifikation des Pi16 Allels war. Mit der globalen genetischen Deletion des LoxP-flankierten Abschnitts von Exon 3 bis 4 konnten wir die Expression des Pi16 Gens komplett unterbinden. Die PI16 Defizienz führte weder im Herz noch in anderen Organen per se zu pathologischen Veränderungen. Zudem war unbekannt, dass PI16 in der gesunden Maus in der kardialen Fibroblastenfraktion enthalten sowie in den Zilien der Epididymis und der Trachea und im Lumen der Schilddrüse lokalisiert ist. Im insuffizienten Herz bestätigten wir eine Akkumulation von PI16, die sich vor allem auf die fibrotischen Bereiche beschränkte. Das lässt Grund zur Annahme, dass die kardiale Funktion von PI16 erst dann offensichtlich wird, wenn man die defizienten Mäuse zukünftig entsprechenden Stressmodellen aussetzt. Das wird zu einem umfassenden Verständnis der kardialen Funktion von PI16 und dessen Potential als therapeutisches Zielmolekül führen.
For a large fraction of the proteins expressed in the human brain only the primary structure is known from the genome project. Proteins conserved in evolution can be studied in genetic models such as Drosophila. In this doctoral thesis monoclonal antibodies (mAbs) from the Wuerzburg Hybridoma library are produced and characterized with the aim to identify the target antigen. The mAb ab52 was found to be an IgM which recognized a cytosolic protein of Mr ~110 kDa on Western blots. The antigen was resolved by two-dimensional gel electrophoresis (2DE) as a single distinct spot. Mass spectrometric analysis of this spot revealed EPS-15 (epidermal growth factor receptor pathway substrate clone 15) to be a strong candidate. Another mAb from the library, aa2, was already found to recognize EPS-15, and comparison of the signal of both mAbs on Western blots of 1D and 2D electrophoretic separations revealed similar patterns, hence indicating that both antigens could represent the same protein. Finally absence of the wild-type signal in homozygous Eps15 mutants in a Western blot with ab52 confirmed the ab52 antigen to be EPS-15. Thus both the mAbs aa2 and ab52 recognize the Drosophila homologue of EPS-15. The mAb aa2, being an IgG, is more suitable for applications like immunoprecipitation (IP). It has already been submitted to the Developmental Studies Hybridoma Bank (DSHB) to be easily available for the entire research community. The mAb na21 was also found to be an IgM. It recognizes a membrane associated antigen of Mr ~10 kDa on Western blots. Due to the membrane associated nature of the protein, it was not possible to resolve it by 2DE and due to the IgM nature of the mAb it was not possible to enrich the antigen by IP. Preliminary attempts to biochemically purify the endogenously expressed protein from the tissue, gave promising results but could not be completed due to lack of time. Thus biochemical purification of the protein seems possible in order to facilitate its identification by mass spectrometry. Several other mAbs were studied for their staining pattern on cryosections and whole mounts of Drosophila brains. However, many of these mAbs stained very few structures in the brain, which indicated that only a very limited amount of protein would be available as starting material. Because these antibodies did not produce signals on Western blots, which made it impossible to enrich the antigens by electrophoretic methods, we did not attempt their purification. However, the specific localization of these proteins makes them highly interesting and calls for their further characterization, as they may play a highly specialized role in the development and/or function of the neural circuits they are present in. The purification and identification of such low expression proteins would need novel methods of enrichment of the stained structures.
Die Amygdala ist ein Kernkomplex, der dicht von serotonergen Afferenzen innerviert wird. Sowohl bei Tieren als auch beim Menschen spielen Interaktionen zwischen dem serotonergen System und der Amygdala bei der Verarbeitung von Reizen, die mit Angst oder Stress assoziiert sind, eine zentrale Rolle. Genetische Variationen im serotonergen System und/oder dauerhafter Stress können dazu führen, dass diese Verarbeitungsprozesse fehlerhaft ablaufen, wodurch Verhaltensanormalitäten bzw. die Entstehung psychiatrischer Erkrankungen begünstigt werden. Die Zielneurone der serotonergen Transmission in der Amygdala, die molekularen Mechanismen möglicher Interaktionen und strukturelle Konsequenzen der Störungen dieser Interaktionen sind jedoch bis zum heutigen Zeitpunkt noch nicht vollständig bekannt. Daher bestand ein Ziel der vorliegenden Arbeit darin, den Einfluss eines Ungleichgewichts im serotonergen System (5-Htt KO) sowie von wiederholtem, sozialem Stress auf die neuronale Morphologie der Amygdala zu analysieren und Zielneurone serotonerger Afferenzen zu identifizieren und zu charakterisieren, um die neuronalen Netzwerke der Emotionsverarbeitung besser verstehen zu können. Um vom 5-Htt–Genotyp abhängige und stressbedingte neuromorphologische Veränderungen zu untersuchen, wurden dreidimensionale Rekonstruktionen von Neuronen der laterobasalen Amygdala von männlichen, adulten Wildtyp (WT)- und 5-Htt KO-Mäusen angefertigt und bezüglich verschiedener morphologischer Parameter ausgewertet. An den Pyramidenzellen wurden nur geringfügige Veränderungen der dendritischen Komplexität, jedoch, im Vergleich zu WT-Mäusen, eine wesentliche Erhöhung der Dornendichte an spezifischen dendritischen Kompartimenten bei gestressten WT-Mäusen, sowie nicht gestressten und gestressten 5-Htt KO-Mäusen nachgewiesen. Im Vergleich zu nicht gestressten WT–Mäusen war die dendritische Dornendichte aller anderen Gruppen gleichermaßen erhöht. Die Sternzelle, zeigten bezüglich der untersuchten Parameter keine morphologischen Veränderungen auf. Eine besondere Subpopulation der Interneurone stellen die NeuropeptidY (NPY)–Neurone der laterobasalen Amygdala dar, da sie in diesen Nuclei anxiolytisch wirken. Es gibt nur wenige Anhaltspunkte darüber, durch welche Systeme NPY–Neurone moduliert werden. Da sowohl NPY–Neurone in der laterobasalen Amygdala als auch das serotonerge System an angstregulierenden Prozessen beteiligt sind, sollte im zweiten Teil der vorliegenden Arbeit untersucht werden, ob es sich bei diesen Neuronen um Zielstrukturen des serotonergen Systems handelt. Mittels licht- und elektronenmikroskopischer Analysen wurden synaptische Kontakte zwischen serotonergen Afferenzen und NPY-immunreaktiven Neuronen in der laterobasalen Amygdala von Ratten verifiziert. Da der funktionelle Einfluss der serotonergen Innervation auf diese Zielneurone von deren Serotoninrezeptor (5-HTR)-Ausstattung abhängt, wurden Koexpressionsanalysen von NPY mRNA mit den mRNAs verschiedener 5-HTR durchgeführt. Die Analysen ergaben, dass NPY mRNA–reaktive Neurone in der laterobasalen Amygdala 5-HT1A und 5-HT2C, jedoch nicht 5-HT3 mRNA koexprimieren. Die in der vorliegenden Arbeit erzielten Resultate liefern neue Erkenntnisse über den Einfluss des serotonergen Systems auf die laterobasale Amygdala von Mäusen und Ratten. Bei den Veränderungen der dendritischen Dornendichte nach sozialen Stresserfahrungen könnte es sich um neuroadaptive bzw. kompensatorische Mechanismen der Pyramidenzellen handeln, die WT-Mäusen eine Anpassung an sich ändernde, negative Umweltbedingungen ermöglicht. Die erhöhte Dornendichte könnte dabei die Ausbildung eines „emotionalen Gedächtnisses“ repräsentieren, das eine flexible Verhaltensantwort auf ein erneutes Auftauchen von Gefahr erlaubt. Eine solche Modulation der Erregbarkeit der laterobasalen Amygdala könnte beispielsweise über eine situationsentsprechende Hemmung des Outputs der Pyramidenzellen durch differentiell aktive inhibitorische Netzwerke erfolgen. Eine differentielle Aktivierung kann z. B. über unterschiedliche Rezeptorausstattungen, wie es in der Subpopulation der NPY–Neurone in der vorliegenden Arbeit nachgewiesen wurde, erfolgen. Das erhöhte angstähnliche Verhalten der 5-Htt KO-Mäuse nach wiederholtem Stress könnte mit der Unfähigkeit zusammenhängen, in entsprechenden Situationen durch Neubildung von Dornen zu reagieren, da die Dornendichte bei diesen Tieren schon unter stressarmen Umweltbedingungen ihr Maximum erreicht hat. Sowohl Fehlfunktionen der neuronalen Plastizität als auch mögliche Fehlfunktionen der differentiellen Inhibierung der Pyramidenzellen durch Interneurone, die durch genetische Variationen und/oder Stress bedingt sein können, könnten eine „offene Tür“ repräsentieren, die zu manifesten Auffälligkeiten im Verhalten bei Tieren führt bzw. auch zur Entstehung bestimmter psychiatrischer Erkrankungen beim Menschen beiträgt.
The acquired immunodeficiency syndrome (AIDS) is currently the most infectious disease worldwide. It is caused by the human immunodeficiency virus (HIV). At the moment there are ~33.3 million people infected with HIV. Sub-Saharan Africa, with ~22.5 million people infected accounts for 68% of the global burden. In most African countries antiretroviral therapy (ART) is administered in limited-resource settings with standardised first- and second-line ART regimens. During this study I analysed the therapy-naïve population of Cape Town, South Africa and Mwanza, Tanzania for any resistance associated mutations (RAMs) against protease inhibitors, nucleoside reverse transcriptase inhibitors and non-nucleoside reverse transcriptase inhibitors. My results indicate that HIV-1 subtype C accounts for ~95% of all circulating strains in Cape Town, South Africa. I could show that ~3.6% of the patient derived viruses had RAMs, despite patients being therapy-naïve. In Mwanza, Tanzania the HIV drug resistance (HIVDR) prevalence in the therapy-naïve population was 14.8% and significantly higher in the older population, >25 years. Therefore, the current WHO transmitted HIVDR (tHIVDR) survey that is solely focused on the transmission of HIVDR and that excludes patients over 25 years of age may result in substantial underestimation of the prevalence of HIVDR in the therapy-naïve population. Based on the prevalence rates of tHIVDR in the study populations it is recommended that all HIV-1 positive individuals undergo a genotyping resistance test before starting ART. I also characterized vif sequences from HIV-1 infected patients from Cape Town, South Africa as the Vif protein has been shown to counteract the antiretroviral activity of the cellular APOBEC3G/F cytidine deaminases. There is no selective pressure on the HIV-1 Vif protein from current ART regimens and vif sequences was used as an evolutionary control. As the majority of phenotypic resistance assays are still based on HIV-1 subtype B, I wanted to design an infectious HIV-1 subtype C proviral molecular clone that can be used for in vitro assays based on circulating strains in South Africa. Therefore, I characterized an early primary HIV-1 subtype C isolate from Cape Town, South Africa and created a new infectious subtype C proviral molecular clone (pZAC). The new pZAC virus has a significantly higher transient viral titer after transfection and replication rate than the previously published HIV-1 subtype C virus from Botswana. The optimized proviral molecular clone, pZAC could be used in future cell culture and phenotypic HIV resistance assays regarding HIV-1 subtype C.
In recent years high-throughput experiments provided a vast amount of data from all areas of molecular biology, including genomics, transcriptomics, proteomics and metabolomics. Its analysis using bioinformatics methods has developed accordingly, towards a systematic approach to understand how genes and their resulting proteins give rise to biological form and function. They interact with each other and with other molecules in highly complex structures, which are explored in network biology. The in-depth knowledge of genes and proteins obtained from high-throughput experiments can be complemented by the architecture of molecular networks to gain a deeper understanding of biological processes. This thesis provides methods and statistical analyses for the integration of molecular data into biological networks and the identification of functional modules, as well as its application to distinct biological data. The integrated network approach is implemented as a software package, termed BioNet, for the statistical language R. The package includes the statistics for the integration of transcriptomic and functional data with biological networks, the scoring of nodes and edges of these networks as well as methods for subnetwork search and visualisation. The exact algorithm is extensively tested in a simulation study and outperforms existing heuristic methods for the calculation of this NP-hard problem in accuracy and robustness. The variability of the resulting solutions is assessed on perturbed data, mimicking random or biased factors that obscure the biological signal, generated for the integrated data and the network. An optimal, robust module can be calculated using a consensus approach, based on a resampling method. It summarizes optimally an ensemble of solutions in a robust consensus module with the estimated variability indicated by confidence values for the nodes and edges. The approach is subsequently applied to two gene expression data sets. The first application analyses gene expression data for acute lymphoblastic leukaemia (ALL) and differences between the subgroups with and without an oncogenic BCR/ABL gene fusion. In a second application gene expression and survival data from diffuse large B-cell lymphomas are examined. The identified modules include and extend already existing gene lists and signatures by further significant genes and their interactions. The most important novelty is that these genes are determined and visualised in the context of their interactions as a functional module and not as a list of independent and unrelated transcripts. In a third application the integrative network approach is used to trace changes in tardigrade metabolism to identify pathways responsible for their extreme resistance to environmental changes and endurance in an inactive tun state. For the first time a metabolic network approach is proposed to detect shifts in metabolic pathways, integrating transcriptome and metabolite data. Concluding, the presented integrated network approach is an adequate technique to unite high-throughput experimental data for single molecules and their intermolecular dependencies. It is flexible to apply on diverse data, ranging from gene expression changes over metabolite abundances to protein modifications in a combination with a suitable molecular network. The exact algorithm is accurate and robust in comparison to heuristic approaches and delivers an optimal, robust solution in form of a consensus module with confidence values. By the integration of diverse sources of information and a simultaneous inspection of a molecular event from different points of view, new and exhaustive insights into biological processes can be acquired.
Type 1 diabetes affects around 0.5% of the population in developed countries and the incidence rates have been rising over the years. The destruction of beta cells is irreversible and the current therapy available to patients only manages the symptoms and does not prevent the associated pathological manifestations. The patients need lifelong therapy and intensive research is being carried out to identify ways to eliminate autoimmune responses directed against pancreatic beta cells and to replace or regenerate beta cells. The work presented herein aimed at analyzing the role of the Th17 T cell subset, characterized by secretion of the pro- inflammatory cytokine IL-17A, in autoimmune diabetes and also at generating a beta cell reporter mouse line in the NOD background, the most widely- used mouse model for type 1 diabetes. We generated IL- 17A knockdown (KD) NOD mice, using RNAi in combination with lentiviral transgenesis. We analyzed diabetes frequency in IL-17A deficient mice and found that the loss of IL-17A did not protect the transgenic mice from diabetes. Based on these observations, we believe that Th17 cells do not play a critical role in type 1 diabetes through the IL-17A pathway, though they might still be involved in the disease process through alternate pathways. We also generated NOD and NOD-SCID mice with a transgene that drives the beta cell specific expression of a luciferase reporter gene. We used a lentiviral construct, which combined a luciferase sequence and a short- hairpin RNA (shRNA) expression cassette, allowing gene- knockdown under the beta cell specific rat insulin promoter (RIP). These mice will be of use in studying beta cell phenotypes resulting from the knockdown of target genes, using non- invasive bioimaging. We believe that the generation of these reporter mouse lines for diabetes studies will prove valuable in future investigations. Furthermore, the demonstration that the loss of IL-17A does not alter susceptibility to type 1 diabetes should help clarify the controversial involvement of Th17 cells in this disease.
