570 Biowissenschaften; Biologie
Refine
Has Fulltext
- yes (139)
Is part of the Bibliography
- yes (139)
Year of publication
- 2019 (139) (remove)
Document Type
- Journal article (78)
- Doctoral Thesis (60)
- Preprint (1)
Keywords
- Tissue Engineering (7)
- Candida albicans (3)
- Genexpression (3)
- In vitro (3)
- Signaltransduktion (3)
- cancer (3)
- leukemic cells (3)
- metagenomics (3)
- 3D Tumormodell (2)
- 3D tissue model (2)
- Annotation (2)
- Bioinformatik (2)
- Blutgerinnung (2)
- Chlamydia trachomatis (2)
- Chronobiologie (2)
- Drosophila (2)
- Drosophila melanogaster (2)
- Ecology (2)
- Fluoreszenzmikroskopie (2)
- Herzmuskelzelle (2)
- Implantat (2)
- Maus (2)
- Mikroskopie (2)
- RNA-seq (2)
- Regenerative Medizin (2)
- Serotonin (2)
- Taufliege (2)
- Vaskularisierung (2)
- Verhalten (2)
- apoptosis (2)
- colorectal cancer (2)
- cytotoxicity (2)
- dSTORM (2)
- evolution (2)
- infection (2)
- ischemic stroke (2)
- microbiome (2)
- optimal drug combination (2)
- rheumatoid arthritis (2)
- ubiquitin (2)
- (hem)ITAM signaling (1)
- 3D (1)
- 3D Modell (1)
- 3D model (1)
- 3D remote sensing (1)
- 3D tumour model (1)
- 3D-Modell (1)
- ABP1 (1)
- ADHD (1)
- ALAN (1)
- AMP-activated kinases (1)
- APOBEC3G (1)
- AUX1 (1)
- Absorbed Doses (1)
- Accurate (1)
- Acids (1)
- Ackerschmalwand (1)
- Adaptation (1)
- Adherens junction (1)
- Administered Activities (1)
- Aggregation (1)
- Algorithmus (1)
- Alkaloid (1)
- Alpine habitats (1)
- Alzheimerkrankheit (1)
- Aminerge Nervenzelle (1)
- Amygdala (1)
- Ancistrocladaceae (1)
- Aneugene (1)
- Angiogenese (1)
- Animales Nervensystem (1)
- Anthropocene (1)
- Anti-infective (1)
- Antigen CD19 (1)
- Apidae (1)
- Apis mellifera (1)
- Apolipoprotein E (1)
- Arabidopsis (1)
- Aspergillus fumigalus (1)
- Associative learning (1)
- Autoaggressionskrankheit (1)
- Automated Image Analysis (1)
- Automatisierung (1)
- Automatisierung der Analyse (1)
- Autophagie <Physiologie> (1)
- Autophagosom (1)
- Autophagozytose (1)
- B-MYB (1)
- BRAF-mutant (1)
- BRAF-mutiert (1)
- Barrier (1)
- Bees (1)
- Behaviour (1)
- Behavioural ecology (1)
- Berberine (1)
- Bildanalyse (1)
- Bildverarbeitung (1)
- Biokinetics (1)
- Biologika (1)
- Biomarker (1)
- Biomaterial (1)
- Biomedical engineering (1)
- Biomedicine (1)
- Biophysics (1)
- Bioreaktor (1)
- Brustkrebs (1)
- C-60 fullerene (1)
- C60 fullerene (1)
- CAR T-Zelltherapie (1)
- C\(_{60}\) fullerene (1)
- Cadherine (1)
- Caenorhabditis elegans (1)
- Callyspongia siphonella (1)
- Cancer Cell (1)
- Cdh13 (1)
- Cell migration (1)
- Cell stainin (1)
- Cellular neuroscience (1)
- Central nervous system (1)
- Channelrhodopsin (1)
- Chondrogenesis (1)
- Chronobiology (1)
- Circular dichroism (1)
- Click Chemie (1)
- CoQ10 (1)
- Cofaktor (1)
- Colonization (1)
- Computational and Systems Biology (1)
- Computer modelling (1)
- Computersimulation (1)
- Confocal microscopy (1)
- Conservation (1)
- Cortex (1)
- CpG (1)
- Cranial sutures (1)
- Cytokine (1)
- Cytologie (1)
- Cytosol (1)
- DC gate (1)
- DLS and AFM measurements (1)
- DNA damage (1)
- DNA-Schäden (1)
- DNS-Doppelstrangbruch (1)
- DNS-Schädigung (1)
- Dendritic cell (1)
- Dendritische Zelle (1)
- Densovirus (1)
- Developmental biology (1)
- Dezellularisierung (1)
- Diagnostic Imaging Exams (1)
- Dickdarmtumor (1)
- Dnaschaden (1)
- Dopamin (1)
- Dopamine (1)
- Dosimetry (1)
- Doxorubicin (1)
- Drug discovery (1)
- Drug targets (1)
- Drug testing (1)
- Dynamics (1)
- E8 symmetry (1)
- EMT (1)
- ERK signaling (1)
- Echinococcosis (1)
- Echinococcus (1)
- Eierstockkrebs (1)
- Einzelmolekülmikroskopie (1)
- Electron Microscopy (1)
- Elektronenmikroskopie (1)
- Embryonic induction (1)
- Embryos (1)
- Endodermis (1)
- Endogene Rhythmik (1)
- Enhancer elements (1)
- Environmental impact (1)
- Epigenetic (1)
- Evolutionary developmental biology (1)
- Evolutionary emergence (1)
- Extracellular matrix (1)
- Eye development (1)
- Eye irritation (1)
- Fear conditioning (1)
- Fgf-signalling (1)
- Flowering plants (1)
- Flowers (1)
- Fluorescence spectroscopy (1)
- Fremdkörpermodell (1)
- G-quadruplex (1)
- Ga-68-labelled Peptides (1)
- Gedächtnis (1)
- Gene by Environment (1)
- Gene expression analysis (1)
- Gene silencing (1)
- Genetik (1)
- Genom (1)
- Genome Annotation (1)
- Genotoxicitiy (1)
- Genotoxizität (1)
- Germline (1)
- Gestational diabetes (1)
- Gestationsdiabetes (1)
- Gewebemodell (1)
- Gliazelle (1)
- Glioblastom (1)
- Glykane (1)
- Haut (1)
- HeLa cells (1)
- Health (1)
- Helicasen (1)
- Hfq (1)
- Hindbrain (1)
- Hippo pathway (1)
- Hippocampus (1)
- Hochauflösende Fluoreszenzmikroskopie (1)
- Host Genome Integrity (1)
- Host-parasite interaction (1)
- Human Herpesvirus 6 (1)
- Humanes Herpesvirus 6 (1)
- Hurwitz theorem (1)
- Hypothalamus (1)
- Hypoxia (1)
- Hypoxie (1)
- Immunologe (1)
- Immunoprecipitation (1)
- Immuntherapie (1)
- Induzierte pluripotente Stammzelle (1)
- Inf-TRAP-Seq (1)
- Inhibitor (1)
- Insect flight (1)
- Insulin (1)
- Invertebrate herbivory (1)
- Isolation (1)
- KDELR2 (1)
- Kardiomyozyt (1)
- Kernspintomografie (1)
- Klassifizierung (1)
- Klastogene (1)
- Knochenimplantat (1)
- Knochenregeneration (1)
- Knorpelbildung (1)
