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- Adenosinrezeptor (12)
- Adhäsion (12)
- Altern (12)
- Arabidopsis (12)
- Arzneimittelüberwachung (12)
- Brain-derived neurotrophic factor (12)
- Click-Chemie (12)
- Comet Assay (12)
- Computer Center University of Wuerzburg (12)
- Dialyse (12)
- Diboren (12)
- Dopamin (12)
- Durchflusscytometrie (12)
- EGFR (12)
- ERP (12)
- Enzym (12)
- FRET (12)
- Fibrose (12)
- Fluorescence (12)
- Gen (12)
- Grundschule (12)
- Hirntumor (12)
- Immunotherapy (12)
- Kinderheilkunde (12)
- Klima (12)
- Klimawandel (12)
- Kohlenstoff-Nanoröhre (12)
- Komplikationen (12)
- Kulturwissenschaften (12)
- Kutikula (12)
- Lymphom (12)
- Magnetische Kernresonanz (12)
- Medienkompetenz (12)
- Metastase (12)
- Modell (12)
- Monitoring (12)
- NFAT (12)
- Nanostrukturiertes Material (12)
- Netzhaut (12)
- Nigeria (12)
- Non-Hodgkin-Lymphom (12)
- Onkogen (12)
- PCR (12)
- PRRT (12)
- Pharmakologie (12)
- Pilzkörper (12)
- Polytrauma (12)
- Quality of Experience (12)
- Quantenchemie (12)
- Rekonstruktion (12)
- Religion (12)
- Replikation (12)
- Rotatorenmanschette (12)
- Ruthenium (12)
- SMN (12)
- SNP (12)
- Schule (12)
- Siliciumorganische Verbindungen (12)
- Spracherwerb (12)
- Supraleitung (12)
- T cell (12)
- Toll-like-Rezeptoren (12)
- Transplantat-Wirt-Reaktion (12)
- Tumor-Nekrose-Faktor <alpha> (12)
- Ubiquitin (12)
- Ultrakurzer Lichtimpuls (12)
- Validierung (12)
- Westafrika (12)
- Zelladhäsion (12)
- adhesion (12)
- antibody (12)
- blood–brain barrier (12)
- carbenes (12)
- catalysis (12)
- clinical trial (12)
- colorectal carcinoma (12)
- complications (12)
- database (12)
- dementia (12)
- drug delivery (12)
- dynamics (12)
- emotions (12)
- endothelium (12)
- genomics (12)
- heart rate (12)
- immunology (12)
- interaction (12)
- lung (12)
- malignant hyperthermia (12)
- measles (12)
- mechanism (12)
- microarray (12)
- microbiome (12)
- molecular beam epitaxy (12)
- molecular biology (12)
- monoclonal antibodies (12)
- neural networks (12)
- neuropathy (12)
- nuclear envelope (12)
- oncology (12)
- optimization (12)
- periodontitis (12)
- phenotype (12)
- plasticity (12)
- pollen (12)
- polymers (12)
- recombination (12)
- review (12)
- screening (12)
- synaptic plasticity (12)
- systematic review (12)
- tinnitus (12)
- topological insulator (12)
- total knee arthroplasty (12)
- trabeculectomy (12)
- tumor microenvironment (12)
- ultrasound (12)
- water oxidation (12)
- ++ (11)
- 3D (11)
- Alzheimer's disease (11)
- Alzheimer’s disease (11)
- Anorexia nervosa (11)
- Antigen CD8 (11)
- Atherosclerosis (11)
- B-Zelle (11)
- Bakterielle Infektion (11)
- Bauchspeicheldrüsenkrebs (11)
- Bioisosterie (11)
- Biosynthese (11)
- Bioverfügbarkeit (11)
- Blazar (11)
- Bruchpilot (11)
- CRISPR/Cas9 (11)
- Chromatin (11)
- Computertomografie (11)
- Dynamik (11)
- Elektronentransfer (11)
- Epidemiologie (11)
- Epidermaler Wachstumsfaktor-Rezeptor (11)
- Flavonoide (11)
- Fließverhalten (11)
- Fotovoltaik (11)
- Genom (11)
- Geschlecht (11)
- Handlungsorientierung (11)
- Hypoxie (11)
- Immuncytochemie (11)
- Immunology (11)
- Japankärpfling (11)
- Knochen (11)
- Knochenersatz (11)
- Kolorektales Karzinom (11)
- Kommunikation (11)
- LASP1 (11)
- LPS (11)
- Laser (11)
- Längsschnittuntersuchung (11)
- MS (11)
- Mais (11)
- Mathematik (11)
- Megakaryozyt (11)
- Mensch-Maschine-Kommunikation (11)
- Mesenchymale Stammzellen (11)
- Mesenchymzelle (11)
- Model (11)
- Multiple Myeloma (11)
- Mutagenität (11)
- NIRS (11)
- Nanodiamant (11)
- Neuropathie (11)
- Niereninsuffizienz (11)
- Operation (11)
- Osteoinduktion (11)
- Parasit (11)
- Parathormon (11)
- Perylenbisimid (11)
- Phylogenie (11)
- Proteaseinhibitor (11)
- Proteasen (11)
- Psychotherapie (11)
- Pulmonale Hypertonie (11)
- Quality of life (11)
- Quanten-Hall-Effekt (11)
- Reperfusion (11)
- Risikofaktoren (11)
- Salmonella (11)
- Schizophrenia (11)
- Schlafapnoe (11)
- Schließzelle (11)
- Schultergelenk (11)
- Schädel-Hirn-Trauma (11)
- Sentinel-1 (11)
- Silicate (11)
- Skala (11)
- Small RNA (11)
- Sprache (11)
- Squark (11)
- Stammzellen (11)
- Sterblichkeit (11)
- Symbiose (11)
- TWEAK (11)
- Training (11)
- Tuberkelbakterium (11)
- USA (11)
- Vasodilatator-stimuliertes Phosphoprotein (11)
- Wahrnehmung (11)
- X-ray crystallography (11)
- Zebrabärbling (11)
- Zelltod (11)
- allergy (11)
- biocompatibility (11)
- biofilm (11)
- cancer therapy (11)
- cancer treatment (11)
- cell adhesion (11)
- chemokines (11)
- cochlear implant (11)
- communication (11)
- coping (11)
- dosimetry (11)
- emotion regulation (11)
- fatigue (11)
- fear (11)
- fluorescence microscopy (11)
- glioblastoma multiforme (11)
- hydrogels (11)
- kinetics (11)
- membrane proteins (11)
- microglia (11)
- molecular dynamics (11)
- nervous system (11)
- neuroendocrine tumor (11)
- next generation sequencing (11)
- oncolytic virus (11)
- psychiatric disorders (11)
- randomized controlled trial (11)
- reactive oxygen species (11)
- relapse (11)
- signaling (11)
- tight junctions (11)
- transport (11)
- university (11)
- Alkohol (10)
- Alps (10)
- Antigen CD28 (10)
- BERA (10)
- BMP-2 (10)
- Bacteria (10)
- Beschichtung (10)
- Bildverarbeitung (10)
- Bioinformatics (10)
- Biokompatibilität (10)
- Borole (10)
- Cadherine (10)
- Calciumphosphate (10)
- Carbene (10)
- Children (10)
- Chiralität <Chemie> (10)
- Chronische Niereninsuffizienz (10)
- Churritisch (10)
- Cochlear-Implantat (10)
- Computational chemistry (10)
- Coronaviren (10)
- Covid-19 (10)
- Cyclo-GMP (10)
- Depressivität (10)
- Diamant (10)
- Digital Humanities (10)
- Dimerisierung (10)
- Dotierung (10)
- Echokardiographie (10)
- Eierstockkrebs (10)
- Elektrochemie (10)
- Elektronenkorrelation (10)
- Englisch (10)
- Enzyminhibitor (10)
- Erbkrankheit (10)
- Ereigniskorreliertes Potenzial (10)
- Erwachsener (10)
- Franken (10)
- Frau (10)
- Funktionalisierung <Chemie> (10)
- Funktionelle Kernspintomografie (10)
- Förderung (10)
- Geldpolitik (10)
- Glatter Krallenfrosch (10)
- Grenzfläche (10)
- HSM-Satztest (10)
- Herzmuskelzelle (10)
- Honigbiene (10)
- Hypophosphatasie (10)
- IL-4 (10)
- Immunfluoreszenz (10)
- Impfstoff (10)
- Induzierte pluripotente Stammzelle (10)
- Interleukin 4 (10)
- Interview (10)
- JNK (10)
- Klassifikation (10)
- Kleinsatellit (10)
- Kniegelenk (10)
- Kohlenstoff (10)
- Komplikation (10)
- Kooperation (10)
- Kultur (10)
- LC-MS (10)
- Ladungstransfer (10)
- Ligand (10)
- Lokalisation (10)
- Magnesiumphosphate (10)
- Magnetismus (10)
- Mathematikunterricht (10)
- Mathematisches Modell (10)
- Medulloblastom (10)
- Metakognition (10)
- Metallocene (10)
- Methylphenidat (10)
- Mikrokerne (10)
- Musik (10)
- NAFLD (10)
- Nahrungserwerb (10)
- Nebennierenrindenkarzinom (10)
- Neolithikum (10)
- Neuropathischer Schmerz (10)
- Niger (10)
- Osteosynthese (10)
- PD-1 (10)
- PD-L1 (10)
- Pain (10)
- Panikstörung (10)
- Parodontitis (10)
- Pathogenität (10)
- Platelets (10)
- Plattenepithelkarzinom (10)
- Prevalence (10)
- Prostaglandine (10)
- Präfrontaler Cortex (10)
- Pädagogik (10)
- Qualitätskontrolle (10)
- RNA interference (10)
- RNA-seq (10)
- RNS-Spleißen (10)
- Radiotherapy (10)
- Remote Sensing (10)
- Rhodium (10)
- Rituximab (10)
- SAR (10)
- Schilddrüse (10)
- Schmalwand <Arabidopsis> (10)
- Silaanaloga (10)
- Silber (10)
- Silicon (10)
- South Africa (10)
- Sprachverstehen (10)
- Staphylococcus (10)
- Starke Kopplung (10)
- Stickstoffmonoxid-Synthase (10)
- Stressreaktion (10)
- Stroke (10)
- Struktur (10)
- T-Lymphozyten (10)
- T-Lymphozyten-Rezeptor (10)
- Tagesrhythmus (10)
- Theologie (10)
- Totalsynthese (10)
- Toxizität (10)
- Transthorakale Echokardiographie (10)
- Treg (10)
- Trypanosomen (10)
- Verwaltungsrecht (10)
- Vitamin D (10)
- Vor- und Frühgeschichte (10)
- Vorschulkind (10)
- Wachstum (10)
- agriculture (10)
- aldosterone (10)
- amino acids (10)
- antimicrobial resistance (10)
- bariatric surgery (10)
- bioinformatics (10)
- biosynthesis (10)
- body size (10)
- boranes (10)
- cell wall (10)
- chronic heart failure (10)
- circadian rhythms (10)
- cognitive impairment (10)
- collagen (10)
- comparison (10)
- complex (10)
- coronary artery disease (10)
- cortisol (10)
- decision-making (10)
- density functional calculations (10)
- dispersal (10)
- eNOS (10)
- education (10)
- efficacy (10)
- forest (10)
- gene therapy (10)
- genome-wide association (10)
- glioma (10)
- global change (10)
- glycine receptor (10)
- human behaviour (10)
- immunomodulation (10)
- induced pluripotent stem cells (10)
- injury (10)
- insect (10)
- knockout (10)
- leaf-cutting ants (10)
- leukemia (10)
- lymph nodes (10)
- mapping (10)
- membrane potential (10)
- metapopulation (10)
- motivation (10)
- near-infrared spectroscopy (10)
- neuroscience (10)
- oncolytic virotherapy (10)
- oxidativer Stress (10)
- quality assurance (10)
- quantification (10)
- regulatorische T-Zellen (10)
- remodeling (10)
- senescence (10)
- silicon (10)
- social interaction (10)
- stem cell transplantation (10)
- subthalamic nucleus (10)
- synapse (10)
- tDCS (10)
- transcriptomics (10)
- transient absorption (10)
- tumors (10)
- vaccination (10)
- vertigo (10)
- walking (10)
- Östrogene (10)
- 3 (9)
- Adenosin (9)
- Adenosine receptors (9)
- Affekt (9)
- Afrika (9)
- Analyse (9)
- Angeregter Zustand (9)
- Antigen CD40 (9)
- Anxiety (9)
- Aorta (9)
- Aortenstenose (9)
- Autoaggressionskrankheit (9)
- Autophagie (9)
- Beyond Standard Model (9)
- Biotransformation (9)
- Biradikal (9)
- Bordetella pertussis (9)
- Borylierung (9)
- Boğazkale (9)
- Butyrat (9)
- CT (9)
- Carcinogenese (9)
- Catalysis (9)
- Cloud Computing (9)
- Deep learning (9)
- Dendritic cells (9)
- Diagnose (9)
- Diborane (9)
- Diskursanalyse (9)
- Dünndarm (9)
- Einzelphotonenemission (9)
- Elektroencephalographie (9)
- Elektronenmikroskopie (9)
- Elektronenstruktur (9)
- Elementarteilchenphysik (9)
- Epithel (9)
- Ernährung (9)
- Europe (9)
- Experimentelle Psychologie (9)
- Extremwertstatistik (9)
- Familie (9)
- Fanconi Anämie (9)
- Fettgewebe (9)
- Fibroblastenwachstumsfaktor (9)
- Fibromyalgie (9)
- Finite-Elemente-Methode (9)
- Frühgeborene (9)
- GIS (9)
- Geistigbehindertenpädagogik (9)
- Gentherapie (9)
- Geoinformationssystem (9)
- Geruchswahrnehmung (9)
- Glioblastoma (9)
- Google Earth Engine (9)
- Grammatik (9)
- Haut (9)
- Herzmuskelkrankheit (9)
- Hethiter (9)
- Hyaluronsäure (9)
- Immunisierung (9)
- Impulsivität (9)
- Innovation (9)
- Instrumentelle Analytik (9)
- Integrine (9)
- Iran (9)
- Isolierung <Chemie> (9)
- Jugendliche (9)
- Juvenile chronische Arthritis (9)
- Kapillarelektrophorese (9)
- Knorpel (9)
- Konflikt (9)
- Koordinationslehre (9)
- Korpus <Linguistik> (9)
- LC-MS/MS (9)
- Landsat (9)
- Leber (9)
- Lehre (9)
- Linguistik (9)
- Löslichkeit (9)
- MAPK (9)
- Macrophage (9)
- Magnetische Resonanz (9)
- Mehrfachbindung (9)
- Mehrsprachigkeit (9)
- Messenger-RNS (9)
- Mice (9)
- Molekulare Erkennung (9)
- Monozyt (9)
- Morphologie (9)
- Mukoviszidose (9)
- Myokardprotektion (9)
- NO (9)
- NSCLC (9)
- Nebennierenrindenkrebs (9)
- Neuroinflammation (9)
- Nierentransplantation (9)
- Oberfläche (9)
- Oberflächenphysik (9)
- Optoelektronik (9)
- Pankreaskarzinom (9)
- Pathogenese (9)
- Persönlichkeit (9)
- Pharmakotherapie (9)
- Photolumineszenzspektroskopie (9)
- Physiologie (9)
- Plasmamembran (9)
- Polycyclische Aromaten (9)
- Protein (9)
- Pseudomonas syringae (9)
- Psychoonkologie (9)
- Quantenmechanik (9)
- Rechenzentrum (9)
- Rechtsvergleich (9)
- Regeneration (9)
- Regionalentwicklung (9)
- Revision (9)
- Sarkoidose (9)
- Satellit (9)
- Schwindel (9)
- Schüttgut (9)
- Spin-Bahn-Wechselwirkung (9)
- Squaraine (9)
- Stathmin (9)
- Stereoselektive Synthese (9)
- Supply Chain Management (9)
- Syntax (9)
- Systembiologie (9)
- Südafrika (9)
- TDM (9)
- Textverstehen (9)
- Therapy (9)
- Tourismus (9)
- Trabekulektomie (9)
- Transplantat (9)
- Transport (9)
- Trypanosoma (9)
- Tumorantigen (9)
- Vakuole (9)
- Vergleich (9)
- Visuelle Aufmerksamkeit (9)
- Wurzel (9)
- Zellkern (9)
- Zentralnervensystem (9)
- Zinkselenid (9)
- actin cytoskeleton (9)
- adolescence (9)
- aggregation (9)
- amyotrophic lateral sclerosis (9)
- antennal lobe (9)
- anxiety disorders (9)
- aromaticity (9)
- astrocytes (9)
- biomechanics (9)
- bladder cancer (9)
- blood brain barrier (9)
- bone cement (9)
- bone marrow (9)
- cardiac surgery (9)
- cardiovascular disease (9)
- cell culture (9)
- cell migration (9)
- central nervous system (9)
- ceramide (9)
- cerebrospinal fluid (9)
- chromatin (9)
- computed tomography (9)
- coronary heart disease (9)
- cuticular hydrocarbons (9)
- cystic fibrosis (9)
- cytotoxic T cells (9)
- decision making (9)
- dendritische Zellen (9)
- dialysis (9)
- earth observation (9)
- electroencephalography (9)
- electron microscopy (9)
- electrophysiology (9)
- energy (9)
- experimental autoimmune encephalomyelitis (9)
- family (9)
- foamy virus (9)
- genes (9)
- gephyrin (9)
- glucose (9)
- high energy physics (9)
- hip (9)
- hyaluronic acid (9)
- hydrogel (9)
- impact (9)
- incidence (9)
- inhibitor (9)
- interferon (9)
- latency (9)
- macrophage (9)
- major depression (9)
- mechanotransduction (9)
- mesenchymale Stammzellen (9)
- methylation (9)
- mitosis (9)
- monitoring (9)
- monoklonale Antikörper (9)
- myocardium (9)
- navigation (9)
- neurogenesis (9)
- optical spectroscopy (9)
- organic chemistry (9)
- organic semiconductors (9)
- pathogens (9)
- pathway (9)
- perfusion (9)
- peripheral nervous system (9)
- permeability (9)
- personality (9)
- pharmacology (9)
- photocatalysis (9)
- platelet activation (9)
- platelet aggregation (9)
- prognostic factors (9)
- quantum dots (9)
- radiation (9)
- rectal cancer (9)
- reveals (9)
- selection (9)
- serum (9)
- sleep (9)
- species richness (9)
- spinal muscular atrophy (9)
- stomata (9)
- systematic uncertainty (9)
- temperature (9)
- tolerance (9)
- transcription factor (9)
- transmission (9)
- type 2 diabetes (9)
- vaccine (9)
- water (9)
- wound healing (9)
- Übersetzung (9)
- ATLAS (8)
- AdS-CFT-Korrespondenz (8)
- Adhärenz (8)
- Aggregat <Chemie> (8)
- Akt (8)
- Aktiver galaktischer Kern (8)
- Algorithmus (8)
- Alltagskultur (8)
- Ancistrocladaceae (8)
- Anthocyane (8)
- Antibiotika (8)
- Antigen (8)
- Aortenklappenersatz (8)
- Arthrose (8)
- Arthroskopie (8)
- Arzneimittelforschung (8)
- Astrophysik (8)
- Asymmetrie (8)
- Asymmetrische Synthese (8)
- Auge (8)
- Autonomer Roboter (8)
- Bauchspeicheldrüse (8)
- Blimp-1 (8)
- Blutdruck (8)
- Blutstammzelle (8)
- Botanik (8)
- Brain (8)
- Burkina Faso (8)
- CD28 (8)
- CD40 (8)
- CD95 (8)
- COMT (8)
- Caenorhabditis elegans (8)
- Camponotus floridanus (8)
- Chlamydia (8)
- Compressed Sensing (8)
- Cross-Section (8)
- Cysteinproteasen (8)
- Cytomegalie-Virus (8)
- DLBCL (8)
- Darm (8)
- Data Mining (8)
- Datenbank (8)
- Degradation (8)
- Demenz (8)
- Deutschunterricht (8)
- Diborene (8)
- Didaktik (8)
- ELISA (8)
- ELISPOT (8)
- ERK (8)
- Elektronenspin (8)
- Elektronenspinresonanz (8)
- Emotionsregulation (8)
- Endothelzellen (8)
- Engagement (8)
- Erbrechen (8)
- Europa (8)
- Exziton-Polariton (8)
- FISH (8)
- Fachdidaktik (8)
- Femtosekundenbereich (8)
- Fibroblasten (8)
- Fitness (8)
- Fluoreszenzspektroskopie (8)
- Forschung (8)
- Fußball (8)
- GABA (8)
- Gammastrahlung (8)
- Gedächtnisleistung (8)
- Geistige Behinderung (8)
- Genotoxicity (8)
- Geschlechtsunterschied (8)
- Gewebe (8)
- Gliom (8)
- Graphen (8)
- HPLC-MS (8)
- Habichtskraut (8)
- Haemophilus influenzae (8)
- Harnwegsinfektion (8)
- Hernie (8)
- Hochbegabung (8)
- Hubbard-Modell (8)
- Humangenetik (8)
- Hymenoptera (8)
- Hypertrophie (8)
- Hüftgelenk (8)
- IL-10 (8)
- Immunantwort (8)
- In vivo (8)
- India (8)
- Indien (8)
- Inklusion (8)
- Keilschrifttext (8)
- Kephalometrie (8)
- Kieferorthopädie (8)
- Kinetik (8)
- Knochenersatzmaterial (8)
- Knockout (8)
- Komposit <Zahnmedizin> (8)
- Kondo-Effekt (8)
- Korrelation (8)
- Kreuzband (8)
- Kristallstruktur (8)
- Landwirtschaft (8)
- Langerhans-Inseln (8)
- Lanthanoide (8)
- Leishmania (8)
- Lernautonomie (8)
- Lower Franconia (8)
- Lumineszenz (8)
- Lungenfibrose (8)
- Machine Learning (8)
- Magenbypass (8)
- Makuladegeneration (8)
- Mass (8)
- Meeresschwämme (8)
- Meiosis (8)
- Meningitis (8)
- Meningokokken (8)
- Metaanalyse (8)
- Methode (8)
- Mikroglia (8)
- Mikrokerntest (8)
- Mikrostruktur (8)
- Mitochondrien (8)
- Mobiler Roboter (8)
- Molekül (8)
- Monozyten (8)
- Multiple myeloma (8)
- Myokarditis (8)
- N-heterocyclic carbenes (8)
- Nachsorge (8)
- Nanodraht (8)
- Naturstoff (8)
- Nervendegeneration (8)
- Neugeborenes (8)
- Neutrino (8)
- Notfallmedizin (8)
- Obesity (8)
- Onkolyse (8)
- Optogenetik (8)
- Organisches Molekül (8)
- Oxylipine (8)
- PONV (8)
- Parton distributions (8)
- Pemphigus (8)
- Peptide (8)
- Periphere arterielle Verschlusskrankheit (8)
- Permeabilität (8)
- Pharmakodynamik (8)
- Photodissoziation (8)
- Photovoltaik (8)
- Platin (8)
- Pollen (8)
- Protein p53 (8)
- Protein-Tyrosin-Kinasen (8)
- Proteinbindung (8)
- Protonen-NMR-Spektroskopie (8)
- Präkonditionierung (8)
- Psychische Störung (8)
- Pump-Probe-Technik (8)
- Quantendynamik (8)
- Quelle (8)
- Quran (8)
- RNA sequencing (8)
- Radikal <Chemie> (8)
- Raf <Biochemie> (8)
- Raf-Kinasen (8)
- Raman spectroscopy (8)
- Rastertunnelmikroskop (8)
- Rechenzentrum Universität Würzburg (8)
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- Lernverhalten (4)
- Leukozyt (4)
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- LiDAR (4)
- Localization (4)
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- Loss of heterozygosity (4)
- Low-density-Lipoproteine (4)
- Lung cancer (4)
- Lungenembolie (4)
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- Lysosom (4)
- MALDI-MS (4)
- MALT (4)
- MCP-1 (4)
- MIBG (4)
- MIZ1 (4)
- MMB (4)
- MMP (4)
- MOLLI (4)
- MPEC (4)
- MPFL (4)
- MR (4)
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- MSCs (4)
- MTUS1 (4)
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- PEG (4)
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- PKA (4)
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Schriftenreihe
- Cultural Animal Studies, Band 3 (24)
- Spezielle Didaktik der Sportarten (2)
- Aesthetische Eigenzeiten, 17 (1)
- Akten des ... Symposiums des Mediävistenverbandes; 13,2 (1)
- Alter Orient und Altes Testament : Sonderreihe Veröffentlichungen zur Kultur und Geschichte des Alten Orients ; 3 (1)
- Aventiuren; 13 (1)
- Berichte aus der Informatik (1)
- Deuterocanonical and Cognate Literature Studies (1)
- Deuterocanonical and Cognate Literature Yearbook (1)
- Epistemata. Reihe Literaturwissenschaft ; 483 (1)
Sonstige beteiligte Institutionen
- VolkswagenStiftung (24)
- Johns Hopkins School of Medicine (18)
- Fraunhofer-Institut für Silicatforschung ISC (8)
- Helmholtz Institute for RNA-based Infection Research (HIRI) (8)
- IZKF Nachwuchsgruppe Geweberegeneration für muskuloskelettale Erkrankungen (7)
- Akademie der Wissenschaften und der Literatur, Mainz (6)
- DFG Forschungsgruppe 2757 / Lokale Selbstregelungen im Kontext schwacher Staatlichkeit in Antike und Moderne (LoSAM) (6)
- Clinical Trial Center (CTC) / Zentrale für Klinische Studien Würzburg (ZKSW) (5)
- Johns Hopkins University School of Medicine (5)
- Universität Leipzig (5)
ResearcherID
- B-1911-2015 (1)
- B-4606-2017 (1)
- C-2593-2016 (1)
- D-1221-2009 (1)
- D-1250-2010 (1)
- D-3057-2014 (1)
- I-5818-2014 (1)
- J-8841-2015 (1)
- M-1240-2017 (1)
- N-2030-2015 (1)