According to a changing environment it is crucial for animals to make experience and learn about it. Sensing, integrating and learning to associate different kinds of modalities enables animals to expect future events and to adjust behavior in the way, expected as the most profitable. Complex processes as memory formation and storage make it necessary to investigate learning and memory on different levels. In this context Drosophila melanogaster represents a powerful model organism. As the adult brain of the fly is still quite complex, I chose the third instar larva as model - the more simple the system, the easier to isolate single, fundamental principles of learning. In this thesis I addressed several kinds of questions on different mechanism of olfactory associative and synaptic plasiticity in Drosophila larvae. I focused on short-term memory throughout my thesis. First, investigating larval learning on behavioral level, I developed a one-odor paradigm for olfactory associative conditioning. This enables to estimate the learnability of single odors, reduces the complexity of the task and simplify analyses of "learning mutants". It further allows to balance learnability of odors for generalization-type experiments to describe the olfactory "coding space". Furthermore I could show that innate attractiveness and learnability can be dissociated and found finally that paired presentation of a given odor with reward increase performance, whereas unpaired presentations of these two stimuli decrease performance, indicating that larva are able to learn about the presence as well as about the absence of a reward. Second, on behavioral level, together with Thomas Niewalda and colleagues we focussed on salt processing in the context of choice, feeding and learning. Salt is required in several physiological processes, but can neither be synthesized nor stored. Various salt concentrations shift the valence from attraction to repulsion in reflexive behaviour. Interestingly, the reinforcing effect of salt in learning is shifted by more than one order of magnitude toward higher concentrations. Thus, the input pathways for gustatory behavior appear to be more sensitive than the ones supporting gustatory reinforcement, which is may be due to the dissociation of the reflexive and the reinforcing signalling pathways of salt. Third, in cooperation with Michael Schleyer we performed a series of behavioral gustatory, olfactory preference tests and larval learning experiments. Based on the available neuroanatomical and behavioral data we propose a model regarding chemosensory processing, odor-tastant memory trace formation and the 'decision' like process. It incorporates putative sites of interaction between olfactory and gustatory pathways during the establishment as well as behavioral expression of odor-tastant memory. We claim that innate olfactory behavior is responsive in nature and suggest that associative conditioned behavior is not a simple substitution like process, but driven more likely by the expectation of its outcome. Fourth, together with Birgit Michels and colleagues we investigated the cellular site and molecular mode of Synapsin, an evolutionarily conserved, presynaptic vesicular phosphoprotein and its action in larval learning. We confirmed a previously described learning impairment upon loss of Synapsin. We localized this Synapsin dependent memory trace in the mushroom bodies, a third-order "cortical" brain region, and could further show on molecular level, that Synapsin is as a downstream element of the AC-cAMP-PKA signalling cascade. This study provides a comprehensive chain of explanation from the molecular level to an associative behavioral change. Fifth, in the main part of my thesis I focused on molecular level on another synaptic protein, the Synapse associated protein of 47kDa (Sap47) and its role in larval behavior. As a member of a phylogenetically conserved gene family of hitherto unknown function. It is localized throughout the whole neuropil of larval brains and associated with presynaptic vesicles. Upon loss of Sap47 larvae exhibit normal sensory detection of the to-be-associated stimuli as well as normal motor performance and basic synaptic transmission. Interestingly, short-term plasticity is distorted and odorant–tastant associative learning ability is reduced. This defect in associative function could be rescued by restoring Sap47 expression. Therefore, this report is the first to suggest a function for Sap47 and specifically argues that Sap47 is required for synaptic as well as for behavioral plasticity in Drosophila larva. This prompts the question whether its homologs are required for synaptic and behavioral plasticity also in other species. Further in the last part of my thesis I contributed to the study of Ayse Yarali. Her central topic was the role of the White protein in punishment and relief learning in adult flies. Whereas stimuli that precede shock during training are subsequently avoided as predictors for punishment, stimuli that follow shock during training are later on approached, as they predict relief. Concerning the loss of White we report that pain-relief learning as well as punishment learning is changed. My contribution was a comparison between wild type and the white1118 mutant larvae in odor-reward learning. It turned out that a loss of White has no effect on larval odorant-tastant learning. This study, regarding painrelief learning provides the very first hints concerning the genetic determinants of this form of learning.
The present work investigated the neural mechanisms underlying cognitive inhibition/thought suppression in Anderson’s and Green’s Think/No-Think paradigm (TNT), as well as different variables influencing these mechanisms at the cognitive, the neurophysiological, the electrophysiological and the molecular level. Neurophysiological data collected with fNIRS and fMRI have added up to the existing evidence of a fronto-hippocampal network interacting during the inhibition of unwanted thoughts. Some evidence has been presented suggesting that by means of external stimulation of the right dlPFC through iTBS thought suppression might be improved, providing further evidence for an implication of this region in the TNT. A combination of fNIRS with ERP has delivered evidence of a dissociation of early condition-independent attentional and later suppression-specific processes within the dlPFC, both contributing to suppression performance. Due to inconsistencies in the previous literature it was considered how stimulus valence would influence thought suppression by manipulating the emotional content of the to-be-suppressed stimuli. Findings of the current work regarding the ability to suppress negative word or picture stimuli have, however, been inconclusive as well. It has been hypothesized that performance in the TNT might depend on the combination of valence conditions included in the paradigm. Alternatively, it has been suggested that inconsistent findings regarding the suppression of negative stimuli or suppression at all might be due to certain personality traits and/or genetic variables, found in the present work to contribute to thought inhibition in the TNT. Rumination has been shown to be a valid predictor of thought suppression performance. Increased ruminative tendencies led to worse suppression performance which, in the present work, has been linked to less effective recruitment of the dlPFC and in turn less effective down-regulation of hippocampal activity during suppression trials. Trait anxiety has also been shown to interrupt thought suppression despite higher, however, inefficient recruitment of the dlPFC. Complementing the findings regarding ruminative tendencies and decreased thought inhibition a functional polymorphism in the KCNJ6 gene, encompassing a G-to-A transition, has been shown to disrupt thought suppression despite increased activation of the dlPFC. Through the investigation of thought suppression at different levels, the current work adds further evidence to the idea that the TNT reflects an executive control mechanism, which is sensitive to alterations in stimulus valence to some extent, neurophysiological functioning as indicated by its sensitivity to iTBS, functional modulations at the molecular level and personality traits, such as rumination and trait anxiety.
Honeybees (Apis mellifera) forage on a great variety of plant species, navigate over large distances to crucial resources, and return to communicate the locations of food sources and potential new nest sites to nest mates using a symbolic dance language. In order to achieve this, honeybees have evolved a rich repertoire of adaptive behaviours, some of which were earlier believed to be restricted to vertebrates. In this thesis, I explore the mechanisms involved in honeybee learning, memory, numerical competence and navigation. The findings acquired in this thesis show that honeybees are not the simple reflex automats they were once believed to be. The level of sophistication I found in the bees’ memory, their learning ability, their time sense, their numerical competence and their navigational abilities are surprisingly similar to the results obtained in comparable experiments with vertebrates. Thus, we should reconsider the notion that a bigger brain automatically indicates higher intelligence.
Attention-deficit/hyperactivity disorder (ADHD) is a genetically complex childhood onset neurodevelopmental disorder which is highly persistent into adulthood. Several chromo-somal regions associated with this disorder were identified previously in genome-wide linkage scans, association (GWA) and copy number variation (CNV) studies. In this work the results of case-control and family-based association studies using a can-didate gene approach are presented. For this purpose, possible candidate genes for ADHD have been finemapped using mass array-based SNP genotyping. The genes KCNIP4, CDH13 and DIRAS2 have been found to be associated with ADHD and, in addition, with cluster B and cluster C personality disorders (PD) which are known to be related to ADHD. Most of the associations found in this work would not withstand correction for multiple testing. However, a replication in several independent populations has been achieved and in conjunction with previous evidence from linkage, GWA and CNV studies, it is assumed that there are true associations between those genes and ADHD. Further investigation of DIRAS2 by quantitative real-time PCR (qPCR) revealed expression in the hippocampus, cerebral cortex and cerebellum of the human brain and a significant increase in Diras2 expression in the mouse brain during early development. In situ hybrid-izations on murine brain slices confirmed the results gained by qPCR in the human brain. Moreover, Diras2 is expressed in the basolateral amygdala, structures of the olfactory system and several other brain regions which have been implicated in the psychopatholo-gy of ADHD. In conclusion, the results of this work provide further support to the existence of a strong genetic component in the pathophysiology of ADHD and related disorders. KCNIP4, CDH13 and DIRAS2 are promising candidates and need to be further examined to get more knowledge about the neurobiological basis of this common disease. This knowledge is essential for understanding the molecular mechanisms underlying the emergence of this disorder and for the development of new treatment strategies.
Ziel der vorliegenden Arbeit war es, zu untersuchen, ob nichtionisierende elektromagnetische Strahlung verschiedener Frequenzbereiche Genomschaden hervorrufen kann. Im Rahmen der vorliegenden Arbeit wurde eine Biomonitoring-Studie zu dieser Thematik konzipiert und durchgeführt. Es wurden 131 Probanden detailliert zu ihrer Mobilfunknutzung befragt. Anschließend wurden Mundschleimhautzellen entnommen und für eine mikroskopische Untersuchung aufbereitet und angefärbt. In den Zellen wurden Mikrokerne und andere Kernanomalien quantifiziert. Es zeigte sich keine Erhöhung der Mikrokernfrequenz in Abhängigkeit von der Dauer der Mobiltelefonnutzung. Auch die anderen abgefragten Parameter hatten keinen Einfluss auf die Höhe des Genomschadens. Als Positivkontrollen wurden vier Patienten, die eine lokale Strahlentherapie (ionisierende Strahlung) erhielten, eingeschlossen. Hier zeigte sich eine deutliche Erhöhung der Mikrokernfrequenz. Um festzustellen, ob die Mikrokerninduktion erst bei höheren Leistungsflussdichten als denen, die beim Mobilfunk verwendet werden, auftritt, wurden in-vitro-Versuche durchgeführt, bei denen verschiedene Zelllinien einer Strahlung von 900 MHz ausgesetzt wurden. Nach Exposition und einer Postinkubationsperiode wurden die Zellen fixiert und die Mikrokernfrequenz bestimmt. Neben den Leistungen wurden hier auch die Expositionszeiten und die Postinkubationsperioden variiert. In keinem Fall konnte eine Erhöhung der Mikrokernfrequenz festgestellt werden. Insgesamt konnte ein Einfluss elektromagnetischer Strahlung auf das Genom weder am Menschen im Rahmen einer Biomonitoring-Studie noch an verschiedenen Zelllinien im Rahmen von in-vitro-Versuchen festgestellt werden. Terahertzstrahlung ist elektromagnetische Strahlung im Bereich von 0,1 bis 10 THz, d. h. sie liegt zwischen Mikrowellen und Infrarotlicht. Derzeit wird sie hauptsächlich für spektroskopische Untersuchungen und zur Qualitätskontrolle im Herstellungs-prozess verschiedener Produkte verwendet. Anwendungen in der Sicherheitstechnik (z. B. Ganzkörperscanner) und in der Medizintechnik (z. B. Bildgebung) stehen kurz vor der Markteinführung bzw. sind bereits etabliert. Diese Anwendungen bringen eine Exposition der betroffenen Menschen mit sich. Außerdem wird an weiteren Techniken wie etwa der Datenübertragung gearbeitet. Die Wirkungen auf biologische Systeme sind im Gegensatz zum Mobilfunkbereich bisher nur unzureichend untersucht. Da bisher keine vollständigen Literaturübersichten vorlagen, wurde eine umfassende Literaturrecherche durchgeführt. Ziel war es, alle bisher durchgeführten Studien zu diesem Thema aufzulisten. Um diese Datenbasis zu verbreitern wurden in-vitro-Versuche bei verschiedenen Frequenzen durchgeführt. Als Strahlungsquellen wurden eine Frequenzvervielfacherkaskade (0,106 THz), ein Rückwärtswellen-Oszillator (0,380 THz) und ein Ferninfrarot-Laser (2,520 THz) eingesetzt. Die Strahlung wurde in einen modifizierten Inkubator geführt, so dass die Expositionen bei definierter Temperatur und konstantem CO2-Gehalt durchgeführt werden konnten. Da Terahertzstrahlung durch Wasser sehr stark absorbiert wird, sind bei einer Exposition des Menschen primär die obersten Hautschichten betroffen. Aus diesem Grund wurden primäre Hautfibroblasten und HaCaT-Zellen, eine Keratinozyten-Zelllinie, als biologische Systeme verwendet. Die Zellen wurden für unterschiedliche Zeitperioden mit verschiedenen Leistungsflussdichten exponiert. Anschließend wurden die Zellen für den Comet Assay aufbereitet und analysiert. Der Comet Assay ist eine Methode zur Quantifizierung von Einzel- und Doppelstrangbrüchen der DNA. Weiterhin wurden die Zellen nach einer Postinkubationsperiode für den Mikrokerntest aufbereitet. Neben unbehandelten Kontrollen und Sham-Expositionen wurden auch Positivkontrollen durchgeführt. Es konnte keine Erhöhung der Anzahl der DNA-Strangbrüche bzw. der Mikrokernfrequenz festgestellt werden. Da bekannt war, dass im Mobilfunkbereich unter bestimmten Bedingungen Störungen der Mitose, nicht aber Erhöhungen der Mikrokernfrequenz, auftreten, wurden Mitosestörungen nach Exposition bei 0,106 THz untersucht. Hierzu wurden AL-Zellen für 30 Minuten exponiert und anschließend ohne Postinkubation direkt fixiert. Analysiert wurden Störungen in allen Phasen der Mitose. Es zeigte sich, dass die Frequenz der Störungen in der Pro- und Metaphase unverändert blieb. Die Störungen in der Ana- und Telophase nahmen dagegen mit steigender Leistungsflussdichte zu. Insgesamt konnte im Terahertzbereich unter den gewählten Expositionsbedingungen kein DNA-Schaden beobachtet werden. Bei 0,106 THz konnten Mitosestörungen als Folge der Exposition gezeigt werden. Der Zusammenhang zwischen diesen Mitosestörungen und DNA-Schäden, insbesondere der Mikrokerninduktion, konnte bisher nicht abschließend geklärt werden und bleibt Gegenstand weiterer Untersuchungen.