- Kongo (1)
- Kraniosynostose (1)
- Krebs (1)
- Krebs <Medizin> (1)
- LC-HRESIMS (1)
- Leaf traits (1)
- Lee Smolin (1)
- Leichte kognitive Beeinträchtigung (1)
- Lernen (1)
- Lernverhalten (1)
- Light sheet microscopy (1)
- Limb development (1)
- Llullaillaco Volcano (1)
- Lung (1)
- Lunge (1)
- Lungenkrebs (1)
- Lungentumor (1)
- Lysosom (1)
- M14 carboxypeptidasses (1)
- MMB complex (1)
- MRI (1)
- Maculinea butterfly (1)
- Magnetic Resonance Imaging (1)
- Mamma carcinoma (1)
- Mammakarzinom (1)
- Markierungen synaptischer Proteine (1)
- Masern-Virus (1)
- Mass spectrometry (1)
- Massenspektrometrie (1)
- Mc4r (1)
- Measles virus (1)
- Megakaryozyt (1)
- Melanom (1)
- Melanoma (1)
- Meniskus (1)
- Meniskusimplantat (1)
- Metabolomics (1)
- Method development (1)
- Methodenentwicklung (1)
- Methylation (1)
- Microbiology and Infectious Disease (1)
- Mikroarray basierte vergleichende Genomhybridisierung (Array-CGH) (1)
- Mitochondria (1)
- Mitose (1)
- Mobile genetic element (1)
- Modell (1)
- Molekulare Zielstrukturen (1)
- Molekulargenetik (1)
- Molekülsystem (1)
- Motor behaviour (1)
- Multiples Myelom (1)
- Myb-MuvB (1)
- Myrmecology (1)
- Myrmica ant non-equilibrium dynamics (1)
- NIR-Spektroskopie (1)
- Nahrungserwerb (1)
- Naphthylisochinolinalkaloide (1)
- Naphthylisoquinoline (1)
- Neisseria gonorrhoeae (1)
- Neisseria meningitidis (1)
- Neural circuits (1)
- Neurodevelopmental Disorder (1)
- Neurogenese (1)
- Neurons (1)
- Next Generation Sequencing (NGS) (1)
- Non-coding RNA (1)
- Nuclear Medicine (1)
- Object recognition (1)
- Optimal foraging (1)
- Optogenetics (1)
- Optogenetik (1)
- Outer membrane proteins (1)
- PDF neurons (1)
- PI3K/mTOR inhibierung (1)
- Parvovirus (1)
- Paternal age and BMI effects (1)
- Patterns and drivers of invertebrate herbivory (1)
- Patterns and drivers of species diversity of phytophagous beetles (1)
- Patterns and drivers of species richness and community biomass of large mammals (1)
- Pediatric Nuclear Medicine (1)
- Pediatric Patients (1)
- Phosphatasen (1)
- Phosphoglykolat-Phosphatase (1)
- Phosphoglykolatphosphatase (1)
- Physiologie (1)
- Phytophthora (1)
- Phytosphingosine (1)
- Pigmentdispergierender Faktor (1)
- Piriformospora indica (1)
- Plant signalling (1)
- Plants (1)
- Plasmamembranorganisation (1)
- Plasmozytom (1)
- Polysaccharide (1)
- Prefrontal cortex (1)
- Premna (1)
- ProQ (1)
- Protein crosslinking (1)
- Protein folding (1)
- Protein interactions (1)
- Proteininteraktionen (1)
- Proteinquervernetzungen (1)
- Proteomics Analysis of Complexes (1)
- Proteotype (1)
- Proteus vulgaris (1)
- Präfrontaler Cortex (1)
- Pseudomonas syringae (1)
- Quantifizierung (1)
- Quantitative Mikroskopie (1)
- R package (1)
- REDD1 (1)
- RNA Sequencing (1)
- RNA metabolism (1)
- RNA protein interactions (1)
- RNA secondary structures (1)
- RNA sequencing (1)
- RNA-Seq (1)
- RNA-seq transcriptome (1)
- RNAi (1)
- RNAlater (1)
- RNS (1)
- Radiation Protection (1)
- Radiation-associated Cancer Risk (1)
- Raphe (1)
- Registrierung <Bildverarbeitung> (1)
- Reiz (1)
- Rescue behaviour (1)
- Research Article (1)
- Resistenz (1)
- Rezeptoren (1)
- Rhizodermis (1)
- Risk Assessment (1)
- SREBP (1)
- SSR42 (1)
- Scarabaeidae (1)
- Schlaganfall (1)
- Schmalwand <Arabidopsis> (1)
- Self-renewal (1)
- Sex chromosome (1)
- Sex determination (1)
- Sexual development and function (1)
- Signalweg (1)
- Skull (1)
- Small RNA (1)
- Small interfering RNAs (1)
- Solution-state NMR (1)
- Somites (1)
- Species delimitation (1)
- Species richness (1)
- Sphingobasen (LCB, LCB-P) (1)
- Sphingobases (1)
- Sphingolipide (1)
- Sphingosine-1-phosphate (1)
- Sphingosine-1-phosphats (1)
- Spred-Proteine (1)
- Stammzelle (1)
- Staphylococcus aureus (1)
- Stechameisen (1)
- Stem cell (1)
- Stem cells (1)
- Stem-cell biotechnology (1)
- Stoffwechsel (1)
- Strains (1)
- Structural elucidation (1)
- Subtercola vilae (1)
- Suspensionskultur (1)
- Syap1 knockout (1)
- Synaptische Vesikel (1)
- TLR3 (1)
- TMEM16F (1)
- TNFR1 (1)
- TRAIL (1)
- TWEAK (1)
- Tc-99m-MAG3 Scans (1)
- Therapeutisches System (1)
- Therapie (1)
- Therapiesimulation (1)
- Thrombo-inflammation (1)
- Thrombosis (1)
- Thrombozyt (1)
- Tiermodell (1)
- Tissue engineering (1)
- Todesdomäne (1)
- Trailmaking Test (1)
- Transcription (1)
- Transcriptomic (1)
- Translation (1)
- Transposable element (1)
- Tumor (1)
- UBXD1 (1)
- UV–Vis (1)
- Ubiquitin (1)
- Ultrashort echo time - UTE (1)
- Ustilago maydis (1)
- Vascularized (1)
- Vaskularisation (1)
- Verticillium (1)
- Vesikel (1)
- Wirkstofftestung (1)
- Wundheilung (1)
- Wurzel (1)
- Wurzelzellschichten (1)
- YAP (1)
- ZFAND1 (1)
- Zebrafish (1)
- Zell Migration (1)
- Zellbiologie (1)
- Zellkultur (1)
- Zellmigration (1)
- Zellschichtspezifische Expression (1)
- Zellteilung (1)
- Zelltod (1)
- Zentralzylinder (1)
- Zinc (1)
- accessory medulla (1)
- accumulation (1)
- acetate (1)
- adalimumab (1)
- adapterprotein (1)
- alternative splicing (1)
- altitudinal gradients (1)
- aminergic neurons (1)
- anakinra (1)
- aneugens (1)
- animal physiology (1)
- antibacterial (1)
- antibiofilm (1)
- anticancer (1)
- antitrypanosomal (1)
- artifacts (1)