The present work deals with the preparation of hydrogels in different size scales for various applications. Thus, macroscopic bulk hydrogels were prepared from differently modified pig gastric mucin (PGM), microgels were made from PGM in combination with hyaluronic acid (HA), as well as from gelatin in combination with poly(ethylene glycol) (PEG), and nanogels were fabricated from poly(glycidol) (PG). According to their size, each hydrogels have different applications. First, it was investigated whether previously existing studies involving the preparation of covalently crosslinked hydrogels via free radical polymerization from bovine submaxillary gland mucin (BSM) could also be carried out with the much cheaper alternative PGM. After this was successfully demonstrated and the hydrogels were systematically investigated for their mechanical properties and biocompatibility, a second hydrogel system was established. Here, PGM was functionalized with allyl glycidyl ether (AGE) and crosslinked in combination with thiolated HA via thiol-ene reaction. These hydrogels were also systematically evaluated and compared with the hydrogels prepared via free radical polymerization. It was confirmed that the more random free radical polymerization leads to more disordered networks than the thiol-ene reaction. In both systems, biocompatibility was demonstrated with both L929 CCL1 murine fibroblasts and human mesenchymal stem cells (hMSCs). Using this knowledge as background and the request to make mucin printable, microgels were prepared via the emulsion technique using the previously established thiol-ene hydrogel precursor solution. Here, applying the recently used photoinitiator 2-hydroxy-4-(2-hydroxyethoxy)-2- methylpropiophenone (Irgacure 2959), which is more soluble in oil than in water, was challenging and did not result in well-crosslinked microgels. Therefore, a third hydrogel system was established, which was based on thiol-ene crosslinked AGE functionalized pig gastric mucin (PGM-AGE)-thiolated hyaluronic acid (HASH) hydrogels and with lithium phenyl-2,4,6- trimethylbenzoylphosphinate (LAP) being used as photoinitiator. Hereby, stably crosslinked microgels could be prepared via the emulsion technique. After the jamming process, which means the extraction of the microgel solution by vacuum, the resulting so-called granular ink could be successfully printed via extrusion-based printing. The widely known challenge of printing living cells was also successfully managed. Cells were encapsulated in the microgels during microgel synthesis. Here, the stirring velocity had to be adjusted to avoid harming the cells during the manufacturing process. The cell-loaded microgels were successfully printed in the same way as the empty microgels in multiple layers resulting in dimensionally stable constructs. Live/dead experiments verified that many viable cells were printable after 24 hours. In the next part of this thesis, microgels were prepared from AGE-functionalized gelatin and thiol-functionalized PEG by the same procedure. Again, cells were incorporated and printed by extrusion-based printing. After the addition of hydroxypropyl-methylcellulose, the right conditions for viable cells and stable constructs were found. The printed constructs were further secondarily crosslinked by immersion in initiator solution after the printing process followed by re-irradiating with light. Hereafter, a strongly increased stability of the constructs could be observed. Microgels for use as cell sensor particles were produced as part of this thesis. Here, microfluidic was applied to prepare microgels with a monodisperse size distribution. After adjusting the oil phase, as well as optimizing the manufacturing parameters to the mucin hydrogel system, the microfluidic setup established by Ilona Paulus in this research group could be used. By setting very fast flow rates, microgels in the size range of cells could be obtained. Furthermore, various parameters affecting the stiffness of the particles were varied. This laid the foundation for follow-up studies within the framework of the SFB TRR225 to be able to produce cellmimicking particles. Further follow-up experiments could include the investigation of hydrogels being based only on mucin, like a crosslinking of thiolated mucin and mucin modified with an allyl function such as the PGM-AGE. Furthermore, the granular mucin ink could serve as a supporting material for other microgels or less stable inks during the printing process and thus expand the field of applicable materials for three dimensional (3D) printing.
Höhergradige Gliome gehören zu den häufigsten malignen Hirntumoren bei
Erwachsenen und gehen mit einer sehr schlechten Prognose einher. Die Patientinnen
und Patienten leiden häufig unter kognitiven Einschränkungen, welche auch auf einen
Integritätsverlust der Weißen Substanz zurückzuführen sind und die Lebensqualität der
Betroffenen stark beeinträchtigen. Um in Zukunft eine Behandlung zu gewährleisten, die
nicht nur das Überleben verlängert, sondern auch den Erhalt der neurokognitiven
Funktionen verbessert, sind zuverlässige Methoden zur Messung von Veränderungen
der neurokognitiven Fähigkeiten in einem frühen Stadium erforderlich. Der direkteste
Weg zur Objektivierung neurokognitiver Eigenschaften sind neuropsychologische Tests.
Wir betrachten das Corpus callosum als eine zuverlässige Struktur zur Identifizierung
der Verschlechterung der Integrität der weißen Substanz. Wir stellten die Hypothese auf,
dass ein Zusammenhang zwischen einer beeinträchtigten strukturellen Integrität in
bestimmten Regionen des Corpus Callosum und neurokognitiven Defiziten bei
Patientinnen und Patienten mit höhergradigem Gliom besteht.
Wir schlossen 25 Patientinnen und Patienten mit höhergradigem Gliom in unsere Studie
ein, die sich präoperativ einer neuropsychologischen Testbatterie und einer MRT mit DTI
Sequenzen unterzogen. Die MRT-Daten wurden mit der Software fsl, Oxford,
verarbeitet. Neuropsychologische Parameter wurden mit der FA in drei Teilen des
Corpus Callosum korreliert: Rostrum bzw. Genu, Truncus und Splenium.
Präoperativ korrelierten die meisten neuropsychologischen Parameter signifikant mit der
FA von mindestens einem Bereich des Corpus Callosum. Höhere FA-Werte wurden mit
besserer Konzentration, Gedächtnis, Schnelligkeit und flüssigem Sprechen in
Verbindung gebracht. Verschiedene Tests untersuchten den gleichen
neuropsychologischen Parameter und korrelierten dann mit der gleichen Region des
Corpus Callosum. So konnten das lexikalische und visuelle Gedächtnis mit dem Genu
und Rostrum in Verbindung gebracht werden, exekutive Funktionen und das
Arbeitsgedächtnis korrelierten mit dem Truncus und die Verarbeitungsgeschwindigkeit
mit dem Splenium. Darüber hinaus stimmte diese Zuordnung mit den Ergebnissen
vorangegangener Studien überein. Wir betrachten Veränderungen der
mikrostrukturellen Integrität der Corpus Callosum als robustes morphologisches Korrelat
für die Untersuchung des neurokognitiven Zustands von Patientinnen und Patienten mit
höhergradigem Gliom.
Cardio- and cerebrovascular diseases (CVDs), such as myocardial infarction and ischemic stroke, are the leading cause of death worldwide, caused by overshooting platelet activation and subsequent thrombus formation. However, at sites of vascular injury this tightly-regulated, multi¬step process is critical to limit blood loss and to prevent bleeding. Anti-platelet agents, such as aspirin or clopidogrel, have been proven to be beneficial in prevention of CVDs, but are associated with an elevated bleeding risk and therefore are often contraindicative.
In recent years, the (hem)ITAM-bearing receptors GPVI and CLEC-2 have been identified as critical regulators of platelet activation and thrombus formation, rendering them promising targets for novel anti-platelet drugs. Yet, they are also involved in a plethora of (patho)physiological processes. Consequently, interference with the (hem)ITAM signaling cascade may lead to severe side-effects. In this context, GPV has previously been identified as a mediator of thrombotic and hemostatic function, while its mode of action remains elusive. Therefore, this thesis focused on the function of GPV in thrombotic and hemostatic processes.
Extensive characterization of GPV-deficient mice as well as generation and analysis of anti-GPV antibodies and mice with a mutation rendering GPV uncleavable by thrombin (Gp5Kin/Kin) revealed an unexpected role of GPV as a central modulator of platelet activation and thrombus formation. Gp5-/- as well as Gp5Kin/Kin mice restored the thrombotic and hemostatic defect in the absence of both (hem)ITAM receptors. The in-house generated monoclonal anti-GPV antibodies 89F12 and 5G2 were found to reproduce the knockout phenotype and extended the thrombus-modulatory role of GPV beyond (hem)ITAM receptors, pointing to a critical role of thrombin-cleaved soluble GPV (sGPV). Surprisingly, recombinant sGPV had a strong antithrombotic effect in in vivo throm¬bosis models as well as in in vitro flow adhesion assays using human or murine blood, without affecting hemostasis. These data establish GPV as a key player in platelet physiology. Although data gained from studies using genetically modified mice cannot always directly be transferred to humans, the findings presented in this thesis may serve as basis for the generation of novel treatment options for bleeding complications (anti-GPV antibodies) and thrombotic diseases (sGPV) with a good safety profile. The newly generated humanized GPV mouse provides a valuable tool to study human GPV in vivo.
A second part of this thesis focused on the analysis of protein kinase C (PKC) ι/λ. PKC family of serine/threonine kinases is involved in several physiological processes regulating platelet activation. However, little is known about atypical PKC isoforms and particularly PKCι/λ has never been studied before in platelets. Therefore, platelet- and megakaryocyte-specific PKCι/λ knockout mice were used to assess its role in platelet function in vitro and in vivo. Surprisingly, PKCι/λ was found to be dispensable for platelet function in thrombosis and hemostasis.
Background
A significant number of oncological patients are heavily burdened by psychosocial stress. Doctors recommending or referring their patients to psycho-oncologists in the course of routine consultations can positively influence psycho-oncological care. The aim of this study was to analyze the frequency and predictors of such recommendations and to examine the use of these services by patients.
Methods
4,020 cancer patients (mean age 58 years; 51% women) were evaluated in a multicenter, cross-sectional study in Germany. Data was gathered about doctors’ referral practices, patients’ utilization of psycho-oncological care services, and disease-related symptoms. The PHQ-9 depression scale and the GAD-7 anxiety scale were used to measure psychological burden. Descriptive data analysis was conducted on the basis of subgroup comparisons and multivariable analysis was done using binary logistical regression.
Results
21.9% of the respondents reported having been given a recommendation or referral for psycho-oncological care by a doctor within the course of their cancer diagnosis and treatment. This comprises 29.5% of the patients identified by screening as being psychologically burdened. Nearly half of the patients who received a recommendation or referral (49.8%) acted on it. Predictors for seeking out psycho-oncological care included: patient desire (OR = 2.0), previous experience with psycho-oncological care (OR = 1.59), and female gender (OR = 1.57). Multivariable analysis indicated that patients’ level of psychological burden (depression, anxiety) had no effect on whether doctors gave them a recommendation or referral.
Conclusions
Along with examining the degree to which patients are burdened (e.g. using screening instruments), determining whether or not patients would like to receive psycho-oncological care is an important aspect of improving referral practices and, by extension, will allow important progress in the field of psycho-oncological care to be made.
Hematopoietic stem cell transplantation (HSCT) has been an effective method for treating a wide range of malignant or non-malignant disorders. In case of an autologous HSCT, patients receive their own stem cells after myeloablation before extraction. Allogeneic HSCT uses stem cells derived from a donor. Despite being associated with a high risk of early and long-term complications, it is often the last curative option. 229 pediatric patients, who between 1 January 2005 and 31 December 2015 received an HSCT at the University Children’s Hospital Wuerzburg, were studied. Correlations between two groups were calculated with the Chi square test or with a 2x2-contingency table. To calculate metric variables, the Mann-Whitney-U-test was used. Survival curves were calculated according to Kaplan and Meier. Significance was assumed for results with a p-value <0.05 (CI (Confident Interval) 95%). We retrospectively analyzed 229 pediatric patients (105 females, 124 males) for early and late complications of allogeneic and autologous hematopoietic stem cell transplantation. Median age at HSCT was seven years. Underlying diseases were leukemia (n = 73), lymphoma (n = 22), solid tumor (n = 65), CNS (central nervous system)- tumor (n = 41), and “other diseases” (n = 28). Survival times, overall survival, and event-free survival were calculated. Of all patients, 80.8% experienced complications of some degree, including mild and transient complications. Allo-HSCT (allogeneic HSCT) carried a significantly higher risk of complications than auto-HSCT (autologous HSCT) (n = 118 vs. n = 67; p = < .001) and the remission rate after allo-HSCT was also higher (58.7% vs. 44,7%; p = .032). Especially infection rates and pulmonary complications are different between auto- and allo-HSCT. Leukemia patients had the highest risk of early and late complications (95,0%; p < .001). Complications within HSCT are major risk factors following morbidity and mortality. In order to detect complications and risk factors early, strict recordings are needed to reduce the rate of complication by recognition and prevention of triggering factors. In the future, these factors should receive greater attention in the planning of HSCT post-transplantation care in order to improve the results of the transplantation and establish protocols to prevent their occurrence.
Objective
Recent preliminary studies indicated a seasonal association of BMI at admission to inpatient treatment for anorexia nervosa (AN), indicating lower BMI in the cold season for restrictive AN. An impaired thermoregulation was proposed as the causal factor, based on findings in animal models of AN. However, findings regarding seasonality of BMI and physical activity levels in the general population indicate lower BMI and higher physical activity in summer than in winter. Therefore, we aimed to thoroughly replicate the findings regarding seasonality of BMI at admission in patients with AN in this study.
Method
AN subtype, age- and gender-standardized BMI scores (BMI-SDS) at admission, mean daily sunshine duration and ambient temperature at the residency of 304 adolescent inpatients with AN of the multi-center German AN registry were analyzed.
Results
A main effect of DSM-5 AN subtype was found (F(2,298) = 6.630, p = .002), indicating differences in BMI-SDS at admission between restrictive, binge/purge and subclinical AN. No main effect of season on BMI-SDS at admission was found (F(1,298) = 4.723, p = .025), but an interaction effect of DSM-5 subtype and season was obtained (F(2,298) = 6.625, p = .001). Post-hoc group analyses revealed a lower BMI-SDS in the warm season for restrictive AN with a non-significant small effect size (t(203.16) = 2.140, p = .033; Hedges′g = 0.28). Small correlations of mean ambient temperature (r = −.16) and daily sunshine duration (r = −.22) with BMI-SDS in restrictive AN were found. However, the data were widely scattered.
Conclusions
Our findings are contrary to previous studies and question the thermoregulatory hypothesis, indicating that seasonality in AN is more complex and might be subject to other biological or psychological factors, for example physical activity or body dissatisfaction. Our results indicate only a small clinical relevance of seasonal associations of BMI-SDS merely at admission. Longitudinal studies investigating within-subject seasonal changes might be more promising to assess seasonality in AN and of higher clinical relevance.
It was previously shown that the estrogen-receptor negative breast cancer cell line MBA-MD-231 expresses high levels of A2B adenosine receptors as the sole adenosine receptor subtype. These receptors couple to both, stimulation of adenylyl cyclase and a Ca2+ signal. In order to establish a potential role of A2B adenosine receptors in tumor growth and development MAPK signaling was investigated in these breast cancer cells. Although it is known that A2B adenosine receptors may stimulate MAPK it was found that in MBA-MD-231 cells ERK1/2 phosphorylation is reduced upon agonist-stimulation of A2B adenosine receptors. This reduction is also triggered by forskolin, but abolished by the PKA inhibitor H89, suggesting an important role for the cAMP-PKA pathway. Likewise, a role for intracellular Ca2+ was established as the Ca2+ chelator 1,2-bis-(o-aminophenoxy)-ethane-N,N,N’,N’-tetraacetic acid, tetraacetoxymethyl ester (BAPTA-AM) abolished the reduction of ERK1/2 phosphorylation triggered by A2B stimulation. It was shown that various pathways downstream from A2B adenosine receptors resulted in a stimulation of MAPK phosphatase-1 (MKP-1) which dephosphorylates phospho ERK1/2, and thus plays a critical role in the regulation of the phosphorylation state of ERK1/2. The reduction of ERK1/2 phosphorylation mediated by A2B adenosine receptors might provide an interesting approach for adjuvant treatment leading to reduced growth of certain tumors expressing the A2B subtype.
We assessed the prevalence, awareness, treatment and control of hypertension in patients with moderate chronic kidney disease (CKD) under nephrological care in Germany. In the German Chronic Kidney Disease (GCKD) study, 5217 patients under nephrology specialist care were enrolled from 2010 to 2012 in a prospective observational cohort study. Inclusion criteria were an estimated glomerular filtration rate (eGFR) of 30–60 mL/min/1.73 m2 or overt proteinuria in the presence of an eGFR>60 mL/min/1.73 m2. Office blood pressure was measured by trained study personnel in a standardized way and hypertension awareness and medication were assessed during standardized interviews. Blood pressure was considered as controlled if systolic < 140 and diastolic < 90 mmHg. In 5183 patients in whom measurements were available, mean blood pressure was 139.5 ± 20.4 / 79.3 ± 11.8 mmHg; 4985 (96.2%) of the patients were hypertensive. Awareness and treatment rates were > 90%. However, only 2456 (49.3%) of the hypertensive patients had controlled blood pressure. About half (51.0%) of the patients with uncontrolled blood pressure met criteria for resistant hypertension. Factors associated with better odds for controlled blood pressure in multivariate analyses included younger age, female sex, higher income, low or absent proteinuria, and use of certain classes of antihypertensive medication. We conclude that blood pressure control of CKD patients remains challenging even in the setting of nephrology specialist care, despite high rates of awareness and medication use.
Exploratory behavior of re-orienting foragers differs from other flight patterns of honeybees
(2018)
Honeybees, Apis mellifera, perform re-orientation flights to learn about the new surroundings of the hive when their hive is transported to a new location. Since the pattern of re-orientation flights has not yet been studied, we asked whether this form of exploratory behavior differs from the well described exploratory orientation flights performed by young honeybees before they start foraging. We also investigated whether the exploratory components of re-orientation flights differ from foraging flights and if so how. We recorded re-orientation flights using harmonic radar technology and compared the patterns and flight parameters of these flights with the first exploratory orientation flights of young honeybees and foraging flights of experienced foragers. Just as exploratory orientation flights of young honeybees, re-orientation flights can be classified into short- and long-range flights, and most short-range re-orientation flights were performed under unfavorable weather conditions. This indicates that bees adapt the flight pattern of their re-orientation and orientation flights to changing weather conditions in a similar way. Unlike exploratory orientation flights, more than one sector of the landscape was explored during a long-range re-orientation flight, and significantly longer flight durations and flight distances were observed. Thus, re-orienting bees explored a larger terrain than bees performing their first exploratory orientation flight. By displacing some bees after their first re-orientation flight, we could demonstrate that a single re-orientation flight seems to be sufficient to learn the new location of the hive. The flight patterns of re-orientation flights differed clearly from those of foraging flights. Thus, re-orientation flights represent a special exploratory behavior that is triggered by a change in the location of the hive.
