Bacterial Predators Possess Unique Membrane Lipid Structures

Bdellovibrio-and-like organisms (BALO) are a phylogenetically diverse group of predatory prokaryotes that consists of the two families Bdellovibrionaceae and Bacteriovoracaceae. We investigated the phospholipid composition of the three important BALO strains Bacteriovorax stolpii (DSM 12778), Bdellovibrio bacteriovorus HD100 (DSM 50701) and Peredibacter starrii (DSM 17039). We confirmed the presence of sphingophosphonolipids in B. stolpii, while we characterized sphingophosphonolipids with a 2-amino-3-phosphonopropanate head group for the first time. In B. bacteriovorus HD100 phosphatidylthreonines were found and, thus, B. bacteriovorus is the second prokaryote investigated so far possessing this rare lipid class. In the third analyzed organism, P. starrii, we observed phosphatidylethanolamine structures with an additional N-glutamyl residue, which form the first reported class of amino acid-containing phosphatidylethanolamines.     

A standardized and biocompatible preparation of aggregate-free amyloid beta peptide for biophysical and biological studies of Alzheimer's disease

We provide a validated and rapid protocol for the solubilization of amyloid β-peptide (Aβ). This procedure involves sequential solubilization using structure-breaking organic solvents hexafluoroisopropanol and DMSO followed by column purification. The low solubility and tendency of Aβ to aggregate considerably impede the in vitro handling and biophysical or biological investigation of Aβ, despite the interest in this peptide because of its implication in Alzheimer's disease. The main advantage of the proposed protocol over others is that it results in standardized aggregate-free Aβ peptide samples that are biocompatible for cell culture studies and yield reproducible aggregation kinetics and cytotoxicities. This three-step protocol also enables the co-solubilization of the longer Aβ42 variant with Aβ40 in ratios relevant to Alzheimer's disease.     

Interlaboratory diagnostic accuracy of a Salmonella specific PCR-based method

A collaborative study involving four European laboratories was conducted to investigate the diagnostic accuracy of a Salmonella specific PCR-based method, which was evaluated within the European FOOD-PCR project (http://www.pcr.dk). Each laboratory analysed by the PCR a set of independent obtained presumably naturally contaminated samples and compared the results with the microbiological culture method. The PCR-based method comprised a preenrichment step in buffered peptone water followed by a thermal cell lysis using a closed tube resin-based method. Artificially contaminated minced beef and whole broiler carcass-rinse resulted in a detection limit of less than 5 cells per 25 g meat or 100 ml broiler rinse. A total of 435 samples from four countries, including pig carcass swabs (n = 285), whole broiler carcass-rinse (n = 25), various raw meat (n = 33), and environmental samples (n = 92) were investigated. The interlaboratory diagnostic accuracy, i.e. diagnostic specificity and sensitivity, was shown to be 97.5%. The co-amplification of an internal amplification control indicated possible inhibitory substances derived from the sample. This work can contribute to the quality assurance of PCR-based diagnostic methods and is currently proposed as international standard document.     

In-situ electrical addressing of one-dimensional gold nanoparticle assemblies

Substrates with 1-dimensional nanosize grooves were prepared using extreme-ultraviolet interference lithography (EUV-IL), wherein gold nanoparticles were self-assembled to form 1-dimensional structures. To measure the electrical properties of gold nanoparticle chains we introduce a novel in-situ measuring method based on nanomanipulator system in a scanning electron microscope. This method comprises enormous versatility for the precisely electrical addressing of low-dimensional nanoscale structures and may even be applied to routinely addressing of structures in the sub-10 nm range.