- artificial light at night (1)
- autoimmune disease (1)
- automatisierte Bildanalyse (1)
- autophagocytosis (1)
- autophagosome (1)
- autophagy (1)
- auxin (1)
- bacteria (1)
- bacterial pathogen (1)
- behavioral plasticity (1)
- bioinformatics tool (1)
- bioink (1)
- biological rhythm (1)
- biological scaffolds (1)
- biological techniques (1)
- biomarker (1)
- biomarker signature (1)
- biomaterial ink (1)
- bioreactor (1)
- biotic interaction (1)
- blood–brain barrier (1)
- bone (1)
- brain (1)
- brain endothelial cell (1)
- burnt-wood (1)
- calcium (1)
- cancer metabolism (1)
- cancer therapy (1)
- carabid beetles (1)
- cardiac tissue (1)
- cardiomyocytes (1)
- cell biology (1)
- cell signalling (1)
- cell therapy (1)
- cell-type specific (1)
- channelrhodopsins (1)
- chlamydia (1)
- chlorophyll fluorescence imaging (1)
- circRNA (1)
- circadian clock (1)
- circadian rhythm (1)
- circular transcriptome sequencing (1)
- classification (1)
- clastogens (1)
- click chemistry (1)
- clinical trial (1)
- closed-loop systems (1)
- co-culture (1)
- coagulation system (1)
- cold adaptation (1)
- comparative genomics (1)
- competition (1)
- composition (1)
- conservation (1)
- cosmology (1)
- crystal growth (1)
- crystallization (1)
- cytokinesis (1)
- cytotoxic (1)
- dead-wood enrichment (1)
- decellularization (1)
- definition (1)
- dendritic cells (1)
- designer cell (1)
- developmental forms (1)
- direct muss spectrometric profiling (1)
- disease modelling (1)
- diversity gradients (1)
- domain-specific language (1)
- dopamine (1)
- doxorubicin (1)
- drivers and patterns of diversity and herbivory (1)
- drug delivery (1)
- drug release (1)
- drug resistance evolution (1)
- earlywood (1)
- ecology (1)
- ecosystem service (1)
- efficient intervention points (1)
- elementary body (1)
- enbrel (1)
- endocytosis (1)
- enercy-richness hypothesis (1)
- energy homeostasis (1)
- enhancer (1)
- enzyme mechanism (1)
- etanercept (1)
- evolutionary genetics (1)
- expansion microscopy (1)
- external stimuli (1)
- extinction dynamics (1)
- fertility (1)
- fibre length (1)
- flash freezing (1)
- fluxosome (1)
- food resources (1)
- forest ecology (1)
- forest fire (1)
- forest management (1)
- friut fly behaviour (1)
- function (1)
- functional analysis (1)
- fungal rhodopsins (1)
- gangliosides and lipid rafts (1)
- gastrointestinal tract (1)
- gene expression analysis (1)
- gene network (1)
- genetic engineering (1)
- genetic recombination (1)
- genome analysis (1)
- genome annotation (1)
- genomics (1)
- glia cells (1)
- global change (1)
- green fluorescence protein (GFP) (1)
- ground-dwelling predators (1)
- growth (1)
- growth ring width (1)
- hemostasis (1)
- heuristics (1)
- hiPSC aggregation (1)
- hochauflösende Fluoreszenzmikroskopie (1)
- honeybee (1)
- honeybees (1)
- iPSC (1)
- ichthyology (1)
- imaging (1)
- imaging PAM (1)
- immunity (1)
- immunocompetent skin (1)
- immunotherapy (1)
- immunotherapy of cancer (1)
- implant (1)
- in situ microscopy (1)
- in vitro (1)
- indole-3-acetic acid (1)
- induced pluripotent stem cells (1)
- infection biology (1)
- inflation (1)
- insect abundance (1)
- insect collection (1)
- integrative management strategy (1)
- intervention point analyzing (1)
- knockout (1)
- kolorektales Karzinom (1)
- land sharing (1)
- land use (1)
- latewood (1)
- learning (1)
- lignan (1)
- localization microscopy (1)
- lowland beech forests (1)
- mRNA (1)
- machine learning (1)
- macrophages (1)
- mating (1)
- mating preference (1)
- measles virus (1)
- mechanisms of disease (1)
- mechanistic modelling (1)
- medaka (1)
- megakaryopoiesis (1)
- melatonin (1)
- memory (1)
- meningitis (1)
- meniscus implant (1)
- meta-analysis (1)
- metabolic adaptation (1)
- metabolic flux (1)
- metabolic modeling (1)
- metabolic modelling (1)
- metabolite profiling (1)
- metabolomic (1)
- metabolomic profiling (1)
- metals (1)
- metaproteomics (1)
- methods (1)
- miR-26 (1)
- microbial rhodopsins (1)
- microbiota (1)
- mitosis (1)
- mitotic gene expression (1)
- mitotic genes (1)
- molecular biology (1)
- multi-photon microscopy (1)
- mutants (1)
- nanocomplex (1)
- native populations (1)
- natural pest control (1)
- neuronal (1)
- next generation sequencing (1)
- next-generation sequencing (1)
- noncoding RNA (1)
- noncovalent complex (1)
- noncovalent nanocomplex (1)
- nuclear envelope (1)
- nuclear export (1)
- oncogenes (1)
- oncology (1)
- oncolysis (1)
- oncolytic vaccinia virus (1)
- oncolytic virus (1)
- optimal drug targeting (1)
- optimal pharmacological modulation (1)
- optimal treatment strategies (1)
- optogenetics (1)
- oxindole alkaloids (1)
- p21-activated kinase Mbt/PAK4 (1)
- p53 (1)
- p97 (1)
- parasexual recombination (1)
- parasite biology (1)
- patch-clamp (1)
- peptidomoics (1)
- pharmacophore map (1)
- phenolic compounds (1)
- phosphorylation (1)
- photodynamic chemotherapy (1)
- piRNA (1)
- pit membrane diameter (1)
- plants (1)
- plant–pathogen interaction (1)
- plasma membrane depolarization (1)
- plasma membrane organization (1)
- platelets (1)
- pollination (1)
- population genetics (1)
- post-fire management (1)
- post-translational modification (1)
