Ligand binding and conformational dynamics of the nicotinamide nucleotide transhydrogenase revealed by hydrogen/deuterium exchange mass spectrometry.

Nicotinamide nucleotide transhydrogenases are integral membrane proteins that utilizes the proton motive force to reduce NADP to NADPH while converting NADH to NAD. Atomic structures of various transhydrogenases in different ligand-bound states have become available, and it is clear that the molecular mechanism involves major conformational changes. Here we utilized hydrogen/deuterium exchange mass spectrometry (HDX-MS) to map ligand binding sites and analyzed the structural dynamics of transhydrogenase.

Short-chain aurachin D derivatives are selective inhibitors of E. coli cytochrome bd-I and bd-II oxidases.

Cytochrome bd-type oxidases play a crucial role for survival of pathogenic bacteria during infection and proliferation. This role and the fact that there are no homologues in the mitochondrial respiratory chain qualify cytochrome bd as a potential antimicrobial target. However, few bd oxidase selective inhibitors have been described so far.

Mechanistic and structural diversity between cytochrome isoforms of .

The treatment of infectious diseases caused by multidrug-resistant pathogens is a major clinical challenge of the 21st century. The membrane-embedded respiratory cytochrome -type oxygen reductase is a critical survival factor utilized by pathogenic bacteria during infection, proliferation and the transition from acute to chronic states. encodes for two cytochrome isoforms that are both involved in respiration under oxygen limited conditions.

The cryo-EM structure of the bd oxidase from M. tuberculosis reveals a unique structural framework and enables rational drug design to combat TB.

New drugs are urgently needed to combat the global TB epidemic. Targeting simultaneously multiple respiratory enzyme complexes of Mycobacterium tuberculosis is regarded as one of the most effective treatment options to shorten drug administration regimes, and reduce the opportunity for the emergence of drug resistance. During infection and proliferation, the cytochrome bd oxidase plays a crucial role for mycobacterial pathophysiology by maintaining aerobic respiration at limited oxygen concentrations.

Reactivation during sleep with incomplete reminder cues rather than complete ones stabilizes long-term memory in humans.

Reactivation by reminder cues labilizes memories during wakefulness, requiring reconsolidation to persist. In contrast, during sleep, cued reactivation seems to directly stabilize memories. In reconsolidation, incomplete reminders are more effective in reactivating memories than complete reminders by inducing a mismatch, i.

Synthesis and Biological Screening of New Lawson Derivatives as Selective Substrate-Based Inhibitors of Cytochrome bo Ubiquinol Oxidase from Escherichia coli.

The respiratory chain of Escherichia coli contains two different types of terminal oxidase that are differentially regulated as a response to changing environmental conditions. These oxidoreductases catalyze the reduction of molecular oxygen to water and contribute to the proton motive force. The cytochrome bo oxidase (cyt bo ) acts as the primary terminal oxidase under atmospheric oxygen levels, whereas the bd-type oxidase is most abundant under microaerobic conditions.

Active site rearrangement and structural divergence in prokaryotic respiratory oxidases.

Cytochrome bd-type quinol oxidases catalyze the reduction of molecular oxygen to water in the respiratory chain of many human-pathogenic bacteria. They are structurally unrelated to mitochondrial cytochrome c oxidases and are therefore a prime target for the development of antimicrobial drugs. We determined the structure of the cytochrome bd-I oxidase by single-particle cryo-electron microscopy to a resolution of 2.

Implementation and first results of a tablet-based assessment referring to patient-reported outcomes in an inpatient cancer care unit.

Background: Inclusion of patient-reported outcomes (PROs) in routine cancer care is of key importance for individualized treatment, shared decision making and patient satisfaction.

Objective: To describe the implementation under routine conditions of an electronic self-administered PRO assessment and comparison of PROs before and after inpatient treatment in oncologic care.

Methods: In a tablet-based survey PROs on symptom burden, global health status/ quality of life (QoL) and health utility were collected twice (at hospital admission and discharge) in an inpatient oncological setting over a 17-month period using the EORTC QLQ-C30 and EQ-5D questionnaires.