Effect of progesterone on Smad signaling and TGF-β/Smad-regulated genes in lung epithelial cells
(2018)
The effect of endogenous progesterone and/or exogenous pre- or postnatal progesterone application on lung function of preterm infants is poorly defined. While prenatal progesterone substitution may prevent preterm birth, in vitro and in vivo data suggest a benefit of postnatal progesterone replacement on the incidence and severity of bronchopulmonary dysplasia (BPD). However, the molecular mechanisms responsible for progesterone’s effects are undefined. Numerous factors are involved in lung development, airway inflammation, and airway remodeling: the transforming growth factor beta (TGF-β)/mothers against decapentaplegic homolog (Smad) signaling pathway and TGF-β-regulated genes, such as connective tissue growth factor (CTGF), transgelin (TAGLN), and plasminogen activator inhibitor-1 (PAI-1). These processes contribute to the development of BPD. The aim of the present study was to clarify whether progesterone could affect TGF-β1-activated Smad signaling and CTGF/transgelin/PAI-1 expression in lung epithelial cells. The pharmacological effect of progesterone on Smad signaling was investigated using a TGF-β1-inducible luciferase reporter and western blotting analysis of phosphorylated Smad2/3 in A549 lung epithelial cells. The regulation of CTGF, transgelin, and PAI-1 expression by progesterone was studied using a promoter-based luciferase reporter, quantitative real-time PCR, and western blotting in the same cell line. While progesterone alone had no direct effect on Smad signaling in lung epithelial cells, it dose-dependently inhibited TGF-β1-induced Smad3 phosphorylation, as shown by luciferase assays and western blotting analysis. Progesterone also antagonized the TGF-β1/Smad-induced upregulation of CTGF, transgelin, and PAI-1 at the promoter, mRNA, and/or protein levels. The present study highlights possible new molecular mechanisms involving progesterone, including inhibition of TGF-β1-activated Smad signaling and TGF-β1-regulated genes involved in BPD pathogenesis, which are likely to attenuate the development of BPD by inhibiting TGF-β1-mediated airway remodeling. Understanding these mechanisms might help to explain the effects of pre- or postnatal application of progesterone on lung diseases of preterm infants.
The role of the subthalamic nucleus in human locomotion is unclear although relevant, given the troublesome management of gait disturbances with subthalamic deep brain stimulation in patients with Parkinson’s disease. We investigated the subthalamic activity and inter-hemispheric connectivity during walking in eight freely-moving subjects with Parkinson’s disease and bilateral deep brain stimulation. In particular, we compared the subthalamic power spectral densities and coherence, amplitude cross-correlation and phase locking value between resting state, upright standing, and steady forward walking. We observed a phase locking value drop in the β-frequency band (≈13-35Hz) during walking with respect to resting and standing. This modulation was not accompanied by specific changes in subthalamic power spectral densities, which was not related to gait phases or to striatal dopamine loss measured with [123I]N-ω-fluoropropyl-2β-carbomethoxy-3β-(4-iodophenyl)nortropane and single-photon computed tomography. We speculate that the subthalamic inter-hemispheric desynchronization in the β-frequency band reflects the information processing of each body side separately, which may support linear walking. This study also suggests that in some cases (i.e. gait) the brain signal, which could allow feedback-controlled stimulation, might derive from network activity.
Information on circulating miRNAs in frontotemporal lobar degeneration is very limited and conflicting results have complicated an interpretation in Alzheimer’s disease thus far. In the present study we I) collected samples from multiple clinical centers across Germany, II) defined 3 homogenous patient groups with high sample sizes (bvFTD n = 48, AD n = 48 and cognitively healthy controls n = 44), III) compared expression levels in both CSF and serum samples and IV) detected a limited set of miRNAs by using a MIQE compliant protocol based on SYBR-green miRCURY assays that have proven reliable to generate reproducible results. We included several quality controls that identified and reduced technical variation to increase the reliability of our data. We showed that the expression levels of circulating miRNAs measured in CSF did not correlate with levels in serum. Using cluster analysis we found expression pattern in serum that, in part, reflects the genomic organization and affiliation to a specific miRNA family and that were specifically altered in bvFTD, AD, and control groups. Applying factor analysis we identified a 3-factor model characterized by a miRNA signature that explained 80% of the variance classifying healthy controls with 97%, bvFTD with 77% and AD with 72% accuracy. MANOVA confirmed signals like miR-320a and miR-26b-5p at BH corrected significance that contributed most to discriminate bvFTD cases with 96% sensitivity and 90% specificity and AD cases with 89% sensitivity and specificity compared to healthy controls, respectively. Correlation analysis revealed that miRNAs from the 3-factor model also correlated with levels of protein biomarker amyloid-beta1-42 and phosphorylated neurofilament heavy chain, indicating their potential role in the monitoring of progressive neuronal degeneration. Our data show that miRNAs can be reproducibly measured in serum and CSF without pre-amplification and that serum includes higher expressed signals that demonstrate an overall better ability to classify bvFTD, AD and healthy controls compared to signals detected in CSF.
Living beings evolved in an environment with cyclic changing conditions where a variety of factors such as light, temperature, or food availability oscillate in a daily 24-h rhythm. Endogenous circadian clocks in addition to controlling daily rhythms, are also thought to serve as an internal reference for measuring day length. This allows animals to adapt to seasonal changes through photoperiodic responses. While these responses are well-documented in insects, the underlying timing mechanisms for day-length discrimination remain incompletely understood. This thesis aimed at the characterization of the circadian clock of a strongly photoperiodic insect, the pea aphid Acyrthosiphon pisum, that allowed us to find putative neuronal connection between the circadian clock and the photoperiodic system of this insect. In the first chapter, we characterized the neuronal organization of aphid clock clusters using antibodies against the clock proteins Period and Cryptochrome. These clusters were found in the dorsal and lateral protocerebrum, and in the lamina and exhibited daily oscillations. Notably, the clusters expressing Cryptochrome showed light-dependent oscillations, indicating their potential role as clock photoreceptors. These Cryptochrome-positive clusters projected towards the pars intercerebralis, a region crucial for photoperiodism in aphids. In the second chapter, we focused on the Pigment-dispersing factor (PDF), the most important clock neuropeptide in insects. We discovered significant changes in the, otherwise highly conserved, insect C-terminal amino acid sequence of the newly identified pdf gene. PDF was identified in the lateral clock neurons, and their terminals in the dorsal protocerebrum close to the insulin-producing cells located in the pars intercerebralis. These terminals showed daily and seasonal variations, suggesting PDF’s involvement in regulating neurohormone release. To further explore the neuroanatomy of the aphid circadian clock and identify clock-related neuropeptides, we conducted transcriptomic analysis, mass spectrometry, and fluorescent immunohistochemistry. We found that the lateral clock neurons expressed various neuropeptides (in particular Allatotropin, FMRFamide, Orcokinin-A and PDF), similar to those in cockroaches involved in light input pathways. The dorsal clock neurons also exhibit neuropeptide immunoreactivity (precisely of Allatostatin A, Diuretic Hormone31, FMRFamide and Myoinhibitory Peptide), supporting their involvement in modulating circadian and seasonal neurohormonal rhythms. Finally, in the fourth chapter, we provide an overview of the putative mechanisms of photoperiodic control in aphids, from the photoreceptors involved in this process to the circadian clock and the neuroendocrine system.
Patients diagnosed with the rare autoimmune disease of Stiff Person Syndrome (SPS) suffer from varying motor symptoms mainly characterized by painful spasms and muscle stiffness. Among patients suffering from Stiff Person spectrum, clinical presentation, course of disease and treatment responses also differ. Regardless of disease severity, which ranges from mild and intermittent motor impairments to the most severe form progressive encephalomyelitis with rigidity and myoclonus (PERM), autoantibodies are the underlying cause. One of the autoantibody targets associated with SPS is the glycine receptor (GlyR). Functional impairment of this protein interferes with inhibitory signal transmission in the central nervous system and subsequently causes motor symptoms. Similar to functional alterations of the GlyR upon autoantibody binding, GlyR function can be altered in patients with mutations in genes encoding GlyR subunits. Such mutations underlie hereditary hyperekplexia. Understanding the GlyR physiology and how different molecular mechanisms contribute to disease pathology is crucial for development of more targeted and effective disease options.
Therefore, novel GlyR β subunit mutations identified in hyperekplexia patients were investigated towards their expression, trafficking and receptor function. The findings suggest that impaired recruitment into functional receptors at the synapses might underlie the functional alterations revealed by electrophysiological recordings for most cases.
To unravel the autoantibody-related pathology causing the highly diverse clinical appearance of the Stiff Person spectrum, antibody binding abilities were studied. Neutralization assays confirmed that presence of the entire target protein, a sub-domain or a short peptide eliminates the autoantibodies from patient samples. Epitope characterization using residue exchanges within the GlyR in cell-based assays uncovered that GlyR autoantibody epitopes are polyclonal and their combination is patient-specific. Tissue-based binding assays emphasized the high variability in autoantibody distribution within spinal cord and brain sections regardless of the patients’ primary diagnosis. The irregular binding patterns among the patient groups of SPS, PERM, epilepsy and ‘others’ reflected the variation in the symptomatic arrangement. Passive transfer of GlyR autoantibodies from patients with different courses and severity of disease similarly revealed variable effects on murine motor and anxiety-related behavior. The detected small effects on motor function and post-mortem analyses indicate glycinergic disorganization and a possible onset of compensatory mechanisms.
Altogether, this study demonstrates that GlyR impairment is patient-specific and of greater variability than expected.
Within this PhD thesis, starting from simple alkene precursors a series of novel boron-doped PAHs were successfully in a sequential one-pot synthetic approach, comprising a hydroboration/borylation cascade as the key step. By applying different postsynthetic reactions, the properties of these boron-doped PAHs were further adjusted, aiming for appealing packing motifs, strong electron-acceptors, and NIR-emitters. The thesis thereby focussed on the synthesis of tailor-made molecules, the investigation of their optical and electronic properties and the discussion on the influence of various factors, e.g. doping pattern, size, shape, and substituents, on these properties.
Fear and anxiety are fundamental emotional states that are critical for survival. These states are characterized by a variety of coordinated responses, including behavioral and autonomic changes, that need to be properly integrated. For the past decades, most studies have separated the behavioral and autonomic elements, generating a gap in understanding their integrative nature. In this thesis, a framework analysis is presented that allows for the integration of cardiac, behavioral, and neuronal readouts in freely moving mice during different emotional states. Furthermore, a growing body of evidence demonstrates that a vital component of these states is the physiological report of bodily states, or interoception, which allows for quick adaptation to changing situations. A set of distinctive interoceptive pathways has been described from the periphery to the brainstem; however, the circuits that process and integrate cardiac interoceptive signals in higher orders are poorly understood. The midbrain periaqueductal gray (PAG) is a region crucially involved in defensive states through its modulation of both, cardiac and behavioral components. Preliminary studies demonstrate an anatomical connection between the major cardiac interoception brainstem area, the nucleus of the solitary tract, and the PAG; however, the functional characterization and the specific neuronal substrates responsible for interoception in this area have not been described. An interesting particularity of the PAG is that the ventro-lateral subcolumn is the highest order of the neuraxis where inhibitory neurons that express the glycine can be found. In the lower brainstem and spinal cord, glycinergic inhibitory neurons have demonstrated a role in processing sensory and autonomic signals from the periphery, raising the question of whether the PAG glycinergic neurons could be involved in integrating cardiac interoceptive signals as part of a defensive state. In this thesis, using virally mediated trans-synaptic retrograde tracing, I showed that glycinergic PAG neurons receive inputs from cardiac regulatory areas in the brainstem and project massively to forebrain and midbrain regions. By employing advanced techniques such as deep brain calcium imaging with a miniaturized microscope and optogenetics, this study provides compelling evidence for the involvement of glycinergic PAG neurons in controlling heart rate and maintaining cardiac macrostate dynamics within physiological levels. The results of the optogenetic manipulation further revealed that a change in the heart rate macrostate caused by the glycinergic PAG neurons leads to anxiety-like behaviors, providing further evidence for the role of these neurons in regulating defensive states. Overall, by unraveling the neural circuitry underlying interoception in the PAG, our study paves the way to better understand fear and anxiety disorders.
This thesis focusses on the synthesis of functional chiral molecules using carbo- or hetero[7]helicenes as a chiral element, combined with multiple helicenes, phthalocyanines, and 1,4-azaborine units. The objective is to achieve properties that surpass those of the parent compounds.
In the first project, an enantiopure, propeller-shaped multi-helicene polycyclic aromatic hydrocarbon containing three (P)-[7]helicene units and three (M)-[5]helicene units was stereospecifically synthesized and can be obtained in gram quantities. Leveraging the configurational stability of [7]helicene and the configurational instability of [5]helicene, we exclusively obtained the most thermodynamically stable enantiomer out of 10 possible enantiomeric pairs. The effects of the multi-helicene structure on optical rotation, UVVis absorption, fluorescence, and electronic circular dichroism (CD) spectroscopy were investigated.1
Building on the success of the first project, the second project used the configurationally stable [7]helicene again. Zinc-[7]helicenocyanine (Zn-7HPc) was stereospecifically synthesized by directly conjugating [7]helicenes with a phthalocyanine (Pc) core. Zn-7HPc demonstrates a CD signal in the near-infrared region, indicating efficient chirality transfer from the helicenes to the Pc core. Zn-7HPc forms stable, discrete homochiral dimers over a wide range of concentrations in tetrahydrofuran and dimethyl sulfoxide, as well as in the solid state. These homochiral dimers are formed even within the racemic mixture due to the interlocking of two homochiral monomers. The large comproportionation constant and the observed intervalence charge transfer band that appeared in spectroelectrochemistry experiments indicate strong communication between the two Pc monomers in the dimer.2
In the third project, aza[7]helicenes were incorporated with a 1,4-azaborine unit, which exhibits a multiple-resonance effect, to achieve narrow-band emission, high fluorescence quantum yield (FL), and a small Stokes shift. These properties are essential for ultrahigh-definition organic light-emitting diodes that emit circularly polarized light (CP-OLEDs). The synthesized series of molecules demonstrate small Stokes shifts (0.06–0.07 eV), exceptionally narrow fluorescence and circularly polarized luminescence bands with small full width at half maximum (FWHM, 17–28 nm, 0.07–0.13 eV), and high FL (72–85%).3
In conclusion, the synthesis of functional chiral molecules based on carbo- or hetero[7]helicenes was successfully achieved. The efficient synthetic strategies and improved properties of these molecules provide valuable insights for further investigations into helicenes with advanced structures and enhanced properties.
Um Schüler:innen mit komplexen Kommunikationsbedürfnissen und sonderpädagogischem Unterstützungsbedarf im Schwerpunkt Geistige Entwicklung in ihrer Kommunikationsentwicklung unterstützen zu können, müssen zunächst ihre kommunikativen Kompetenzen eingeschätzt werden. Diese Kompetenzen können jedoch je nach Kommunikationspartner:in und Kontext erheblich variieren. Die Umweltabhängigkeit kommunikativer Kompetenzen sowie methodische Herausforderungen bei der Diagnostik kommunikativer Kompetenzen führen zu der Frage, wie Eltern, Lehrkräfte und andere Kommunikationspartner:innen die kommunikativen Kompetenzen dieser Schüler:innen einschätzen, welche Gemeinsamkeiten und Unterschiede zwischen den Einschätzungen bestehen und wie diese erklärt werden können.
Mittels empirischer Daten eines mehrperspektivisch angelegten Fragebogens (N = 357) im Kontext des Forschungsprojektes SFGE II (Baumann et al., 2021) konnten signifikante Unterschiede zwischen der Einschätzung der Eltern und der Lehrkräfte bei vier der acht untersuchten Items zur Einschätzung der kommunikativen Kompetenzen nachgewiesen werden. Die unjustierte Interraterreliabilitätsanalyse konnte einen Einfluss der Familiensprache, der Diagnose sowie des Grades der Intelligenzminderung auf die Höhe der Reliabilität zwischen Eltern und Lehrkräften nachweisen. Die deskriptive Analyse von fünf Fallbeispielen aus zwei weiteren bayerischen Schulen mit sonderpädagogischem Schwerpunkt Geistige Entwicklung untersuchte die Einschätzungen weiterer Kommunikationspartner:innen und betonte vor allem die Bedeutung der UK-Expertise der Kommunikationspartner:innen sowie den Einfluss der aktuell genutzten Kommunikationsformen der Schüler:innen.
Mit den Ergebnissen dieser Studie liegt erstmals ein empirischer Beleg für die unterschiedlichen Einschätzungen kommunikativer Kompetenzen zwischen Eltern und Lehrkräften von kaum und nicht lautsprachlich kommunizierenden Schüler:innen im sonderpädagogischen Schwerpunkt Geistige Entwicklung vor. Die umfassenden Analysen ermöglichen differenzierte Einblicke in das Einschätzungsverhalten verschiedener Kommunikationspartner:innen, liefern Hinweise zur Erklärung übereinstimmender sowie unterschiedlicher Einschätzungen und verweisen auf die Bedeutung von Mehrperspektivität im Kontext von UK-Diagnostik.
Axon growth, a fundamental process of neuron development, is regulated by both intrinsic and external guidance signals. Impairment of axon growth and maintenance is implicated in the pathogenesis of neurodegenerative disorders such as Amyotrophic Lateral Sclerosis and Alzheimer’s disease (AD). Axon growth is driven by several post-transcriptional RNA processing mechanisms, including alternative splicing, polyadenylation, subcellular localization, and translation. These mechanisms are controlled by RNA-binding proteins (RBPs) through interacting with their target RNAs in a sequence-dependent manner. In this study, we investigate the cytosolic functions of two neuronal RBPs, Ptbp2 and hnRNP R, which are essential for axon growth in motoneurons.
Polypyrimidine tract binding protein 2 (Ptbp2) contributes to neuronal differentiation and axonogenesis by modulating different splicing programs to adjust the level of proteins involved in these processes. While the nuclear functions of Ptbp2 in alternative splicing have been studied in more detail, the cytosolic roles of Ptbp2 associated with axon growth have remained elusive. In the first part of the study, we show that Ptbp2 is present in cytosolic fractions of motoneurons including axons and axon terminals. Depletion of Ptbp2 impairs axon growth and growth cone maturation in cultured embryonic mouse motoneurons. Moreover, Ptbp2 knockdown affects the level of piccolo protein in the growth cone of cultured motoneurons. We detect Ptbp2 as a top interactor of the 3' UTR of the Hnrnpr transcript encoding the RBP hnRNP R. This interaction results in axonal localization of and thereby local translation of Hnrnpr mRNA in motoneurons. Consequently, axonal synthesis of hnRNP R was diminished upon depletion of Ptbp2 in motoneurons. We present evidence that Ptbp2 through cooperation with translation factor eIF5A2 controls hnRNP R synthesis. Additionally, we observe that re-expression of hnRNP R in Ptbp2-deficient motoneurons rescued axon growth defect while Ptbp2 overexpression failed to normalize the axon elongation defect observed in hnRNP R-deficient motoneurons. Our findings pinpoint axonal synthesized hnRNP R as a mediator of Ptbp2 functions in axon growth.
In the second part of this study, we identify hnRNP R binds to the 3' UTR of microtubule-associated tau (Mapt) transcript encoding tau protein and regulates the axonal translocation and translation of Mapt mRNA. Tau protein has a central role in neuronal microtubule assembly and stability. However, in AD, the accumulation of abnormally hyperphosphorylated tau protein leads to axon outgrowth defects. Loss of hnRNP R reduces axonal tau protein but not the total level of tau. We observe that the brains of 5xFAD mice, as a mouse model of AD, deficient for hnRNP R contain lower phospho-tau and amyloid-β plaques. Likewise, Neurons treated with blocking antisense oligonucleotides (ASO) to prevent binding of hnRNP R to Mapt mRNA show reduced axonal Mapt mRNA and consequently newly synthesized tau protein levels. We show that blocking Mapt mRNA transport to axons impairs axon elongation. Our data thus suggest that reducing tau levels selectively in axons, a major subcellular site of tangle formation, might represent a novel therapeutic approach for the treatment of AD.
In this work, two techniques, based on the established method of pump--probe spectroscopy were used to investigate the properties of molecular systems in the liquid phase within the visible spectral wavelength range.
The first technique is standard transient absorption (TA) spectroscopy which was applied to a diazo-precursor to identify the formation of a biradical in an inert solvent after UV excitation. With the combination of EPR spectroscopy and quantum chemical calculations, the formation of a biradical in an unpolar and non-protic solvent was proven. Besides, in the presence of air or a polar and protic solvent, the biradical reacts ultrafast to various side products.
The second technique is time-resolved circular dichroism (TRCD) spectroscopy, which was performed in two different ways. The first approach based on a pulse-enantiomer (PE) setup, where an initially circularly polarized pulse was split into two pulses, of which one was mirrored under normal incidence, to flip its polarization. The result was two pulses with mirrored polarization states that propagate collinearly to the sample as left and right circularly polarized probe pulses. The alignment procedure as well as the drawbacks of this setup are described in detail.
However, a new TRCD setup was built that used a polarization grating to get left and right circularly polarized pulses. With the experiences of working with the PE setup, the new TRCD setup could be optimized so that TRCD spectra of a chiral squaraine polymer could be measured. With the help of quantum chemical calculations, the signals were assigned to exciton dynamics that describe spatial and energetic rearrangements of the excitation energy. The alignment and the measurement procedures to perform TRCD spectroscopy with the new setup are described in detail for future experiments.
Parkinson’s disease (PD) is a neurodegenerative disorder characterized by progressive loss of dopaminergic neurons in the substantia nigra of the human brain, leading to depletion of dopamine production. Dopamine replacement therapy remains the mainstay for attenuation of PD symptoms. Nonetheless, the potential benefit of current pharmacotherapies is mostly limited by adverse side effects, such as drug-induced dyskinesia, motor fluctuations and psychosis. Non-dopaminergic receptors, such as human A2A adenosine receptors, have emerged as important therapeutic targets in potentiating therapeutic effects and reducing the unwanted side effects. In this study, new chemical entities targeting both human A2A adenosine receptor and dopamine D2 receptor were designed and evaluated. Two computational methods, namely support vector machine (SVM) models and Tanimoto similarity-based clustering analysis, were integrated for the identification of compounds containing indole-piperazine-pyrimidine (IPP) scaffold. Subsequent synthesis and testing resulted in compounds 5 and 6, which acted as human A2A adenosine receptor binders in the radioligand competition assay (Ki = 8.7–11.2 μM) as well as human dopamine D2 receptor binders in the artificial cell membrane assay (EC50 = 22.5–40.2 μM). Moreover, compound 5 showed improvement in movement and mitigation of the loss of dopaminergic neurons in Drosophila models of PD. Furthermore, in vitro toxicity studies on compounds 5 and 6 did not reveal any mutagenicity (up to 100 μM), hepatotoxicity (up to 30 μM) or cardiotoxicity (up to 30 μM).
Background
Alveolar echinococcosis (AE) is a lethal zoonosis caused by the metacestode larva of the tapeworm Echinococcus multilocularis. The infection is characterized by tumour-like growth of the metacestode within the host liver, leading to extensive fibrosis and organ-failure. The molecular mechanisms of parasite organ tropism towards the liver and influences of liver cytokines and hormones on parasite development are little studied to date.