Platelet activation and adhesion results in thrombus formation that is essential for normal hemostasis, but can also cause irreversible vessel occlusion leading to myocardial infarction or stroke. The C-type lectin-like receptor 2 (CLEC-2) was recently identified to be expressed on the platelet surface, however, a role for this receptor in hemostasis and thrombosis had not been demonstrated. In the current study, the involvement of CLEC-2 in platelet function and thrombus formation was investigated using mice as a model system. In the first part of the thesis, it was found that treatment of mice with a newly generated monoclonal antibody against murine CLEC-2 (INU1) led to the complete and highly specific loss of the receptor in circulating platelets (a process termed “immunodepletion”). CLEC-2-deficient platelets were completely unresponsive to the CLEC-2-specific agonist rhodocytin, whereas activation induced by all other tested agonists was unaltered. This selective defect translated into severely decreased platelet aggregate formation under flow ex vivo; and in vivo thrombosis models revealed impaired stabilization of formed thrombi with enhanced embolization. Consequently, CLEC-2 deficiency profoundly protected mice from occlusive arterial thrombus formation. Furthermore, variable bleeding times in INU1-treated mice indicated a moderate hemostatic defect. This reveals for the first time that CLEC-2 significantly contributes to thrombus stability in vitro and in vivo and plays a crucial role in hemostasis and arterial thrombosis. Thus, CLEC-2 represents a potential novel anti-thrombotic target that can be functionally inactivated in vivo. This in vivo down-regulation of platelet surface receptors might be a promising approach for future anti-thrombotic therapy. The second part of the work investigated the effect of double-immunodepletion of the immunoreceptor tyrosine-based activation motif (ITAM)- and hemITAM-coupled receptors, platelet glycoprotein (GP) VI and CLEC-2, on hemostasis and thrombosis using a combination of the GPVI- and CLEC-2-specific antibodies, JAQ1 and INU1, respectively. Isolated targeting of either GPVI or CLEC-2 in vivo did not affect expression or function of the respective other receptor. However, simultaneous treatment with both antibodies resulted in the sustained loss of GPVI and CLEC-2 signaling in platelets, while leaving other activation pathways intact. In contrast to single deficiency of either receptor, GPVI/CLEC-2 double-deficient mice displayed a dramatic hemostatic defect. Furthermore, this treatment resulted in profound impairment of arterial thrombus formation that far exceeded the effects seen in single-depleted animals. Importantly, similar results were obtained in Gp6-/- mice that were depleted of CLEC-2 by INU1-treatment, demonstrating that this severe bleeding phenotype was not caused by secondary effects of combined antibody treatment. These data suggest that GPVI and CLEC-2 can be independently or simultaneously down-regulated in platelets in vivo and reveal an unexpected functional redundancy of the two receptors in hemostasis and thrombosis. Since GPVI and CLEC-2 have intensively been discussed as potential anti-thrombotic targets, these results may have important implications for the development of novel, yet save anti-GPVI or anti-CLEC-2-based therapies.
Investigation on Distinct Roles of Smad Proteins in Mediating Bone Morphogenetic Proteins Signals
(2011)
Bone morphogenetic proteins (BMPs) belong to the transforming growth factor-β (TGF-β) superfamily and play important roles in numerous biological events in the development of almost all multi-cellular organisms. Dysregulated BMP signaling is the underlying causes of numerous heritable and non-heritable human diseases including cancer. The vast range of biological responses induced by BMPs converges on three closely related Smad proteins that convey intracellular signals from BMP receptors to the nucleus. The specificity of BMP signaling has been intensively investigated at the level of ligand-receptor interactions, but how the different Smad proteins contribute to differential signals elicited by BMPs remains unclear. In this work, we investigated the BMP/Smad signaling in different aspects. In search for an appropriate fluorescence reporter in zebrafish, we compared different photo-switchable proteins and found EosFP the best candidate this model system for its fast maturation and fluorescence intensity. We modified and created appropriate vectors enabling Tol2-transposon based trangenesis in zebrafish, with which transgenic zebrafish lines were generated. We combined fluorescence protein tagging with high resolution microscopy and investigate the dynamics of Smad proteins in model system zebrafish. We observed that Smad5 undergoes nucleo-translocation as BMP signal transmitter during zebrafish gastrulation. We explored the Smad involvement during myogenic-to-osteogenic conversion of C2C12 cell line induced by BMP4. We created transient loss-of-function of Smads by siRNA-mediated knockdowns and analyzed the effects on these coupled yet distinct procedures by quantitative real-time PCR and terminal marker staining. We found that different Smad-complex stoichiometry might be responsible for distinct cellular signals elicited by BMPs.
Dilated cardiomyopathy (DCM) represents an important subgroup of patients suffering from heart failure. The disease is supposed to be associated with autoimmune mechanisms in about one third of the cases. In the latter patients functionally active conformational autoantibodies directed against the second extracellular loop of the β1-adrenergic receptor (AR, β1ECII-aabs) have been detected. Such antibodies chronically stimulate the β1-AR thereby inducing the adrenergic signaling cascade in cardiomyocytes, which, in the long run, contributes to heart failure progression. We analyzed the production of cAMP after aab-mediated β1-AR activation in vitro using a fluorescence resonance energy transfer (FRET) assay. This assay is based on HEK293 cells stably expressing human β1-AR as well as the cAMP-sensor Epac1-camps. The assay showed a concentration-dependent increase in intracellular cAMP upon stimulation with the full agonist (-) isoproterenol. This response was comparable to results obtained in isolated adult murine cardiomyocytes and was partially blockable by a selective β1-AR antagonist. In the same assay poly- and monoclonal anti-β1ECII-abs (induced in different animals) could activate the adrenergic signaling cascade, whereas isotypic control abs had no effect on intracellular cAMP levels. Using the same method, we were able to detect functionally activating aabs in the serum of heart failure patients with ischemic and hypertensive heart disease as well as patients with DCM, but not in sera of healthy control subjects. In patients with DCM we observed an inverse correlation between the stimulatory potential of anti-β1-aabs and left ventricular pump function. To adopt this assay for the detection of functionally activating anti-β1ECII-aabs in clinical routine we attempted to establish an automated large-scale approach. Neither flow cytometry nor FRET detection with a fluorescence plate reader provided an acceptable signal-to-noise ratio. It was possible to detect (-) isoproterenol in a concentration-dependent manner using two different FRET multiwell microscopes. However, due to focus problems large-scale detection of activating anti-β1ECII-abs could not be implemented. Neutralization of anti-β1-aabs with the corresponding epitope-mimicking peptides is a possible therapeutic approach to treat aab-associated autoimmune DCM. Using our FRET assay we could demonstrate a reduction in the stimulatory potential of anti-β1ECII-abs after in vitro incubation with β1ECII-mimicking peptides. Cyclic (and to a lesser extent linear) peptides in 40-fold molar excess acted as efficient ab-scavengers in vitro. Intravenously injected cyclic peptides in a rat model of DCM also neutralized functionally active anti-β1ECII-abs efficiently in vivo. For a detailed analysis of the receptor-epitope targeted by anti-β1ECII-abs we used sequentially alanine-mutated β1ECII-mimicking cyclic peptides. Our data revealed that the disulfide bridge between the cysteine residues C209 and C215 of the human β1-AR appears essential for the formation of the ab-epitope. Substitution of further amino acids relevant for ab-binding in the cyclic scavenger peptide by alanine reduced its affinity to the ab and the receptor-activating potential was blocked less efficiently. In contrast, the non-mutant cyclic peptide almost completely blocked ab-induced receptor activation. Using this ala-scan approach we were able to identify a “NDPK”-epitope as essential for ab binding to the β1ECII. In summary, neutralization of conformational activating anti-β1ECII-(a)abs by cyclic peptides is a plausible therapeutic concept in heart failure that should be further exploited based on the here presented data.
Die Funktionalität β1- und β2-adrenerger Rezeptoren wird durch Polymorphismen in ihrer kodierenden Region moduliert. Wir haben uns die Technik des Fluoreszenz-Resonanz- Energie-Transfers (FRET) zu Nutze gemacht, um den Einfluss der am häufigsten vorkommenden Polymorphismen (Ser49Gly und Gly389Arg im β1AR, Arg16Gly und Gln27Glu im β2AR) auf die Rezeptorkonformation nach Aktivierung zu untersuchen. Dafür wurden FRET-Sensoren für die beiden βAR-Subtypen mit einem gelb-fluoreszierenden Protein (YFP) sowie einem cyan-fluoreszierenden Protein (CFP oder Cerulean) in der dritten intrazellulären Schleife bzw. am C-Terminus verwendet. Nach Stimulierung der βARSensoren konnte die Aktivierung der polymorphen Rezeptorvarianten in lebenden Zellen in Echtzeit untersucht werden. Dabei behielten die FRET-Sensoren sowohl die Bindungsaffinitäten der nativen Rezeptoren als auch eine intakte Funktionalität hinsichtlich der Bildung von sekundären Botenstoffen. Der Vergleich der Aktivierungskinetiken der verschieden polymorphen Varianten des β1AR und β2AR ergab keine signifikanten Unterschiede nach einer einmaligen Stimulation. Es zeigte sich jedoch, dass Rezeptorpolymorphismen die Aktivierungskinetik vorstimulierter βAR erheblich beeinflussen. So konnten wir im Vergleich zur ersten Aktivierung eine schnellere Aktivierung der Gly16-Varianten des β2AR sowie des Gly49Arg389-β1AR feststellen, während die Arg16-β2AR-Variante und der Ser49Gly389-β1AR dagegen bei einer wiederholten Stimulation langsamer aktiviert wurden. Diese Ergebnisse lassen auf ein "Rezeptorgedächtnis" schließen, das spezifisch für bestimmte polymorphe Rezeptorvarianten ist und eine βAR-Subtyp-spezische Ausprägung zeigt. Die Ausbildung der unterschiedlichen Aktivierungskinetiken hing von der Interaktion des Rezeptors mit löslichen intrazellulären Faktoren ab und bedurfte einer Phosphorylierung intrazellulärer Serin- und Threonin-Reste durch G-Protein-gekoppelte Rezeptorkinasen. Die Interaktion mit löslichen intrazellulären Faktoren scheint für den β1AR weniger stark ausgeprägt zu sein als für den β2AR. Die cAMP-Produktion war für die schneller werdenden, “hyperfunktionellen” Gly16-β2ARVarianten signifikant um mehr als 50% höher im Vergleich zur “hypofunktionellen” Arg16- Variante. Die unterschiedliche Funktionalität spiegelte sich im Therapieausgang bei Tokoysepatientinnen wider, dessen Erfolg mit dem Arg16Gly Polymorphismus verknüpft war. Die Daten implizieren eine intrinsische, polymorphismusabhängige Eigenschaft der βAR, die die Aktivierungskinetik der Rezeptoren bei wiederholten Stimulationen determiniert. Diese könnte auch für die zwischen Individuen variierende Ansprechbarkeit auf β-Agonisten und β-Blocker mitverantwortlich sein.
Thrombus formation at sites of vascular lesions is a dynamic process that requires a defined series of molecular events including the action of platelet adhesion/activation receptors, intracellular signal transduction, cytoskeletal rearrangements and activation of plasma coagulation factors. This process is essential to limit post-traumatic blood loss but may also contribute to acute thrombotic diseases such as myocardial infarction and stroke. With the help of genetically modified mice and the use of specific protein inhibitors and receptordepleting antibodies, the work presented in this thesis identified novel mechanisms underlying thrombus formation in hemostasis and thrombosis. In the first part of the study, it was shown that von Willebrand Factor (vWF) binding to glycoprotein (GP)Iba is critical for the formation of stable pathological thrombi at high shear rates, suggesting GPIba as an attractive pharmacological target for antithrombotic therapy. The subsequent analysis of recently generated phospholipase (PL)D1-deficient mice identified this enzyme, whose role in platelet function had been largely unknown, as a potential target protein downstream of GPIba. This was based on the finding that PLD1- deficient mice displayed severely defective GPIba-dependent thrombus stabilization under high shear conditions in vitro and in vivo without affecting normal hemostasis. The second part of the thesis characterizes the functional relevance of the immunoreceptor tyrosine-based activation motif (ITAM)-bearing collagen receptor GPVI and the recently identified hemITAM-coupled C-type lectin-like receptor 2 (CLEC-2) for in vivo thrombus formation. Genetic- and antibody-induced GPVI deficiency was found to similarly protect mice from arterial vessel occlusion in three different thrombosis models. These results confirmed GPVI as a promising antithrombotic target and revealed that antibody-treatment had no obvious off-target effects on platelet function. Similarly, immunodepletion of CLEC-2 by treating mice with the specific antibody INU1 resulted in markedly impaired thrombus growth and stabilization under flow in vitro and in vivo. Furthermore, it could be demonstrated that double-immunodepletion of GPVI and CLEC-2 resulted in severely decreased arterial thrombus formation accompanied by dramatically prolonged bleeding times. These data revealed an unexpected redundant function of the two receptors for in vivo thrombus formation and might have important implications for the potential development of anti-GPVI and anti-CLEC-2 antithrombotic agents. The third part of the thesis provides the first functional analysis of megakaryocyte- and platelet-specific RhoA knockout mice. RhoA-deficient mice displayed a defined signaling defect in platelet activation, leading to a profound protection from arterial thrombosis andand ischemic brain infarction, but at the same time also strongly increased bleeding times. These findings identified the GTPase as an important player for thrombus formation in hemostasis and thrombosis. Based on the previous proposal that the coagulation factor (F)XII might represent an ideal target for safe antithrombotic therapy without causing bleeding side effects, the last part of this thesis assesses the antithrombotic potential of the newly generated FXIIa inhibitor rHAInfestin- 4. It was found that rHA-Infestin-4 injection into mice resulted in virtually abolished arterial thrombus formation but no change in bleeding times. Moreover, rHA-Infestin-4 was similarly efficient in a murine model of ischemic stroke, suggesting that the inhibitor might be a promising agent for effective and safe therapy of cardio- and cerebrovascular diseases.
