Advanced EPI-X4 Derivatives Covalently Bind Human Serum Albumin Resulting in Prolonged Plasma Stability

Advanced derivatives of the Endogenous Peptide Inhibitor of CXCR4 (EPI-X4) have shown therapeutic efficacy upon topical administration in animal models of asthma and dermatitis. Here, we studied the plasma stability of the EPI-X4 lead compounds WSC02 and JM#21, using mass spectrometry to monitor the chemical integrity of the peptides and a functional fluorescence-based assay to determine peptide function in a CXCR4-antibody competition assay. Although mass spectrometry revealed very rapid disappearance of both peptides in human plasma within seconds, the functional assay revealed a significantly higher half-life of 9 min for EPI-X4 WSC02 and 6 min for EPI-X4 JM#21. Further analyses demonstrated that EPI-X4 WSC02 and EPI-X4 JM#21 interact with low molecular weight plasma components and serum albumin. Albumin binding is mediated by the formation of a disulfide bridge between Cys10 in the EPI-X4 peptides and Cys34 in albumin. These covalently linked albumin–peptide complexes have a higher stability in plasma as compared with the non-bound peptides and retain the ability to bind and antagonize CXCR4. Remarkably, chemically synthesized albumin-EPI-X4 conjugates coupled by non-breakable bonds have a drastically increased plasma stability of over 2 h. Thus, covalent coupling of EPI-X4 to albumin in vitro before administration or in vivo post administration may significantly increase the pharmacokinetic properties of this new class of CXCR4 antagonists.     

LXRα Regulates oxLDL-Induced Trained Immunity in Macrophages

Reprogramming of metabolic pathways in monocytes and macrophages can induce a proatherosclerotic inflammatory memory called trained innate immunity. Here, we have analyzed the role of the Liver X receptor (LXR), a crucial regulator of metabolism and inflammation, in oxidized low-density lipoprotein (oxLDL)-induced trained innate immunity. Human monocytes were incubated with LXR agonists, antagonists, and oxLDL for 24 h. After five days of resting time, cells were restimulated with the TLR-2 agonist Pam3cys. OxLDL priming induced the expression of LXRα but not LXRβ. Pharmacologic LXR activation was enhanced, while LXR inhibition prevented the oxLDL-induced inflammatory response. Furthermore, LXR inhibition blocked the metabolic changes necessary for epigenetic reprogramming associated with trained immunity. In fact, enrichment of activating histone marks at the IL-6 and TNFα promotor was reduced following LXR inhibition. Based on the differential expression of the LXR isoforms, we inhibited LXRα and LXRβ genes using siRNA in THP1 cells. As expected, siRNA-mediated knock-down of LXRα blocked the oxLDL-induced inflammatory response, while knock-down of LXRβ had no effect. We demonstrate a specific and novel role of the LXRα isoform in the regulation of oxLDL-induced trained immunity. Our data reveal important aspects of LXR signaling in innate immunity with relevance to atherosclerosis formation.     

Determination of mercury methylation potentials in the water column of lakes across Canada

A stable isotope technique was used to trace the formation of methylmercury in lake water incubation assays at in situ conditions in five lakes across Canada. Methylation activity was only detected in the anoxic hypolimnia of lakes. The stable isotope was methylated at varying rates between lakes and depths within lakes ranging from 0.56%/day to 14.8%/day. A peak in methylation potential was typically observed just below the oxycline, which decreased with increasing depth. The depth and rates of methylation potential changed seasonally with no methylation activity occurring after fall turnover. A decrease in the sulfate concentration was concomitant with the zone of mercury methylation potential indicating the likely involvement of sulfate reducing bacteria in the methylation process. A simple correlation test between DOC concentrations and methylation rates indicated a positive relationship (r2=0.62; p=0.006; n=27). The demethylation rate constant in the anoxic hypolimnia was less than 0.12 d(-1).     

A paradoxical flame-retardant effect of nitrates in ATH-filled ethylene–vinyl acetate copolymer

In the course of a study of metal salts as flame retardants, it was surprisingly found that metal nitrates reduced the flammability of ATH-filled ethylene–vinyl acetate copolymer (EVA). The limiting oxygen index (LOI) of ATH-filled EVA was increased by the nitrates in the order of Cu(NO₃)₂·3H₂O > (NH₄)₂Ce(NO₃)₆ > Zn(NO₃)₂·6H₂O > Fe(NO₃)₃·9H₂O > Al(NO₃)₃·9H₂O > NaNO₃. The effects were not caused by the water of hydration. All metal nitrates except NaNO₃ reached a UL 94 V-2 rating at 3 phr. Based on TGA, DSC, FTIR, and gas detection, the proposed mechanism of the flame-retardant effect of nitrates is the oxidative degradation of the polymer to produce noncombustible products (CO₂ and nitrogen oxides) at a rate sufficient to interfere with the normal combustion process despite the exothermicity of the oxidative degradation. It is possible that surface carboxylic acid structures also contribute to the flame-retardant effect. © 1995 John Wiley & Sons, Inc.     