Methodology/Principal findings
We show that the E. multilocularis larval stage expresses three members of the fibroblast growth factor (FGF) receptor family with homology to human FGF receptors. Using the Xenopus expression system we demonstrate that all three Echinococcus FGF receptors are activated in response to human acidic and basic FGF, which are present in the liver. In all three cases, activation could be prevented by addition of the tyrosine kinase (TK) inhibitor BIBF 1120, which is used to treat human cancer. At physiological concentrations, acidic and basic FGF significantly stimulated the formation of metacestode vesicles from parasite stem cells in vitro and supported metacestode growth. Furthermore, the parasite’s mitogen activated protein kinase signalling system was stimulated upon addition of human FGF. The survival of metacestode vesicles and parasite stem cells were drastically affected in vitro in the presence of BIBF 1120.
Conclusions/Significance
Our data indicate that mammalian FGF, which is present in the liver and upregulated during fibrosis, supports the establishment of the Echinococcus metacestode during AE by acting on an evolutionarily conserved parasite FGF signalling system. These data are valuable for understanding molecular mechanisms of organ tropism and host-parasite interaction in AE. Furthermore, our data indicate that the parasite’s FGF signalling systems are promising targets for the development of novel drugs against AE.
Peptidergic signaling from clock neurons regulates reproductive dormancy in Drosophila melanogaster
(2019)
With the approach of winter, many insects switch to an alternative protective developmental program called diapause. Drosophila melanogaster females overwinter as adults by inducing a reproductive arrest that is characterized by inhibition of ovarian development at previtellogenic stages. The insulin producing cells (IPCs) are key regulators of this process, since they produce and release insulin-like peptides that act as diapause-antagonizing hormones. Here we show that in D. melanogaster two neuropeptides, Pigment Dispersing Factor (PDF) and short Neuropeptide F (sNPF) inhibit reproductive arrest, likely through modulation of the IPCs. In particular, genetic manipulations of the PDF-expressing neurons, which include the sNPF-producing small ventral Lateral Neurons (s-LNvs), modulated the levels of reproductive dormancy, suggesting the involvement of both neuropeptides. We expressed a genetically encoded cAMP sensor in the IPCs and challenged brain explants with synthetic PDF and sNPF. Bath applications of both neuropeptides increased cAMP levels in the IPCs, even more so when they were applied together, suggesting a synergistic effect. Bath application of sNPF additionally increased Ca2+ levels in the IPCs. Our results indicate that PDF and sNPF inhibit reproductive dormancy by maintaining the IPCs in an active state.
Circadian clocks coordinate time-of-day-specific metabolic and physiological processes to maximize organismal performance and fitness. In addition to light and temperature, which are regarded as strong zeitgebers for circadian clock entrainment, metabolic input has now emerged as an important signal for clock entrainment and modulation. Circadian clock proteins have been identified to be substrates of O-GlcNAcylation, a nutrient sensitive post-translational modification (PTM), and the interplay between clock protein O-GlcNAcylation and other PTMs is now recognized as an important mechanism by which metabolic input regulates circadian physiology. To better understand the role of O-GlcNAcylation in modulating clock protein function within the molecular oscillator, we used mass spectrometry proteomics to identify O-GlcNAcylation sites of PERIOD (PER), a repressor of the circadian transcriptome and a critical biochemical timer of the Drosophila clock. In vivo functional characterization of PER O-GlcNAcylation sites indicates that O-GlcNAcylation at PER(S942) reduces interactions between PER and CLOCK (CLK), the key transcriptional activator of clock-controlled genes. Since we observe a correlation between clock-controlled daytime feeding activity and higher level of PER O-GlcNAcylation, we propose that PER(S942) O-GlcNAcylation during the day functions to prevent premature initiation of circadian repression phase. This is consistent with the period-shortening behavioral phenotype of per(S942A) flies. Taken together, our results support that clock-controlled feeding activity provides metabolic signals to reinforce light entrainment to regulate circadian physiology at the post-translational level. The interplay between O-GlcNAcylation and other PTMs to regulate circadian physiology is expected to be complex and extensive, and reach far beyond the molecular oscillator.
The Sterol Regulatory Element Binding Proteins (SREBPs) are basic-helix-loop-helix transcription regulators that control the expression of sterol biosynthesis genes in higher eukaryotes and some fungi. Surprisingly, SREBPs do not regulate sterol biosynthesis in the ascomycete yeasts (Saccharomycotina) as this role was handed off to an unrelated transcription regulator in this clade. The SREBPs, nonetheless, expanded in fungi such as the ascomycete yeasts Candida spp., raising questions about their role and evolution in these organisms. Here we report that the fungal SREBPs diversified their DNA binding preferences concomitantly with an expansion in function. We establish that several branches of fungal SREBPs preferentially bind non-palindromic DNA sequences, in contrast to the palindromic DNA motifs recognized by most basic-helix-loop-helix proteins (including SREBPs) in higher eukaryotes. Reconstruction and biochemical characterization of the likely ancestor protein suggest that an intrinsic DNA binding promiscuity in the family was resolved by alternative mechanisms in different branches of fungal SREBPs. Furthermore, we show that two SREBPs in the human commensal yeast Candida albicans drive a transcriptional cascade that inhibits a morphological switch under anaerobic conditions. Preventing this morphological transition enhances C. albicans colonization of the mammalian intestine, the fungus’ natural niche. Thus, our results illustrate how diversification in DNA binding preferences enabled the functional expansion of a family of eukaryotic transcription regulators.
Understanding the genetic mechanisms underlying segregation of phenotypic variation through successive generations is important for understanding physiological changes and disease risk. Tracing the etiology of variation in gene expression enables identification of genetic interactions, and may uncover molecular mechanisms leading to the phenotypic expression of a trait, especially when utilizing model organisms that have well-defined genetic lineages. There are a plethora of studies that describe relationships between gene expression and genotype, however, the idea that global variations in gene expression are also controlled by genotype remains novel. Despite the identification of loci that control gene expression variation, the global understanding of how genome constitution affects trait variability is unknown. To study this question, we utilized Xiphophorus fish of different, but tractable genetic backgrounds (inbred, F1 interspecies hybrids, and backcross hybrid progeny), and measured each individual’s gene expression concurrent with the degrees of inter-individual expression variation. We found, (a) F1 interspecies hybrids exhibited less variability than inbred animals, indicting gene expression variation is not affected by the fraction of heterozygous loci within an individual genome, and (b), that mixing genotypes in backcross populations led to higher levels of gene expression variability, supporting the idea that expression variability is caused by heterogeneity of genotypes of cis or trans loci. In conclusion, heterogeneity of genotype, introduced by inheritance of different alleles, accounts for the largest effects on global phenotypical variability.
Invasion of epithelial cells by Salmonella enterica requires expression of genes located in the pathogenicity island I (SPI-1). The expression of SPI-1 genes is very tightly regulated and activated only under specific conditions. Most studies have focused on the regulatory pathways that induce SPI-1 expression. Here, we describe a new regulatory circuit involving CRP-cAMP, a widely established metabolic regulator, in silencing of SPI-1 genes under non-permissive conditions. In CRP-cAMP-deficient strains we detected a strong upregulation of SPI-1 genes in the mid-logarithmic growth phase. Genetic analyses revealed that CRP-cAMP modulates the level of HilD, the master regulator of Salmonella invasion. This regulation occurs at the post-transcriptional level and requires the presence of a newly identified regulatory motif within the hilD 3’UTR. We further demonstrate that in Salmonella the Hfq-dependent sRNA Spot 42 is under the transcriptional repression of CRP-cAMP and, when this transcriptional repression is relieved, Spot 42 exerts a positive effect on hilD expression. In vivo and in vitro assays indicate that Spot 42 targets, through its unstructured region III, the 3’UTR of the hilD transcript. Together, our results highlight the biological relevance of the hilD 3’UTR as a hub for post-transcriptional control of Salmonella invasion gene expression.
Emerging pathogens are a major threat to public health, however understanding how pathogens adapt to new niches remains a challenge. New methods are urgently required to provide functional insights into pathogens from the massive genomic data sets now being generated from routine pathogen surveillance for epidemiological purposes. Here, we measure the burden of atypical mutations in protein coding genes across independently evolved Salmonella enterica lineages, and use these as input to train a random forest classifier to identify strains associated with extraintestinal disease. Members of the species fall along a continuum, from pathovars which cause gastrointestinal infection and low mortality, associated with a broad host-range, to those that cause invasive infection and high mortality, associated with a narrowed host range. Our random forest classifier learned to perfectly discriminate long-established gastrointestinal and invasive serovars of Salmonella. Additionally, it was able to discriminate recently emerged Salmonella Enteritidis and Typhimurium lineages associated with invasive disease in immunocompromised populations in sub-Saharan Africa, and within-host adaptation to invasive infection. We dissect the architecture of the model to identify the genes that were most informative of phenotype, revealing a common theme of degradation of metabolic pathways in extraintestinal lineages. This approach accurately identifies patterns of gene degradation and diversifying selection specific to invasive serovars that have been captured by more labour-intensive investigations, but can be readily scaled to larger analyses.
Once biological systems are modeled by regulatory networks, the next step is to include external stimuli, which model the experimental possibilities to affect the activity level of certain network’s nodes, in a mathematical framework. Then, this framework can be interpreted as a mathematical optimal control framework such that optimization algorithms can be used to determine external stimuli which cause a desired switch from an initial state of the network to another final state. These external stimuli are the intervention points for the corresponding biological experiment to obtain the desired outcome of the considered experiment. In this work, the model of regulatory networks is extended to controlled regulatory networks. For this purpose, external stimuli are considered which can affect the activity of the network’s nodes by activation or inhibition. A method is presented how to calculate a selection of external stimuli which causes a switch between two different steady states of a regulatory network. A software solution based on Jimena and Mathworks Matlab is provided. Furthermore, numerical examples are presented to demonstrate application and scope of the software on networks of 4 nodes, 11 nodes and 36 nodes. Moreover, we analyze the aggregation of platelets and the behavior of a basic T-helper cell protein-protein interaction network and its maturation towards Th0, Th1, Th2, Th17 and Treg cells in accordance with experimental data.
Flagellin hypervariable region determinessymbiotic properties of commensalEscherichia coli strains
(2019)
Escherichia coli represents a classical intestinal gram-negative commensal. Despite this commensalism, different E. coli strains can mediate disparate immunogenic properties in a given host. Symbiotic E. coli strains such as E. coli Nissle 1917 (EcN) are attributed beneficial properties, e.g., promotion of intestinal homeostasis. Therefore, we aimed to identify molecular features derived from symbiotic bacteria that might help to develop innovative therapeutic alternatives for the treatment of intestinal immune disorders. This study was performed using the dextran sodium sulphate (DSS)-induced colitis mouse model, which is routinely used to evaluate potential therapeutics for the treatment of Inflammatory Bowel Diseases (IBDs). We focused on the analysis of flagellin structures of different E. coli strains. EcN flagellin was found to harbor a substantially longer hypervariable region (HVR) compared to other commensal E. coli strains, and this longer HVR mediated symbiotic properties through stronger activation of Toll-like receptor (TLR)5, thereby resulting in interleukin (IL)-22–mediated protection of mice against DSS-induced colitis. Furthermore, using bone-marrow–chimeric mice (BMCM), CD11c+ cells of the colonic lamina propria (LP) were identified as the main mediators of these flagellin-induced symbiotic effects. We propose flagellin from symbiotic E. coli strains as a potential therapeutic to restore intestinal immune homeostasis, e.g., for the treatment of IBD patients.
Dmrt1 is a highly conserved transcription factor, which is critically involved in regulation of gonad development of vertebrates. In medaka, a duplicate of dmrt1—acting as master sex-determining gene—has a tightly timely and spatially controlled gonadal expression pattern. In addition to transcriptional regulation, a sequence motif in the 3′ UTR (D3U-box) mediates transcript stability of dmrt1 mRNAs from medaka and other vertebrates. We show here that in medaka, two RNA-binding proteins with antagonizing properties target this D3U-box, promoting either RNA stabilization in germ cells or degradation in the soma. The D3U-box is also conserved in other germ-cell transcripts, making them responsive to the same RNA binding proteins. The evolutionary conservation of the D3U-box motif within dmrt1 genes of metazoans—together with preserved expression patterns of the targeting RNA binding proteins in subsets of germ cells—suggest that this new mechanism for controlling RNA stability is not restricted to fishes but might also apply to other vertebrates.
It is widely accepted for humans and higher animals that vision is an active process in which the organism interprets the stimulus. To find out whether this also holds for lower animals, we designed an ambiguous motion stimulus, which serves as something like a multi-stable perception paradigm in Drosophila behavior. Confronted with a uniform panoramic texture in a closed-loop situation in stationary flight, the flies adjust their yaw torque to stabilize their virtual self-rotation. To make the visual input ambiguous, we added a second texture. Both textures got a rotatory bias to move into opposite directions at a constant relative angular velocity. The results indicate that the fly now had three possible frames of reference for self-rotation: either of the two motion components as well as the integrated motion vector of the two. In this ambiguous stimulus situation, the flies generated a continuous sequence of behaviors, each one adjusted to one or another of the three references.
Blinatumomab is a first-in-class immunotherapy based on the bispecific T-cell engager (BiTE®) immune-oncology platform, which redirects CD3+ T cells to kill CD19+ target cells. The objective of this analysis was to describe the correlation between B- and T-cell kinetics and response to blinatumomab in patients with relapsed or refractory (r/r) non-Hodgkin lymphoma (NHL). The clinical efficacy of treatment with blinatumomab in patients with r/r NHL was recently investigated in a phase 1 dose-escalation and expansion trial (NCT00274742) wherein 76 patients received blinatumomab by continuous intravenous infusion at various doses (0.5–90 μg/m2/day). B-Cell depletion and expansion of CD3+, CD4+, and CD8+ T cells was analyzed in patients stratified per clinical response (complete response [CR], n = 16; partial response [PR], stable disease [SD], or progressive disease [PD], n = 54) for at least 4 weeks (additional 4 weeks after clinical benefit) from the date of administration of blinatumomab until dose-limiting toxicity or PD. B-cell depletion kinetics were faster in patients who had a CR than in patients who did not have a complete response (PR, SD, or PD). T-cell expansion (T-cell counts exceeding the baseline level on day 22) was more pronounced in patients with CR than in patients without CR. T-cell expansion in patients with CR correlated with increased T-cell counts of both CD4+ and CD8+ T cells compared with patients without CR. Patients with r/r NHL who achieved a CR had faster B-cell depletion and increased expansion of CD3+, CD4+, and CD8+ T cells than patients who did not achieve a CR.
In response to cardiac injury, increased activity of the hexosamine biosynthesis pathway (HBP) is linked with cytoprotective as well as adverse effects depending on the type and duration of injury. Glutamine-fructose amidotransferase (GFAT; gene name gfpt) is the rate-limiting enzyme that controls flux through HBP. Two protein isoforms exist in the heart called GFAT1 and GFAT2. There are conflicting data on the relative importance of GFAT1 and GFAT2 during stress-induced HBP responses in the heart.
Using neonatal rat cardiac cell preparations, targeted knockdown of GFPT1 and GFPT2 were performed and HBP activity measured. Immunostaining with specific GFAT1 and GFAT2 antibodies was undertaken in neonatal rat cardiac preparations and murine cardiac tissues to characterise cell-specific expression. Publicly available human heart single cell sequencing data was interrogated to determine cell-type expression. Western blots for GFAT isoform protein expression were performed in human cardiomyocytes derived from induced pluripotent stem cells (iPSCs).
GFPT1 but not GFPT2 knockdown resulted in a loss of stress-induced protein O-GlcNAcylation in neonatal cardiac cell preparations indicating reduced HBP activity. In rodent cells and tissue, immunostaining for GFAT1 identified expression in both cardiac myocytes and fibroblasts whereas immunostaining for GFAT2 was only identified in fibroblasts. Further corroboration of findings in human heart cells identified an enrichment of GFPT2 gene expression in cardiac fibroblasts but not ventricular myocytes whereas GFPT1 was expressed in both myocytes and fibroblasts. In human iPSC-derived cardiomyocytes, only GFAT1 protein was expressed with an absence of GFAT2.
In conclusion, these results indicate that GFAT1 is the primary cardiomyocyte isoform and GFAT2 is only present in cardiac fibroblasts. Cell-specific isoform expression may have differing effects on cell function and should be considered when studying HBP and GFAT functions in the heart.
Proper maternal care is an essential factor of reproductive success in mammals, involving a repertoire of behaviors oriented toward the feeding and care of the offspring. Among the neurotransmitters involved in the initiation of these behaviors, serotonin (5-HT) seems to play an important role. Here we compared pup-oriented maternal behaviors in mice with constitutive 5-HT depletion, the tryptophan hydroxylase 2-knock-out (Tph2-KO) and the Pet1-KO mice. We report that the only common pup-oriented defect in these 2 hyposerotoninergic models is a defective nursing in parturient mice and altered nursing-like (crouching) behavior in virgin mice, while pup retrieval defects are only present in Tph2-KO. Despite a normal mammary gland development and milk production, the defect in appropriate nursing is responsible for severe growth retardation and early lethality of pups born to hyposerotonergic dams. This nursing defect is due to acute rather constitutive 5-HT depletion, as it is reproduced by adult knockdown of Tph2 in the dorsal raphe nucleus in mothers with a prior normal maternal experience. We conclude that 5-HT innervation from the dorsal raphe is required for both the initiation and maintenance of a normal nursing behavior. Our findings may be related to observations of reduced maternal/infant interactions in human depression.
Background
The new subgroup screening tool “subscreen” aims to understand the unclear and complex association between socioeconomic status (SES) and childhood allergy. This software R package has been successfully used in clinical trials but not in large population-based studies.
Objective
To screen and identify subgrouping factors explaining their impact on the association between SES and respiratory allergies in childhood and youth.
Methods
Using the national German childhood and youth survey dataset (KiGGS Wave 2), we included 56 suspected subgrouping factors to investigate the association between SES (low vs. high) and allergic rhinitis and/or asthma in an exploratory manner. The package enabled a comprehensive overview of odds ratios when considering the SES impact per subgroup and analogously all disease proportions per subgroup.
Result
Among the 56 candidate factors, striking subgrouping factors were identified; e.g., if mothers were younger and in the low SES group, their children had a higher risk of asthma. In addition children of the teen’s age were associated with increased risks in the low SES group. For the crude proportions, factors such as (parental) smoking or having had no “contact with farm animals” were identified as strong risk factors for rhinitis.
Significance
The “subscreen” package enabled the detection of notable subgroups for further investigations exemplarily for similar epidemiological research questions.
The development of crop varieties that are resistant to lodging is a top priority for breeding programmes. Herein, we characterize the rye mutant ´Stabilstroh’ (‘stable straw’) possessing an exceptional combination of high lodging resistance, tall posture and high biomass production. Nuclear magnetic resonance imaging displayed the 3-dimensional assembly of vascular bundles in stem. A higher number of vascular bundles and a higher degree of their incline were the features of lodging-resistant versus lodging-prone lines. Histology and electron microscopy revealed that stems are fortified by a higher proportion of sclerenchyma and thickened cell walls, as well as some epidermal invaginations. Biochemical analysis using Fourier-transform infrared spectroscopy and inductively coupled plasma-optical emission spectrometry further identified elevated levels of lignin, xylan, zinc and silicon as features associated with high lodging resistance. Combined effects of above features caused superior culm stability. A simplistic mathematical model showed how mechanical forces distribute within the stem under stress. Main traits of the lodging-resistant parental line were heritable and could be traced back to the genetic structure of the mutant. Evaluation of lodging-resistant wheat ‘Babax’ (‘Baviacora’) versus contrasting, lodging-prone, genotype ´Pastor´ agreed with above findings on rye. Our findings on mechanical stability and extraordinary culm properties may be important for breeders for the improvement of lodging resistance of tall posture cereal crops.
According to the updated International Myeloma Working Group criteria, smoldering multiple myeloma (SMM) is an asymptomatic plasma cell disorder characterized by an M-component >3 g/dL, bone marrow plasma cell infiltration >10% and <60%, and absence of any myeloma-defining event. Active multiple myeloma is preceded by SMM, with a median time to progression of approximately 5 years. Cases of SMM range from the extremes of “monoclonal gammopathy of undetermined significance-like”, in which patients never progress during their lifetimes, to “early multiple myeloma”, in which transformation into symptomatic disease, based on genomic evolution, may be rapid and devastating. Such a “split personality” makes the prognosis and management of individual patients challenging, particularly with regard to the identification and possible early treatment of high-risk SMM. Outside of clinical trials, the conventional approach to SMM generally remains close observation until progression to active multiple myeloma. However, two prospective, randomized trials have recently demonstrated a significant clinical benefit in terms of time to progression, and of overall survival in one of the two studies, for some patients with higher-risk SMM treated with lenalidomide ± dexamethasone, raising the question of whether such an approach should be considered a new standard of care. In this paper, experts from the European Myeloma Network describe current biological and clinical knowledge on SMM, focusing on novel insights into its molecular pathogenesis, new prognostic scoring systems proposed to identify SMM patients at higher risk of early transformation, and updated results of completed or ongoing clinical trials. Finally, some practical recommendations for the real-life management of these patients, based on Delphi consensus methodology, are provided.
The latency of spontaneous eye blinks marks relevant visual and auditory information processing
(2021)
Eye blinks are influenced by external sensory and internal cognitive factors, as mainly shown in the visual domain. In previous studies, these factors corresponded to the time period of task-relevant sensory information and were often linked to a motor response. Our aim was to dissociate the influence of overall sensory input duration, task-relevant information duration, and the motor response to further understand how the temporal modulation of blinks compares among sensory modalities.
Using a visual and an auditory temporal judgment task, we found that blinks were suppressed during stimulus presentation in both domains and that the overall input length had a significant positive relationship with the length of this suppression (i.e., with the latency of the first blink after stimulus onset). Importantly, excluding the influence of the overall sensory input duration we could show that the duration of task-relevant input had an additional influence on blink latency in the visual and the auditory domain. Our findings further suggest that this influence was not based on sensory input but on top–down processes. We could exclude task difficulty and the timing of the motor response as driving factors in the blink modulation.
Our results suggest a sensory domain–independent modulation of blink latencies, introduced by changes in the length of the task-relevant, attended period. Therefore, not only do blinks mark the timing of sensory input or the preparation of the motor output, but they can also act as precise indicators of periods of cognitive processing.
Background
Sudden unexpected infant death (SUID) continues to be a major contributor to infant mortality in the United States. The objective was to analyze time trends in SUID and their association with immunization coverage.