Desert ants of the genus Cataglyphis have become model systems for the study of insect navigation. An age-related polyethism subdivides their colonies into interior workers and short-lived light-exposed foragers. While foraging in featureless and cluttered terrain over distances up to several hundred meters, the ants are able to precisely return back to their often inconspicuous nest entrance. They accomplish this enormous navigational performance by using a path integration system - including a polarization compass and an odometer - as their main navigational means in addition to landmark-dependent orientation and olfactory cues. C. fortis, being the focus of the present thesis, is endemic to the salt flats of western North Africa, which are completely avoided by other Cataglyphis species. The fact that Cataglyphis ants undergo a behavioral transition associated with drastically changing sensory demands makes these ants particularly interesting for studying synaptic plasticity in visual and olfactory brain centers. This thesis focuses on plastic changes in the mushroom bodies (MBs) - sensory integration centers supposed to be involved in learning and memory presumably including landmark learning - and in synaptic complexes belonging to the lateral accessory lobe (LAL) known to be a relay station in the polarization processing pathway. To investigate structural synaptic plasticity in the MBs of C. fortis, synaptic complexes (microglomeruli, MG) in the visual (collar) and olfactory (lip) input regions of the MB calyx were immunolabeled and their pre- and postsynaptic profiles were quantified. The results show that a volume increase of the MB calyx during behavioral transition is associated with a decrease of MG number - an effect called pruning - in the collar and, less pronounced, in the lip that goes along with dendritic expansion in MB intrinsic Kenyon cells. Light-exposure of dark-reared ants of different age classes revealed similar effects and dark-reared ants age-matched to foragers had MG numbers comparable to those of interior workers. The results indicate that the enormous structural synaptic plasticity of the MB calyx collar is primarily driven by visual experience rather than by an internal program. Ants aged artificially for up to one year expressed a similar plasticity indicating that the system remains flexible over the entire life-span. To investigate whether light-induced synaptic reorganization is reversible, experienced foragers were transferred back to darkness with the result that their MBs exhibit only some reverse-type characteristics, in particular differences in presynaptic synapsin expression. To investigate the structure of large synaptic complexes in the LAL of C. fortis and to detect potential structural changes, pre- and postsynaptic profiles in interior workers and foragers were immunolabeled and quantified by using confocal imaging and 3D-reconstruction. The results show that these complexes consist of postsynaptic processes located in a central region that is surrounded by a cup-like presynaptic profile. Tracer injections identified input and output tracts of the LAL: projection neurons from the anterior optic tubercle build connections with neurons projecting to the central complex. The behavioral transition is associated with an increase by ~13% of synaptic complexes suggesting that the polarization pathway may undergo some sort of calibration process. The structural features of these synaptic contacts indicate that they may serve a fast and reliable signal transmission in the polarization vision pathway. Behavioral analyses of C. fortis in the field revealed that the ants perform exploration runs including pirouette-like turns very close to the nest entrance for a period of up to two days, before they actually start their foraging activity. During these orientation runs the ants gather visual experience and might associate the nest entrance with specific landmarks or get entrained to other visual information like the polarization pattern, and, concomitantly adapt their neuronal circuitries to the upcoming challenges. Moreover, the pirouettes may serve to stimulate and calibrate the neuronal networks involved in the polarization compass pathway. Video recordings and analyses demonstrate that light experience enhanced the ants’ locomotor activity after three days of exposure. The fact that both the light-induced behavioral and neuronal changes in visual brain centers occur in the same time frame suggests that there may be a link between structural synaptic plasticity and the behavioral transition from interior tasks to outdoor foraging. Desert ants of the genus Cataglyphis possess remarkable visual navigation capabilities, but also employ olfactory cues for detecting nest and food sites. Using confocal imaging and 3D-reconstruction, potential adaptations in primary olfactory brain centers were analyzed by comparing the number, size and spatial arrangement of olfactory glomeruli in the antennal lobe of C. fortis, C. albicans, C. bicolor, C. rubra, and C. noda. Workers of all Cataglyphis species have smaller numbers of glomeruli compared to those of more olfactory-guided Formica species - a genus closely related to Cataglyphis - and to those previously found in other olfactory-guided ant species. C. fortis has the lowest number of glomeruli compared to all other species, but possesses a conspicuously enlarged glomerulus that is located close to the antennal nerve entrance. Males of C. fortis have a significantly smaller number of glomeruli compared to female workers and queens and a prominent male-specific macroglomerulus likely to be involved in sex pheromone communication. The behavioral significance of the enlarged glomerulus in female workers remains elusive. The fact that C. fortis inhabits microhabitats that are avoided by all other Cataglyphis species suggests that extreme ecological conditions may not only have resulted in adaptations of visual capabilities, but also in specializations of the olfactory system. The present thesis demonstrates that Cataglyphis is an excellent candidate for studying the neuronal mechanisms underlying navigational features and for studying neuronal plasticity associated with the ant’s lifelong flexibility of individual behavioral repertoires.
Early-life stress has been shown to influence the development of the brain and to increase the risk for psychiatric disorders later in life. Furthermore, variation in the human serotonin transporter (5-HTT, SLC6A4) gene is suggested to exert a modulating effect on the association between early-life stress and the risk for depression. At the basis of these gene x environment (G x E) interactions, epigenetic mechanisms, such as DNA-methylation, seem to represent the primary biological processes mediating early-life programming for stress susceptibility or resilience, respectively. The exact molecular mechanisms however remain to be elucidated, though. In the present study, we used two different stress paradigms to assess the molecular mechanisms mediating the relationship between early-life stress and disorders of emotion regulation later in life. First, a 5-Htt x prenatal stress (PS) paradigm was applied to investigate whether the effects of PS are dependent on the 5-Htt genotype. For this purpose, the effects of PS on cognition and anxiety- / depression-related behavior were examined using a maternal restraint stress paradigm of PS in C57BL/6 wild-type (WT) and heterozygous 5-Htt deficient (5-Htt+/-) mice. Additionally, in female offspring, a genome-wide hippocampal gene expression and DNA methylation profiling was performed using the Affymetrix GeneChip® Mouse Genome 430 2.0 Array and the AffymetrixGeneChip® Mouse Promoter 1.0R Array. Some of the resulting candidate genes were validated by quantitative real-time PCR. Further, the gene expression of these genes was measured in other brain regions of the PS animals as well as in the hippocampus of offspring of another, 5-Htt x perinatal stress (PeS) paradigm, in which pregnant and lactating females were stressed by an olfactory cue indicating infanticide. To assess resilience to PS and PeS, correlation studies between gene expression and behaviour were performed based on an initial performance-based LIMMA analysis of the gene expression microarray. 5-Htt+/- offspring of the PS paradigm showed enhanced memory performance and signs of reduced anxiety as compared to WT offspring. In contrast, exposure of 5-Htt+/- mice to PS was associated with increased depression-like behavior, an effect that tended to be more pronounced in female offspring. Further, 5-Htt genotype, PS and their interaction differentially affected the expression and DNA methylation of numerous genes and related pathways within the female hippocampus. Specifically, MAPK and neurotrophin signaling were regulated by both the 5-Htt+/- genotype and PS exposure, whereas cytokine and Wnt signaling were affected in a 5-Htt genotype x PS manner, indicating a gene x environment interaction at the molecular level. The candidate genes of the expression array could be validated and their expression patterns were partly consistent in the prefrontal cortex and striatum. Furthermore, the genotype effect of XIAP associated factor 1 (Xaf1) was also detected in the mice of the PeS paradigm. Concerning resilience, we found that the expression of growth hormone (Gh), prolactin (Prl) and fos-induced growth factor (Figf) were downregulated in WTPS mice that performed well in the forced swim test (FST). At the same time, the results indicated that Gh and Prl expression correlated positively with adrenal weight, whereas Figf expression correlated positively with basal corticosteron concentration, indicating an intricate relationship between depression-like behavior, hippocampal gene expression and the hypothalamo-pituitary-adrenal (HPA) axis activity. Correlation studies in the PeS animals revealed a link between Gh / Prl expression and anxiety-like behavior. In conclusion, our data suggest that although the 5-Htt+/- genotype shows clear adaptive capacity, 5-Htt+/- mice, particularly females, appear to be more vulnerable to developmental stress exposure when compared to WT offspring. Moreover, hippocampal gene expression and DNA methylation profiles suggest that distinct epigenetic mechanisms at the molecular level mediate the behavioral effects of the 5-Htt genotype, PS exposure, and their interaction. Further, resilience to early-life stress might be conferred by genes whose expression is linked to HPA axis function.
PTPN22 encodes the lymphoid tyrosine phosphatase Lyp that can dephosphorylate Lck, ZAP-70 and Fyn to attenuate TCR signaling. A single-nucleotide polymorphism (C1858T) causes a substitution from arginine (R) to tryptophan (W) at 620 residue (R620W). Lyp-620W has been confirmed as a susceptible allele in multiple autoimmune diseases, including type 1 diabetes (T1D). Several independent studies proposed that the disease-associated allele is a gain-of-function variant. However, a recent report found that in human cells and a knockin mouse containing the R620W homolog that Ptpn22 protein degradation is accelerated, indicating Lyp-620W is a loss-of-function variant. Whether Lyp R620W is a gain- or loss-of-function variant remains controversial. To resolve this issue, we generated two lines (P2 and P4) of nonobese diabetic (NOD) mice in which Ptpn22 can be inducibly silenced by RNAi. We found long term silencing of Ptpn22 increased spleen cellularity and regulatory T (Treg) cell numbers, replicating the effect of gene deletion reported in the knockout (KO) B6 mice. Notably, Ptpn22 silencing also increased the reactivity and apoptotic behavior of B lymphocytes, which is consistent with the reduced reactivity and apoptosis of human B cells carrying the alleged gain-of-function PTPN22 allele. Furthermore, loss of Ptpn22 protected P2 KD mice from spontaneous and Cyclophosphamide (CY) induced diabetes. Our data support the notion that Lyp-620W is a gain-of-function variant. Moreover, Lyp may be a valuable target for the treatment of autoimmune diseases.
Escherichia coli Nissle 1917 (EcN) gehört zu den am besten untersuchten und charakterisierten probiotischen Bakterienstämmen. Seit Beginn des letzten Jahrhunderts wird er als Medikament eingesetzt, um verschiedene Darmerkrankungen wie z.B. Diarrhöe, entzündliche Darmerkrankungen und Verstopfung zu behandeln. Die Flagelle des EcN vermittelt Beweglichkeit und kann die Produktion von humanem β-Defensin 2 (hBD2) durch Epithelzellen induzieren. Somit ist dieses Organell direkt in die probiotische Funktion des EcN involviert. Es konnte gezeigt werden, dass die Flagellen anderer Bakterien, wie z.B. dem probiotischen Stamm Bacillus cereus CH oder den pathogenen Stämmen Pseudomonas aeruginosa und Clostridium difficile, die Adhäsion an intestinalen Mucus, welcher von Epithelzellen sekretiert wird, vermitteln. Allerdings blieb unklar, welcher Teil der Flagelle an welche Mucuskomponente bindet. Die Fähigkeit effizient an Wirtgewebe zu adhärieren wird als wichtiges Attribut eines probiotischen Stammes angesehen. Ex vivo Adhäsionsstudien mit Kryoschnitten humaner Darmbiopsien haben gezeigt, dass die Flagelle des EcN in die effiziente Adhäsion an humanes Darmgewebe involviert sein muss. Aus diesem Grund wurde in dieser Arbeit die Funktion der Flagelle des EcN als Adhäsin untersucht. Zunächst wurde die hyperflagellierte Variante EcN ATHF isoliert und durch verschiedene Experimente, z.B. Schwärmagartests und Elektronenmikroskopie, charakterisiert. Weitere ex vivo Adhäsionsstudien mit EcN ATHF zeigten eine höhere Adhäsionseffizienz dieser hyperflagellierten Variante und bestätigten damit die Rolle der Flagelle bei der effizienten Adhäsion von EcN an die Kryoschnitte der humanen Darmbiopsien. Interessanterweise fungierte die Flagelle in in vitro Studien mit den humanen Epithelzellen Caco-2 und T24 nicht als Adhäsin. Diese Unterschiede zwischen den in vitro und ex vivo Studien führten zu der Annahme, dass die Flagelle des EcN in vivo die Adhäsion an Mucus vermittelt, welcher von den Caco-2- und T24-Zellen nicht produziert wird, aber in den Kryoschnitten der Darmbiopsien nachgewiesen wurde. Diese Vermutung wurde durch in vitro Adhäsionsstudien mit der Mucin-produzierenden Epithelzelllinie LS174-T bestätigt, da die Flagellen für eine effektive Adhäsion an diese Zellen essentiell waren. Zudem reduzierte die Präinkubation flagellierter EcN-Stämme mit Mucin2 ihre Adhäsionseffizienz an Kryoschnitte humaner Darmbiopsien. Um die direkte Interaktion zwischen Flagellen des EcN Wildtyps und Mucus zu zeigen, wurde ein ELISA etabliert. Es konnte eine direkte konzentrationsabhängige Interaktion zwischen isolierten Flagellen des EcN Wildtyps und Mucin2, bzw. humanem Mucus (Kolon) beobachtet werden. Interessanterweise konnte keine Interaktion zwischen isolierten Flagellen des EcN Wildtyps und murinem Mucus (Duodenum, Ileum, Caecum, Colon) festgestellt werden. Dies weist darauf hin, dass die Mucuszusammensetzung zwischen verschiedenen Spezies variiert. Verschiedene Kohlenhydrate, welche bekannte Mucusbestandteile sind, wurden auf ihre Interaktion mit der Flagelle von EcN getestet und Gluconat wurde als ein Rezeptor identifiziert. Die Präinkubation isolierter Flagellen mit Gluconat reduzierte ihre Interaktion mit Mucin2, bzw. humanem Mucus signifikant. Zudem wurde die oberflächenexponierte Domäne D3 des Flagellins, der Hauptuntereinheit der Flagelle, als möglicher Interaktionspartner von Mucin2, bzw. humanem Mucus ausgeschlossen. Flagellen, die aus einer Domäne D3 Deletionsmutante isoliert wurden, zeigten sogar eine effizientere Bindung an Mucin2, bzw. humanen Mucus. Weiterhin konnte gezeigt werden, dass Änderungen des pH-Wertes signifikante Effekte auf die Interaktion zwischen Mucus und isolierten Flagellen hatten, vermutlich aufgrund von Konformationsänderungen. Zusammenfassend wurde in dieser Arbeit die Flagelle als neues und scheinbar wichtigstes Adhäsin in vivo für den probiotischen Stamm EcN identifiziert. Hierfür wurden sowohl eine hyperflagellierte Variante, eine ΔfliC Mutante, sowie der dazugehörige komplementierte Stamm verwendet. EcN ist zudem der erste probiotische Stamm für den eine direkte Bindung der Flagellen an humanen Mucus nachgewiesen werden konnte. Die Mucuskomponente Gluconat konnte dabei als wichtiger Rezeptor identifiziert werden. Da einige pathogene Bakterien ihre Flagelle zur Adhäsion an Wirtsgewebe nutzen, könnte dieses Organell EcN dazu befähigen, mit Pathogenen um die erfolgreiche Kolonisierung des Darms zu konkurrieren, was als wichtige Eigenschaft eines Probiotikums betrachtet wird.
Based on genetic association and functional imaging studies, reduced function of tryptophan hydroxylase-2 (TPH2) has been shown to be critically involved in the pathophysiology of anxiety-disorders and depression. In order to elucidate the impact of a complete neuronal 5-HT deficiency, mice with a targeted inactivation of the gene encoding Tph2 were generated. Interestingly, survival of Tph2-/- mice, the formation of serotonergic neurons and the pathfinding of their projections was not impaired. Within this thesis, I investigated the influence of 5-HT deficiency on the γ-amino butyric acid (GABA) system. The GABAergic system is implicated in the pathophysiology of anxiety disorders. Therefore, measurement of GABA concentrations in different limbic brain regions was carried out. These measurements were combined with immunohistochemical estimation of GABAergic cell subpopulations in the dorsal hippocampus and amygdala. In Tph2-/- mice GABA concentrations were increased exclusively in the dorsal hippocampus. In heterozygous Tph2+/- mice concentrations of GABA were increased in the amygdala compared to Tph2-/- and wt control mice, while the reverse was found in the prefrontal cortex. The changes in GABA concentrations were accompanied by altered cell density of GABAergic neurons within the basolateral complex of the amygdala and parvalbumin (PV) neurons of the dorsal hippocampus and by adaptational changes of 5-HT receptors. Thus, adaptive changes during the development on the GABA system may reflect altered anxiety-like and depressive-like behavior in adulthood. Moreover, chronic mild stress (CMS) rescues the depressive-like effects induced by 5-HT deficiency. In contrast, 5-HT is important in mediating an increased innate anxiety-like behavior under CMS conditions. This is in line with a proposed dual role of 5-HT acting through different mechanisms on anxiety and depressive-like behavior, which is influenced by gene-environment interaction effects. Further research is needed to disentangle these complex networks in the future.
