Functional Coupling of Slack Channels and P2X3 Receptors Contributes to Neuropathic Pain Processing

The sodium-activated potassium channel Slack (K_Na1.1, Slo2.2, or Kcnt1) is highly expressed in populations of sensory neurons, where it mediates the sodium-activated potassium current (I_KNa) and modulates neuronal activity. Previous studies suggest that Slack is involved in the processing of neuropathic pain. However, mechanisms underlying the regulation of Slack activity in this context are poorly understood. Using whole-cell patch-clamp recordings we found that Slack-mediated I_KNa in sensory neurons of mice is reduced after peripheral nerve injury, thereby contributing to neuropathic pain hypersensitivity. Interestingly, Slack is closely associated with ATP-sensitive P2X3 receptors in a population of sensory neurons. In vitro experiments revealed that Slack-mediated I_KNa may be bidirectionally modulated in response to P2X3 activation. Moreover, mice lacking Slack show altered nocifensive responses to P2X3 stimulation. Our study identifies P2X3/Slack signaling as a mechanism contributing to hypersensitivity after peripheral nerve injury and proposes a potential novel strategy for treatment of neuropathic pain.     

Preclinical Evaluation of 99mTc-ZHER2:41071, a Second-Generation Affibody-Based HER2-Visualizing Imaging Probe with a Low Renal Uptake

Radionuclide imaging of HER2 expression in tumours may enable stratification of patients with breast, ovarian, and gastroesophageal cancers for HER2-targeting therapies. A first-generation HER2-binding affibody molecule [^99mTc]Tc-ZHER2:V2 demonstrated favorable imaging properties in preclinical studies. Thereafter, the affibody scaffold has been extensively modified, which increased its melting point, improved storage stability, and increased hydrophilicity of the surface. In this study, a second-generation affibody molecule (designated ZHER2:41071) with a new improved scaffold has been prepared and characterized. HER2-binding, biodistribution, and tumour-targeting properties of [^99mTc]Tc-labelled ZHER2:41071 were investigated. These properties were compared with properties of the first-generation affibody molecules, [^99mTc]Tc-ZHER2:V2 and [^99mTc]Tc-ZHER2:2395. [^99mTc]Tc-ZHER2:41071 bound specifically to HER2 expressing cells with an affinity of 58 ± 2 pM. The renal uptake for [^99mTc]Tc-ZHER2:41071 and [^99mTc]Tc-ZHER2:V2 was 25–30 fold lower when compared with [^99mTc]Tc-ZHER2:2395. The uptake in tumour and kidney for [^99mTc]Tc-ZHER2:41071 and [^99mTc]Tc-ZHER2:V2 in SKOV-3 xenografts was similar. In conclusion, an extensive re-engineering of the scaffold did not compromise imaging properties of the affibody molecule labelled with ^99mTc using a GGGC chelator. The new probe, [^99mTc]Tc-ZHER2:41071 provided the best tumour-to-blood ratio compared to HER2-imaging probes for single photon emission computed tomography (SPECT) described in the literature so far. [^99mTc]Tc-ZHER2:41071 is a promising candidate for further clinical translation studies.     

Process design and modelling methods for automated handling and draping strategies for composite components

Fibre-reinforced-thermoplastics (FRT) have excellent weight-specific properties compared to conventional engineering materials. However, a wider dissemination of this technology into existing plant technologies is restrained by the low degree of automation. Complex FRT component geometries pose special challenges to gripper design and handling strategies in automated preform processes regarding limp material behaviour and fast cooling time. The preform quality is influenced by the component geometry, reinforcing fabric, and preform process. This paper presents the development of an automated handling and draping strategy, which is validated by finite-element-analysis and experimental testing to meet the requirements of large-scale preforming processes for complex geometries.     