Methods
The number of deaths and live births per year and per state (1992–2015) was obtained from the Centers for Disease Control and Prevention (CDC). We calculated infant mortality rates (i.e., deaths below one year of age) per 1000 live births for SUID. We obtained data on immunization in children aged 19–35 months with three doses or more of diphtheria-tetanus-pertussis (3+ DTP), polio (3+ Polio), and Haemophilus influenzae type b (3+ Hib) as well as four doses or more of DTP (4+ DTP) from the National Immunization Survey, and data on infant sleep position from the Pregnancy Risk Assessment Monitoring System (PRAMS) Study. Data on poverty and race were derived from the Current Population and American Community Surveys of the U.S. Census Bureau. We calculated mean SUID mortality rates with 95% confidence interval (CI) as well as the annual percentage change using breakpoint analysis. We used Poisson regression with random effects to examine the dependence of SUID rates on immunization coverage, adjusting for sleep position and poverty (1996–2015). In a second model, we additionally adjusted for race (2000–2015).
Results
Overall, SUID mortality decreased in the United States. The mean annual percent change was − 9.6 (95% CI = − 10.5, − 8.6) between 1992 and 1996, and − 0.3 (95% CI = − 0.4, − 0.1) from 1996 onwards. The adjusted rate ratios for SUID mortality were 0.91 (95% CI = 0.80, 1.03) per 10% increase for 3+ DTP, 0.88 (95% CI = 0.83, 0.95) for 4+ DTP, 1.00 (95% CI = 0.90, 1.10) for 3+ polio, and 0.95 (95% CI = 0.89, 1.02) for 3+ Hib. After additionally adjusting for race, the rate ratios were 0.76 (95% CI = 0.67, 0.85) for 3+ DTP, 0.83 (95% CI = 0.78, 0.89) for 4+ DTP, 0.81 (95% CI = 0.73, 0.90) for 3+ polio, and 0.94 (95% CI = 0.88, 1.00) for 3+ Hib.
Conclusions
SUID mortality is decreasing, and inversely related to immunization coverage. However, since 1996, the decline has slowed down.
Aims
Despite advances in interventional treatment strategies, atrial fibrillation (AF) remains associated with significant morbidity and mortality. Fibrotic atrial myopathy (FAM) is a main factor for adverse outcomes of AF-ablation, but complex to diagnose using current methods. We aimed to derive a scoring system based entirely on easily available clinical parameters to predict FAM and ablation-success in everyday care.
Methods
In this multicenter, prospective study, a new risk stratification model termed AF-SCORE was derived in 220 patients undergoing high-density left-atrial(LA) voltage-mapping to quantify FAM. AF-SCORE was validated for FAM in an external mapping-validation cohort (n = 220) and for success following pulmonary vein isolation (PVI)-only (without adjunctive left- or right atrial ablations) in an external outcome-validation cohort (n = 518).
Results
FAM was rare in patients < 60 years (5.4%), but increased with ageing and affected 40.4% (59/146) of patients ≥ 60 years. Sex and AF-phenotype had additional predictive value in older patients and remained associated with FAM in multivariate models (odds ratio [OR] 6.194, p < 0.0001 for ≥ 60 years; OR 2.863, p < 0.0001 for female sex; OR 41.309, p < 0.0001 for AF-persistency). Additional clinical or diagnostic variables did not improve the model. AF-SCORE (+ 1 point for age ≥ 60 years and additional points for female sex [+ 1] and AF-persistency [+ 2]) showed good discrimination to detect FAM (c-statistic 0.792) and predicted arrhythmia-freedom following PVI (74.3%, 54.7% and 45.5% for AF-SCORE ≤ 2, 3 and 4, respectively, and hazard ratio [HR] 1.994 for AF-SCORE = 3 and HR 2.866 for AF-SCORE = 4, p < 0.001).
Conclusions
Age, sex and AF-phenotype are the main determinants for the development of FAM. A low AF-SCORE ≤ 2 is found in paroxysmal AF-patients of any age and younger patients with persistent AF irrespective of sex, and associated with favorable outcomes of PVI-only. Freedom from arrhythmia remains unsatisfactory with AF-SCORE ≥ 3 as found in older patients, particularly females, with persistent AF, and future studies investigating adjunctive atrial ablations to PVI-only should focus on these groups of patients.
Context
The adrenal cortex produces specific steroid hormones including steroid sulfates such as dehydroepiandrosterone sulfate (DHEAS), the most abundant steroid hormone in the human circulation. Steroid sulfation involves a multistep enzyme machinery that may be impaired by inborn errors of steroid metabolism. Emerging data suggest a role of steroid sulfates in the pathophysiology of adrenal tumors and as potential biomarkers.
Evidence Acquisition
Selective literature search using “steroid,” “sulfat*,” “adrenal,” “transport,” “mass spectrometry” and related terms in different combinations.
Evidence Synthesis
A recent study highlighted the tissue abundance of estrogen sulfates to be of prognostic impact in adrenocortical carcinoma tissue samples using matrix-assisted laser desorption ionization mass spectrometry imaging. General mechanisms of sulfate uptake, activation, and transfer to substrate steroids are reasonably well understood. Key aspects of this pathway, however, have not been investigated in detail in the adrenal; these include the regulation of substrate specificity and the secretion of sulfated steroids. Both for the adrenal and targeted peripheral tissues, steroid sulfates may have relevant biological actions beyond their cognate nuclear receptors after desulfation. Impaired steroid sulfation such as low DHEAS in Cushing adenomas is of diagnostic utility, but more comprehensive studies are lacking. In bioanalytics, the requirement of deconjugation for gas-chromatography/mass-spectrometry has precluded the study of steroid sulfates for a long time. This limitation may be overcome by liquid chromatography/tandem mass spectrometry.
Conclusions
A role of steroid sulfation in the pathophysiology of adrenal tumors has been suggested and a diagnostic utility of steroid sulfates as biomarkers is likely. Recent analytical developments may target sulfated steroids specifically.
Background:
Currently, health care systems worldwide are challenged with providing care to an increasing number of migrants, refugees, and displaced persons. In this article, we report on disease burden and drug prescription patterns in a large refugee cohort in Germany.
Methods:
We conducted a cross-sectional study of anonymized medical records including demographic data, diagnoses, and drug prescriptions in two refugee reception centres between 2015 and 2019. Refugees and migrants received medical assistance exclusively through the on-site clinics. Thus, this study represents all medical visits of the housed residents.
Results:
In total, n = 15531 diagnoses from n = 4858 patients in a cohort of n = 10431 accommodated refugees were recorded. N = 11898 medications were prescribed. Overall, 29.8% of all refugees sought medical attention. Half of the patients were female (49.6%), the average age was 23.8 years (SD [standard deviation] 17.0, min 0, max 81), and 41.5% were minors (<18 years). Most patients had Middle Eastern or Northern African origin (63.9%). The largest proportion of diagnoses belonged to the ICD (International Statistical Classification of Diseases and Related Health Problems) category "R" (miscellaneous, 33.5%), followed by diseases of the respiratory system (category "J", 16.5%), or the musculoskeletal system (category "M", 7.1%). Non-steroidal anti-inflammatory drugs were most frequently prescribed.
Conclusions:
This analysis in two large refugee centres in Germany shows that about one third of refugees seek medical attention upon initial arrival. Complaints are manifold, with a high prevalence of respiratory infections.
A principal hires an agent to provide a verifiable service. Initially, the agent can exert unobservable effort to reduce his disutility from providing the service. If the agent is free to waive his right to quit, he may voluntarily sign a contract specifying an inefficiently large service level, while there are insufficient incentives to exert effort. If the agent’s right to quit is inalienable, the underprovision of effort may be further aggravated, but the service level is ex post efficient. Overall, it turns out that the total surplus can be larger when agents are not permitted to contractually waive their right to quit work. Yet, we also study an extension of our model in which even the agent can be strictly better off when the parties have the contractual freedom to waive the agent’s right to quit.
Interventions to promote physical activity (PA) in childcare centers have been shown to increase children’s PA levels; moreover, a growing number of evidence-based best practice guidelines exist for this setting. However, there is a lack of knowledge on the facilitators of and barriers to the successful implementation of PA guidelines and interventions. We used Cooperative Planning to improve capabilities for PA in childcare centers. This qualitative study aimed to explore childcare center directors’ views on the Cooperative Planning process and identify the facilitators of and barriers to its implementation. We conducted guided semi-structured interviews with the directors of nine childcare centers after completion of the 12-month Cooperative Planning process. The interviews were recorded, transcribed and analyzed using qualitative content analysis with inductive category development. Facilitators and barriers were systematized according to the Consolidated Framework for Implementation Research (CFIR). Cooperative Planning was regarded as being helpful for structuring the process and involving all team members. Several facilitators within the CFIR domains inner setting (structural characteristics, networks and communications, implementation climate), outer setting (support from parents and provider), characteristics of individuals (intrinsic motivation of the staff) and process (individual drivers) were identified. The reported barriers included structural characteristics (e.g. lack of time), networks and communications (e.g. team conflicts) and characteristics of individuals (e.g. lack of willingness to accept change). Several contextual and interpersonal factors seem to influence the extent to which a Cooperative Planning process can be implemented by a childcare center’s team. Future research is needed to evaluate the strategies needed to overcome the identified barriers.
Activity in the healthy brain relies on a concerted interplay of excitation (E) and inhibition (I) via balanced synaptic communication between glutamatergic and GABAergic neurons. A growing number of studies imply that disruption of this E/I balance is a commonality in many brain disorders; however, obtaining mechanistic insight into these disruptions, with translational value for the patient, has typically been hampered by methodological limitations. Cadherin-13 (CDH13) has been associated with autism and attention-deficit/hyperactivity disorder. CDH13 localizes at inhibitory presynapses, specifically of parvalbumin (PV) and somatostatin (SST) expressing GABAergic neurons. However, the mechanism by which CDH13 regulates the function of inhibitory synapses in human neurons remains unknown. Starting from human-induced pluripotent stem cells, we established a robust method to generate a homogenous population of SST and MEF2C (PV-precursor marker protein) expressing GABAergic neurons (iGABA) in vitro, and co-cultured these with glutamatergic neurons at defined E/I ratios on micro-electrode arrays. We identified functional network parameters that are most reliably affected by GABAergic modulation as such, and through alterations of E/I balance by reduced expression of CDH13 in iGABAs. We found that CDH13 deficiency in iGABAs decreased E/I balance by means of increased inhibition. Moreover, CDH13 interacts with Integrin-β1 and Integrin-β3, which play opposite roles in the regulation of inhibitory synaptic strength via this interaction. Taken together, this model allows for standardized investigation of the E/I balance in a human neuronal background and can be deployed to dissect the cell-type-specific contribution of disease genes to the E/I balance.
Quantitative high-confidence human mitochondrial proteome and its dynamics in cellular context
(2021)
Mitochondria are key organelles for cellular energetics, metabolism, signaling, and quality control and have been linked to various diseases. Different views exist on the composition of the human mitochondrial proteome. We classified >8,000 proteins in mitochondrial preparations of human cells and defined a mitochondrial high-confidence proteome of >1,100 proteins (MitoCoP). We identified interactors of translocases, respiratory chain, and ATP synthase assembly factors. The abundance of MitoCoP proteins covers six orders of magnitude and amounts to 7% of the cellular proteome with the chaperones HSP60-HSP10 being the most abundant mitochondrial proteins. MitoCoP dynamics spans three orders of magnitudes, with half-lives from hours to months, and suggests a rapid regulation of biosynthesis and assembly processes. 460 MitoCoP genes are linked to human diseases with a strong prevalence for the central nervous system and metabolism. MitoCoP will provide a high-confidence resource for placing dynamics, functions, and dysfunctions of mitochondria into the cellular context.
Histones control gene expression by regulating chromatin structure and function. The posttranslational modifications (PTMs) on the side chains of histones form the epigenetic landscape, which is tightly controlled by epigenetic modulator enzymes and further recognized by so-called reader domains. Histone microarrays have been widely applied to investigate histone–reader interactions, but not the transient interactions of Zn2+-dependent histone deacetylase (HDAC) eraser enzymes. Here, we synthesize hydroxamic acid-modified histone peptides and use them in femtomolar microarrays for the direct capture and detection of the four class I HDAC isozymes. Follow-up functional assays in solution provide insights into their suitability to discover HDAC substrates and inhibitors with nanomolar potency and activity in cellular assays. We conclude that similar hydroxamic acid-modified histone peptide microarrays and libraries could find broad application to identify class I HDAC isozyme-specific substrates and facilitate the development of isozyme-selective HDAC inhibitors and probes.
The study of gene expression in fungi has typically relied on measuring transcripts in populations of cells. A major disadvantage of this approach is that the transcripts’ spatial distribution and stochastic variation among individual cells within a clonal population is lost. Traditional fluorescence in situ hybridization techniques have been of limited use in fungi due to poor specificity and high background signal. Here, we report that in situ hybridization chain reaction (HCR), a method that employs split-initiator probes to trigger signal amplification upon mRNA-probe hybridization, is ideally suited for the imaging and quantification of low-abundance transcripts at single-cell resolution in the fungus Candida albicans. We show that HCR allows the absolute quantification of transcripts within a cell by microscopy as well as their relative quantification by flow cytometry. mRNA imaging also revealed the subcellular localization of specific transcripts. Furthermore, we establish that HCR is amenable to multiplexing by visualizing different transcripts in the same cell. Finally, we combine HCR with immunostaining to image specific mRNAs and proteins simultaneously within a single C. albicans cell. The fungus is a major pathogen in humans where it can colonize and invade mucosal surfaces and most internal organs. The technical development that we introduce, therefore, paves the way to study the patterns of expression of pathogenesis-associated C. albicans genes in infected organs at single-cell resolution.
Animal pollinators are globally threatened by anthropogenic land use change and agricultural intensification. The yield of many food crops is therefore negatively impacted because they benefit from biotic pollination. This is especially the case in the tropics. For instance, fruit set of Coffea arabica has been shown to increase by 10–30% in plantations with a high richness of bee species, possibly influenced by the availability of surrounding forest habitat. Here, we performed a global literature review to (1) assess how much animal pollination enhances coffee fruit set, and to (2) examine the importance of the amount of forest cover, distance to nearby forest and forest canopy density for bee species richness and coffee fruit set. Using a systematic literature review, we identified eleven case studies with a total of 182 samples where fruit set of C. arabica was assessed. We subsequently gathered forest data for all study sites from satellite imagery. We modelled the effects of open (all forest with a canopy density of ≥25%), closed (≥50%) and dense (≥75%) forests on pollinator richness and fruit set of coffee. Overall, we found that animal pollination increases coffee fruit set by ~18% on average. In only one of the case studies, regression results indicate a positive effect of dense forest on coffee fruit set, which increased with higher forest cover and shorter distance to the forest. Against expectations, forest cover and distance to open forest were not related to bee species richness and fruit set. In summary, we provide strong empirical support for the notion that animal pollinators increase coffee fruit set. Forest proximity had little overall influence on bee richness and coffee fruit set, except when farms were surrounded by dense tropical forests, potentially because these may provide high-quality habitats for bees pollinating coffee. We, therefore, advocate that more research is done to understand the biodiversity value of dense forest for pollinators, notably assessing the mechanisms underlying the importance of forest for pollinators and their pollination services.
The pandemic spread of an infectious disease poses a plethora of challenges to society, clinicians, health care providers and regulating authorities. In order to mount a rapid response and to provide hope in a potentially catastrophic situation as the current COVID-19 pandemic, emergency plans, regulations and funding strategies have to be developed on regional, national and international levels. The speed needed to establish rapid response programs is challenged by the dynamics of the spread of the disease, the concurrent and competing development of different and potentially more effective treatment options, and not the least by regulatory uncertainty. Convalescent plasma, that is plasma collected from patients who have recovered from COVID-19 infections, has emerged as one of the first potential treatment options in the absence of drugs or vaccines with proven efficacy against SARS-CoV-2. The societal aspects of convalescent plasma and the public awareness gave an additional boost to the rapid employment of convalescent plasma donation platforms immediately after the SARS-CoV-2 outbreak. At the same time, uncertainty remains as to the efficacy of convalescent plasma. With evidence mostly limited to empirical reports, convalescent plasma has been used for decades for the prophylaxis and treatment of various infectious diseases. Clinical trials have addressed different infectious agents, stages of disease, target groups of patients and yielded sometimes inconclusive results. The aim of this short review is to delineate the regulatory background for the emergency use of convalescent plasma in the USA, in the European Union and in Germany, and the transition to the setting of clinical trials. In addition, we describe observations made in the process of collecting COVID-19 convalescent plasma (herein referred to as CCP), and formulate proposals to further improve the framework for rapid responses in future emergency situations.
Anxiolytic drugs often have sedative effects that impair the ability to drive. Our double-blind, randomized crossover trial investigated the effect of Silexan, a non-sedating, anxiolytic herbal medicinal product, on driving performance in healthy volunteers. Part 1 aimed at demonstrating equivalence between 80 mg/d Silexan and placebo. Part 2 was performed to demonstrate superiority of 160 and 320 mg Silexan over 1 mg lorazepam and included a placebo arm for assay sensitivity. Driving performance was assessed in a validated, alcohol-calibrated simulator test. The primary outcome was the standard deviation of the lane position (SDLP). Secondary outcomes included driving errors and sleepiness. Fifty and 25 subjects were randomized in Parts 1 and 2, respectively. In Part 1, Silexan 80 mg was confirmed to be equivalent to placebo after single administration (equivalence range: δ = ±2 cm). The 95% confidence interval (CI) for the SDLP marginal mean value difference Silexan–placebo for single administration was −1.43; +1.38 and thus similar to the 95% CI of −1.45; +0.79 cm for 7 days’ multiple dosing. In Part 2, 95% CIs for SDLP marginal mean value differences to lorazepam were −8.58; −5.42 cm for Silexan 160 mg and −8.65; −5.45 cm for 320 mg (p < 0.001). Confirmatory results were supported by secondary outcomes, where results for Silexan were comparable to placebo and more favorable than for lorazepam. The study demonstrates that single doses of up to 320 mg Silexan and multiple doses of 80 mg/d have no adverse effect on driving performance.
A novel time-resolved pump–probe spectroscopic approach that enables to keep high resolution in both the time and energy domain, nanosecond excitation–picosecond ionization–picosecond infrared probe (ns–ps–ps TRIR) spectroscopy, has been applied to the trans-4-methylformanilide–water (4MetFA–W) cluster. Water migration dynamics from the CO to the NH binding site in a peptide linkage triggered by photoionization of 4MetFA–W is directly monitored by the ps time evolution of IR spectra, and the presence of an intermediate state is revealed. The time evolution is analyzed by rate equations based on a four-state model of the migration dynamics. Time constants for the initial to the intermediate and hot product and to the final product are obtained. The acceleration of the dynamics by methyl substitution and the strong contribution of intracluster vibrational energy redistribution in the termination of the solvation dynamics is suggested. This picture is well confirmed by the ab initio on-the-fly molecular dynamics simulations. Vibrational assignments of 4MetFA and 4MetFA–W in the neutral (S0 and S1) and ionic (D0) electronic states measured by ns IR dip and electron-impact IR photodissociation spectroscopy are also discussed prior to the results of time-resolved spectroscopy.
Background
To characterise the longitudinal dynamics of C-reactive protein (CRP) and Procalcitonin (PCT) in a cohort of hospitalised patients with COVID-19 and support antimicrobial decision-making.
Methods
Longitudinal CRP and PCT concentrations and trajectories of 237 hospitalised patients with COVID-19 were modelled. The dataset comprised of 2,021 data points for CRP and 284 points for PCT. Pairwise comparisons were performed between: (i) those with or without significant bacterial growth from cultures, and (ii) those who survived or died in hospital.
Results
CRP concentrations were higher over time in COVID-19 patients with positive microbiology (day 9: 236 vs 123 mg/L, p < 0.0001) and in those who died (day 8: 226 vs 152 mg/L, p < 0.0001) but only after day 7 of COVID-related symptom onset. Failure for CRP to reduce in the first week of hospital admission was associated with significantly higher odds of death. PCT concentrations were higher in patients with COVID-19 and positive microbiology or in those who died, although these differences were not statistically significant.
Conclusions
Both the absolute CRP concentration and the trajectory during the first week of hospital admission are important factors predicting microbiology culture positivity and outcome in patients hospitalised with COVID-19. Further work is needed to describe the role of PCT for co-infection. Understanding relationships of these biomarkers can support development of risk models and inform optimal antimicrobial strategies.
The alkaline comet assay, or single cell gel electrophoresis, is one of the most popular methods for assessing DNA damage in human population. One of the open issues concerning this assay is the identification of those factors that can explain the large inter-individual and inter-laboratory variation. International collaborative initiatives such as the hCOMET project - a COST Action launched in 2016 - represent a valuable tool to meet this challenge. The aims of hCOMET were to establish reference values for the level of DNA damage in humans, to investigate the effect of host factors, lifestyle and exposure to genotoxic agents, and to compare different sources of assay variability. A database of 19,320 subjects was generated, pooling data from 105 studies run by 44 laboratories in 26 countries between 1999 and 2019. A mixed random effect log-linear model, in parallel with a classic meta-analysis, was applied to take into account the extensive heterogeneity of data, due to descriptor, specimen and protocol variability. As a result of this analysis interquartile intervals of DNA strand breaks (which includes alkali-labile sites) were reported for tail intensity, tail length, and tail moment (comet assay descriptors). A small variation by age was reported in some datasets, suggesting higher DNA damage in oldest age-classes, while no effect could be shown for sex or smoking habit, although the lack of data on heavy smokers has still to be considered. Finally, highly significant differences in DNA damage were found for most exposures investigated in specific studies. In conclusion, these data, which confirm that DNA damage measured by the comet assay is an excellent biomarker of exposure in several conditions, may contribute to improving the quality of study design and to the standardization of results of the comet assay in human populations.
Background
While it is now apparent clinical sequelae (long COVID) may persist after acute COVID-19, their nature, frequency and aetiology are poorly characterised. This study aims to regularly synthesise evidence on long COVID characteristics, to help inform clinical management, rehabilitation strategies and interventional studies to improve long-term outcomes.
Methods
A living systematic review. Medline, CINAHL (EBSCO), Global Health (Ovid), WHO Global Research on COVID-19 database, LitCovid and Google Scholar were searched till 17 March 2021. Studies including at least 100 people with confirmed or clinically suspected COVID-19 at 12 weeks or more post onset were included. Risk of bias was assessed using the tool produced by Hoy et al. Results were analysed using descriptive statistics and meta-analyses to estimate prevalence.
