A Detailed Analysis of the Morphology of Fibrils of Selectively Mutated Amyloid β (1–40)

A small library of rationally designed amyloid β [Aβ(1–40)] peptide variants is generated, and the morphology of their fibrils is studied. In these molecules, the structurally important hydrophobic contact between phenylalanine 19 (F19) and leucine 34 (L34) is systematically mutated to introduce defined physical forces to act as specific internal constraints on amyloid formation. This Aβ(1–40) peptide library is used to study the fibril morphology of these variants by employing a comprehensive set of biophysical techniques including solution and solid‐state NMR spectroscopy, AFM, fluorescence correlation spectroscopy, and XRD. Overall, the findings demonstrate that the introduction of significant local physical perturbations of a crucial early folding contact of Aβ(1–40) only results in minor alterations of the fibrillar morphology. The thermodynamically stable structure of mature Aβ fibrils proves to be relatively robust against the introduction of significantly altered molecular interaction patterns due to point mutations. This underlines that amyloid fibril formation is a highly generic process in protein misfolding that results in the formation of the thermodynamically most stable cross‐β structure.     

Distinguishing two isomeric mephedrone substitutes with selective reagent ionisation mass spectrometry (SRI‐MS)

The isomers 4‐methylethcathinone and N‐ethylbuphedrone are substitutes for the recently banned drug mephedrone. We find that with conventional proton transfer reaction mass spectrometry (PTR‐MS), it is not possible to distinguish between these two isomers, because essentially for both substances, only the protonated molecules are observed at a mass‐to‐charge ratio of 192 (C₁₂H₁₈NO⁺). However, when utilising an advanced PTR‐MS instrument that allows us to switch the reagent ions (selective reagent ionisation) from H₃O⁺ (which is commonly used in PTR‐MS) to NO⁺, O₂⁺ and Kr⁺, characteristic product (fragment) ions are detected: C₄H₁₀N⁺ (72 Da) for 4‐methylethcathinone and C₅H₁₂N⁺ (86 Da) for N‐ethylbuphedrone; thus, selective reagent ionisation MS proves to be a powerful tool for fast detection and identification of these compounds. Copyright © 2013 John Wiley & Sons, Ltd.     

Rapid characterization of dry cured ham produced following different PDOs by proton transfer reaction time of flight mass spectrometry (PTR-ToF-MS)

In the present study, the recently developed proton transfer reaction time of flight mass spectrometry (PTR-ToF-MS) technique was used for the rapid characterization of dry cured hams produced according to 4 of the most important Protected Designations of Origin (PDOs): an Iberian one (Dehesa de Extremadura) and three Italian ones (Prosciutto di San Daniele, Prosciutto di Parma and Prosciutto Toscano). In total, the headspace composition and respective concentration for nine Spanish and 37 Italian dry cured ham samples were analyzed by direct injection without any pre-treatment or pre-concentration. Firstly, we show that the rapid PTR-ToF-MS fingerprinting in conjunction with chemometrics (Principal Components Analysis) indicates a good separation of the dry cured ham samples according to their production process and that it is possible to set up, using data mining methods, classification models with a high success rate in cross validation. Secondly, we exploited the higher mass resolution of the new PTR-ToF-MS, as compared with standard quadrupole based versions, for the identification of the exact sum formula of the mass spectrometric peaks providing analytical information on the observed differences. The work indicates that PTR-ToF-MS can be used as a rapid method for the identification of differences among dry cured hams produced following the indications of different PDOs and that it provides information on some of the major volatile compounds and their link with the implemented manufacturing practices such as rearing system, salting and curing process, manufacturing practices that seem to strongly affect the final volatile organic profile and thus the perceived quality of dry cured ham.     

Extending the dynamic range of proton transfer reaction time-of-flight mass spectrometers by a novel dead time correction

Proton transfer reaction time-of-flight mass spectrometry (PTR-TOF-MS) allows for very fast simultaneous monitoring of volatile organic compounds (VOCs) in complex environments. In several applications, food science and food technology in particular, peaks with very different intensities are present in a single spectrum. For VOCs, the concentrations range from the sub-ppt all the way up to the ppm level. Thus, a large dynamic range is necessary. In particular, high intensity peaks are a problem because for them the linear dependency of the detector signal on VOC concentration is distorted. In this paper we present, test with real data, and discuss a novel method which extends the linearity of PTR-TOF-MS for high intensity peaks far beyond the limit allowed by the usual analytical correction methods such as the so-called Poisson correction. Usually, raw data can be used directly without corrections with an intensity of up to about 0.1 ions/pulse, and the Poisson correction allows the use of peaks with intensities of a few ions/pulse. Our method further extends the linear range by at least one order of magnitude. Although this work originated from the necessity to extend the dynamic range of PTR-TOF-MS instruments in agro-industrial applications, it is by no means limited to this area, and can be implemented wherever dead time corrections are an issue.     

Direct determination of glucose, lactate and triglycerides in blood serum by a tunable quantum cascade laser-based mid-IR sensor

This work reports on a compact sensor for fast and reagent-free point-of-care determination of glucose, lactate and triglycerides in blood serum based on a tunable (1030–1230 cm^?1) external-cavity quantum cascade laser (EC-QCL). For simple and robust operation a single beam set-up was designed and only thermoelectric cooling was used for the employed laser and detector. Full computer control of analysis including liquid handling and data analysis facilitated routine measurements. A high optical pathlength (>100 μm) is a prerequisite for robust measurements in clinical practice. Hence, the optimum optical pathlength for transmission measurements in aqueous solution was considered in theory and experiment. The experimentally determined maximum signal-to-noise ratio (SNR) was around 140 μm for the QCL blood sensor and around 50 μm for a standard FT-IR spectrometer employing a liquid nitrogen cooled mercury cadmium telluride (MCT) detector. A single absorption spectrum was used to calculate the analyte concentrations simultaneously by using a partial-least-squares (PLS) regression analysis. Glucose was determined in blood serum with a prediction error (RMSEP) of 6.9 mg/dl and triglycerides with an error of cross-validation (RMSECV) of 17.5 mg/dl in a set of 42 different patients. In spiked serum samples the lactate concentration could be determined with an RMSECV of 8.9 mg/dl.     