Malaria and HIV are among the most important global health problems of our time and together are responsible for approximately 3 million deaths annually. These two diseases overlap in many regions of the world including sub-Saharan Africa, Southeast Asia and South America, leading to a higher risk of co-infection. In this study, we generated and characterized hybrid molecules to target P. falciparum and HIV simultaneously for a potential HIV/malaria combination therapy. Hybrid molecules were synthesized by covalent fusion between azidothymidine (AZT) and dihydroartemisinin (DHA), tetraoxane or chloroquine (CQ); and a small library was generated and tested for antiviral and antimalarial activity. Our data suggest that dihyate is the most potent molecule in vitro, with antiplasmodial activity comparable to that of DHA (IC50 = 26 nM, SI > 3000), a moderate activity against HIV (IC50 = 2.9 µM; SI > 35) and safe to HeLa cells at concentrations used in the assay (CC50 > 100 µM). Pharmacokinetic studies further revealed that dihyate is metabolically unstable and is cleaved following an O-dealkylation once in contact with cytochrome P450 enzymes. The later further explains the uneffectiveness of dihyate against the CQ-sensitive P. berghei N strain in mice when administered by oral route at 20 mg/kg. Here, we report on a first approach to develop antimalarial/anti-HIV hybrid molecules and future optimization efforts will aim at producing second generation hybrid molecules to improve activity against HIV as well as compound bioavailability. With the emergence of resistant parasites against all the counterpart drugs of artemisinin derivatives used in artemisinin based combination therapies (ACTs), the introduction of antibiotics in the treatment of malaria has renewed interest on the identification of antibiotics with potent antimalarial properties. In this study we also investigated the antiplasmodial potential of thiostrepton and derivatives, synthesized using combinations of tail truncation, oxidation, and addition of lipophilic thiols to the terminal dehydroamino acid. We showed that derivatives SS231 and SS234 exhibit a better antiplasmodial activity (IC50 = 1 µM SI > 59 and SI > 77 respectively) than thiostrepton (IC50 = 8.95 µM, SI = 1.7). The antiplasmodial activity of these derivatives was observed at concentrations which are not hemolytic and non-toxic to human cell lines. Thiostrepton and derivatives appeared to exhibit transmission blocking properties when administered at their IC50 or IC90 concentrations and our data also showed that they attenuate proteasome activity of Plasmodium, which resulted in an accumulation of ubiquitinated proteins after incubation with their IC80 concentrations. Our results indicate that the parasite’s proteasome could be an attractive target for therapeutic intervention. In this regard, thiostrepton derivatives are promising candidates by dually acting on two independent targets, the proteasome and the apicoplast, with the capacity to eliminate both intraerythrocytic asexual and transmission stages of the parasite. To further support our findings, we evaluated the activity of a new class of antimalarial and proteasome inhibitors namely peptidyl sulfonyl fluorides on gametocyte maturation and analogues AJ34 and AJ38 were able to completely suppress gametocytogenesis at IC50 concentrations (0.23 µM and 0.17 µM respectively) suggesting a strong transmission blocking potential. The proteasome, a major proteolytic complex, responsible for the degradation and re-cycling of non-functional proteins has been studied only indirectly in P. falciparum. In addition, an apparent proteasome-like protein with similarity to bacterial ClpQ/hslV threonine-peptidases was predicted in the parasite. Antibodies were generated against the proteasome subunits alpha type 5 (α5-SU), beta type 5 (β5-SU) and pfhslV in mice and we showed that the proteasome is expressed in both sexual and asexual blood stages of P. falciparum, where they localize in the nucleus and in the cytoplasm. However, expression of PfhslV was only observed in trophozoites and shizonts. The trafficking of the studied proteasome subunits was further investigated by generating parasites expressing GFP tagged proteins. The expression of α5-SU-GFP in transgenic parasite appeared to localize abundantly in the cytoplasm of all blood stages, and no additional information was obtained from this parasite line. In conclusion, our data highlight two new tools towards combination therapy. Hybrid molecules represent promising tools for the cure of co-infected individuals, while very potent antibiotics with a wide scope of activities could be useful in ACTs by eliminating resistant parasites and limiting transmission of both, resistances and disease.
SUMMARY Mast cell activation in allergic and inflammatory disease causes increased vascular permeability and edema. This thesis identifies a paracrine mechanism, by which heparin released from intracellular granules, is involved in mast cell-evoked alteration of endothelial barrier function in vivo. Negatively charged heparin initiated factor XII-driven contact activation. Activated factor XII triggered the formation of the inflammatory mediator bradykinin in plasma. Congenital deficiency and pharmacological targeting of factor XII and kinin B2 receptor provided protection from mast cell-heparin-induced leukocyte-endothelial adhesion and hypotension in rats and mice. Intravital laser scanning microscopy and tracer measurements showed that heparin increased leakage with fluid extravasation in skin microvessels in mice. Deficiency in factor XII or kinin B2 receptor conferred resistance to heparin-induced skin edema and largely protected mice from endothelial barrier dysfunction, caused by allergen-induced mast cell activation and anaphylactic reactions. In contrast, heparin and mast cell activation caused excessive edema formation in mice, deficient in the major inhibitor of factor XII, C1 esterase inhibitor. Hereditary angioedema patients, lacking C1 esterase inhibitor, suffered from allergeninduced edema. The data indicate that mast cell-heparin-initiated bradykinin formation plays a fundamental role in defective barrier function of pathological mast cell-mediated inflammation, hypotension and edema formation.
Autoimmune diseases, unwanted overshooting immune responses against self antigens, are due to an imbalance in immunity and tolerance. Although negatively impacting cancer prognosis, myeloid derived suppressor cells (MDSC), with their potent suppressive capabilities, might be applicable in a more beneficial light when applied in to autoimmunity. As previous shown MDSC have protective roles in Experimental Autoimmune Encephalomyelitis (EAE) (Zhu et al., 2007), the established inducible mouse model for the autoimmune disease multiple sclerosis (MS). This decrease in disease severity indicates in vitro generated immature myeloid cells (IMC) from bone marrow (BM) as precursors of MDSC are promising candidates for cellular therapy. Important to any cellular therapy by adoptive transfer, the major questions regarding IMC efficacy was addressed within the thesis. This thesis attempts to elucidate how IMC operate in EAE. This thesis defines the factors within the autoimmune microenvironment that lead to the activation of MDSC, where IMC home once delivered in vivo, and the protective mechanisms BMIMC employ. To emulate BM cells when they first enter circulation through the blood, IMC were injected intravenously (i.v.). IMC are protective with no regard to the various routes delivered (i.v., i.p.). They protect to a lesser extent when pre-activated before injection. IMC suppress by causing a delay and/or by decreasing the severity of the disease via a mechanism yet determined. To understand the migration pattern of IMC after i.v. injection, in vivo kinetics experiments employing bioluminescence imaging were performed. This techinique allows for whole in vivo mouse imaging daily, allowing the tracking of cell migration over days within a single mouse. During steady-state, BMIMC circulate and appear to accumulate in the spleen by day 4 after injection, whereas they alternatively home to inflammatory sites (immunization site), draining lymph nodes, and the spleen within mice with low grade EAE. Visualization of CMDiI-labelled BMIMC by fluorescence microscopy could locate IMC injected cells outside the white pulp, as they were colocalizing in the regions stained with CD169 or outside, but not within the follicles of spleens on day 4. Consistant with these findings, the attempt to analyze the phenotype of these cells by flow cytometry was problematic as these cells seem to adhere strongly to collagen also indicating the cells are located in the collagenous area of the marginal zone and the red pulp.To determine factors influencing MDSC activation, we utilized different stimuli through a high throughput method detecting release of nitric oxide (NO). Extracts from yeast, fungi, and bacteria were observed to activate MDSC to produce nitric oxide. Surprisingly, material mimicking viral DNA (CpG) and RNA (poly I:C), and several self glycolipids, could not activate the MDSC to produce NO. Upon attempts to understand synergistic effects between microbial pathogens and host cytokines, IFNg was determined to boost the signal of pathogen stimuli, whereas IL17, another cytokine which causes pathology during EAE, and IFNb, a drug used in therapy to treat MS, did not cause any additional effects. Activation of MDSC was determined by the microbial pathogens components LPS, curdlan, and zymosan, to induce upregulation of B7H1 on the cell surface. MDSC did not increase any co-stimulatory markers, such as CD40, CD80, CD86, CD70, or the co-inhibitory marker, PDL2. On day 1 after EAE induction, endogenous MDSC populations when stimulated showed an increase in B7H1 expression and a downregulation of CD80. After further analysis, these cells were concluded to be mostly granulocytic cells (Ly6G+). As the B7H1 ligand PD1 is upregulated in chronic diseases and correlates to an exhausted phenotype, the PD1 : B7H1 interaction was a good candidate for the mechanism our cells may employ for their suppressive capacity. To investigate this interaction, fixed BM-IMC deficient in B7H1 were incubated with restimulated memory T cells. IMC deficient in B7H1 resulted in a significant loss of T cell suppression, as compared to the wildtype control BMIMC. To assess this interaction in vivo, we injected wildtype (WT) and B7H1-/- IMC into mice followed by induction of EAE to assess whether B7H1 mediated this suppression. The lack of B7H1 did not alter their suppressive capacity under these conditions, contrary to other findings which have described this interaction to be important in their suppressive capacity when administered post EAE induction (Ioannou et al., 2012). Interestingly, EAE mice pre-treated with IMC had similar amounts of cytokine production in the CNS after restimulation. Spleens from IMC injected mice had increased amounts of Arg-1 suggesting suppression is via oxidation or recruitment by soluble mediators may lead to this protection. We speculate this may inhibit T cell reactivation in the CNS.
Plant-derived natural products and their analogs continue to play an important role in the discovery of new drugs for the treatment of human diseases. Potentially promising representatives of secondary metabolites are the naphthylisoquinoline alkaloids, which show a broad range of activities against protozoan pathogens, such as plasmodia, leishmania, and trypanosoma. Due to the increasing resistance of those pathogens against current therapies, highly potent novel agents are still urgently needed. Thus, it is worthy to discover new naphthylisoquinoline alkaloids hopefully with pronounced bioactivities by isolation from plants or by synthesis. The naphthylisoquinoline alkaloids are biosynthetically related to another class of plant-derived products, the naphthoquinones, some of which have been recently found to display excellent anti-multiple myeloma activities without showing any cytotoxicities on normal blood cells. Multiple myeloma still remains incurable, although remissions may be induced with co-opted therapeutic treatments. Therefore, more potent naphthoquinones are urgently required, and can be obtained by isolation from plants or by synthesis. In detail, the results in this thesis are listed as follows: 1) Isolation and characterization of naphthylisoquinoline alkaloids from the stems of a Chinese Ancistrocladus tectorius species. Nine new naphthylisoquinoline alkaloids, named ancistectorine A1 (60), N-methylancistectorine A1 (61), ancistectorine A2 (62a), 5-epi-ancistectorine A2 (62b), 4'-O-demethylancistectorine A2 (63), ancistectorine A3 (64), ancistectorine B1 (65), ancistectorine C1 (66), and 5-epi-ancistrolikokine D (67) were isolated from the Chinese A. tectorius and fully characterized by chemical, spectroscopic, and chiroptical methods. Furthermore, the in vitro anti-infectious activities of 60-62 and 63-66 have been tested. Three of the metabolites, 61, 62a, and 62b, exhibited strong antiplasmodial activities against the strain K1 of P. falciparum without showing significant cytotoxicities. With IC50 values of 0.08, 0.07, and 0.03 μM, respectively, they were 37 times more active than the standard chloroquine (IC50 = 0.26 μM). Moreover, these three compounds displayed high antiplasmodial selectivity indexes ranging from 100 to 3300. According to the TDR/WHO guidelines, they could be considered as lead compounds. In addition, seven alkaloids, 69-74 (structures not shown here), were isolated from A. tectorius that were known, but new to the plant, together with another fourteen known compounds (of these, only the structures of the three main alkaloids, 5a, 5b, and 78 are shown here), which had been previously found in the plant. The three metabolites ancistrocladine (5a), hamatine (5b), and (+)-ancistrocline (78) were found to show no or moderate activities against the MM cell lines. 2) Isolation and characterization of naphthylisoquinoline alkaloids from the root bark of a new, botanically yet undescribed Congolese Ancistrocladus species. An unprecedented dimeric Dioncophyllaceae-type naphthylisoquinoline alkaloid, jozimine A2 (84), as first recognized by G. Bauckmann from an as yet undescribed Ancistrocladus species, was purified and characterized as part of this thesis. Its full structural assignment was achieved by spectroscopic and chiroptical methods, and further confirmed by an X-ray diffraction analysis, which had never succeeded for any other dimeric naphthylisoquinoline alkaloids before. Structurally, the dimer is composed of two identical 4'-O-demethyldioncophylline A halves, coupled through a sterically hindered central axis at C-3',3'' of the two naphthalene moieties. Pharmacologically, jozimine A2 (84) showed an extraordinary antiplasmodial activity (IC50 = 1.4 nM) against the strain NF54 of P. falciparum. Beside jozimine A2 (85), another new alkaloid, 6-O-demethylancistrobrevine C (84), and four known ones, ancistrocladine (5a), hamatine (5b), ancistrobrevine C (86), and dioncophylline A (6) were isolated from the Ancistrocladus species, the latter in a large quantity (~500 mg), showing that the plant produces Ancistrocladaceae-type, mixed-Ancistrocladaceae/Dioncophyllaceae-type, and Dioncophyllaceae-type naphthyl- isoquinoline alkaloids. Remarkably, it is one of the very few plants, like A. abbreviatus, and A. barteri, that simultaneously contain typical representatives of all the above three classes of alkaloids. 