- posttranscriptional control (1)
- programmed cell death (1)
- programmierter Zelltod (1)
- protected forests (1)
- protein processing (1)
- proteomics (1)
- puberty (1)
- resonance theory (1)
- restriction factors (1)
- reticulate body (1)
- risk factors (1)
- sample storage (1)
- saproxylic organisms (1)
- secreted effectors (1)
- sentinel prey (1)
- short neuropeptide F (1)
- signalling (1)
- single-molecule tracking (1)
- skin model (1)
- sleep (1)
- small RNA (1)
- small intestinal submucosa scaffold (1)
- small-cell lung cancer (1)
- sodium (1)
- soil fauna (1)
- species richness (1)
- species‐area hypothesis (1)
- sphingolipids (1)
- sporidia (1)
- stem Cells (1)
- stem cells (1)
- storage-pool diseases (1)
- stromal vascular fraction (1)
- structure (1)
- structured illumination microscopy (1)
- super-resolution fluorescence microscopy (1)
- super-resolution microscopy (1)
- superresolution (1)
- suppressor cells (1)
- suppressor mutation (1)
- survival analysis (1)
- synapses (1)
- synergistic effect (1)
- synthetic biology (1)
- systematic affiliation (1)
- systematic drug targeting (1)
- targeted therapies (1)
- targeted therapy (1)
- temperature‐mediated resource exploitation hypothesis (1)
- temperature‐richness hypothesis (1)
- therapy simulation (1)
- thrombosis (1)
- time lag (1)
- tisindoline (1)
- tissue engineering (1)
- transcription (1)
- transcriptome (1)
- transcriptomics (1)
- transient dynamics (1)
- tree cavities (1)
- trypanosomes (1)
- tumour (1)
- tumour-necrosis factors (1)
- tyloses (1)
- ubiquitin ligase (1)
- ubiquitylation (ubiquitination) (1)
- unmanaged broadleaved forests (1)
- uptake (1)
- vaccinia (1)
- vascularization (1)
- vertical and radial variation (1)
- vessel lumen diameter (1)
- vessel wall resident stem cells (1)
- virotherapy (1)
- virus (1)
- whole-genome sequencing (1)
- wood anatomy (1)
- wound healing (1)
- yH2AX-Foci (1)
- zielgerichtete Behandlung (1)
- zielgerichtete Therapien (1)
Institute
- Theodor-Boveri-Institut für Biowissenschaften (68)
- Graduate School of Life Sciences (33)
- Julius-von-Sachs-Institut für Biowissenschaften (10)
- Lehrstuhl für Tissue Engineering und Regenerative Medizin (9)
- Rudolf-Virchow-Zentrum (8)
- Fakultät für Biologie (4)
- Institut für Hygiene und Mikrobiologie (4)
- Institut für Molekulare Infektionsbiologie (4)
- Institut für Pharmakologie und Toxikologie (4)
- Institut für Virologie und Immunbiologie (3)
Sonstige beteiligte Institutionen
Since its first experimental implementation in 2005, single-molecule localization microscopy (SMLM) emerged as a versatile and powerful imaging tool for biological structures with nanometer resolution. By now, SMLM has compiled an extensive track-record of novel insights in sub- and inter- cellular organization.\\
Moreover, since all SMLM techniques rely on the analysis of emission patterns from isolated fluorophores, they inherently allocate molecular information $per$ $definitionem$.\\
Consequently, SMLM transitioned from its origin as pure high-resolution imaging instrument towards quantitative microscopy, where the key information medium is no longer the highly resolved image itself, but the raw localization data set.\\
The work presented in this thesis is part of the ongoing effort to translate those $per$ $se$ molecular information gained by SMLM imaging to insights into the structural organization of the targeted protein or even beyond. Although largely consistent in their objectives, the general distinction between global or segmentation clustering approaches on one side and particle averaging or meta-analyses techniques on the other is usually made.\\
During the course of my thesis, I designed, implemented and employed numerous quantitative approaches with varying degrees of complexity and fields of application.\\ \\
In my first major project, I analyzed the localization distribution of the integral protein gp210 of the nuclear pore complex (NPC) with an iterative \textit{k}-means algorithm. Relating the distinct localization statistics of separated gp210 domains to isolated fluorescent signals led, among others, to the conclusion that the anchoring ring of the NPC consists of 8 homo-dimers of gp210.\\
This is of particular significance, both because it answered a decades long standing question about the nature of the gp210 ring and it showcased the possibility to gain structural information well beyond the resolution capabilities of SMLM by crafty quantification approaches.\\ \\
The second major project reported comprises an extensive study of the synaptonemal complex (SNC) and linked cohesin complexes. Here, I employed a multi-level meta-analysis of the localization sets of various SNC proteins to facilitate the compilation of a novel model of the molecular organization of the major SNC components with so far unmatched extend and detail with isotropic three-dimensional resolution.\\
In a second venture, the two murine cohesin components SMC3 and STAG3 connected to the SNC were analyzed. Applying an adapted algorithm, considering the disperse nature of cohesins, led to the realization that there is an apparent polarization of those cohesin complexes in the SNC, as well as a possible sub-structure of STAG3 beyond the resolution capabilities of SMLM.\\ \\