Results
A total of 39 studies were included: 32 cohort, 6 cross-sectional and 1 case–control. Most showed high or moderate risk of bias. None were set in low-income countries and few included children. Studies reported on 10 951 people (48% female) in 12 countries. Most included previously hospitalised people (78%, 8520/10 951). The longest mean follow-up time was 221.7 (SD: 10.9) days post COVID-19 onset. Over 60 physical and psychological signs and symptoms with wide prevalence were reported, most commonly weakness (41%; 95% CI 25% to 59%), general malaise (33%; 95% CI 15% to 57%), fatigue (31%; 95% CI 24% to 39%), concentration impairment (26%; 95% CI 21% to 32%) and breathlessness (25%; 95% CI 18% to 34%). 37% (95% CI 18% to 60%) of patients reported reduced quality of life; 26% (10/39) of studies presented evidence of reduced pulmonary function.
Conclusion
Long COVID is a complex condition with prolonged heterogeneous symptoms. The nature of studies precludes a precise case definition or risk evaluation. There is an urgent need for prospective, robust, standardised, controlled studies into aetiology, risk factors and biomarkers to characterise long COVID in different at-risk populations and settings.
PROSPERO registration number CRD42020211131.
Lungfishes belong to lobe-fined fish (Sarcopterygii) that, in the Devonian period, ‘conquered’ the land and ultimately gave rise to all land vertebrates, including humans1,2,3. Here we determine the chromosome-quality genome of the Australian lungfish (Neoceratodus forsteri), which is known to have the largest genome of any animal. The vast size of this genome, which is about 14× larger than that of humans, is attributable mostly to huge intergenic regions and introns with high repeat content (around 90%), the components of which resemble those of tetrapods (comprising mainly long interspersed nuclear elements) more than they do those of ray-finned fish. The lungfish genome continues to expand independently (its transposable elements are still active), through mechanisms different to those of the enormous genomes of salamanders. The 17 fully assembled lungfish macrochromosomes maintain synteny to other vertebrate chromosomes, and all microchromosomes maintain conserved ancient homology with the ancestral vertebrate karyotype. Our phylogenomic analyses confirm previous reports that lungfish occupy a key evolutionary position as the closest living relatives to tetrapods4,5, underscoring the importance of lungfish for understanding innovations associated with terrestrialization. Lungfish preadaptations to living on land include the gain of limb-like expression in developmental genes such as hoxc13 and sall1 in their lobed fins. Increased rates of evolution and the duplication of genes associated with obligate air-breathing, such as lung surfactants and the expansion of odorant receptor gene families (which encode proteins involved in detecting airborne odours), contribute to the tetrapod-like biology of lungfishes. These findings advance our understanding of this major transition during vertebrate evolution.
The benefits of cochlear implantation in children with severe hearing impairments are widely known; however, there is no consensus regarding which minimal outcome measurements (MOMs) should be used to determine outcomes in this population with pediatric cochlear implant (CI). Therefore, the authors aim to propose a MOM test battery for pediatric CI recipients that can facilitate international multi-center research and collaboration. A pediatric MOM test battery was developed and agreed-upon by members of the HEARRING group across 30 expert clinics in the field of hearing implantation. The MOM test battery was chosen based on a literature search that focused on outcome measurements applied in clinical trials involving children with a hearing implant. Members of the HEARRING group were then asked to evaluate each of the pediatric MOM tests used. The final pediatric MOM test battery was defined for different chronological age categories (six weeks–18 years) at different suggested test intervals. The test battery includes objective hearing measurements, aided and unaided audiometry, speech perception tests in quiet and in noise, subjective hearing assessments, assessment of language development, and mental and motor development. This study presents a consensus on a MOM test battery for pediatric CI recipients that was agreed upon by members of the HEARRING group. This test battery should allow for international multi-center research to be able to extend and share evidence that will guide future clinical practice and research efforts in pediatric populations with CI.
The study presented in the following verifies some assumptions of the novel ‘unsafe world’ model of selective mutism (SM). According to this model, SM is a stress reaction to situations erroneously experienced via cognition without awareness as ‘unsafe’. It assumes a high sensitivity to unsafety, whereby the nervous system triggers dissociation or freeze mode at relatively low thresholds. We examine whether there is a correlation between SM, sensory-processing sensitivity and dissociation. We compared a sample of 28 children and adolescents with SM (mean age 12.66 years; 18 females) to 33 controls without SM (mean age 12.45 years; 21 females). Both groups were compared using a medical history sheet, the ‘Selective Mutism Questionnaire’ (SMQ), a ‘Checklist for Speaking Behaviour’ (CheckS), the ‘Highly Sensitive Person Scale’ (HSPS), the ‘Child Dissociative Checklist’ (CDC), the ‘Adolescent Dissociative Experience Scale’ (A-DES) and the ‘Social Phobia and Anxiety Inventory for Children’ (SPAIK). Appropriate parametric and non-parametric tests were conducted to examine differences between groups. The results indicate that sensory-processing sensitivity was significantly higher in the group of children and adolescents with SM [X2(1) = 7.224, p = 0.0007; d = 1.092]. Furthermore, dissociative symptoms were more common in children and adolescents with SM than in controls [F(1, 33) = 13.004, p = 0.001; d = 0.986]. The results indicate that sensory-processing sensitivity and dissociation are important factors of SM that may hold important implications for the treatment.
A search for heavy long-lived multicharged particles is performed using the ATLAS detector at the LHC. Data with an integrated luminosity of 36.1 fb(-1) collected in 2015 and 2016 from proton-proton collisions at root s = 13 TeV are examined. Particles producing anomalously high ionization, consistent with long-lived massive particles with electric charges from vertical bar q vertical bar = 2e to vertical bar q vertical bar = 7e, are searched for. No events are observed, and 95% confidence level cross-section upper limits are interpreted as lower mass limits for a Drell-Yan production model. Multicharged particles with masses between 50 and 980-1220 GeV (depending on their electric charge) are excluded.
As the treatment of effluents containing the antibiotic drug sulfadiazine (SZ) is one of the challenging problems in the field of environmental chemistry, it is essential to determine the concentration of SZ by a rapid and accurate method and then find a suitable method to degrade the assayed products into harmless chemicals. The color of the charge transfer (CT) complexes developed from the reaction of SZ with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), chloranilic acid (CHL) and picric acid (PA) was used to determine the concentration of SZ at 528, 510 and 410 nm, respectively. The Lambert–Beer's law is obeyed in the ranges of 6.80–68.06, 13.61–136.12 and 6.80–27.22 μg mL\(^{−1}\) for DDQ, CHL and PA complexes. The photolysis of SZ → DDQ in presence of sodium nitrite at 256 nm leads to faster degradation of SZ compared with the control experiments. This was simply spectrophotometrically followed by a decrease in the intensity of the CT band. The effect of some additives such as oxalic acid, and hematite nano particles was studied. For comparison, other π-acceptor reagents such as CHL and PA were used. About 80% of SZ is degraded in 45 min upon the illumination of SZ → DDQ at 256 nm, whereas 90 min is required in the case of CHL and PA to attain the same degradation limit.
Background: Cellular glucose uptake may involve either non-concentrative glucose carriers of the GLUT family or Na\(^+\)-coupled glucose-carrier SGLT1, which accumulates glucose against glucose gradients and may thus accomplish cellular glucose uptake even at dramatically decreased extracellular glucose oncentrations. SGLT1 is not only expressed in epithelia but as well in tumour cells and immune cells. Immune cell functions strongly depend on their metabolism, therefore we hypothesized that deficiency of SGLT1 modulates the defence against bacterial infection. To test this hypothesis, we infected wild type mice and gene targeted mice lacking functional SGLT1 with Listeria monocytogenes.
Methods: SGLT1 deficient mice and wild type littermates were infected with 1x10\(^4\) CFU Listeria monocytogenes intravenously. Bacterial titers were determined by colony forming assay, SGLT1, TNF-α, IL-6 and IL-12a transcript levels were determined by qRT-PCR, as well as SGLT1 protein abundance and localization by immunohistochemistry.
Results: Genetic knockout of SGLT1 (Slc5a1\(^{–/–}\) mice) significantly compromised bacterial clearance following Listeria monocytogenes infection with significantly enhanced bacterial load in liver, spleen, kidney and lung, and significantly augmented hepatic expression of TNF-α and IL-12a. While all wild type mice survived, all SGLT1 deficient mice died from the infection.
Conclusions: SGLT1 is required for bacterial clearance and host survival following murine Listeria infection.
Staphylococcus aureus (S. aureus) infections are a major clinical problem and range from mild skin and soft-tissue infections to severe and even lethal infections such as pneumonia, endocarditis, sepsis, osteomyelitis, and toxic shock syndrome. Toxins that are released from S. aureus mediate many of these effects. Here, we aimed to identify molecular mechanisms how α-toxin, a major S. aureus toxin, induces inflammation. Methods: Macrophages were isolated from the bone marrow of wildtype and acid sphingomyelinase-deficient mice, stimulated with S. aureus α-toxin and activation of the acid sphingomyelinase was quantified. The subcellular formation of ceramides was determined by confocal microscopy. Release of cathepsins from lysosomes, activation of inflammasome proteins and formation of Interleukin-1β (IL-1β) and Tumor Necrosis Factor-α (TNF-α) were analyzed by western blotting, confocal microscopy and ELISA. Results: We demonstrate that S. aureus α-toxin activates the acid sphingomyelinase in ex vivo macrophages and triggers a release of ceramides. Ceramides induced by S. aureus α-toxin localize to lysosomes and mediate a release of cathepsin B and D from lysosomes into the cytoplasm. Cytosolic cathepsin B forms a complex with Nlrc4. Treatment of macrophages with α-toxin induces the formation of IL-1β and TNF-α. These events are reduced or abrogated, respectively, in cells lacking the acid sphingomyelinase and upon treatment of macrophages with amitriptyline, a functional inhibitor of acid sphingomyelinase. Pharmacological inhibition of cathepsin B prevented activation of the inflammasome measured as release of IL-1β, while the formation of TNF-α was independent of cathepsin B. Conclusion: We demonstrate a novel mechanism how bacterial toxins activate the inflammasome and mediate the formation and release of cytokines: S. aureus α-toxin triggers an activation of the acid sphingomyelinase and a release of ceramides resulting in the release of lysosomal cathepsin B and formation of pro-inflammatory cytokines.
Previous studies have shown that ingroup/outgroup membership influences individual’s fairness considerations. However, it is not clear yet how group membership influences brain activity when a recipient evaluates the fairness of asset distribution. In this study, subjects participated as recipients in an Ultimatum Game with alleged members of both an experimentally induced ingroup and outgroup. They either received extremely unequal, moderately unequal, or equal offers from proposers while electroencephalogram was recorded. Behavioral results showed that the acceptance rates for unequal offers were higher when interacting with ingroup partners than with outgroup partners. Analyses of event related potentials revealed that proposers’ group membership modulated offer evaluation at earlier processing stages. Feedback-related negativity was more negative for extremely and moderately unequal offers compared to equal offers in the ingroup interaction whereas it did not show differential responses to different offers in the outgroup interaction. Analyses of event related oscillations revealed that the theta power (4–6 Hz) was larger for moderately unequal offers than equal offers in the ingroup interaction whereas it did not show differential responses to different offers in the outgroup interaction. Thus, early mechanisms of fairness evaluation are strongly modulated by the ingroup/outgroup membership of the interaction partner.
Heat-assisted magnetic recording (HAMR) is often considered the next major step in the storage industry: it is predicted to increase the storage capacity, the read/write speed and the data lifetime of future hard disk drives. However, despite more than a decade of development work, the reliability is still a prime concern. Featuring an inherently fragile surface-plasmon resonator as a highly localized heat source, as part of a near-field transducer (NFT), the current industry concepts still fail to deliver drives with sufficient lifetime. This study presents a method to aid conventional NFT-designs by additional grazing-incidence laser illumination, which may open an alternative route to high-durability HAMR. Magnetic switching is demonstrated on consumer-grade CoCrPt perpendicular magnetic recording media using a green and a near-infrared diode laser. Sub-500 nm magnetic features are written in the absence of a NFT in a moderate bias field of only μ0H = 0.3 T with individual laser pulses of 40 mW power and 50 ns duration with a laser spot size of 3 μm (short axis) at the sample surface – six times larger than the magnetic features. Herein, the presence of a nanoscopic object, i.e., the tip of an atomic force microscope in the focus of the laser at the sample surface, has no impact on the recorded magnetic features – thus suggesting full compatibility with NFT-HAMR.
A system-wide understanding of cellular function requires knowledge of all functional interactions between the expressed proteins. The STRING database aims to collect and integrate this information, by consolidating known and predicted protein–protein association data for a large number of organisms. The associations in STRING include direct (physical) interactions, as well as indirect (functional) interactions, as long as both are specific and biologically meaningful. Apart from collecting and reassessing available experimental data on protein–protein interactions, and importing known pathways and protein complexes from curated databases, interaction predictions are derived from the following sources: (i) systematic co-expression analysis, (ii) detection of shared selective signals across genomes, (iii) automated text-mining of the scientific literature and (iv) computational transfer of interaction knowledge between organisms based on gene orthology. In the latest version 10.5 of STRING, the biggest changes are concerned with data dissemination: the web frontend has been completely redesigned to reduce dependency on outdated browser technologies, and the database can now also be queried from inside the popular Cytoscape software framework. Further improvements include automated background analysis of user inputs for functional enrichments, and streamlined download options. The STRING resource is available online, at http://string-db.org/.
The Kryptolebias marmoratus is unique because it is the only selffertilizing hermaphroditic vertebrate, known to date. It primarily reproduces by internal self-fertilization in a mixed ovary/testis gonad. Here, we report on a high-quality genome assembly for the K. marmoratus South Korea (SK) strain highlighting the diversity and distribution of transposable elements (TEs). We find that K. marmoratus genome maintains number and composition of TEs. This can be an important genomic attribute promoting genome recombination in this selfing fish, while, in addition to a mixed mating strategy, it may also represent a mechanism contributing to the evolutionary adaptation to ecological pressure of the species. Future work should help clarify this point further once genomic information is gathered for other taxa of the family Rivulidae that do not self-fertilize. We provide a valuable genome resource that highlights the potential impact of TEs on the genome evolution of a fish species with an uncommon life cycle.
Climate change and associated extreme weather events are a threat not only for agricultural
yields but the plant kingdom in general. Therefore, there is a great necessity to better
understand the plants' intrinsic mechanisms to combat heat stress. The plant heat stress
response already has been investigated in many studies, including the role of HSFA1
transcription factors as the central regulators. Other aspects such as the initial perception of
heat and the role of heat-induced changes in plant metabolism are rather unknown.
In this thesis, the natural variation of 250 different accessions of Arabidopsis thaliana was
investigated regarding the temperature-dependent accumulation of raffinose and
triacylglycerols. A connection between these phenotypes and respective genotypes was
established using genome-wide association studies. As a result, the candidate gene
TREHALOSE-6-PHOSPHATE SYNTHASE 1 (TPS1), was identified. Enzymatic TPS1 is responsible
for the synthesis of trehalose 6-phosphate (T6P), which serves as an indicator and regulator
of sucrose homeostasis.
Subsequent analyses using tps1 tilling mutants demonstrated a link between T6P metabolism
and an increased accumulation of various soluble carbohydrates and starch, including
raffinose both under control conditions and during heat exposure. Furthermore, the mutant
lines displayed enhanced thermotolerance and survival rates following long-term heat stress.
Transcriptome analyses, however, did not show any difference in the regulation of canonical
heat stress-associated genes. Instead, genes related to photosynthesis were overrepresented
among the differentially upregulated genes in tps1 tilling lines during heat exposure. In this
work, a direct connection of T6P signaling, sucrose homeostasis, and thermotolerance is
shown for the first time.
In a second project, two Arabidopsis thaliana accessions (Oberursel-0, accession ID: 7276;
Nieps-0, accession ID: 7268) showing distinct capacities to acquire short-term
thermotolerance were compared to identify the putative causative regulators or mechanisms
that lead to the different levels of thermotolerance.
An examination of the transcriptomes of 7268 and 7276 showed that several hundreds of
genes were already differentially regulated within 10 minutes of exposure to 32 °C or 34 °C.
Among these, several genes associated with sulfur metabolism were more highly induced in
the more thermotolerant accession 7268. However, experimental as well as genetic
manipulation of sulfur availability and metabolism did not result in altered thermotolerance.
In addition to sulfur-related genes, most of the canonical heat stress-associated genes were
more highly expressed in 7268 than in 7276. While we could not identify a causative regulator
or mechanism of differential thermotolerances, the data strongly suggests that 7268 either
has a higher overall sensitivity, i.e., the heat stress response is initiated at lower temperatures,
or stronger overall heat stress response when exposed to a certain elevated temperature.
Ziel:
Die mediokarpale Teilarthrodese (MKTA) des Handgelenks ist eine sehr häufig durchgeführte Rettungsoperation, wenn intolerable Schmerzen aufgrund eines KK ein operatives Vorgehen erforderlich machen. Zahlreiche Studien beschreiben eine deutliche Beschwerdelinderung durch die MKTA, aber keine vollständige Schmerzfreiheit. Die Ursachen hierfür sind vielfältig, unter anderem kommt eine Pisotriquetralarthrose in Be-tracht. Die Entstehung einer solchen wird durch die Handwurzelfehlstellung beim KK begünstigt [8]. In dieser Arbeit wurde der Einfluss einer PT-Arthrose auf das mittel- bis langfristige Ergebnis einer MKTA untersucht. Des Weiteren wurde untersucht, inwie-fern sich eine PT-Arthrose nach einer MKTA entwickeln kann, sofern diese nicht bereits zum Operationszeitpunkt bestand.
Methode:
Es wurden 48 Personen, die zwischen 2004 und 2016 eine MKTA erhielten und deren Status hinsichtlich einer PT-Arthrose zum OP-Zeitpunkt durch eine Schnitt-bilddiagnostik analysiert werden konnte, in die Studie eingeschlossen. Zum Zeitpunkt der MKTA hatten 25 Patienten eine ausgeprägte PT-Arthrose und 23 Patienten keine PT-Arthrose. Die Patienten wurden durchschnittlich 75 Monate postoperativ klinisch und radiologisch nachuntersucht. Es wurde der Krimmer-Score und der DASH-Score erfasst. Ferner wurden klinische Untersuchungsparameter erhoben sowie die Handkraft und Handgelenksbeweglichkeit gemessen. Arthrose-Zeichen im pisotriquetralen und radiolunären Gelenk wurden durch Röntgenaufnahmen des Handgelenks in zwei Ebe-nen und einer Pisiformen-Zielaufnahme beurteilt.
Ergebnis:
Es zeigte sich, dass eine PT-Arthrose keinen negativen Einfluss auf das Er-gebnis einer MKTA ausübt. Darüber hinaus entwickelten einige Patienten auch nach einer MKTA eine PT-Arthrose, sodass die veränderte Biomechanik durch die Operation keinen protektiven Faktor hierfür darstellt hat. Als klinischer Test erwies sich der Schmerz am Pisiforme bei passivem Überstrecken des Handgelenks als aussagekräftigs-ter Untersuchungsparameter hinsichtlich des Vorhandenseins einer PT-Arthrose.
Diskussion:
Selbst bei einer radiologisch nachgewiesenen PT-Arthrose kann ein KK ausschließlich mit einer MKTA behandelt werden. Halten nach einer MKTA ulnopalma-re Beschwerden am Handgelenk an oder treten neu auf, muss ätiologisch eine PT-Arthrose in Erwägung gezogen, abgeklärt und gegebenenfalls behandelt werden.
The implementation of high‐throughput sequencing (HTS) technologies in research and diagnostic laboratories has linked many new genes to rare bleeding, thrombotic, and platelet disorders (BTPD), and revealed multiple genetic variants linked to those disorders, many of them being of uncertain pathogenicity when considering the accepted evidence (variant consequence, frequency in control datasets, number of reported patients, prediction models, and functional assays). The sequencing effort has also resulted in resources for gathering disease‐causing variants associated with specific genes, but for BTPD, such well‐curated databases exist only for a few genes. On the other hand, submissions by individuals or diagnostic laboratories to the variant database ClinVar are hampered by the lack of a submission process tailored to capture the specific features of hemostatic diseases. As we move toward the implementation of HTS in the diagnosis of BTPD, the Scientific and Standardization Committee for Genetics in Thrombosis and Haemostasis has developed and tested a REDCap‐based interface, aimed at the community, to submit curated genetic variants for diagnostic‐grade BTPD genes. Here, we describe the use of the interface and the initial submission of 821 variants from 30 different centers covering 14 countries. This open‐access variant resource will be shared with the community to improve variant classification and regular bulk data transfer to ClinVar.
Precise quantitative information about the molecular architecture of synapses is essential to understanding the functional specificity and downstream signaling processes at specific populations of synapses. Glycine receptors (GlyRs) are the primary fast inhibitory neurotransmitter receptors in the spinal cord and brainstem. These inhibitory glycinergic networks crucially regulate motor and sensory processes. Thus far, the nanoscale organization of GlyRs underlying the different network specificities has not been defined. Here, we have quantitatively characterized the molecular arrangement and ultra-structure of glycinergic synapses in spinal cord tissue using quantitative super-resolution correlative light and electron microscopy. We show that endogenous GlyRs exhibit equal receptor-scaffold occupancy and constant packing densities of about 2000 GlyRs µm-2 at synapses across the spinal cord and throughout adulthood, even though ventral horn synapses have twice the total copy numbers, larger postsynaptic domains, and more convoluted morphologies than dorsal horn synapses. We demonstrate that this stereotypic molecular arrangement is maintained at glycinergic synapses in the oscillator mouse model of the neuromotor disease hyperekplexia despite a decrease in synapse size, indicating that the molecular organization of GlyRs is preserved in this hypomorph. We thus conclude that the morphology and size of inhibitory postsynaptic specializations rather than differences in GlyR packing determine the postsynaptic strength of glycinergic neurotransmission in motor and sensory spinal cord networks.
The advances in cancer immunotherapy come with several obstacles, limiting its widespread use and benefits so far only to a small subset of patients. One of the underlying challenges remains to be the lack of representative nonclinical models that translate to human immunity and are able to predict clinical efficacy and safety outcomes. In recent years, immunocompetent Cancer-on-Chip models emerge as an alternative human-based platform that enables the integration and manipulation of complex tumor microenvironment. In this review, we discuss novel opportunities offered by Cancer-on-Chip models to advance (mechanistic) immuno-oncology research, ranging from design flexibility to multimodal analysis approaches. We then exemplify their (potential) applications for the research and development of adoptive cell therapy, immune checkpoint therapy, cytokine therapy, oncolytic virus, and cancer vaccines.