3) Semi-synthesis of jozimine A2 (85), 3'-epi-85, jozimine A3 (93) and other alkaloids from dioncophylline A (6). The dimeric naphthylisoquinoline alkaloids, jozimine A2 (85) and 3'-epi-85, constitute rewarding synthetic targets for a comparative analysis of their antiplasmodial activities and for a further confirmation of the assigned absolute configurations of the isolated natural product of 85. They were semi-synthesized in a four-step reaction sequence from dioncophylline A (6) in cooperation with T. Büttner. The key step was a biomimetic phenol-oxidative dimerization at C-3' of the N,O-dibenzylated derivative of 89 by utilizing Pb(OAc)4. This is the first time that the synthesis of such an extremely sterically hindered (four ortho-substituents) naphthylisoquinoline alkaloid – with three consecutive biaryl axes! – has been successfully achieved. A novel dimeric naphthylisoquinoline, jozimine A3 (93), bearing a 6',6''-central biaryl axis, was semi-synthesized from 5'-O-demethyldioncophylline A (90) by a similar biomimetic phenol-oxidative coupling reaction as a key step, by employing Ag2O. HPLC analysis with synthetic reference material of 3'-epi-85 and 93 for co-elution revealed that these two alkaloids clearly are not present in the crude extract of the Ancistrocladus species from which jozimine A2 (85) was isolated. This evidences that jozimine A2 (85) is very specifically biosynthesized by the plant with a high regio- and stereoslectivity. Remarkably, the two synthetic novel dimeric naphthylisoquinoline alkaloids 3'-epi-85 and 93 were found to display very good antiplasmodial activities, albeit weaker than that of the natural and semi-synthetic product 85. Additionally, the two compounds 3'-epi-85 and 93 possessed high or moderate selectivity indexes, which were much lower than that of 85. However, they can still be considered as new lead structures. Two unprecedented oxidative products of dioncophylline A, the diastereomeric dioncotetralones A (94a) and B (94b), were synthesized from dioncophylline A (6) in a one-step reaction. Remarkably, the aromatic properties in the “naphthalene” and the “isoquinoline” rings of 94a and 94b are partially lost and the “biaryl” axis has become a C,C-double bond, so that the two halves are nearly co-planar to each other, which has never been found among any natural or synthetic naphthylisoquinoline alkaloid. Their full structural characterization was accomplished by spectroscopic methods and quantum-chemical CD calculations (done by Y. Hemberger). The presumed reaction mechanism was proposed in this thesis. In addition, one of the two compounds, 94a, exhibited a highly antiplasmodial activity (IC50 = 0.09 μM) with low cytotoxicity, and thus, can be considered as a new promising lead structure. Its 2'-epi-isomer, 94b, was inactive, evidencing a significant effect of chirality on the bioactivity. Of a number of naphthylisoquinoline alkaloids tested against the multiple-myeloma cell lines, the three compounds, dioncophylline A (6), 4'-O-demethyldioncophylline A (89), and 5'-O-demethyldioncophylline A (90) showed excellent activities, even much stronger than dioncoquinones B (10), C (102), the epoxide 175, or the standard drug melphalan. 4) Isolation and characterization of bioactive naphthoquinones from cell cultures of Triphyophyllum peltatum. Three new naphthoquinones, dioncoquinones C (102), D (103), and E (104), the known 8-hydroxydroserone (105), which is new to this plant, and one new naphthol dimer, triphoquinol A (107), were isolated from cell cultures of T. peltatum in cooperation with A. Irmer. Dioncoquinone C (102) showed an excellent activity against the MM cells, very similar to that of the previously found dioncoquinone B (10), without showing any inhibitory effect on normal cells. The other three naphthoquinones, 103105, were inactive or only weakly active. 5) Establishment of a new strategy for a synthetic access to dioncoquinones B (10) and C (102) on a large scale for in vivo experiments and for the synthesis of their analogs for first SAR studies. Before the synthesis of dioncoquinone B (10) described in this thesis, two synthetic pathways had previously been established in our group. The third approach described here involved the preparation of the joint synthetic intermediate 42 with the previous two routes. The tertiary benzamide 135 was ortho-deprotonated by using s-BuLi/TMEDA, followed by transmetallation with MgBr2▪2Et2O, and reaction with 2-methylallyl bromide (139). It resulted in the formation of ortho-allyl benzamide 140, which was cyclized by using methyl lithium to afford the naphthol 42. This strategy proved to be the best among the established three approaches with regard to its very low number of steps and high yields. By starting with 136, this third strategy yielded the related bioactive natural product, dioncoquinone C (102), which was accessed by total synthesis for the first time. To identify the pharmacophore of the antitumoral naphthoquinones, a library of dioncoquinone B (10) and C (102) analogs were synthesized for in vitro testing. Among the numerous naphthoquinones tested, the synthetic 7-O-demethyldioncoquinone C (or 7-O-hydroxyldioncoquinone B) (145), constitutes another promising basic structure to develop a new anti-MM agent. Furthermore, preliminary SAR results evidence that the three hydroxy functions at C-3, C-5, and C-6 are essential for the biological properties as exemplarily shown through the compounds 10, 102, and 145. All other mixed OH/OMe- or completely OMe-substituted structures were entirely inactive. By a serendipity the expoxide 175 was found to display the best anti-MM activity of all the tested isolated metabolites from T. peltatum, the synthesized naphthoquinones, and their synthetic intermediates. Toxic effects of 175 on normal cells were not observed, in contrast to the high toxicities of all other epoxides. Thus, the anti-MM activity of 175 is of high selectivity. The preliminary SAR studies revealed that the 6-OMe group in 175 is required, thus differed with the above described naphthoquinones (where 6-OH is a requisite in 10, 102, and 145), which evidenced potentially different modes of action for these two classes of compounds. 6) The first attempted total synthesis of the new naturally occurring triphoquinone (187a), which was recently isolated from the root cultures of T. peltatum in our group. A novel naphthoquinone-naphthalene dimer, 187a (structure shown in Chapter 10), was isolated in small quantities from the root cultures of T. peltatum. Thus, its total synthesis was attempted for obtaining sufficient amounts for selected biotestings. The key step was planned to prepare the extremely sterically hindered (four ortho-substituents) binaphthalene 188 by a coupling reaction between the two 2-methylnaphthalene derivatives. Test reactions involving a system of two simplified 2-methylnaphthylboron species and 2-methylnaphthyl bromide proved the Buchwald ligand as most promising. The optimized conditions were then applied to the two true - highly oxygenated - coupling substrates, between the 2-methylnaphthylboron derivatives 210, 211, 213, or 214 and the 2-methylnaphthyl iodides (or bromides) 215 (206), 215 (206), 212 (205), or 212 (205), respectively. Unfortunately, this crucial step failed although various bases and solvent systems were tested. This could be due to the high electron density of the two coupling substrates, both bearing strongly OMOM/OMe-donating function groups. Therefore, a more powerful catalyst system or an alternative synthetic strategy must be explored for the total synthesis of 187a. 7) Phytochemical investigation of the Streptomyces strain RV-15 derived from a marine sponge. Cyclodysidins A-D (216-219), four new cyclic lipopeptides with a- and ß-amino acids, were isolated from the Streptomyces strain RV15 derived from a marine sponge by Dr. U. Abdelmohsen. Their structures were established as cyclo-(ß-AFA-Ser-Gln-Asn-Tyr-Asn-Ser-Thr) by spectroscopic analysis using 2D NMR techniques and CID-MS/MS in the course of this thesis. In conclusion, the present work contributes to the discovery of novel antiplasmodial naphthylisoquinoline alkaloids and antitumoral naphthoquinones, which will pave the way for future studies on these two classes of compounds.
Das Y-Box-bindende Protein 1 (YB-1) ist ein Vertreter der hochkonservierten Familie eukaryotischer Kälteschockproteine und ein DNA/RNA-bindendes Protein. In Abhängigkeit von seiner Lokalisation übernimmt es Aufgaben bei der DNA-Transkription oder mRNA-Translation. YB-1 ist ein potentielles Onkogen beim Multiplen Myelom (MM), dass in primären MM-Zellen exprimiert ist. Für die funktionellen Untersuchungen von YB-1 in der vorliegenden Arbeit wurden humane Myelomzelllinien (HMZL) verwendet, die als in vitro Modell dieser malignen B Zell-Erkrankung dienen. Aufgrund der potentiellen Expression von YB-1 im Zellkern und/oder Zytoplasma von HMZL, wurde zunächst die Lokalisation des Proteins bestimmt. Es konnte gezeigt werden, dass YB 1 in den HMZL ausschließlich im Zytoplasma lokalisiert ist. Eine Translokation von YB-1 in den Nukleus kann durch die Serin-Phosphorylierung (Aminosäure 102) in der Kälteschockdomäne induziert werden. Die analysierten Myelomzelllinien zeigen jedoch kein nukleäres YB 1 und keine S102-Phosphorylierung. Diese Ergebnisse stützen die These, dass die Regulation der mRNA-Translation im Zytoplasma die vorherrschende Funktion von YB-1 beim MM ist. YB-1 könnte über diesen Mechanismus seine anti-apoptotische Wirkung vermitteln und die MM-Zellen vor genotoxischem Stress schützen. Um YB-1-regulierte mRNAs zu identifizieren wurden YB 1-Immunpräzipitationen mit zwei HMZL, einer Maus-Plasmozytomzelllinie und einem primären Maus-Plasmazelltumor durchgeführt. Zu den YB-1-gebundenen mRNAs gehören Translationsfaktoren und ribosomale Proteine, die eine starke Beteiligung von YB-1 beim RNA-Metabolismus bestätigen. In der vorliegenden Arbeit wurden spezifisch zwei mRNA-Kandidaten untersucht, die für den malignen Phänotyp von MM-Zellen wichtig sein können: das translationell kontrollierte Tumorprotein TCTP und MYC. Sowohl TCTP als auch MYC wurden bereits in Zusammenhang mit der Proliferation und Apoptose-Resistenz von malignen Zellen beschrieben. Die immunhistochemische Untersuchung der Knochenmarkbiopsien von MM-Patienten ergab eine gute Ko-Expression von YB-1 und TCTP in intramedullären MM-Zellen, während MYC erst in extramedullärem MM-Tumormaterial verstärkt mit der hohen YB 1-Expression korreliert. Die funktionellen Analysen der Arbeit haben gezeigt, dass YB 1 für die Translation der TCTP- und MYC-mRNA essentiell ist. Es kontrolliert die Verteilung dieser mRNAs zwischen translationell aktiven und inaktiven messenger Ribonukleoprotein-Partikeln. Die shRNA-vermittelte Reduktion von YB-1 führte zur Hemmung der TCTP- und MYC-Translation in der Phase der Initiation. Um den Einfluss der Kandidaten auf das Überleben der HMZL zu untersuchen, wurden proteinspezifische Knockdown-Experimente durchgeführt. Beim shRNA-vermittelten TCTP-Knockdown konnten keine Auswirkungen auf die Proliferation oder Viabilität von MM-Zellen beobachtet werden. Im Gegensatz dazu ist MYC für das Überleben und Wachstum der HMZL ausschlaggebend, denn der MYC-Knockdown induzierte Apoptose. Wie beim YB 1-Knockdown war ein Anstieg der Caspase-Aktivität und der Zusammenbruch des mitochondrialen Membranpotentials in den HMZL nachweisbar. Da es beim MYC-Knockdown gleichzeitig zur einer Reduktion der YB 1-Protein- und mRNA-Expression kam, wurde der Einfluss von MYC auf die Transkription des YB-1-Gens untersucht. Mit Hilfe von embryonalen Mausfibroblasten, die ein induzierbares MYC als Transgen besitzen, konnte gezeigt werden, dass die Aktivierung von MYC mit einer Zunahme der YB-1-mRNA einher geht. YB-1 ist somit ein direktes Zielgen des Transkriptionsfaktors MYC. Die Ergebnisse der vorliegenden Arbeit haben zum ersten Mal ein gegenseitiges regulatorisches Netzwerk aufgezeigt, in dem YB 1 transkriptionell durch MYC reguliert wird und YB-1 für die Translation der MYC-mRNA essentiell ist. Die Ko-Expression beider Proteine trägt zum Wachstum und Überleben von malignen Plasmazellen bei.
Dendritic cell-based vaccination is a well established technique for preventive and therapeutic instruction of the immune system where conservative vaccine formulations fail to cure or prevent diseases, respectively. Efficiency of this technique already was demonstrated in infectious diseases as well as for cancer in animal or human studies. Well controlled manipulation and antigen-loading of immature DC is most beneficial to this technique. But, time-consuming and cost-extensive procedures for preparation of DC precursors, expansion and stimulation of DC and inpatient administration are big disadvantages regarding vaccine development for pandemic infectious diseases that occur mainly in underdeveloped countries. Therefore vaccines are needed that are pathogen-tailored and able to induce equal immune responses as their DC-based vaccine models. For vaccination against Leishmania parasites such a DC-based vaccine is feasible and its efficacy to induce protective Th1-based immune responses was already demonstrated in several animal studies. But, one of our own studies indicated supportive activity of host cells exceeding the allocation of T cells to become activated by transferred DC. IL-12, an important cytokine for the induction of Th1-related immune responses, has to be produced by host cells. Therefore, the aim of this study was to investigate the mechanism of BMDC-based vaccination with regard to simplification of the vaccine formulation. Key questions that have been addressed are: Which cells process the information that is transferred by the injected DC and what are the key components of this information? Further more, it was looked at whether altered vaccine formulations are able to induce protective immunity and whether they share equal molecular mechanisms. The current paradigm of BMDC-based vaccination proposes direct interaction of transferred BMDC with host T cells. These BMDC have to be antigen-loaded for stimulation via antigen-peptide-MHC molecule-complexes and they have to be activated for proper co-stimulation of T cells. Here, this study demonstrates that neither activation for co-stimulation nor direct interaction with adequate MHC molecules is needed for the induction of protective immunity against infection with Leishmania-parasites. Disrupted antigen-loaded BMDC are able to induce protective immunity in BALB/c mice without pre-stimulation via CpG ODN. Beyond, if BMDC were used with a different MHC-background than recipient mice then the vaccine still would be efficient in terms of reduction of footpad swelling and parasite load in draining lymph nodes. Even more, DC-specific features are no key component that leads to protective immunity as vaccination with disrupted antigen-loaded MΦ shows equal properties than before mentioned vaccine formulations. Further more, it was found that host DC play a major role in transforming the incoming signal, received from transferred antigen-loaded DC, into Th1-related stimuli and Leishmania-antigen-specific T cell activation. Suspensions of disrupted antigen-loaded DC resemble a combination of laid off soluble molecules together with exosome-like vesicles that formed after disruption of membranes. Here it was shown that separation of the membranous and soluble fractions and subsequent transfer into BALB/c mice will lead to protection of these mice against infection with L. major promastigotes only if the membranous fraction is used as vaccine. More, this vaccine formulation takes advantage of easy storage at -80°C with no need of fresh production. This clearly demonstrates that the immunity-inducing principle of disrupted DC-based vaccination lies within the membrane enclosed fraction. On a molecular level, disrupted antigen-loaded DC induce Th1-related cytokines during vaccination and as response on pathogen encounter. In vivo assays revealed IL-12 production and antigen-specific T cell proliferation among splenocytes that were stimulated with disrupted antigen-loaded DC. Splenocytes of accordingly vaccinated mice produce tremendous amounts of IFNγ after stimulation with Leishmania parasites. In summary, disrupted antigen-loaded BMDC fulfil all characteristics of DC-based vaccination against Leishmania major. But, while purification of membranes of antigen-loaded DC and subsequent transfer to BALB/c mice leads to control of the disease in the animal model, only slight levels of Th1-related cytokines are seen in the in vivo assays. Whether this points towards a loss of vaccine activity on unseen levels or unknown sites where Th1-related immunity is induced by both, complete solution and purified membranes, still has to be determined.