Other minor projects connected to localization quantification included the study of plasma membrane glycans regarding their overall localization distribution and particular homogeneity as well as the investigation of two flotillin proteins in the membrane of bacteria, forming clusters of distinct shapes and sizes.\\ \\
Finally, a novel approach to three-dimensional SMLM is presented, employing the precise quantification of single molecule emitter intensities. This method, named TRABI, relies on the principles of aperture photometry which were improved for SMLM.\\
With TRABI it was shown, that widely used Gaussian fitting based localization software underestimates photon counts significantly. This mismatch was utilized as a $z$-dependent parameter, enabling the conversion of 2D SMLM data to a virtual 3D space. Furthermore it was demonstrated, that TRABI can be combined beneficially with a multi-plane detection scheme, resulting in superior performance regarding axial localization precision and resolution.\\
Additionally, TRABI has been subsequently employed to photometrically characterize a novel dye for SMLM, revealing superior photo-physical properties at the single-molecule level.\\
Following the conclusion of this thesis, the TRABI method and its applications remains subject of diverse ongoing research.
Obwohl Pflanzenwurzeln mit einer Vielzahl von Pathogenen in Kontakt kommen, sind induzierbare Abwehrreaktionen der Wurzel bisher kaum beschrieben. Aufgrund der konzentrischen Zellschicht-Organisation der Wurzel wird angenommen, dass bei einer Immunantwort in jeder Zellschicht ein spezifisches genetisches Programm aktiviert wird. Eine Überprüfung dieser Hypothese war bisher wegen methodischen Limitierungen nicht möglich. Die zellschichtspezifische Expression Epitop-markierter ribosomaler Proteine erlaubt eine Affinitätsaufreinigung von Ribosomen und der assoziierten mRNA. Diese Methodik, als TRAP (Translating Ribosome Affinity Purification) bezeichnet, ermöglicht die Analyse des Translatoms und wurde dahingehend optimiert, pflanzliche Antworten auf Befall durch bodenbürtige Mikroorganismen in Rhizodermis, Cortex, Endodermis sowie Zentralzylinder spezifisch zu lokalisieren. Die Genexpression in der Arabidopsis-Wurzel nach Inokulation mit drei Bodenorganismen mit unterschiedlichen Lebensweisen wurde vergleichend betrachtet: Piriformospora indica kann als mutualistischer Pilz pflanzliches Wachstum und Erträge positiv beeinflussen, wohingegen der vaskuläre Pilz Verticillium longisporum für erhebliche Verluste im Rapsanbau verantwortlich ist und der hemibiotrophe Oomycet Phytophthora parasitica ein breites Spektrum an Kulturpflanzen befällt und Ernten zerstört. Für die Interaktionsstudien zwischen Arabidopsis und den Mikroorganismen während ihrer biotrophen Lebensphase wurden sterile in vitro-Infektionssysteme etabliert und mittels TRAP und anschließender RNA-Sequenzierung eine zellschichtspezifische, genomweite Translatomanalyse durchgeführt (Inf-TRAP-Seq). Dabei zeigten sich massive Unterschiede in der differentiellen Genexpression zwischen den Zellschichten, was die Hypothese der zellschichtspezifischen Antworten unterstützt. Die Antworten nach Inokulation mit pathogenen bzw. mutualistischen Mikroorganismen unterschieden sich ebenfalls deutlich, was durch die ungleichen Lebensweisen begründbar ist. Durch die Inf-TRAP-Seq Methodik konnte z.B. im Zentralzylinder der Pathogen-infizierten Wurzeln eine expressionelle Repression von positiven Regulatoren des Zellzyklus nachgewiesen werden, dagegen in den mit P. indica besiedelten Wurzeln nicht. Dies korrelierte mit einer Pathogen-induzierten Inhibition des Wurzelwachstums, welche nicht nach Inokulation mit P. indica zu beobachten war. Obwohl keines der drei Mikroorganismen in der Lage ist, den Zentralzylinder direkt zu penetrieren, konnte hier eine differentielle Genexpression detektiert werden. Demzufolge ist ein Signalaustausch zu postulieren, über den äußere und innere Zellschichten miteinander kommunizieren. In der Endodermis konnten Genexpressionsmuster identifiziert werden, die zu einer Verstärkung der Barriere-Funktionen dieser Zellschicht führen. So könnte etwa durch Lignifizierungsprozesse die Ausbreitung der Mikroorganismen begrenzt werden. Alle drei Mikroorganismen lösten besonders im Cortex die Induktion von Genen für die Biosynthese Trp-abhängiger, antimikrobieller Sekundärmetaboliten aus. Die biologische Relevanz dieser Verteilungen kann nun geklärt werden. Zusammenfassend konnten in dieser Dissertation erstmals die durch Mikroorganismen hervorgerufenen zellschichtspezifischen Antworten der pflanzlichen Wurzel aufgelöst werden. Vergleichende bioinformatische Analyse dieses umfangreichen Datensatzes ermöglicht nun, gezielt testbare Hypothesen zu generieren. Ein Verständnis der zellschichtspezifischen Abwehrmaßnahmen der Wurzel ist essentiell für die Entwicklung neuer Strategien zur Ertragssteigerung und zum Schutz von Nutzpflanzen gegen Pathogene in der Landwirtschaft.