Die Sturzflut von Braunsbach – Ingenieuranalyse der Gebäudeschäden

The flash flood of Braunsbach – engineering analysis of the building damage At the end of May to the beginning of June, a series of extreme rainfall events occurred in the southern part of Germany, which triggered some severe flash floods with significant building damage in the affected municipalities. The devastating flash flood of May 29, 2016 in Braunsbach in the district Schwäbisch Hall in Baden‐Württemberg caused heavy structural damage to some of the approximately 130 affected buildings. The paper gives an overview of the building damages documented immediately after the event. The damages cases were classified and assessed using the evaluation system of the EDAC‐flood damage model developed at the Earthquake Damage Analysis Center (EDAC) of the Bauhaus‐Universität Weimar. The analysis illustrates the particularities of the damage patterns due to flash floods compared to conventional river floods.     

Electron energy loss spectroscopy for analysis of inhaled ultrafine particles in rat lungs

Epidemiologic studies have associated cardiovascular morbidity and mortality with ambient particulate air pollution. Particles smaller than 100 nm in diameter (ultrafine particles) are present in the urban atmosphere in very high numbers yet at very low mass concentration. Organs beyond the lungs are considered as targets for inhaled ultrafine particles, whereby the route of particle translocation deeper into the lungs is unclear. Five rats were exposed to aerosols of ultrafine titanium dioxide particles of a count median diameter of 22 nm (geometric standard deviation, GSD 1.7) for 1 hour. The lungs were fixed by intravascular perfusion of fixatives immediately thereafter. TiO(2) particles in probes of the aerosol as well as in systematic tissue samples were analyzed with a LEO 912 transmission electron microscope equipped with an energy filter for elemental microanalysis. The characteristic energy loss spectra were obtained by fast spectrum acquisition. Aerosol particles as well as those in the lung tissue were unambiguously identified by electron energy loss spectroscopy. Particles were mainly found as small clusters with a rounded shape. Seven percent of the particles in the lung tissue had a needle-like shape. The size distribution of the cluster profiles in the tissue had a count median diameter of 29 nm (GSD 1.7), which indicates no severe clustering or reshaping of the originally inhaled particles. Electron energy loss spectroscopy and related analytical methods were found to be suitable to identify and localize ultrafine titanium dioxide particles within chemically fixed and resin-embedded lung tissue.     

Ca15(CBN)6(C2)2O- a new cubic structure type containing CBN-anions together with acetylide and oxo anions

Ca₁₅(CBN)₆(C₂)₂O contains CBN⁴⁻ anions as well as C₂⁻ units and isolated O²⁻ anions. The compound is obtained by heating a stoichiometric mixture of CaO, C and BN with an excess of Ca in sealed niobium ampoules to 1270 K. It crystallizes in the space group Ia d with a = 1656.84(9) pm. Preparation, crystal structure, NMR and IR-spectroscopic properties are discussed.     

Predicting and tuning physicochemical properties in lead optimization: amine basicities

This review describes simple and useful concepts for predicting and tuning the pK(a) values of basic amine centers, a crucial step in the optimization of physical and ADME properties of many lead structures in drug-discovery research. The article starts with a case study of tricyclic thrombin inhibitors featuring a tertiary amine center with pK(a) values that can be tuned over a wide range, from the usual value of around 10 to below 2 by (remote) neighboring functionalities commonly encountered in medicinal chemistry. Next, the changes in pK(a) of acyclic and cyclic amines upon substitution by fluorine, oxygen, nitrogen, and sulfur functionalities, as well as carbonyl and carboxyl derivatives are systematically analyzed, leading to the derivation of simple rules for pK(a) prediction. Electronic and stereoelectronic effects in cyclic amines are discussed, and the emerging computational methods for pK(a) predictions are briefly surveyed. The rules for tuning amine basicities should not only be of interest in drug-discovery research, but also to the development of new crop-protection agents, new amine ligands for organometallic complexes, and in particular, to the growing field of amine-based organocatalysis.     

Compatibilisation of PPE/SAN blends by triblock terpolymers: Correlation between block terpolymer composition, morphology and properties

Immiscible blends of poly(2,6-dimethyl-1,4-phenylene ether) (PPE) and poly(styrene-co-acrylonitrile) (SAN) with a weight composition of 60/40 were compatibilised by polystyrene-block-polybutadiene-block-poly(methyl methacrylate) triblock terpolymers (SBM) using a two-stage melt-processing approach. In order to investigate the influence of the SBM composition on the compatibilisation efficiency, the block lengths of the triblock terpolymers were systematically varied. The resulting morphological features of the blend systems as function of SBM composition and processing parameters are correlated with the resulting thermal and thermo-mechanical properties. In the ideal case, SBM should be located at the interface as PS is miscible with PPE while PMMA is miscible with SAN. The elastomeric middle block as an immiscible component should remain at the interface. This particular morphological arrangement is known as the ‘raspberry morphology’. A detailed TEM analysis of the blend morphologies following initial extrusion-compounding revealed a high compatibilisation efficiency of the SBM types with equal lengths of the end blocks and, furthermore, the desired raspberry morphology was achieved. In contrast, high PS contents in comparison to the other blocks led to a pronounced micelle formation in the PPE phase. Further evaluation of the blend structures following injection-moulding indicated that the morphologies remain relatively stable during this second melt-processing step. A detailed thermal analysis of all blend systems supports the interpretation of the observed morphological features. The fundamental correlation between SBM composition and blend morphology established in this study opens the door for the controlled development of interfacial properties of such compatibilised PPE/SAN blends during melt-processing.