Dissociative Electron Attachment to β‐Alanine

A detailed study on dissociative electron attachment (DEA) to β‐alanine (βA) in the gas phase is presented. Ion yields as a function of the incident electron energy from about 0 to 15 eV have been measured for most of the fragments. As for all α‐amino acids, the main reaction corresponds to the loss of a hydrogen atom, although many other fragments have been observed that involved more complex bond cleavages. Threshold energies have been calculated by using the G4(MP2) method for various decomposition reactions. Fragmentation pathways were also investigated to measure metastable decays of the intermediate fragment anion (βA?H)^? by using the mass‐analyzed ion kinetic energy (MIKE) scan technique. Comparisons with α‐alanine and other amino acids are made when relevant.     

Ruthenium-benzocrownether complexes: Synthesis, structures, catalysis and immobilisation in ionic liquids

A facile pathway to [RuCl₂(η⁶-benzocrownether)]₂ complexes is described and crystal structures of the complexes [RuCl₂(η⁶-benzo-15-crown-5)]₂ and [RuCl₂(η⁶-dibenzo-18-crown-6)]₂ are reported. The complexes were derivatised with (1S,2R)-2-amino-1,2-diphenylethanol and evaluated in the enantioselective transfer-hydrogenation of acetophenone. The effect of complexation of different alkaline metals (Na, K, Cs) within the crown on the selectivity and reaction rate was studied. Interaction of a sulfonated phosphine ligand with the crown was probed by NOESY-NMR and utilisation of the crownether to serve as an anchor for catalyst immobilisation was investigated.     

Twistor spaces and spinors over loop spaces

In this paper two natural twistor spaces over the loop space of a Riemannian manifold are constructed and their equivalence is shown in the Kählerian case. This relies on a detailed study of frame bundles of loop spaces on the one hand and, on the other hand, on an explicit local trivialization of the Atiyah operator family [defined in Atiyah (SMF 131:43–59, 1985)] associated to a loop space. We relate these constructions to the Dixmier-Douady obstruction class against the existence of a string structure, as well as to pseudo - line bundle gerbes in the sense of Brylinski (Loop spaces, characteristic classes and geometric quantization. Birkhäuser, Basel, 1993).     

Hybrid Chelator-Based PSMA Radiopharmaceuticals: Translational Approach

(1) Background: Prostate-specific membrane antigen (PSMA) has been extensively studied in the last decade. It became a promising biological target in the diagnosis and therapy of PSMA-expressing cancer diseases. Although there are several radiolabeled PSMA inhibitors available, the search for new compounds with improved pharmacokinetic properties and simplified synthesis is still ongoing. In this study, we developed PSMA ligands with two different hybrid chelators and a modified linker. Both compounds have displayed a promising pharmacokinetic profile. (2) Methods: DATA^5m.SA.KuE and AAZTA⁵.SA.KuE were synthesized. DATA^5m.SA.KuE was labeled with gallium-68 and radiochemical yields of various amounts of precursor at different temperatures were determined. Complex stability in phosphate-buffered saline (PBS) and human serum (HS) was examined at 37 °C. Binding affinity and internalization ratio were determined in in vitro assays using PSMA-positive LNCaP cells. Tumor accumulation and biodistribution were evaluated in vivo and ex vivo using an LNCaP Balb/c nude mouse model. All experiments were conducted with PSMA-11 as reference. (3) Results: DATA^5m.SA.KuE was synthesized successfully. AAZTA⁵.SA.KuE was synthesized and labeled according to the literature. Radiolabeling of DATA^5m.SA.KuE with gallium-68 was performed in ammonium acetate buffer (1 M, pH 5.5). High radiochemical yields (>98%) were obtained with 5 nmol at 70 °C, 15 nmol at 50 °C, and 60 nmol (50 µg) at room temperature. [⁶⁸Ga]Ga-DATA^5m.SA.KuE was stable in human serum as well as in PBS after 120 min. PSMA binding affinities of AAZTA⁵.SA.KuE and DATA^5m.SA.KuE were in the nanomolar range. PSMA-specific internalization ratio was comparable to PSMA-11. In vivo and ex vivo studies of [¹⁷⁷Lu]Lu-AAZTA⁵.SA.KuE, [⁴⁴Sc]Sc-AAZTA⁵.SA.KuE and [⁶⁸Ga]Ga-DATA^5m.SA.KuE displayed specific accumulation in the tumor along with fast clearance and reduced off-target uptake. (4) Conclusions: Both KuE-conjugates showed promising properties especially in vivo allowing for translational theranostic use.     

Properization: constructing proper scoring rules via Bayes acts

Scoring rules serve to quantify predictive performance. A scoring rule is proper if truth telling is an optimal strategy in expectation. Subject to customary regularity conditions, every scoring rule can be made proper, by applying a special case of the Bayes act construction studied by Grünwald and Dawid (Ann Stat 32:1367–1433, 2004) and Dawid (Ann Inst Stat Math 59:77–93, 2007), to which we refer as properization. We discuss examples from the recent literature and apply the construction to create new types, and reinterpret existing forms, of proper scoring rules and consistent scoring functions. In an abstract setting, we formulate sufficient conditions under which Bayes acts exist and scoring rules can be made proper.