Die NFκB-Signalwege, Apoptose und Nekroptose sind essentielle Prozesse in der Immunantwort. Außerdem sind diese Signalwege Teil der Regulation von Zelldifferenzierung, -proliferation, -tod und Entzündungsreaktionen. Dabei wird zuerst der Rezeptor (TNFR1 oder TRAILR 1/2) aktiviert, die rekrutierten DD-Adapterproteine TRADD, FADD und RIPK1 leiten dann die entsprechende Signalkaskade weiter und bestimmen durch ihre Zusammenwirkung, ob der NFκB-Signalweg, Apoptose oder Nekroptose induziert wird.
TNFR1 und TRAILR 1/2 benötigen die DD-Adapterproteine TRADD, FADD und RIPK1 für die Zelltodinduktion, deren konkrete Bedeutung in Bezug auf Rezeptor-Spezifität, Zusammenwirken und Relevanz allerdings noch unklar ist. Um das Zusammenspiel dieser Proteine besser zu verstehen, wurden in dieser Arbeit Nekroptose-kompetente RIPK3-exprimierende HeLa-Zellen verwendet, bei denen die DD-Adapterproteine FADD, TRADD und RIPK1 einzeln oder in Kombination von zweien ausgeknockt wurden. Es stellte sich heraus, dass RIPK1 essentiell für die TNFR1- und TRAILR 1/2-vermittelte Nekroptose-Induktion ist, doch RIPK1 alleine, d.h. ohne FADD- oder TRADD-Mitbeteiligung, nur bei der TNFR1-Nekroptose-Induktion ausreicht. Wiederum inhibiert TRADD die TNFR1- und TRAILR 1/2-induzierte Nekroptose. RIPK1 und TRADD sind aber unverzichtbar für die NFκB-Aktivierung durch TNFR1 oder TRAILR 1/2 und spielen eine wichtige Rolle bei TNFR1-induzierter Apoptose. Andererseits ist FADD alleine ausreichend für die TRAILR 1/2-bezogene Caspase-8 Aktivierung. Zudem ist FADD notwendig für die TRAIL-induzierte NFκB-Signalaktivierung. In Abwesenheit von FADD und TRADD vermittelt RIPK1 die TNF-induzierte Caspase-8 Aktivierung. FADD wird für die TRAIL-induzierte Nekroptose benötigt, aber gegenläufig wirkt die TNF-induzierte Nektroptose in einer Caspase-8 abhängigen und unabhängigen Weise. Zudem sensitiviert TWEAK die TNF- und TRAIL-induzierte Nekroptose.
Zusammenfassend wurde in dieser Arbeit die Auswirkung von TNFR1 und TRAILR 1/2 auf die Aktivierung der unterschiedlichen Signalkaskaden untersucht. Des Weiteren wurde gezeigt, in welcher Weise sich das Zusammenspiel von TRADD, FADD und RIPK1 auf die Induktion von NFκB, Apoptose und Nekroptose auswirkt.
Aim: While elevational gradients in species richness constitute some of the best depicted patterns in ecology, there is a large uncertainty concerning the role of food resource availability for the establishment of diversity gradients in insects. Here, we
analysed the importance of climate, area, land use and food resources for determining diversity gradients of dung beetles along extensive elevation and land use gradients on Mt. Kilimanjaro, Tanzania.
Location: Mt. Kilimanjaro, Tanzania.
Taxon: Scarabaeidae (Coleoptera).
Methods: Dung beetles were recorded with baited pitfall traps at 66 study plots along a 3.6 km elevational gradient. In order to quantify food resources for the dung beetle community in form of mammal defecation rates, we assessed mammalian diversity and biomass with camera traps. Using a multi‐model inference framework and path analysis, we tested the direct and indirect links between climate, area, land use and mammal defecation rates on the species richness and abundance of dung beetles.
Results: We found that the species richness of dung beetles declined exponentially with increasing elevation. Human land use diminished the species richness of functional groups exhibiting complex behaviour but did not have a significant influence on total species richness. Path analysis suggested that climate, in particular temperature and to a lesser degree precipitation, were the most important predictors of dung beetle species richness while mammal defecation rate was not supported as a predictor variable.
Main conclusions: Along broad climatic gradients, dung beetle diversity is mainly limited by climatic factors rather than by food resources. Our study points to a predominant role of temperature‐driven processes for the maintenance and origination of species diversity of ectothermic organisms, which will consequently be subject to ongoing climatic changes.
Sphingobasen bilden das Grundgerüst und die Ausgangsbausteine für die Biosynthese von Sphingolipiden. Während komplexere Sphingolipide einen wichtigen Bestandteil von eukaryotischen Membranen bilden, sind Sphingobasen, die auch als long-chain bases (LCBs) bezeichnet werden, als Signalmoleküle bei zellulären Prozessen in Eukaryoten bekannt. Im tierischen System wurden antagonistische Effekte von nicht-phosphorylierten Sphingobasen (LCBs) und ihren phosphorylierten Gegenstücken (LCB-Ps) bei vielen Zellfunktionen, insbesondere der Apoptose, nachgewiesen und die zugrundeliegenden Signalwege umfassend aufgeklärt. Im Gegensatz dazu sind in Pflanzen weniger Belege für einen antagonistischen Effekt und mögliche Signaltransduktionsmechanismen bekannt. Für eine regulatorische Funktion von Sphingobasen beim programmierten Zelltod (PCD) in Pflanzen existieren mehrere Hinweise: (I) Mutationen in Genen, die den Sphingobasen-Metabolismus betreffen, führen zum Teil zu spontanem PCD und veränderten Zelltodreaktionen. (II) Die Gehalte von LCBs sind bei verschiedenen Zelltod-auslösenden Bedingungen erhöht. (III) Nekrotrophe Pathogene produzieren Toxine, wie Fumonisin B1 (FB1), die mit dem Sphingolipid-Metabolismus der Wirtspflanze interferieren, was wiederum die Ursache für den dadurch ausgelösten PCD darstellt. (IV) Die Behandlung von Pflanzen mit LCBs, nicht aber mit LCB-Ps, führt zu Zelltod.