Background
In the phase 3 ALCYONE study, daratumumab plus bortezomib/melphalan/prednisone (D-VMP) versus bortezomib/melphalan/prednisone (VMP) significantly improved progression-free survival (PFS) and overall survival (OS) in transplant-ineligible, newly diagnosed multiple myeloma (NDMM) patients. We present a subgroup analysis of ALCYONE by patient frailty status.
Patients and Methods
Frailty assessment was performed retrospectively using age, Charlson comorbidity index, and baseline Eastern Cooperative Oncology Group performance status score. Patients were classified as fit (0), intermediate (1), or frail (≥2); a nonfrail category combined fit and intermediate patients.
Results
Among randomized patients (D-VMP, n = 350; VMP, n = 356), 391 (55.4%) were nonfrail (D-VMP, 187 [53.4%]; VMP, 204 [57.3%]) and 315 (44.6%) were frail (163 [46.6%]; 152 [42.7%]). After 40.1-months median follow-up, nonfrail patients had longer PFS and OS than frail patients, but benefits of D-VMP versus VMP were maintained across subgroups: PFS nonfrail (median, 45.7 vs. 19.1 months; hazard ratio [HR], 0.36; P < .0001), frail (32.9 vs. 19.5 months; HR, 0.51; P < .0001); OS nonfrail (36-month rate, 83.6% vs. 74.5%), frail (71.4% vs. 59.0%). Improved greater than or equal to complete response and minimal residual disease (10−5)-negativity rates were observed for D-VMP versus VMP across subgroups. The 2 most common grade 3/4 treatment-emergent adverse events were neutropenia (nonfrail: 39.2% [D-VMP] and 42.4% [VMP]; frail: 41.3% and 34.4%) and thrombocytopenia (nonfrail: 32.8% and 36.9%; frail: 36.9% and 39.1%).
Conclusion
Our findings support the clinical benefit of D-VMP in transplant-ineligible NDMM patients enrolled in ALCYONE, regardless of frailty status.
Oligophenyleneethynylenes (OPEs) are prominent building blocks with exciting optical and supramolecular properties. However, their generally small spectroscopic changes upon aggregation make the analysis of their self-assembly challenging, especially in the absence of additional hydrogen bonds. Herein, by investigating a series of OPEs of increasing size, we have unravelled the role of the conjugation length on the self-assembly properties of OPEs.
We investigate a scenario inspired by natural supersymmetry, where neutrino data is explained within a low-scale seesaw scenario. For this the minimal supersymmetric Standard Model is extended by adding light right-handed neutrinos and their superpartners, the R-sneutrinos. Moreover, we consider the lightest neutralinos to be Higgsino-like. We first update a previous analysis and assess to which extent does existing LHC data constrain the allowed slepton masses. Here we find scenarios where sleptons with masses as low as 175 GeV are consistent with existing data. However, we also show that the upcoming run will either discover or rule out sleptons with masses of 300 GeV, even for these challenging scenarios. We then take a scenario which is on the borderline of observability of the upcoming LHC run assuming a luminosity of 300 fb(-1). We demonstrate that a prospective international e(+)e(-) linear collider with a center of mass energy of 1 TeV will be able to discover sleptons in scenarios which are difficult for the LHC. Moreover, we also show that a measurement of the spectrum will be possible within 1-3 percent accuracy.
The occlusal design plays a decisive role in the fabrication of dental restorations. Dentists and dental technicians depend on mechanical simulations of mandibular movement that are as accurate as possible, in particular, to produce interference-free yet chewing-efficient dental restorations. For this, kinetic data must be available, i.e., movements and deformations under the influence of forces and stresses. In the present study, so-called functional data were collected from healthy volunteers to provide consistent information for proper kinetics. For the latter purpose, biting and chewing forces, electrical muscle activity and jaw movements were registered synchronously, and individual magnetic resonance tomograms (MRI) were prepared. The acquired data were then added to a large complex finite element model of the complete masticatory system using the functional information obtained and individual anatomical geometries so that the kinetics of the chewing process and teeth grinding could be realistically simulated. This allows developing algorithms that optimize computer-aided manufacturing of dental prostheses close to occlusion. In this way, a failure-free function of the dental prosthesis can be guaranteed and its damage during usage can be reduced or prevented even including endosseous implants.
Background
A basic requirement for artificial intelligence (AI)–based image analysis systems, which are to be integrated into clinical practice, is a high robustness. Minor changes in how those images are acquired, for example, during routine skin cancer screening, should not change the diagnosis of such assistance systems.
Objective
To quantify to what extent minor image perturbations affect the convolutional neural network (CNN)–mediated skin lesion classification and to evaluate three possible solutions for this problem (additional data augmentation, test-time augmentation, anti-aliasing).
Methods
We trained three commonly used CNN architectures to differentiate between dermoscopic melanoma and nevus images. Subsequently, their performance and susceptibility to minor changes (‘brittleness’) was tested on two distinct test sets with multiple images per lesion. For the first set, image changes, such as rotations or zooms, were generated artificially. The second set contained natural changes that stemmed from multiple photographs taken of the same lesions.
Results
All architectures exhibited brittleness on the artificial and natural test set. The three reviewed methods were able to decrease brittleness to varying degrees while still maintaining performance. The observed improvement was greater for the artificial than for the natural test set, where enhancements were minor.
Conclusions
Minor image changes, relatively inconspicuous for humans, can have an effect on the robustness of CNNs differentiating skin lesions. By the methods tested here, this effect can be reduced, but not fully eliminated. Thus, further research to sustain the performance of AI classifiers is needed to facilitate the translation of such systems into the clinic.
Natural and cryptic peptides dominate the immunopeptidome of atypical teratoid rhabdoid tumors
(2021)
Background
Atypical teratoid/rhabdoid tumors (AT/RT) are highly aggressive CNS tumors of infancy and early childhood. Hallmark is the surprisingly simple genome with inactivating mutations or deletions in the SMARCB1 gene as the oncogenic driver. Nevertheless, AT/RTs are infiltrated by immune cells and even clonally expanded T cells. However, it is unclear which epitopes T cells might recognize on AT/RT cells.
Methods
Here, we report a comprehensive mass spectrometry (MS)-based analysis of naturally presented human leukocyte antigen (HLA) class I and class II ligands on 23 AT/RTs. MS data were validated by matching with a human proteome dataset and exclusion of peptides that are part of the human benignome. Cryptic peptide ligands were identified using Peptide-PRISM.
Results
Comparative HLA ligandome analysis of the HLA ligandome revealed 55 class I and 139 class II tumor-exclusive peptides. No peptide originated from the SMARCB1 region. In addition, 61 HLA class I tumor-exclusive peptide sequences derived from non-canonically translated proteins. Combination of peptides from natural and cryptic class I and class II origin gave optimal representation of tumor cell compartments. Substantial overlap existed with the cryptic immunopeptidome of glioblastomas, but no concordance was found with extracranial tumors. More than 80% of AT/RT exclusive peptides were able to successfully prime CD8+ T cells, whereas naturally occurring memory responses in AT/RT patients could only be detected for class II epitopes. Interestingly, >50% of AT/RT exclusive class II ligands were also recognized by T cells from glioblastoma patients but not from healthy donors.
Conclusions
These findings highlight that AT/RTs, potentially paradigmatic for other pediatric tumors with a low mutational load, present a variety of highly immunogenic HLA class I and class II peptides from canonical as well as non-canonical protein sources. Inclusion of such cryptic peptides into therapeutic vaccines would enable an optimized mapping of the tumor cell surface, thereby reducing the likelihood of immune evasion.
Background
The human leucocyte antigen (HLA) complex controls adaptive immunity by presenting defined fractions of the intracellular and extracellular protein content to immune cells. Understanding the benign HLA ligand repertoire is a prerequisite to define safe T-cell-based immunotherapies against cancer. Due to the poor availability of benign tissues, if available, normal tissue adjacent to the tumor has been used as a benign surrogate when defining tumor-associated antigens. However, this comparison has proven to be insufficient and even resulted in lethal outcomes. In order to match the tumor immunopeptidome with an equivalent counterpart, we created the HLA Ligand Atlas, the first extensive collection of paired HLA-I and HLA-II immunopeptidomes from 227 benign human tissue samples. This dataset facilitates a balanced comparison between tumor and benign tissues on HLA ligand level.
Methods
Human tissue samples were obtained from 16 subjects at autopsy, five thymus samples and two ovary samples originating from living donors. HLA ligands were isolated via immunoaffinity purification and analyzed in over 1200 liquid chromatography mass spectrometry runs. Experimentally and computationally reproducible protocols were employed for data acquisition and processing.
Results
The initial release covers 51 HLA-I and 86 HLA-II allotypes presenting 90,428 HLA-I- and 142,625 HLA-II ligands. The HLA allotypes are representative for the world population. We observe that immunopeptidomes differ considerably between tissues and individuals on source protein and HLA-ligand level. Moreover, we discover 1407 HLA-I ligands from non-canonical genomic regions. Such peptides were previously described in tumors, peripheral blood mononuclear cells (PBMCs), healthy lung tissues and cell lines. In a case study in glioblastoma, we show that potential on-target off-tumor adverse events in immunotherapy can be avoided by comparing tumor immunopeptidomes to the provided multi-tissue reference.
Conclusion
Given that T-cell-based immunotherapies, such as CAR-T cells, affinity-enhanced T cell transfer, cancer vaccines and immune checkpoint inhibition, have significant side effects, the HLA Ligand Atlas is the first step toward defining tumor-associated targets with an improved safety profile. The resource provides insights into basic and applied immune-associated questions in the context of cancer immunotherapy, infection, transplantation, allergy and autoimmunity. It is publicly available and can be browsed in an easy-to-use web interface at https://hla-ligand-atlas.org .
Background
We aimed to define the clinical and variant spectrum and to provide novel molecular insights into the DHX30-associated neurodevelopmental disorder.
Methods
Clinical and genetic data from affected individuals were collected through Facebook-based family support group, GeneMatcher, and our network of collaborators. We investigated the impact of novel missense variants with respect to ATPase and helicase activity, stress granule (SG) formation, global translation, and their effect on embryonic development in zebrafish. SG formation was additionally analyzed in CRISPR/Cas9-mediated DHX30-deficient HEK293T and zebrafish models, along with in vivo behavioral assays.
Results
We identified 25 previously unreported individuals, ten of whom carry novel variants, two of which are recurrent, and provide evidence of gonadal mosaicism in one family. All 19 individuals harboring heterozygous missense variants within helicase core motifs (HCMs) have global developmental delay, intellectual disability, severe speech impairment, and gait abnormalities. These variants impair the ATPase and helicase activity of DHX30, trigger SG formation, interfere with global translation, and cause developmental defects in a zebrafish model. Notably, 4 individuals harboring heterozygous variants resulting either in haploinsufficiency or truncated proteins presented with a milder clinical course, similar to an individual harboring a de novo mosaic HCM missense variant. Functionally, we established DHX30 as an ATP-dependent RNA helicase and as an evolutionary conserved factor in SG assembly. Based on the clinical course, the variant location, and type we establish two distinct clinical subtypes. DHX30 loss-of-function variants cause a milder phenotype whereas a severe phenotype is caused by HCM missense variants that, in addition to the loss of ATPase and helicase activity, lead to a detrimental gain-of-function with respect to SG formation. Behavioral characterization of dhx30-deficient zebrafish revealed altered sleep-wake activity and social interaction, partially resembling the human phenotype.
Conclusions
Our study highlights the usefulness of social media to define novel Mendelian disorders and exemplifies how functional analyses accompanied by clinical and genetic findings can define clinically distinct subtypes for ultra-rare disorders. Such approaches require close interdisciplinary collaboration between families/legal representatives of the affected individuals, clinicians, molecular genetics diagnostic laboratories, and research laboratories.
Chemicals reactivating epigenetically silenced genes target diverse classes of enzymes, including DNMTs, HDACs, HMTs and BET protein family members. They can strongly influence the expression of genes and endogenous retroviral elements with concomitant dsRNA synthesis and massive transcription of LTRs. Chemicals reactivating gene expression may cause both beneficial effects in cancer cells and may be hazardous by promoting carcinogenesis. Among chemicals used in medicine and commerce, only a small fraction has been studied with respect to their influence on epigenetic silencing. Screening of chemicals reactivating silent genes requires adequate systems mimicking whole-genome processes. We used a HeLa TSA-inducible cell population (HeLa TI cells) obtained by retroviral infection of a GFP-containing vector followed by several rounds of cell sorting for screening purposes. Previously, the details of GFP epigenetic silencing in HeLa TI cells were thoroughly described. Herein, we show that the epigenetically repressed gene GFP is reactivated by 15 agents, including HDAC inhibitors–vorinostat, sodium butyrate, valproic acid, depsipeptide, pomiferin, and entinostat; DNMT inhibitors–decitabine, 5-azacytidine, RG108; HMT inhibitors–UNC0638, BIX01294, DZNep; a chromatin remodeler–curaxin CBL0137; and BET inhibitors–JQ-1 and JQ-35. We demonstrate that combinations of epigenetic modulators caused a significant increase in cell number with reactivated GFP compared to the individual effects of each agent. HeLa TI cells are competent to metabolize xenobiotics and possess constitutively expressed and inducible cytochrome P450 mono-oxygenases involved in xenobiotic biotransformation. Thus, HeLa TI cells may be used as an adequate test system for the extensive screening of chemicals, including those that must be metabolically activated. Studying the additional metabolic activation of xenobiotics, we surprisingly found that the rat liver S9 fraction, which has been widely used for xenobiotic activation in genotoxicity tests, reactivated epigenetically silenced genes. Applying the HeLa TI system, we show that N-nitrosodiphenylamine and N-nitrosodimethylamine reactivate epigenetically silenced genes, probably by affecting DNA methylation.
Josephson junctions based on three-dimensional topological insulators offer intriguing possibilities to realize unconventional 𝑝-wave pairing and Majorana modes. Here, we provide a detailed study of the effect of a uniform magnetization in the normal region: We show how the interplay between the spin-momentum locking of the topological insulator and an in-plane magnetization parallel to the direction of phase bias leads to an asymmetry of the Andreev spectrum with respect to transverse momenta. If sufficiently large, this asymmetry induces a transition from a regime of gapless, counterpropagating Majorana modes to a regime with unprotected modes that are unidirectional at small transverse momenta. Intriguingly, the magnetization-induced asymmetry of the Andreev spectrum also gives rise to a Josephson Hall effect, that is, the appearance of a transverse Josephson current. The amplitude and current phase relation of the Josephson Hall current are studied in detail. In particular, we show how magnetic control and gating of the normal region can enable sizable Josephson Hall currents compared to the longitudinal Josephson current. Finally, we also propose in-plane magnetic fields as an alternative to the magnetization in the normal region and discuss how the planar Josephson Hall effect could be observed in experiments.
Polarized Z bosons from the decay of a Higgs boson produced in association with two jets at the LHC
(2021)
Investigating the polarization of weak bosons provides an important probe of the scalar and gauge sector of the Standard Model. This can be done in the Higgs decay to four leptons, whose Standard-Model leading-order amplitude enables to generate polarized observables from unpolarized ones via a fully-differential reweighting method. We study the Z-boson polarization from the decay of a Higgs boson produced in association with two jets, both in the gluon-fusion and in the vector-boson fusion channel. We also address the possibility of extending the results of this work to higher orders in perturbation theory.
Vitamin D is considered to play an important role in musculoskeletal health. It’s classical function is the regulation of calcium and phosphate homeostasis, thus ensuring a balanced bone metabolism that is characterised by an equal amount of bone resorption and bone formation. In the past decades, a plethora of pre-clinical and clinical studies reporting on potential health-beneficial properties of vitamin D have emerged. Moreover, there is an abundance of reports highlighting vitamin D deficiency and insufficiency in patients with almost innumerable diseases. Further, it is estimated that more than one billion people globally are affected by insufficient vitamin D levels. As such, research on vitamin D has been particularly popular over the past years. In orthopaedics and traumatology, most studies describe favourable effects of vitamin D in general. However, the relative importance of vitamin D is oftentimes debated. In this narrative review of the literature, we consider first, the properties of vitamin D and how vitamin D, vitamin D deficiency and the vitamin D receptor (VDR) impact on musculoskeletal health. Secondly, we provide an overview of studies reporting the prevalence of vitamin D deficiency in traumatology and diverse orthopaedic diseases including bone oncology. Lastly, we emphasise recent findings and touch on future perspectives in vitamin D research.
Background
Intensive Care Resources are heavily utilized during the COVID-19 pandemic. However, risk stratification and prediction of SARS-CoV-2 patient clinical outcomes upon ICU admission remain inadequate. This study aimed to develop a machine learning model, based on retrospective & prospective clinical data, to stratify patient risk and predict ICU survival and outcomes.
Methods
A Germany-wide electronic registry was established to pseudonymously collect admission, therapeutic and discharge information of SARS-CoV-2 ICU patients retrospectively and prospectively. Machine learning approaches were evaluated for the accuracy and interpretability of predictions. The Explainable Boosting Machine approach was selected as the most suitable method. Individual, non-linear shape functions for predictive parameters and parameter interactions are reported.
Results
1039 patients were included in the Explainable Boosting Machine model, 596 patients retrospectively collected, and 443 patients prospectively collected. The model for prediction of general ICU outcome was shown to be more reliable to predict “survival”. Age, inflammatory and thrombotic activity, and severity of ARDS at ICU admission were shown to be predictive of ICU survival. Patients’ age, pulmonary dysfunction and transfer from an external institution were predictors for ECMO therapy. The interaction of patient age with D-dimer levels on admission and creatinine levels with SOFA score without GCS were predictors for renal replacement therapy.
Conclusions
Using Explainable Boosting Machine analysis, we confirmed and weighed previously reported and identified novel predictors for outcome in critically ill COVID-19 patients. Using this strategy, predictive modeling of COVID-19 ICU patient outcomes can be performed overcoming the limitations of linear regression models.
Trial registration “ClinicalTrials” (clinicaltrials.gov) under NCT04455451.
Seit Beginn des neuen Jahrtausends haben Tablets in der medizinischen Lehre zunehmend an Bedeutung gewonnen. In dieser Studie wurden ein Tablet-basiertes und ein Dozierenden-zentriertes Lehrformat im Rahmen eines Radiologie-Seminars verglichen. Ziel war es, vergleichende Erkenntnisse über selbsteingeschätzten und objektiven Lernzuwachs und empfundene didaktische Qualität zu erlangen und mögliche Einflüsse des Charismas der Dozierenden sowie der studentischen Technikaffinität auf diese Variablen zu untersuchen.
Von n=366 Studierenden wurden über drei Semester hinweg Daten zu studentischer Technikaffinität generell sowie didaktischer Qualität und Zufriedenheit mit den Seminaren abhängig vom Lehrformat erhoben. Im letzten Studiensemester nahmen die Studierenden zudem eine Selbsteinschätzung ihres Lernzuwachses vor und legten ein Wissens- und Bildinterpretationstestat sowie eine Einschätzung des Charismas der Dozierenden ab. Mittels Maximum-Likelihood Faktorenanalyse erfolgte eine Strukturaufklärung der didaktischen Qualität und des Charismas. Pearson-Korrelationskoeffizienten wurden zur Untersuchung möglicher Korrelationen zwischen den Variablen berechnet. Ein Strukturregressionsmodell des Charismas der Dozierenden, der studentischen Technikaffinität und der didaktischen Qualität wurde erstellt und daraus eine lineare Regressionsgleichung abgeleitet.
Dem Tablet-basierten Lehrformat wurden signifikant höhere Werte in didaktischer Qualität und studentischer Zufriedenheit zugeschrieben. Das Dozierenden-zentrierte Format schnitt hingegen in selbsteingeschätztem und objektivem Lernzuwachs signifikant besser ab. Ausschließlich im Tablet-basierten Unterricht korrelierte das Charisma der Dozierenden mit dem selbsteingeschätzten Lernzuwachs. Die studentische Technikaffinität wirkte sich in diesem Format stärker auf die didaktische Qualität aus. Zudem wurden gute Organisation, eindeutige Lernziele und adäquate Lernformvariation als bedeutsame Faktoren identifiziert.
Diese Studie hebt die Relevanz des dozentischen Charismas und der studentischen Technikaffinität für die Sicherstellung hoher didaktischer Qualität in Tablet-basiertem Unterricht hervor. Sie zeigt zudem die Wichtigkeit der Kongruenz von Lernzielen, Lehrkonzept und Prüfungsmethode dieses Lehrformats auf, um hohe Studienleistungen sicherzustellen. Die untersuchten Faktoren sollten bei Entwurf digitaler Lehrkonzepte über spezifische Messinstrumente objektiviert und bei Bedarf geschult oder angepasst werden, um eine erfolgreiche Integration digitaler Medien in die medizinische Lehre zu gewährleisten.
The seasonal snow cover in the European Alps plays a crucial role in the region's climate, ecology, and economy. It affects the local climate through its high albedo, protects permafrost, provides habitats, and acts as a water reservoir that feeds European rivers. However, these functions are threatened by climate change. Analyzing snow cover dynamics is essential to predict future developments and assess related ecological and economic impacts.
This study explores the potential of long Earth Observation (EO) time series for modeling and predicting the snow line elevation (SLE) in the Alps. Based on approximately 15,000 Landsat satellite images, SLE time series were generated for the years 1985 to 2022. Various univariate forecasting models were evaluated, with the best results achieved by Random Forests, Telescope, and Seasonal ARIMA. A newly developed approach combines the best models into a robust ensemble, achieving an average Nash-Sutcliffe efficiency (NSE) of 0.8 in catchments with strong seasonal signals.
Forecasts for 2030 indicate significant upward shifts in the SLE, particularly in the Western and Southern Alps. Given the variability in results, a multivariate modeling approach using climate variables is recommended to improve prediction accuracy. This study lays the groundwork for future models that could potentially project SLE dynamics through the end of the 21st century under various climate scenarios, which is highly relevant for climate policy in the Alpine region.