Critical conditions for onset of mould growth under varying climate conditions

A performance-based service life design format based on climatic exposure on one hand and “resistance” of materials against mould growth on the other hand, is presented in this paper. A limit state for onset of mould growth is defined as the occurrence of traces of mould observed by microscopy. A dose–response model is proposed by which onset of mould growth can be predicted for an arbitrary climate history of combined relative humidity ? and temperature T. The model is calibrated and verified against a comprehensive set of experimental data published by Viitanen et al. [Viitanen H, Ritschkoff A-C. Mould growth in pine and spruce sapwood in relation to air humidity and temperature. Uppsala: Swedish University of Agricultural Sciences, Department of Forest Products; 1991. Report No. 221, 49 p.; Viitanen H. Modelling the time factor in the development of mould fungi – effect of critical humidity and temperature conditions in pine and spruce sapwood. Holzforschung 1997;51(1):6–14; Viitanen H. Modelling the time factor in the development of brown rot decay in pine and spruce sapwood – the effect of critical humidity and temperature conditions. Holzforschung 1997;51(2):99–106; Viitanen H, Bjurman J. Mould growth on wood under fluctuating humidity conditions. Material und Organismen 1995;29(1):27–46] describing mould development on spruce and pine sapwood as a function of climatic exposure. The model is applied to predict time to onset of mould growth under natural outdoor climate (under shelter) as well as mould development in building attics and in crawl space foundations. The predicted response shows reasonable agreement with experimental observations and proven experience, although biological processes of this type display great variability. The results show that a generally applicable, quantitative model can be used as a powerful tool for moisture safe design in practice. The model is designed to facilitate continuous improvement of prediction capability by further laboratory testing of various materials under specified climate conditions. In combination with currently available building physics software the model is suitable for moisture safe design of wood-based components in the building envelope.     

A mechanistic study of electrodeposition of bismuth telluride on stainless steel substrates

Miniaturization of Bi₂Te₃ compounds is of great interest in semiconductor industries due to their distinct anisotropic thermoelectric properties at room temperature. The aim of the present work was to investigate the mechanism of the electrodeposition of Bi₂Te₃ compounds on stainless steel substrates and relate the morphology and composition of the resulting deposits to experimental parameters. Cyclic voltammetry (CV) experiments in acidic solutions containing Bi³⁺ and/or HTeO₂⁺ ions show that the deposition potential for the Bi₂Te₃ compound is more positive than either of the single elements alone. A detailed mechanism of the co-deposition was obtained by varying the concentrations of the two elements and evaluating the corresponding morphological and compositional changes of the deposits. The results show that the deposition of Te is kinetically hindered and that Bi deposition plays a major role during the co-deposition.     

Cell‐Selective,Apoptosis‐inducing Rhodium(III) Crown Thiaether Complexes

Half‐sandwich rhodium(III) polypyridyl (pp) complexes with the metal atom capped by the facial crown thiaether 1,4,7‐trithiacyclononane [9]aneS₃ represent a promising class of apoptosis‐inducing potent cytostatic agents. The necrotic damage caused by the complexes is negligible. In vitro cytotoxicity assays with the human cancer cell lines MCF‐7 and HT‐29 and immortalized HEK‐293 cells indicate that the dicationic κ²N(imino) complexes [([9]aneS₃)RhCl(pp)]²⁺ are much more active than monocationic complexes [([9]aneS₃)RhCl₂(L)]⁺ (L=imidazole, CH₃CN). Whereas the κ²N(amino) complex [([9]aneS₃)RhCl(piperazine)]²⁺ is inactive, replacing piperazine with the structurally analogous κ²S (thiaether) ligand 1,4‐dithiane restores cytotoxicity as evidenced by IC₅₀ values in the range 8.1‐11.6 μM . Spectroscopic (CD, UV/Vis, NOESY) and viscosity measurements indicate that the active dppz complex 8 (IC₅₀ values: 4.7–8.9 μM ) exhibits strong intercalative binding towards DNA whereas the even more potent bipyrimidine complex 9 (IC₅₀ values: 0.6–1.9 μM ) causes no alteration of the duplex B conformation. Weaker intercalative binding is observed for the dpq complex 7 . A comparative annexin V–propidium iodide binding assay with lymphoma (BJAB) cells and healthy leukocytes demonstrates that the cytotoxic activity of complex 8 and particularly complex 9 is highly selective towards the malignant cells.     