In dieser Arbeit wurde die Rolle von Sphingobasen in der pflanzlichen Zelltodreaktion untersucht, wobei der Fokus auf der Überprüfung der Hypothese eines antagonistischen, Zelltod-hemmenden Effekts von LCB-Ps lag. Anhand von Leitfähigkeit-basierten Messungen bei Blattscheiben von Arabidopsis thaliana wurde der durch Behandlung mit LCBs und separater oder gleichzeitiger Zugabe von LCB-Ps auftretende Zelltod bestimmt. Mit dieser Art der Quantifizierung wurde der an anderer Stelle publizierte inhibierende Effekt von LCB-Ps auf den LCB-induzierten Zelltod nachgewiesen. Durch parallele Messung der Spiegel der applizierten Sphingobasen im Gewebe mittels HPLC-MS/MS konnte dieser Antagonismus allerdings auf eine reduzierte Aufnahme der LCB bei Anwesenheit der LCB-P zurückgeführt werden, was auch durch eine zeitlich getrennte Behandlung mit den Sphingobasen bestätigt wurde. Darüber hinaus wurde der Einfluss einer exogenen Zugabe von LCBs und LCB-Ps auf den durch Pseudomonas syringae induzierten Zelltod von A. thaliana untersucht. Für LCB-Ps wurde dabei kein Zelltod-hemmender Effekt beobachtet, ebenso wenig wie ein Einfluss von LCB-Ps auf den PCD, der durch rekombinante Expression und Erkennung eines Avirulenzproteins in Arabidopsis ausgelöst wurde. Für LCBs wurde dagegen eine direkte antibakterielle Wirkung im Zuge der Experimente mit P. syringae gezeigt, die den in einer anderen Publikation beschriebenen inhibierenden Effekt von LCBs auf den Pathogen-induzierten Zelltod in Pflanzen relativiert.
In weiteren Ansätzen wurden Arabidopsis-Mutanten von Enzymen des Sphingobasen-Metabolismus (LCB-Kinase, LCB-P-Phosphatase, LCB-P-Lyase) hinsichtlich veränderter in-situ-Spiegel von LCBs/LCB-Ps funktionell charakterisiert. Der Phänotyp der Mutanten gegenüber Fumonisin B1 wurde zum einen anhand eines Wachstumstests mit Keimlingen und zum anderen anhand des Zelltods von Blattscheiben bestimmt und die dabei akkumulierenden Sphingobasen quantifiziert. Die Sensitivität der verschiedenen Linien gegenüber FB1 korrelierte eng mit den Spiegeln der LCBs, während hohe Gehalte von LCB-Ps alleine nicht in der Lage waren den Zelltod zu verringern. In einzelnen Mutanten konnte sogar eine Korrelation von stark erhöhten LCB-P-Spiegeln mit einer besonderen Sensitivität gegenüber FB1 festgestellt werden.
Die Ergebnisse der vorliegenden Arbeit stellen die Hypothese eines antagonistischen Effekts von phosphorylierten Sphingobasen beim pflanzlichen Zelltod in Frage. Stattdessen konnte in detaillierten Analysen der Sphingobasen-Spiegel die positive Korrelation der Gehalte von LCBs mit dem Zelltod gezeigt werden. Die hier durchgeführten Experimente liefern damit nicht nur weitere Belege für die Zelltod-fördernde Wirkung von nicht-phosphorylierten Sphingobasen, sondern tragen zum Verständnis der Sphingobasen-Homöostase und des Sphingobasen-induzierten PCD in Pflanzen bei.
The nuclear envelope serves as important messenger RNA (mRNA) surveillance system. In yeast and human, several control systems act in parallel to prevent nuclear export of unprocessed mRNAs. Trypanosomes lack homologues to most of the involved proteins and their nuclear mRNA metabolism is non-conventional exemplified by polycistronic transcription and mRNA processing by trans-splicing. We here visualized nuclear export in trypanosomes by intra- and intermolecular multi-colour single molecule FISH. We found that, in striking contrast to other eukaryotes, the initiation of nuclear export requires neither the completion of transcription nor splicing. Nevertheless, we show that unspliced mRNAs are mostly prevented from reaching the nucleus-distant cytoplasm and instead accumulate at the nuclear periphery in cytoplasmic nuclear periphery granules (NPGs). Further characterization of NPGs by electron microscopy and proteomics revealed that the granules are located at the cytoplasmic site of the nuclear pores and contain most cytoplasmic RNA-binding proteins but none of the major translation initiation factors, consistent with a function in preventing faulty mRNAs from reaching translation. Our data indicate that trypanosomes regulate the completion of nuclear export, rather than the initiation. Nuclear export control remains poorly understood, in any organism, and the described way of control may not be restricted to trypanosomes.
In mammals, anucleate platelets circulate in the blood flow and are primarily responsible for maintaining functional hemostasis. Platelets are generated in the bone marrow (BM) by megakaryocytes (MKs), which mainly reside directly next to the BM sinusoids to release proplatelets into the blood. MKs originate from hematopoietic stem cells and are thought to migrate from the endosteal to the vascular niche during their maturation, a process, which is, despite being intensively investigated, still not fully understood.
Long-term intravital two photon microscopy (2PM) of MKs and vasculature in murine bone marrow was performed and mean squared displacement analysis of cell migration was performed. The MKs exhibited no migration, but wobbling-like movement on time scales of 3 h. Directed cell migration always results in non-random spatial distribution. Thus, a computational modelling algorithm simulating random MK distribution using real 3D light-sheet fluorescence microscopy data sets was developed. Direct comparison of real and simulated random MK distributions showed, that MKs exhibit a strong bias to vessel-contact. However, this bias is not caused by cell migration, as non-vessel-associated MKs were randomly distributed in the intervascular space. Furthermore, simulation studies revealed that MKs strongly impair migration of other cells in the bone marrow by acting as large-sized obstacles. MKs are thought to migrate from the regions close to the endosteum towards the vasculature during their maturation process. MK distribution as a function of their localization relative to the endosteal regions of the bones was investigated by light sheet fluorescence microscopy (LSFM). The results show no bone-region dependent distribution of MKs. Taken together, the newly established methods and obtained results refute the model of MK migration during their maturation.