Imprinted genes play important roles in brain development. As the neural developmental capabilities of human parthenogenetic embryonic stem cells (hpESCs) with only a maternal genome were not assessed in great detail, hence here the potential of hpESCs to differentiate into various neural subtypes was determined. In addition DNA methylation and expression of imprinted genes upon neural differentiation was also investigated. The results demonstrated that hpESC-derived neural stem cells (hpNSCs) showed expression of NSC markers Sox1, Nestin, Pax6, and Musashi1 (MS1), the silencing of pluripotency genes (Oct4, Nanog) and the absence of activation of neural crest (Snai2, FoxD3) and mesodermal (Acta1) markers. Moreover, confocal images of hpNSC cultures exhibited ubiquitous expression of NSC markers Nestin, Sox1, Sox2 and Vimentin. Differentiating hpNSCs for 28 days generated neural subtypes with neural cell type-specific morphology and expression of neuronal and glial markers, including Tuj1, NeuN, Map2, GFAP, O4, Tau, Synapsin1 and GABA. hpNSCs also responded to region-specific differentiation signals and differentiated into regional phenotypes such as midbrain dopaminergic- and motoneuron-type cells. hpESC-derived neurons showed typical neuronal Na+/K+ currents in voltage clamp mode, elicited multiple action potentials with a maximum frequency of 30 Hz. Cell depicted a typical neuron-like current pattern that responded to selective pharmacological blockers of sodium (tetrodotoxin) and potassium (tetraethylammonium) channels. Furthermore, in hpESCs and hpNSCs the majority of CpGs of the differentially methylated regions (DMRs) KvDMR1 were methylated whereas DMR1 (H19/Igf2 locus) showed partial or complete absence of CpG methylation, which is consistent with a parthenogenetic (PG) origin. Upon differentiation parent-of-origin-specific gene expression was maintained in hpESCs and hpNSCs as demonstrated by imprinted gene expression analyses. Together this shows that despite the lack of a paternal genome, hpNSCs are proficient in differentiating into glial- and neuron-type cells, which exhibit electrical activity similar to newly formed neurons. Moreover, maternal-specific gene expression and imprinting-specific DNA-methylation are largely maintained upon neural differentiation. hpESCs are a means to generate histocompatible and disease allele-free ESCs. Additionally, hpESCs are a unique model to study the influence of imprinting on neurogenesis.
Cutaneous leishmaniasis is endemic in tropical and subtropical regions of the world. Effective vaccination strategies are urgently needed because of the emergence of drug-resistant parasites and severe side effects of chemotherapy. The research group of Heidrun Moll previously established a DC-based vaccination strategy to induce complete and long-lasting immunity to experimental leishmaniasis using LmAg-loaded and CpG ODN-activated DC as a vaccine carrier. Prevention of tissue damages at the site of L. major inoculation can be achieved if the BALB/c mice were systemically given LmAg-loaded BMDC that had been exposed to CpG ODN. The interest in further exploring the role of IL-4 aroused as previous studies allowed establishing that IL-4 was involved in the redirection of the immune response towards a type 1 profile. Thus, wt BALB/c mice or DC-specific CD11ccreIL-4Rα-/lox BALB/c mice were given either wt or IL-4Rα-deficient LmAg-loaded BMDC exposed or not to CpG ODN prior to inoculation of 2 x 105 stationary phase L. major promastigotes into the BALB/c footpad. The results provide evidence that IL4/IL-4Rα-mediated signaling in the vaccinating DC is required to prevent tissue damages at the site of L. major inoculation, as properly conditioned wt DC but not IL-4Rα-deficient DC were able to confer resistance. Furthermore, uncontrolled L. major population size expansion was observed in the footpad and the footpad draining LN in CD11ccreIL-4Rα-/lox mice immunized with CpG ODN-exposed LmAg-loaded IL-4Rα-deficient DC, indicating the influence of IL-4R-mediated signaling in host DC to control parasite replication. In addition, no footpad damage was observed in BALB/c mice that were systemically immunized with LmAg-loaded wt DC doubly exposed to CpG ODN and recombinant IL-4. Discussing these findings allow the assumption that triggering the IL4/IL4Rα signaling pathway could be a precondition when designing vaccines aimed to prevent damaging processes in tissues hosting intracellular microorganisms.
Der Tumornekrosefaktor (TNF) entfaltet seine vielfältigen biologischen Aktivitäten durch die Stimulation der beiden TNF-Rezeptoren TNFR1 und TNFR2. Die TNFR1-vermittelte Signaltransduktion ist in vielen Details gut verstanden, wohingegen die TNFR2-vermittelte Signaltransduktion bis heute kaum untersucht ist. Mit Hilfe einer in unserer Gruppe entwickelten hochaktiven TNFR2-spezifischen TNF-Variante sowie einer bereits länger bekannten TNFR1-spezifischen TNF-Variante wurde in dieser Arbeit die TNF-Signaltransduktion insbesondere im Mutiplen Myelom untersucht. Mit Hilfe der beiden TNF-Varianten konnte gezeigt werden, dass die alleinige Stimulation des TNFR2 die Aktivierung des alternativen NFkappaB-Signalweges vermittelt, wohingegen TNFR1 nicht dazu in der Lage ist. So zeigte sich im Einklang mit der inhibitorischen Funktion des Adapterproteins TRAF2 in der Signaltransduktion des alternativen NFkappaB-Signalweges, dass die TNFR2-Stimulation in einer TRAF2-Depletion resultiert. Dies führt weiterhin zur Akkumulation von NIK und der Prozessierung von p100 zu seiner aktiven Form p52, den klassischen biochemisch nachweisbaren Ereignissen der Aktivierung des alternativen NFkappaB-Signalweges. Aufgrund der Rolle des NFkappaB-Systems im Multiplen Myelom (MM) und der stimulierenden Wirkung des TNFR1 und TNFR2 auf das NFkappaB-System wurde die Expression und Funktion dieser beiden Rezeptoren auf Myelomzelllinien untersucht. Insbesondere wurde analysiert, welchen Effekt eine spezifische Stimulation der beiden TNF-Rezeptoren auf die apoptotische Sensitivität von Myelomzellen hat. Mit einer Ausnahme wiesen alle untersuchten Myelomzelllinien eine eindeutige TNFR2-Oberflächenexpression auf, die TNFR1-Expression hingegen war heterogen. Die TNFR1-Stimulation in den TNFR1-positiven Zelllinien zeigte keinen wesentlichen Einfluss auf die Zellviabilität. Allerdings resultierte eine Vorstimulation mit TNF in einer gesteigerten Sensitivität für den CD95L-induzierten Zelltod, schützte aber gleichzeitig vor der TRAIL-vermittelten Induktion der Apoptose. Der gegenläufige Effekt der TNF-Vorstimulation auf den CD95L- und TRAIL-induzierten Zelltod konnte auf die Hochregulation der CD95-Oberflächenexpression und der gesteigerten Expression des antiapoptotischen cFLIPLong-Proteins zurückgeführt werden. Beide Effekte basieren auf der TNF-induzierten Aktivierung des klassischen NFkappaB-Signalweges. Im CD95L-induzierten Zelltod überkompensierte die Induktion der CD95-Expression offensichtlich die Hochregulation von cFLIPLong und resultierte in gesteigertem Zelltod. Der TRAIL-induzierte Zelltod hingegen wurde durch die TNF-Vorstimulation abgeschwächt, da hier lediglich die durch den klassischen NFkappaB-Signalweg vermittelte gesteigerte Expression des antiapoptotischen cFLIPLong eine Rolle spielte. Desweiteren zeigten die Analysen in dieser Arbeit, dass die TNFR2-Stimulation zu einer Depletion von TRAF2 und z. B. in JJN3-Zellen zu einer Sensitivierung für den TNFR1-induzierten Zelltod führte. Die Ergebnisse dieser Arbeit zeigten in der Summe somit, dass das TNF-TNFR-Signaling durch verschiedene Mechanismen Einfluss auf den Ausgang der extrinsischen Apoptoseinduktion hat, und dass der Effekt von TNF auf das Überleben von MM-Zellen kontextabhängig ist.
Die Anzahl neurologischer Erkrankungen bei denen Autoantikörper gegen zentralnervöse An-tigene bekannt sind, hat in den letzten Jahren deutlich zugenommen. Allerdings gibt es nur für wenige dieser Erkrankungen hinreichende experimentelle Belege für eine pathogene Wir-kung der Autoantikörper. Zwei dieser Erkrankungen wurden im Rahmen dieser Arbeit näher untersucht: die Juvenile Neuronale Zeroid-Lipofuszinose (JNCL) mit Autoantikörpern gegen die 65 kD Isoform der Glutamatdecarboxylase und das Stiff Person Syndrom (SPS) mit Auto-antikörpern gegen Amphiphysin. Die phänotypische Charakterisierung der cln3 knockout-Maus, einem Mausmodell für die JNCL, zeigte eine progressive Verschlechterung der motorischen und koordinativen Fä-higkeiten, eingeschränktes reizbedingtes Lernen und gesteigertes angstähnliches Verhalten. Diese Symptome ähneln denen der humanen Erkrankung. Elektrophysiologisch konnte eine Antikörper-induzierte zerebelläre Dysfunktion identifiziert werden, die einer verminderten lokalen GABAergen Hemmung zugeordnet wird. Eine Reduktion der Antiköperproduktion im Tiermodell durch eine Depletion der Plasmazellen durch den Proteseinhibitor Bortezomib hatte einen positiven Effekt auf die Krankheitsentwicklung. Im zweiten experimentellen Teil der Arbeit wurde der Einfluss von Autoantikörpern gegen Amphiphysin von Patienten mit SPS auf die synaptische Transmission untersucht. Es zeigte sich hierbei in Patch-Clamp Experimenten eine Störung der GABAergen Übertragung v.a. bei hochfrequenter Stimulation, was im Einklang mit dem vermuteten Antikörper-induzierten Endozytosedefekt steht. Passiver Transfer von humanen Autoantikörpern gegen Amphiphysin induzierte angst-ähnliches Verhalten in Ratten, einem weiteren Kernsymptom des SPS. Aktive Immunisierung gegen Amphiphysin und anschließende Öffnung der Blut-Hirn-Schranke in Mäusen führte zu einer subklinischen Veränderung der Reflexverarbeitung von Ia Afferenzen auf Motoneurone im Rückenmark der Mäuse. Insgesamt konnten in zwei Erkrankungen des ZNS autoimmune Mechanismen identifi-ziert werden, die zu einer Antikörper-induzierten Fehlregulation der zentralen synaptischen Transmission führen. Diese Ergebnisse können wegweisend sein auch für die Erforschung der Pathophysiologie anderer Antikörper-assoziierte Erkrankungen des ZNS.
Cancer is one of the leading causes of death all over the world. Malnutrition and toxic contaminations of food with substances such as mycotoxins have been thought to account for a high percentage of cancers. However, human diet can deliver both mutagens and components that decrease the cancer risk. Genomic damage could be reduced by food components through different mechanisms such as scavenging of reactive oxygen species. In the first part of this study we tried to investigate the effects of patulin and resveratrol on DNA stability in V79 cells. Patulin is a mycotoxin, which is frequently found in spoiled apples and other fruits. The WHO has established a safety level of 50 µg/L, which is indeed not observed by all manufacturers. The acute toxicity of patulin in high concentrations is well known, however its potential carcinogenicity is still a matter of debate. Therefore we wanted to investigate further steps in the mechanism of patulin-induced genotoxicity. Patulin caused the formation of micronuclei and nucleoplasmic bridges in a dose-dependent manner. Further analysis revealed that patulin induced both kinetochore-negative and positive micronuclei. Time course of incubation indicate a new mechanism for patulin-induced nucleoplasmic bridge formation. We hypothized a mechanism via cross-linking of DNA, which was confirmed by a modified version of comet assay. Incubations of cells with patulin led to an increased number of multinucleated cells and multipolar mitoses. Cell cytometry revealed a G2 arrest by patulin, which might explain the amplification of centrosomes and patulin-induced aneuploidy. Patulin cause a dose-dependent DNA damage in comet assay which was influenced by the cellular GSH content. However, an induction of oxidative stress was just seen with higher concentrations of patulin. Levels of cellular glutathione were increased after 24 h incubation indicating an adaptive response to patulin-induced stress. There is growing interest in polyphenols such as resveratrol which have shown many positive effects on human health. The beneficial properties are partially attributed to their ability to scavenge reactive oxygen species. Co-incubation of V79 cells with patulin and 10 µM of the antioxidant resveratrol led to a slight reduction of micronucleus frequency compared to cells which were just treated with patulin. However, in higher concentrations resveratrol themselves caused the formation of micronuclei in V79 cells. Kinetochore analysis indicated only clastogenic properties for resveratrol but no disturbance of mitosis. The antioxidant properties of resveratrol were shown in ferric reducing antioxidant power (FRAP) assay. However, in cellular system resveratrol in higher concentrations revealed also prooxidative properties, as shown in 2,7-dichlordihydrofluorescein (DCF) assay. The increased level of glutathione after resveratrol treatment might reflect an adaptive response to resveratrol-induced oxidative stress. For the second part of this thesis we investigated the effects of an anthocyanin-rich grape extract on hypertensive Ren-2 rats. Ren-2 rats are an accepted genetically modified rat model for the investigation of hypertension and increased oxidative stress. We divided 23 female Ren-2 rats into three groups. One group was fed with an anthocyanin-rich Dacapo grape extract, one group was treated with the angiotensin converting enzyme (ACE) inhibitor ramipril and the third group was kept without medication during the experiment. After one week untreated group showed a clear increase in systolic and diastolic blood pressure compared to the ramipril treated rats. This was in part attenuated in the animals fed with anthocyanin-rich Dacapo grape extract. Effects on blood pressure were also reflected in an increased thirst of untreated and extract fed animals. Comet assay with cells of kidney and liver revealed a slight protective impact of Dacapo extract on DNA damage compared to the other groups. Similar results were obtained after evaluation of ɣ-H2AX-staining of kidney and heart sections. However, in the small intestine oppositional effects were seen, indicating an increased number of double strand breaks probably due to the high local concentration of polyphenols after oral ingestion. Antioxidative properties of the extract were shown in FRAP assay. However, this effect was not reflected in an increased antioxidative capacity in serum or a protective impact in the dihydroethidium (DHE) assay. The extract showed protective effects on DNA damage in comet assay and ɣ-H2AX-staining, but was not able to reduce hypertension back to the control level of ramipril treated animals. High local concentrations could also result in an increased damage of the affected tissue. Therefore, the administration of such concentrated compounds should be handled with care.