The role of GABA in the expression of updated information through the reconsolidation process in humans.

Consolidated memory can be again destabilized by the presentation of a memory cue (reminder) of the previously acquired information. During this process of labilization/restabilization memory traces can be either impaired, strengthened or updated in content. Here, we study if a consolidated memory can be updated by linking one original cue to two different outcomes and whether this process was modulated by the GABAergic system.

Implementation of a mobile inpatient quality of life (QoL) assessment for oncology nursing.

Purpose: Cancer patients suffer from a variety of symptoms, but little is known about changes during hospitalization and symptom burden at discharge. We implemented an electronic quality of life (QoL) assessment used by the nursing team in routine inpatient care. Feasibility, acceptance, and the course of QoL were investigated.

Activation of Mitochondrial Complex II-Dependent Respiration Is Beneficial for α-Synucleinopathies.

Parkinson's disease and dementia with Lewy bodies are major challenges in research and clinical medicine world-wide and contribute to the most common neurodegenerative disorders. Previously, specific mitochondrial polymorphisms have been found to enhance clearance of amyloid-β from the brain of APP-transgenic mice leading to beneficial clinical outcome. It has been discussed whether specific mitochondrial alterations contribute to disease progression or even prevent toxic peptide deposition, as seen in many neurodegenerative diseases.

A novel glucokinase activator modulates pancreatic islet and hepatocyte function.

The glucose-sensing enzyme glucokinase (GK) plays a key role in glucose metabolism. We report here the effects of a novel glucokinase activator, LY2121260. The activator enhanced GK activity via binding to the allosteric site located in the hinge region of the enzyme.

Epilepsy and sports participation.

Epilepsy is a common disease found in 2% of the population, affecting both young and old. Unfortunately, epileptics have previously been discouraged from participation in physical activity and sports for fear of inducing seizures or increasing seizure frequency. Despite a shift in medical recommendations toward encouraging rather than restricting participation, the stigma remains and epileptics continue to be less active than the general population.

Protein phosphorylation in alveolar macrophages after stimulation with heavy metal-coated silica particles.

The aim of our study was to determine agonist- and time-dependent phosphorylation of distinct proteins in bovine alveolar macrophages after activation with quartz dusts and heavy metal-coated silica particles. Silica particles were coated with vanadium oxide or cadmium oxide. Separation of proteins was achieved by one-dimensional and optimized horizontal two-dimensional polyacrylamide gel electrophoresis.

Hsp27 phosphorylation is induced in alveolar macrophages exposed to CdO-coated silica particles.

Protein phosphorylation in bovine alveolar macrophages (BAM) activated by quartz dusts and metal oxide-coated silica particles was investigated by means of two-dimensional electrophoresis (2D-PAGE) and densitometric 32[P]-phosphate image analysis. BAM activity was monitored by determining generated superoxide anions and hydrogen peroxide. In vitro stimulation of BAM with cadmium oxide-coated silica particles (LiC-CdO) resulted in characteristic time-dependent changes in 2D-PAGE spot patterns, that were similar to the effects induced by 4ss-phorbol-12-myristate-13-acetate (PMA).

Nanogram detection of phospholipids on thin-layer chromatograms.

An extremely sensitive method for the quantitative determination of phospholipids on silica gel-precoated thin-layer chromatography plates is reported. The procedure is based on the successive application of two lipid spray reagents. Firstly, phospholipid chromatograms are sprayed with the lipophilic fluorochrome 1,6-diphenyl-1,3,5-hexatriene, followed by application of the phosphorus-specific molybdenum blue (Dittmer-Lester) reagent.

Protein kinase C activity and phosphoprotein pattern in stimulated alveolar macrophages.

Bovine alveolar macrophages (BAM) were stimulated with quartz dusts, metal oxide-coated silica particles, and zymosan. To investigate the role of protein kinase C (PKC) in the mechanism of agonist-induced activation 12-O-tetradecanoyl phorbol 13-acetate (TPA), staurosporine, and the PKC specific inhibitor GF 109203X were applied. PKC activity was determined by means of a continuous fluorescence assay [1].