Ischemia reperfusion (I/R) injury is a frequent complication of cerebral ischemic stroke, where brain tissue damage occurs despite successful recanalization. Platelets, endothelial cells and immune cells have been demonstrated to affect the progression of I/R injury in experimental mouse models 24 h after recanalization. However, the underlying Pathomechanisms, especially in the first hours after recanalization, are poorly understood.
Here, LSFM, 2PM and complemental advanced image analysis workflows were established for investigation of platelets, the vasculature and neutrophils in ischemic brains. Quantitative analysis of thrombus formation in the ipsilateral and contralateral hemispheres at different time points revealed that platelet aggregate formation is minimal during the first 8 h after recanalization and occurs in both hemispheres. Considering that maximal tissue damage already is present at this time point, it can be concluded that infarct progression and neurological damage do not result from platelet aggregated formation. Furthermore, LSFM allowed to confirm neutrophil infiltration into the infarcted hemisphere and, here, the levels of endothelial cell marker PECAM1 were strongly reduced. However, further investigations must be carried out to clearly identify the role of neutrophils and the endothelial cells in I/R injury.
Zinc (Zn2+) can modulate platelet and coagulation activation pathways, including fibrin formation. Here, we studied the (patho)physiological consequences of abnormal platelet Zn2+ storage and release. To visualize Zn2+ storage in human and mouse platelets, the Zn2+ specific fluorescent dye FluoZin3 was used. In resting platelets, the dye transiently accumulated into distinct cytosolic puncta, which were lost upon platelet activation. Platelets isolated from Unc13d−/− mice, characterized by combined defects of α/δ granular release, showed a markedly impaired Zn2+ release upon activation. Platelets from Nbeal2−/− mice mimicking Gray platelet syndrome (GPS), characterized by primarily loss of the α-granule content, had strongly reduced Zn2+ levels, which was also confirmed in primary megakaryocytes. In human platelets isolated from patients with GPS, Hermansky-Pudlak Syndrome (HPS) and Storage Pool Disease (SPD) altered Zn2+ homeostasis was detected. In turbidity and flow based assays, platelet-dependent fibrin formation was impaired in both Nbeal2−/− and Unc13d−/− mice, and the impairment could be partially restored by extracellular Zn2+. Altogether, we conclude that the release of ionic Zn2+ store from secretory granules upon platelet activation contributes to the procoagulant role of Zn2+ in platelet-dependent fibrin formation.
C60 fullerene as an effective nanoplatform of alkaloid Berberine delivery into leukemic cells
(2019)
A herbal alkaloid Berberine (Ber), used for centuries in Ayurvedic, Chinese, Middle-Eastern, and native American folk medicines, is nowadays proved to function as a safe anticancer agent. Yet, its poor water solubility, stability, and bioavailability hinder clinical application. In this study, we have explored a nanosized carbon nanoparticle—C60 fullerene (C60)—for optimized Ber delivery into leukemic cells. Water dispersions of noncovalent C60-Ber nanocomplexes in the 1:2, 1:1, and 2:1 molar ratios were prepared. UV–Vis spectroscopy, dynamic light scattering (DLS), and atomic force microscopy (AFM) evidenced a complexation of the Ber cation with the negatively charged C60 molecule. The computer simulation showed that π-stacking dominates in Ber and C\(_{60}\) binding in an aqueous solution. Complexation with C\(_{60}\) was found to promote Ber intracellular uptake. By increasing C\(_{60}\) concentration, the C\(_{60}\)-Ber nanocomplexes exhibited higher antiproliferative potential towards CCRF-CEM cells, in accordance with the following order: free Ber < 1:2 < 1:1 < 2:1 (the most toxic). The activation of caspase 3/7 and accumulation in the sub-G1 phase of CCRF-CEM cells treated with C\(_{60}\)-Ber nanocomplexes evidenced apoptosis induction. Thus, this study indicates that the fast and easy noncovalent complexation of alkaloid Ber with C\(_{60}\) improved its in vitro efficiency against cancer cells.
Synergy of chemo- and photodynamic therapies with C\(_{60}\) Fullerene-Doxorubicin nanocomplex
(2019)
A nanosized drug complex was explored to improve the efficiency of cancer chemotherapy, complementing it with nanodelivery and photodynamic therapy. For this, nanomolar amounts of a non-covalent nanocomplex of Doxorubicin (Dox) with carbon nanoparticle C\(_{60}\) fullerene (C\(_{60}\)) were applied in 1:1 and 2:1 molar ratio, exploiting C\(_{60}\) both as a drug-carrier and as a photosensitizer. The fluorescence microscopy analysis of human leukemic CCRF-CEM cells, in vitro cancer model, treated with nanocomplexes showed Dox’s nuclear and C\(_{60}\)'s extranuclear localization. It gave an opportunity to realize a double hit strategy against cancer cells based on Dox's antiproliferative activity and C\(_{60}\)'s photoinduced pro-oxidant activity. When cells were treated with 2:1 C\(_{60}\)-Dox and irradiated at 405 nm the high cytotoxicity of photo-irradiated C\(_{60}\)-Dox enabled a nanomolar concentration of Dox and C\(_{60}\) to efficiently kill cancer cells in vitro. The high pro-oxidant and pro-apoptotic efficiency decreased IC\(_{50}\) 16, 9 and 7 × 10\(^3\)-fold, if compared with the action of Dox, non-irradiated nanocomplex, and C\(_{60}\)'s photodynamic effect, correspondingly. Hereafter, a strong synergy of therapy arising from the combination of C\(_{60}\)-mediated Dox delivery and C\(_{60}\) photoexcitation was revealed. Our data indicate that a combination of chemo- and photodynamic therapies with C\(_{60}\)-Dox nanoformulation provides a promising synergetic approach for cancer treatment.