On the physics of power,energy and economics of renewable electric energy sources - Part II

Renewable Energy Technologies (RETs) are often recognized as less competitive than traditional electric energy conversion systems. Obstacles with renewable electric energy conversion systems are often referred to the intermittency of the energy sources [1] and the relatively high maintenance cost. However, due to an intensified discourse on climate change and its effects, it has from a societal point of view, become more desirable to adopt and install CO₂ neutral power plants. Even if this has increased the competitiveness of RETs in a political sense, the new goals for RET installations must also be met with economical viability. We propose that the direction of technical development, as well as the chosen technology in new installations, should not primarily be determined by policies, but by the basic physical properties of the energy source and the associated potential for inexpensive energy production. This potential is the basic entity that drives the payback of the investment of a specific RET power plant. With regard to this, we argue that the total electric energy conversion system must be considered if effective power production is to be achieved, with focus on the possible number of full loading hours and the Degree of Utilization [2]. This will increase the cost efficiency and economical competitiveness of RET investments, and could enhance faster diffusion of new innovations and installations without over-optimistic subsidies. This paper elaborates on the overall problem of the economy of renewable electric energy conversion systems by studying the interface between physics, engineering and economy reported for RET power plants in different scientific publications. The core objective is to show the practical use of the Degree of Utilization and how the concept is crucial for the design and economical optimization disregarding subsidies. The results clearly indicate that the future political regulative frameworks should consider the choice of renewable energy source since this strongly affects the economical output from the RET power plants.     

Spectroscopic and in vitro Investigations of Fe2+/α-Ketoglutarate-Dependent Enzymes Involved in Nucleic Acid Repair and Modification

The activation of molecular oxygen for the highly selective functionalization and repair of DNA and RNA nucleobases is achieved by α-ketoglutarate (α-KG)/iron-dependent dioxygenases. Of special interest are the human homologues AlkBH of Escherichia coli EcAlkB and ten-eleven translocation (TET) enzymes. These enzymes are involved in demethylation or dealkylation of DNA and RNA, although additional physiological functions are continuously being found. Given their importance, studying enzyme-substrate interactions, turnover and kinetic parameters is pivotal for the understanding of the mode of action of these enzymes. Diverse analytical methods, including X-ray crystallography, UV/Vis absorption, electron paramagnetic resonance (EPR), circular dichroism (CD) and nuclear magnetic resonance (NMR) spectroscopy have been employed to study the changes in the active site and the overall enzyme structure upon substrate, cofactor, and inhibitor addition. Several methods are now available to assess the activity of these enzymes. By discussing limitations and possibilities of these techniques for EcAlkB, AlkBH and TET we aim to give a comprehensive synopsis from a bioinorganic point-of-view, addressing researchers from different disciplines working in the highly interdisciplinary and rapidly evolving field of epigenetic processes and DNA/RNA repair and modification.     

Production of the Carboxylate Reductase from Nocardia otitidiscaviarum in a Soluble,Active Form for in vitro Applications

Accessing aldehydes from carboxylate moieties is often a challenging task. In this regard, carboxylate reductases (CARs) are promising catalysts provided by nature that are able to accomplish this task in just one step, avoiding over-reduction to the alcohol product. However, the heterologous expression of CARs can be quite difficult due to the excessive formation of insoluble protein, thus hindering further characterization and application of the enzyme. Here, the heterologous production of the carboxylate reductase from Nocardia otitidiscaviarum (NoCAR) was optimized by a combination of i) optimized cultivation conditions, ii) post-translational modification with a phosphopantetheinyl transferase and iii) selection of an appropriate expression strain. Especially, the selection of Escherichia coli tuner cells as host had a strong effect on the final 110-fold increase in the specific activity of NoCAR. This highly active NoCAR was used to reduce sodium benzoate to benzaldehyde, and it was successfully assembled with an in vitro regeneration of ATP and NADPH, being capable of reducing about 30 mM sodium benzoate with high selectivity in only 2 h of reaction.