Im experimentellen Ansatz sollte mithilfe der CRISPR-Cas9-Methode eine gerichtete ALK1-Rezeptor-Eliminierung in myoblastischen C2C12-Zellen durchgeführt werden. Nach erfolgreicher Klonierung der jeweiligen, für den Typ-I-Rezeptor ALK1-kodierenden, gRNA-Sequenzen in die Puro- und GFP-CRISPR-Plasmide gelang der mittels Lipofektion durchgeführte Transfer der vier klonierten Plasmide in die C2C12-Zellen. Parallel aufgetaut wurden C2C12*ALK2- sowie ALK3-Knockout-Zelllinien, welche zuvor durch die Masterandin L. Wiesmann, ebenfalls mithilfe der CRISPR-Cas9-Methode, induzierte Knockouts der jeweiligen Rezeptoren ALK2 sowie ALK3 enthielten. Anschließend erfolgte die Puromycin-Selektion der mit den Puro-Klonen transfizierten C2C12*ALK1-3, ALK1-4-, ALK2- sowie ALK3-KO-Zellpopulationen. Die Zellen der C2C12*ALK1-3-KO-Population überlebten die Selektion trotz erneuter Durchführung der Transfektion sowie Selektion nicht. Somit erfolgte die Kultivierung der verbliebenen Zellen der C2C12*ALK1 4-, C2C12*ALK2- sowie C2C12*ALK3-KO-Population. Anschließend galt es zu untersuchen, wie responsiv die einzelne KO-Zelle für verschiedene Liganden ist. Im Rahmen der Durchführung differenter, zellbasierter Versuche wie der qPCR, des Western Blots und des ALP-Assays wirkten verschiedene BMPs auf die KO-Populationen ein. Somit konnten die BMP-induzierten, nachfolgenden Ereignisse wie die mRNA-Expression, die SMAD-Phosphorylierung sowie die Induktion der ALP-Expression innerhalb der KO-Populationen genauer betrachtet werden.
Es ist allgemein bekannt, dass ALK1 sowohl bei der Angiogenese als auch bei der kardio-vaskulären Homöostase eine wichtige Rolle übernimmt. ALK1 ist vermutlich für die Gefäßneubildung in manchen Tumoren verantwortlich und auch die vaskuläre Erkrankung „Hereditäre hämorrhagische Teleangiektasie (HHT)“ steht im Zusammenhang mit einer Mutation des ALK1-Rezeptorgens. BMP9 beeinflusst als ALK1-bindender Ligand neben der Tumorentwicklung und der Angiogenese auch die osteogene Differenzierung mesenchymaler Stammzellen. Im Hinblick auf zukünftige Versuche sind daher weitere, noch aussagekräftigere Ergebnisse erstrebenswert, allerdings unter der Verwendung von ausschließlich homozygoten KO-Zelllinien. Weitere Erkenntnisse über die Rolle des ALK1-Rezeptors in BMP-vermittelter Signaltransduktion könnten für therapeutische Ansätze bei der Behandlung von vaskulären Erkrankungen und Tumorprogression sowie bei der Förderung der Knochenregeneration und -heilung hilfreich sein.
Zusammenfassend lässt sich sagen, dass diese prospektive Studie sowohl den prognostischen als auch den prädiktiven Nutzen eines Tumormarker-Abfalls von CA19- 9 auf eine Induktionschemotherapie bei Patienten mit LAPC bestätigt. Nicht die Ausgangswerte des Tumormarkers vor Therapie sollten zur Planung des weiteren Vorgehens herangezogen werden, sondern die Werte nach Abschluss der Induktionschemotherapie bzw. der Abfall unter der Induktionschemotherapie. Das biochemische Ansprechen von CA19-9 ist ein bedeutender Indikator für den Behandlungserfolg und verbessert die diagnostische Genauigkeit bei der Auswahl von Patienten für eine chirurgische Exploration. Sowohl die Betrachtung des biochemischen Ansprechens allein als auch in Kombination mit dem radiologischen Ansprechen verbessert Sensitivität und Spezifität hinsichtlich einer möglichen R0-Resektion. Kombiniert man bei Patienten mit einer SD in Woche 16 ein gutes biochemisches Ansprechen (Rückgang von CA19-9 > 55%), so wird eine Sensitivität von 100% hinsichtlich der möglichen R0-Resektion erreicht. Zur Abschätzung des Gesamtüberlebens von Patienten mit LAPC sollten dagegen eher die absoluten Werte von CA19-9 nach Induktionschemotherapie verwendet werden und nicht der relative Rückgang des Markers. So zeigen Patienten, welche in Woche 16 einen CA19-9 Wert < 50 U/ml aufweisen mit 27,8 Monaten das beste mediane Gesamtüberleben.
Die Entstehung kollinearer und nicht-kollinearer Spinstrukturen wird auf verschiedene magnetische Wechselwirkungen zurückgeführt. Für Anwendungen in der Medizin und in der Datenspeicherung ist es notwendig zu verstehen, unter welchen Parametern Frustrationen auftreten, um diese entweder zu vermeiden oder zu nutzen. In dieser Arbeit werden kollineare und nicht-kollineare Spinstrukturen auf zwei verschiedenen Materialsystemen untersucht. Das erste Materialsystem besteht aus drei atomaren Lagen Mangan auf einer (001) Oberfläche eines Wolfram-Einkristalls und das zweite Materialsystem enthält Mangan, welches verbunden mit Sauerstoff kettenförmig auf einer (001) Oberfläche eines Iridium-Einkristalls vorliegt.
Spinpolarisierte Rastertunnelmikroskopie (SP-RTM)-Messungen und -Simulationen der
dreilagigen, pseudomorphen Manganoberfläche ergeben eine nicht-kollineare Spinstruktur. Dichtefunktionaltheorie (DFT)-Berechnungen legen eine kollineare ↑↓↓-
Spinkonstellation nahe. Unter Berücksichtigung der chiralen biquadratischen Paarwechselwirkung befinden sich konische Spinspiralen mit kleinem Öffnungswinkel nah an dem energetisch niedrigsten Zustand. Spinaufgelöste DFT-Berechnungen sind abhängig von der genäherten, geometrischen Relaxation der atomaren Struktur. Kombinierte SP-RTM-Methoden weisen auf einem dreilagigen Materialsystem Spinspiralen nach und zufolge der DFT ist der kollineare bzw. nicht-kollineare Zustand des Systems durch den Abstand seiner Lagen bedingt.
SP-RTM-Messungen auf den Manganoxidketten weisen je nach Präparation eine kollineare antiferromagnetische (AFM) oder eine nicht-kollineare Spinstruktur nach. Zudem wird präsentiert, dass sich diese Spinstrukturen durch zwei verschiedene Sauerstoffdrücke und die Zufuhr von Wärme während der Präparation ineinander umschalten lassen. Durch niederenergetische Elektronenbeugung mit variabler Spannung werden zwei atomare Strukturen bestimmt, welche sich durch ihren Oxidationsgrad unterscheiden. Die nicht-kollineare Spinstruktur ist bereits in der Fachliteratur als 120° chirale Spinspirale, verursacht durch die Dzyaloshinskii-Moriya-verstärkte Ruderman-Kittel-Kasuya-Yosida (RKKY)-Wechselwirkung, bekannt. Nach aktuellen, kollinearen DFT-Berechnungen ist die kollineare Spinstruktur als AFM entlang der Ketten und als ferromagnetische Kopplung zwischen den Ketten ermittelt. Aufgrund des Nachweises eines höheren Oxidationsgrades wird eine stärkere RKKY-Austauschwechselwirkung auf der Basis der Heisenberg-Austauschwechselwirkung vermutet. Hier korreliert die Entstehung kollinearer oder nicht-kollinearer Spinstrukturen mit dem Oxidationsgrad.
The TRAF-binding receptor CD40 belongs to the TNFR superfamily and is broadly expressed on healthy cells, mainly on antigen-presenting cells, but also on other immune cells and non-immune cells. CD40 is bound by its ligand CD40L, which is essential for a wide range of immunological responses by inducing or inhibiting different pathways that are essential for a variety of cellular processes, including immune activation and maturation. (1,2) Dysregulated CD40 signalling has been implicated in inflammatory diseases, such as hyper-IgM syndrome, psoriasis, and cancer. (3–6) Due to its broad expression across various tumour types, it can serve as a tumour-associated antigen and has therefore been proposed as a target for antibodies for cancer treatment. (2,7,8)
Agonistic anti-CD40 antibodies have been demonstrated to induce anti-tumoural immune responses as well as therapeutic immunity. (2) Furthermore, prolonged stimulation of CD40 in tumour cells in vitro has been shown to decrease proliferation, increase expression of cytotoxic TNFSFLs and induce apoptosis. (9,10) Their effect on anti-tumoral responses has been well studied and anti-tumoral responses by DC maturation and suppression of malignant growth of B-cells have been confirmed and were found to induce cell death in tumours in vitro. (11–14)
Many agonistic anti-CD40 antibodies specifically have been reported to require secondary crosslinking by binding to either activating or inhibitory FcγRs to be agonistic in vitro, while in vivo studies have indicated inhibitory FcƴR2B expression as critical factor. (15–17) However, FcƴR independent agonism has also been reported for anti-CD40 antibodies. (18,19) While agonistic anti-CD40 IgG1, IgG3 and IgG4 antibodies have been shown to display FcƴR dependent agonism, agonistic anti-CD40 IgG2 antibodies have shown to display FcƴR independent agonism. Conversion of anti-CD40 IgG1 antibodies into IgG2 has also been shown to convert the antibody’s agonism into FcƴR independent agonism. (20)
To overcome FcƴR dependency, bispecific antibody fusion proteins containing a scFv as anchoring domain allowing for crosslink independent of FcƴR binding have been designed before. This approach has been found to display strong agonism for other antibody fusion proteins when bound to both targets, with response levels resembling that of FcƴR bound antibodies. (21,22)
The relevance of antibody isotype and idiotype for FcƴR-dependent agonism as well as the relevance of valency and antibody oligomerization for FcƴR-independent agonism were investigated in this study on a panel of different anti-CD40 antibodies. Several clinically investigated anti-CD40 antibodies (ADC-1013(23), APX005M(24), ChiLob7.4(25) and CP-870,893(26)) and one preclinical antibody (G28.5(27,28)) were considered. Selected antibodies were then cloned onto an IgG1, IgG1(N297A), IgG2 and IgG4 backbone. The IgG1(N297A) isotype is an IgG1 antibody with a point mutation (N297A) that is known to strongly reduce binding to FcƴR1, while reducing the binding affinity to FcƴR2B to undetectable levels. (29,30) In this work it is demonstrated that the investigated anti-CD40 antibody variants across different isotypes activate both the classical and alternative NFκB pathway by stimulating U2OS cells in an FcƴR dependent manner. Stimulation in the presence of both human FcƴRs as well as murine FcƴRs resulted in CD40 stimulation. A difference in binding competition was observed for the various anti-CD40 IgG1 antibodies, but no indication of a CRD-dependent mechanism responsible for their agonistic activity was found. Moreover, this FcƴR dependency could be overcome by creation of tetravalent antibody fusion proteins.
CRISPR-Cas systems are a versatile tool in genetic engineering because they can be easily reprogrammed to cut a specific chromosomal region or RNA transcript. The choice of nuclease, gRNA design, and target region all influence targeting efficiency, so the appropriate CRISPR components should be chosen depending on the desired application. This thesis examines factors that influence targeting in both DNA- and RNA-targeting CRISPR systems. Chapter 1 discusses the importance of target RNA abundance in shaping the immunity of type VI CRISPR systems. In bacteria, the Cas13 nuclease is known to degrade RNA specifically and non-specifically, leading to cell growth arrest, also known as dormancy. In this chapter, the factors that determine dormancy are investigated by targeting genome- and plasmid-encoded transcripts in E. coli. The observations are extended to a gRNA library targeting the entire coding genome and gRNA design rules are extrapolated. Finally, the role of Cas13 in defense is investigated by testing how the system behaves during viral infection or plasmid transformation. Chapter 2 also looks at the factors that characterize targeting efficiency, but focuses on the Cas12a DNA-targeting system in K. pneumoniae. The ultimate goal is to develop CRISPR antimicrobials as alternatives to antibiotics to eliminate multidrug-resistant and hypervirulent bacteria. Several nucleases are tested for antimicrobial activity, the Cas12a nuclease is selected and the same gRNAs are used against different strains to understand the robustness of the method. Rules for gRNA design are also investigated by looking at secondary structure and testing a gRNA library across several genomic regions in two different strains. This information is used to develop a machine-learning algorithm to predict gRNA activity. In addition, the CRISPR-Cas systems are also packaged in a T7-like phage with engineered tail fibers and delivered to K. pneumoniae. While Chapter 2 uncovers various factors that improve targeting efficiency, Chapter 3 aims to reduce targeting by the Cas9 and Cas12a nucleases to favor homology-directed repair for genome editing in E. coli. Targeting is slowed down so that some copies of the chromosomes remain intact, allowing the bacterium to survive and integrate the desired edit. To reduce targeting, different gRNA formats or nuclease variations are used, gRNA expression is modulated, or gRNAs with attenuated targeting are designed. Attenuated gRNAs are tested to introduce point mutations as well as whole gene deletions and substitutions, and the method is extended to Klebsiella oxytoca and Klebsiella pneumoniae, where it is applied to block transcription of an antibiotic resistance gene in the genome, restoring sensitivity to ampicillin. Overall, this work discusses how changing the CRISPR components alters the outcome of targeting and highlights strategies to achieve efficient or attenuated targeting depending on the desired application.
Variabilität der Penetranz und klinischen Manifestation der autosomal dominanten Osteopetrose Typ II
(2024)
Die autosomal dominante Osteopetrose Typ II ist eine seltene sklerosierende Skeletterkrankung, die durch heterozygote Varianten im CLCN7-Gen verursacht wird. Die klinische Manifestation umfasst ein breites Symptomspektrum. Charakteristisch sind die unvollständige Penetranz und eine hohe Variabilität der Phänotyp-Expression. Beide Phänomene führen wir am ehesten auf eine monoallelische Inaktivierung zurück.
Diese Studie war eine retrospektive Datenauswertung, die 14 ADO II-Betroffene mit klinischer Manifestation der Erkrankung sowie 5 PatientInnen mit Carrier-Status eingeschlossen hat. Diese wurden im Rahmen einer ausführlichen Diagnostik untersucht, bei der Parameter im Hinblick auf den Zustand des Skelettsystems, körperliche Funktionen und Symptome sowie die gesundheitsbezogene Lebensqualität und der Leidensdruck erfasst wurden. Diese Variablen wurden zwischen der Betroffenen- und Carrier-Gruppe verglichen, wobei die Ergebnisse humangenetischer Untersuchungen berücksichtigt wurden, welche Gene umfassten, deren Produkte im Knochenstoffwechsel eine Rolle spielen.
In 6 Stammbäumen und einem sporadischen Fall konnten 5 verschiedene heterozygote CLCN7-Varianten festgestellt werden. Zusätzlich wurden ALPL- und LRP5-Varianten entdeckt. Der Einfluss dieser zusätzlichen Varianten auf die Penetranz und die klinische Manifestation des ADO II-Phänotyps wurden diskutiert.
Es konnte festgestellt werden, dass Betroffene in allen betrachteten Bereichen, d.h. klinisch, osteodensitometrisch und laborchemisch krankheitstypische Veränderungen zeigten, wohingegen Carrier keinerlei Auffälligkeiten aufwiesen.
Die nach Alter stratifizierte Auswertung der Funktionstestung und Lebensqualität offenbarte mit zunehmendem Alter stärkere Einbußen in der Funktionsfähigkeit und der gesundheitsbezogenen Lebensqualität bei Betroffenen gegenüber den Carriern. Eine Zunahme der Krankheitssymptome und eine Verschlechterung des Gesundheitszustandes im Laufe des Lebens könnten demnach angenommen werden.
Hematopoietic stem cell transplantation (HSCT) is a promising therapy for various malignancies and immune deficiency diseases, but it is often associated with graft versus host disease (GvHD), a life-threatening complication arising from immunological incompatibility between donor T cells and host tissues. Current standard therapies for GvHD involve the use of calcineurin inhibitors (CNIs) such as cyclosporine A (CsA) and tacrolimus (FK506), which effectively suppress T cell activation and proliferation. However, these drugs also impair the graft versus leukemia (GvL) effect, which is the advantageous ability of donor T cells to eliminate malignant cells.
Our previous studies demonstrated that the selective deletion of one or two members of the nuclear factor of activated T cells (NFAT) transcription factor family in donor T cells effectively prevented harmful GvHD without compromising GvL activity. This finding highlighted the potential of NFAT as a therapeutic target for GvHD.
In this study, we developed and evaluated novel treatment strategies that specifically target NFAT during allogeneic HSCT. We focused on the development of small molecules that mimic the PxIxIT motif of NFAT, thereby competitively inhibiting its binding to CN (CN) without affecting CN phosphatase activity. We identified two promising candidates, C17 and MRD37, and evaluated their efficacy in inhibiting NFAT and suppressing pro-inflammatory cytokine production. Among these molecules, MRD37 demonstrated the highest potency in selectively inhibiting NFAT at a sub-IC50 concentration without compromising the functional capacity of regulatory T cells (Tregs) in vitro. Furthermore, we demonstrated that MRD37 could effectively protect mice from major mismatch GvHD in vivo. This protection was initially predicted to be due to the enhanced presence of Tregs and Tr1-type cells but when pretreated T cells devoid of Tregs were transplanted it unraveled an additional increase of Th2-like cytokine release. Finally, our in vitro studies on human T cells confirmed that MRD37 could specifically inhibit NFAT while preserving the Treg population, suggesting its potential as a novel therapeutic strategy for GvHD.
Our findings provide compelling evidence for the development of MRD37 as promising alternative to CNIs in mitigating GvHD.
Analytic integration of soft and collinear radiation in factorised QCD cross sections at NNLO
(2021)
Within the framework of local analytic sector subtraction, we present the full analytic integration of double-real and real-virtual local infrared counterterms that enter NNLO QCD computations with any number of massless final-state partons. We show that a careful choice of phase-space mappings leads to simple analytic results, including non-singular terms, that can be obtained with conventional integration techniques.
Background:
Internalizing disorders are the most common psychiatric problems observed among youth in Canada. Sadly, youth with internalizing disorders often avoid seeking clinical help and rarely receive adequate treatment. Current methods of assessing internalizing disorders usually rely on subjective symptom ratings, but internalizing symptoms are frequently underreported, which creates a barrier to the accurate assessment of these symptoms in youth. Therefore, novel assessment tools that use objective data need to be developed to meet the highest standards of reliability, feasibility, scalability, and affordability. Mobile sensing technologies, which unobtrusively record aspects of youth behaviors in their daily lives with the potential to make inferences about their mental health states, offer a possible method of addressing this assessment barrier.
Objective:
This study aims to explore whether passively collected smartphone sensor data can be used to predict internalizing symptoms among youth in Canada.
Methods:
In this study, the youth participants (N=122) completed self-report assessments of symptoms of anxiety, depression, and attention-deficit hyperactivity disorder. Next, the participants installed an app, which passively collected data about their mobility, screen time, sleep, and social interactions over 2 weeks. Then, we tested whether these passive sensor data could be used to predict internalizing symptoms among these youth participants.
Results:
More severe depressive symptoms correlated with more time spent stationary (r=0.293; P=.003), less mobility (r=0.271; P=.006), higher light intensity during the night (r=0.227; P=.02), and fewer outgoing calls (r=−0.244; P=.03). In contrast, more severe anxiety symptoms correlated with less time spent stationary (r=−0.249; P=.01) and greater mobility (r=0.234; P=.02). In addition, youths with higher anxiety scores spent more time on the screen (r=0.203; P=.049). Finally, adding passively collected smartphone sensor data to the prediction models of internalizing symptoms significantly improved their fit.
Conclusions:
Passively collected smartphone sensor data provide a useful way to monitor internalizing symptoms among youth. Although the results replicated findings from adult populations, to ensure clinical utility, they still need to be replicated in larger samples of youth. The work also highlights intervention opportunities via mobile technology to reduce the burden of internalizing symptoms early on.
The neglected zoonotic disease alveolar echinococcosis (AE) is caused by the metacestode stage of the tapeworm parasite Echinococcus multilocularis. MicroRNAs (miRNAs) are small non-coding RNAs with a major role in regulating gene expression in key biological processes. We analyzed the expression profile of E. multilocularis miRNAs throughout metacestode development in vitro, determined the spatial expression of miR-71 in metacestodes cultured in vitro and predicted miRNA targets. Small cDNA libraries from different samples of E. multilocularis were sequenced. We confirmed the expression of 37 miRNAs in E. multilocularis being some of them absent in the host, such as miR-71. We found a few miRNAs highly expressed in all life cycle stages and conditions analyzed, whereas most miRNAs showed very low expression. The most expressed miRNAs were miR-71, miR-9, let-7, miR-10, miR-4989 and miR-1. The high expression of these miRNAs was conserved in other tapeworms, suggesting essential roles in development, survival, or host-parasite interaction. We found highly regulated miRNAs during the different transitions or cultured conditions analyzed, which might suggest a role in the regulation of developmental timing, host-parasite interaction, and/or in maintaining the unique developmental features of each developmental stage or condition. We determined that miR-71 is expressed in germinative cells and in other cell types of the germinal layer in E. multilocularis metacestodes cultured in vitro. MiRNA target prediction of the most highly expressed miRNAs and in silico functional analysis suggested conserved and essential roles for these miRNAs in parasite biology. We found relevant targets potentially involved in development, cell growth and death, lifespan regulation, transcription, signal transduction and cell motility. The evolutionary conservation and expression analyses of E. multilocularis miRNAs throughout metacestode development along with the in silico functional analyses of their predicted targets might help to identify selective therapeutic targets for treatment and control of AE.
Agricultural biodiversity and associated ecosystem functions are declining at alarming rates due to widespread land use intensification. They can only be maintained through targeted landscape management that supports species with different habitat preferences, dispersal capacities and other functional traits that determine their survival. However, we need better understanding whether short-term measures can already improve functional diversity in European agroecosystems.
We investigated spatio-temporal responses of bees (solitary bees, bumblebees and honey bees), hoverflies, carabid beetles and spiders to newly established grassland strips in Lower Austria over 3 years, and along a distance gradient to old grasslands. Specifically, we asked if new grasslands, compared to old grasslands and cereal fields, serve as temporal dispersal habitat or corridor, and how species-specific traits affect dispersal patterns. Using a trait-based functional diversity approach, we investigated year and distance effects for nine selected key traits per taxon (e.g. body size, feeding guild and habitat preferences).
Our results show that the functional diversity of predators and pollinators (i.e. functional richness and evenness), as well as community-weighted means of selected key traits in new grasslands significantly differed from adjacent cereal fields, but only slowly adjusted to adjacent old grasslands. These effects significantly decreased with increasing distance to old grasslands for carabids and spiders, but not for mobile bees and hoverflies.
Synthesis and applications. Over 3 years, newly established grassland strips supported larger sized and actively foraging/hunting species in the agricultural landscape. Adjacent crops likely benefit from such measures through enhanced functional diversity and related ecosystem services. However, our results also suggest that 3-year period is too short to enhance the occurrence of pollinators and epigeic predators in new grasslands. Agri-environment measures need to be complemented by the conservation of permanent habitats to effectively maintain species and functional diversity. Our findings should be acknowledged by European policy and agricultural decision makers for the design of more effective agri-environment schemes, taking into account trait-dependent species responses to land use change.