Streptococcus pneumoniae (Pneumococcus) is one of the leading causes of childhood meningitis,pneumonia and sepsis. Despite the availability of childhood vaccination programs and antimicrobial agents, childhood pneumococcal meningitis is still a devastating illness with mortality rates among the highest of any cause of bacterial meningitis. Especially in low-income countries, where medical care is less accessible, mortality rates up to 50 % have been reported. In surviving patients, neurological sequelae, including hearing loss, focal neurological deficits and cognitive impairment, is reported in 30 to 50 %. Growing resistance of pneumococci towards conventional antibiotics emphasize the need for effective therapies and development of effective vaccines against Streptococcus pneumoniae. One major virulence factor of Streptococcus pneumoniae is the protein toxin Pneumolysin (PLY). PLY belongs to a family of structurally related toxins, the so-called cholesterol-dependent cytolysins (CDCs). Pneumolysin is produced by almost all clinical isolates of the bacterium. It is expressed during the late log phase of bacterial growth and gets released mainly through spontaneous autolysis of the bacterial cell. After binding to cholesterol in the host cell membranes, oligomerization of up to 50 toxin monomers and rearrangement of the protein structure, PLY forms large pores, leading to cell lysis in higher toxin concentrations. At sub-lytic concentrations, however, PLY mediates several other effects, such as activation of the classic complement pathway and the induction of apoptosis. First experiments with pneumococcal strains, deficient in pneumolysin, showed a reduced virulence of the organism, which emphasizes the contribution of this toxin to the course of bacterial meningitis and the urgent need for the understanding of the multiple mechanisms leading to invasive pneumococcal disease. The aim of this thesis was to shed light on the contribution of pneumolysin to the course of the disease as well as to the mental illness patients are suffering from after recovery from pneumococcal meningitis. Therefore, we firstly investigated the effects of sub-lytic pneumolysin concentrations onto primary mouse neurons, transfected with a GFP construct and imaged with the help of laser scanning confocal microscopy. We discovered two major morphological changes in the dendrites of primary mouse neurons: The formation of focal swellings along the dendrites (so-called varicosities) and the reduction of dendritic spines. To study these effects in a more complex system, closer to the in vivo situation, we established a reproducible method for acute brain slice culturing. With the help of this culturing method, we were able to discover the same morphological changes in dendrites upon challenge with sub-lytic concentrations of pneumolysin. We were able to reverse the seen alterations in dendritic structure with the help of two antagonists of the NMDA receptor, connecting the toxin´s mode of action to a non-physiological stimulation of this subtype of glutamate receptors. The loss of dendritic spines (representing the postsynapse) in our brain slice model could be verified with the help of brain slices from adult mice, suffering from pneumococcal meningitis. By immunohistochemical staining with an antibody against synapsin I, serving as a presynaptic marker, we were able to identify a reduction of synapsin I in the cortex of mice, infected with a pneumococcal strain which is capable of producing pneumolysin. The reduction of synapsin I was higher in these brain slices compared to mice infected with a pneumococcal strain which is not capable of producing pneumolysin, illustrating a clear role for the toxin in the reduction of dendritic spines. The fact that the seen effects weren´t abolished under calcium free conditions clarifies that not only the influx of calcium through the pneumolysin-pore is responsible for the alterations. These findings were further supported by calcium imaging experiments, where an inhibitor of the NMDA receptor was capable of delaying the time point, when the maximum of calcium influx upon PLY challenge was reached. Additionally, we were able to observe the dendritic beadings with the help of immunohistochemistry with an antibody against MAP2, a neuron-specific cytoskeletal protein. These observations also connect pneumolysin´s mode of action to excitotoxicity, as several studies mention the aggregation of MAP2 in dendritic beadings in response to excitotoxic stimuli. All in all, this is the first study connecting pneumolysin to excitotoxic events, which might be a novel chance to tie in other options of treatment for patients suffering from pneumococcal meningitis.
In cultured motoneurons of a mouse model for the motoneuron disease spinal muscular atrophy (SMA), reduced levels of the protein SMN (survival of motoneurons) cause defects in axonal growth. This correlates with reduced β-actin mRNA and protein in growth cones, indicating that anterograde transport and local translation of β-actin mRNA are crucial for motoneuron function. However, direct evidence that indeed local translation is a physiological phenomenon in growth cones of motoneurons was missing. Here, a lentiviral GFP-based reporter construct was established to monitor local protein synthesis of β-actin mRNA. Time-lapse imaging of fluorescence recovery after photobleaching (FRAP) in living motoneurons revealed that β-actin is locally translated in the growth cones of embryonic motoneurons. Interestingly, local translation of the β-actin reporter construct was differentially regulated by different laminin isoforms, indicating that laminins provide extracellular cues for the regulation of local translation in growth cones. Notably, local translation of β-actin mRNA was deregulated when motoneurons of a mouse model for type I SMA (Smn-/-; SMN2) were analyzed. In situ hybridization revealed reduced levels of β-actin mRNA in the axons of Smn-/-; SMN2 motoneurons. The distribution of the β-actin mRNA was not modified by different laminin isoforms as revealed by in situ hybridization against the mRNA of the eGFP encoding element of the β-actin reporter. In case of the mRNA of α-actin and γ-actin isoforms, the endogenous mRNA did not localize to the axons and the localization pattern was not affected by the SMN levels expressed in the cell. Taken together our findings suggest that regulation of local translation of β-actin in growth cones of motoneurons critically depends on laminin signaling and the amount of SMN protein. Embryonic stem cell (ESC)-derived motoneurons are an excellent in vitro system to sort out biochemical and cellular pathways which are defective in neurodegenerative diseases like SMA. Here, a protocol for the differentiation and antibody-mediated enrichment of ESC-derived motoneurons is presented, which was optimized during the course of this study. Notably, this study contributes the production and purification of highly active recombinant sonic hedgehog (Shh), which was needed for the efficient differentiation of mouse ESCs to motoneurons. ESC-derived motoneurons will now offer high amounts of cellular material to allow the biochemical identification of disease-relevant molecular components involved in regulated local protein synthesis in axons and growth cones of motoneurons.
RKIP reguliert Proteinkinasen der Signaltransduktionskaskaden von G Protein-gekoppelten Rezeptoren, der Raf/MEK/ERK-MAPK, des Transkriptionsfaktors NFκB und von GSK3β. Unklar war bisher, wie die spezifische Interaktion von RKIP mit seinen mannigfaltigen Interaktionspartnern ermöglicht und reguliert wird. Raf1 und GRK2 sind die einzigen bekannten direkten Interaktionspartner von RKIP und wurden deshalb gewählt, um die zugrundeliegenden molekularen Mechanismen dieser Interaktion genauer zu untersuchen. In dieser Arbeit wurde gezeigt, dass RKIP nach PKC-vermittelter Phosphorylierung von Serin153 dimerisiert und dass diese Dimerisierung für die RKIP/Raf1-Dissoziation und die RKIP/GRK2-Interaktion essentiell ist. Co-Immunpräzipitationsexperimente mit einer phosphorylierungsdefizienten Mutante zeigten, dass für diese Dimerisierung die Phosphorylierung von beiden RKIP-Molekülen notwendig ist. Als Dimerinteraktionsfläche wurden die Aminosäuren 127-146 von RKIP identifiziert, da das Peptid RKIP127-146 die Dimerisierung von RKIP spezifisch und effizient hemmte. Um die Bedeutung dieser phosphorylierungsinduzierten Dimerisierung von RKIP für seine Interaktion mit Raf1 und GRK2 zu untersuchen, wurden eine phosphomimetische Mutante (RKIPSK153/7EE) und eine Mutante von RKIP generiert, welche bereits unphosphoryliert dimerisiert (RKIP∆143-6). Folgende Ergebnisse legen nahe, dass die Dimerisierung von RKIP für die spezifische Interaktion mit Raf1 bzw. GRK2 entscheidend ist: (i) Die Dimerisierung von phosphoryliertem RKIP ging mit der Dissoziation von RKIP und Raf1 und der Assoziation von RKIP und GRK2 einher; (ii) die Mutanten RKIPSK153/7EE und RKIP∆143-6, die bereits in unstimulierten Zellen eine starke Dimerisierung zeigten, hatten eine höhere Affinität zu GRK2 als zu Raf1; (iii) die Hemmung der RKIP-Dimerisierung interferierte nur mit der RKIP/GKR2- aber nicht mit der RKIP/Raf1-Interaktion; (iv) in vitro und in Mausherzen konnte ein RKIP- und GRK2-immunreaktiver Komplex nachgewiesen werden; (v) Untersuchungen zur RKIP-vermittelten Hemmung der Kinaseaktivität von GRK2 und Raf implizierten, dass dimerisiertes RKIP nur die Aktivität von GRK2, nicht aber von Raf hemmt. Diese Arbeit zeigt, dass die phosphorylierungsinduzierte Dimerisierung von RKIP die spezifische Interaktion von RKIP mit Raf1 und GRK2 koordiniert. Die Aufklärung dieses Mechanismus erweitert unser Verständnis der spezifischen Interaktion von Kinasen mit ihren Regulatorproteinen.
Die Stimulation des CD95-Todesrezeptors durch seinen natürlichen membranständigen Li-ganden CD95L führt zur kontextabhängigen Aktivierung von sowohl apoptotischen als auch nicht-apoptotischen Signalwegen. Durch Proteolyse wird aus dem membranständigen CD95L löslicher trimerer CD95L freigesetzt. Die Bindung von löslichem trimerem CD95L an CD95 ist nicht ausreichend, um die CD95-Signaltransduktion effizient zu stimulieren. Die Fähigkeit von löslichen CD95L-Trimeren CD95-vermittelte Signalwege robust zu aktivieren kann jedoch durch Oligomerisierung und artifizielle Immobilisierung an eine Oberfläche drastisch gesteigert werden. In dieser Arbeit wurde zunächst bestätigt, dass nur oligomere CD95L-Varianten, die z.B. durch Antikörpervernetzung von N-terminal getaggten rekombinanten CD95L-Varianten oder durch eine gentechnisch erzwungene Hexamerisierung von CD95L-Molekülen erhalten wur-den, in der Lage sind, effizient apoptotische und nicht-apoptotische Signalwege zu aktivieren. Ferner zeigte sich dann, dass die Bindung von löslichen CD95L-Trimeren nicht ausreichend ist, um die Translokation von CD95-Molekülen in detergenzunlösliche „Lipid Raft“- Membrandomänen zu stimulieren. Die „Lipid Raft“-Translokation ist ein zentrales Ereignis bei der CD95-Aktivierung und vor allem für die Induktion der Apoptose bedeutsam. Dabei ist ein selbstverstärkender Prozess aus Caspase-8-Aktivierung und „Lipid Raft“-Assoziation des CD95 von Bedeutung. Um die Interaktion von CD95 und CD95L mit Hilfe von hoch sensitiven zellulären Bindungs-studien analysieren zu können, wurden in dieser Arbeit desweiteren CD95L-Fusionsproteine entwickelt und hergestellt, an welche N-terminal eine Gaussia princeps Luziferase (GpL)- Reporterdomäne gekoppelt ist. So konnte mit den GpL-CD95L-Fusionsproteinen gezeigt werden, dass die Oligomerisierung von CD95L-Trimeren keinen Effekt auf die Ligandenbele-gung des CD95 hat. Dies spricht dafür, dass die höhere spezifische Aktivität von oligomeri-sierten CD95L-Trimeren nicht auf einer Aviditäts-vermittelten Zunahme der apparenten Affi-nität beruht, sondern dies deutet darauf hin, dass die sekundäre Aggregation von sich initial bildenden trimeren CD95L-CD95-Komplexen eine entscheidende Rolle in der CD95-Aktivierung spielt. Durch Scatchard-Analysen zeigte sich ferner, dass trimerer CD95L mit mindestens zwei zellulären Bindungsstellen unterschiedlicher Affinität interagiert. Bindungs-studien mit löslichen monomeren und trimeren GpL-CD95-Rezeptoren an membranständigen CD95L, als auch Inhibitionsstudien ergaben, dass trimerer CD95 weitaus besser an CD95L bindet. Dies legt nahe, dass es sich bei den zuvor beobachteten hoch- und niederaffinen Bindungsstellen für CD95L um monomere bzw. prä-assemblierte CD95-Moleküle handelt. Die GpL-CD95L-Fusionsproteine wurden auch genutzt, um die CD95-Translokation in „Lipid Rafts“ zu analysieren. So wurde trimerer GpL-CD95L als „Tracer“ zur Markierung von inaktiven CD95-Molekülen eingesetzt. Nach Aktivierung der übrigen freien CD95-Moleküle mit hoch aktivem hexameren Fc-CD95L konnte eine Zunahme der inaktiven GpL-CD95L-markierten Rezeptoren in „Lipid Rafts“ beobachtet werden. Offensichtlich stimulieren also aktivierte CD95-Moleküle in „trans“ die Ko-Translokation inaktiver CD95-Rezeptoren in „Lipid Rafts“. Dies bestätigte sich auch in Experimenten mit Transfektanten, die einen chimären CD40-CD95-Rezeptor exprimieren. Letzterer ist nach Stimulation mit CD40L in der Lage, intrazellu-läre CD95-vermittelte Signalwege zu aktivieren. Die Aktivierung von CD95-assoziierten Sig-nalwegen durch Stimulation von endogenem CD95 in CD40-CD95-Transfektanten resultierte nun in der Ko-Translokation von unstimulierten CD40-CD95-Rezeptoren in „Lipid Rafts“. Vice versa zeigte sich die Ko-Translokation von endogenem CD95 nach spezifischer Aktivierung des chimären CD40-CD95-Rezeptors. Schlussendlich erwiesen sich eine funktionsfähige Todesdomäne und die Aktivierung der Caspase-8 als essentiell für die „Lipid Raft“-Assoziation von aktivierten CD95-Molekülen und auch für die durch diese Rezeptorspezies induzierte Ko-Translokation von inaktiven Rezeptoren in „Lipid Rafts“.
The probiotic Escherichia coli strain Nissle 1917 (EcN) is one of the few probiotics licensed as a medication in several countries. Best documented is its effectiveness in keeping patients suffering from ulcerative colitis (UC) in remission. This might be due to its ability to induce the production of human beta defensin 2 (HBD2) in a flagellin-dependent way in intestinal epithelial cells. In contrast to ulcerative colitis, for Crohn´s disease (CD) convincing evidence is lacking that EcN might be clinically effective, most likely due to the genetically based inability of sufficient defensin production in CD patients. As a first step in the development of an alternative approach for the treatment of CD patients, EcN strains were constructed which were able to produce human alpha-defensin 5 (HD5) or beta-defensin 2 (HBD2). For that purpose codon-optimized defensin genes encoding either the proform with the signal sequence or the mature form of human alpha defensin 5 (HD5) or the gene encoding HBD2 with or without the signal sequence were cloned in an expression vector plasmid under the control of the T7 promoter. Synthesis of the encoded defensins was shown by Western blots after induction of expression and lysis of the recombinant EcN strains. Recombinant mature HBD2 with an N-terminal His-tag could be purified by Ni-column chromatography and showed antimicrobial activity against E. coli, Salmonella enterica serovar Typhimurium and Listeria monocytogenes. In a second approach, that part of the HBD2-gene which encodes mature HBD2 was fused with yebF gene. The resulting fusion protein YebFMHBD2 was secreted from the encoding EcN mutant strain after induction of expression. Presence of YebFMHBD2 in the medium was not the result of leakage from the bacterial cells, as demonstrated in the spent culture supernatant by Western blots specific for ß-galactosidase and maltose-binding protein. The dialyzed and concentrated culture supernatant inhibited the growth of E. coli, Salmonella enterica serovar Typhimurium and Listeria monocytogenes in radial diffusion assays as well as in liquid coculture. This demonstrates EcN to be a suitable probiotic E. coli strain for the production of certain defensins.
Upon oncogenic stress, the tumor suppressor Arf can induce irreversible cell cycle arrest or apoptosis, depending on the oncogenic insult. In this study, it could be shown that Arf interacts with Myc and the Myc-associated zinc-finger protein Miz1 to facilitate repression of genes involved in cell adhesion. Formation of a DNA-binding Arf/Myc/Miz1 complex disrupts interaction of Miz1 with its coactivator nucleophosmin and induces local heterochromatinisation, causing cells to lose attachment and undergo anoikis. The assembly of the complex relies on Myc, which might explain why high Myc levels trigger apoptosis and not cell cycle arrest in the Arf response. This mechanism could play an important role in eliminating cells harboring an oncogenic mutation. Arf furthermore induces sumoylation of Miz1 at a specific lysine by repressing the desumoylating enzyme Senp3. A sumoylation-deficient mutant of Miz1 however does not show phenotypic differences under the chosen experimental conditions. Myc can also be modified by Sumo by multisumoylation at many different lysines, which is unaffected by Arf. The exact mechanism and effect of this modification however stays unsolved.