Nitric Oxide Signalling in Descending Vasa Recta after Hypoxia/Re-Oxygenation

Reduced renal medullary oxygen supply is a key factor in the pathogenesis of acute kidney injury (AKI). As the medulla exclusively receives blood through descending vasa recta (DVR), dilating these microvessels after AKI may help in renoprotection by restoring renal medullary blood flow. We stimulated the NO-sGC-cGMP signalling pathway in DVR at three different levels before and after hypoxia/re-oxygenation (H/R). Rat DVR were isolated and perfused under isobaric conditions. The phosphodiesterase 5 (PDE5) inhibitor sildenafil (10⁻⁶ mol/L) impaired cGMP degradation and dilated DVR pre-constricted with angiotensin II (Ang II, 10⁻⁶ mol/L). Dilations by the soluble guanylyl cyclase (sGC) activator BAY 60-2770 as well as the nitric oxide donor sodium nitroprusside (SNP, 10⁻³ mol/L) were equally effective. Hypoxia (0.1% O₂) augmented DVR constriction by Ang II, thus potentially aggravating tissue hypoxia. H/R left DVR unresponsive to sildenafil, yet sGC activation by BAY 60-2770 effectively dilated DVR. Dilation to SNP under H/R is delayed. In conclusion, H/R renders PDE5 inhibition ineffective in dilating the crucial vessels supplying the area at risk for hypoxic damage. Stimulating sGC appears to be the most effective in restoring renal medullary blood flow after H/R and may prove to be the best target for maintaining oxygenation to this vulnerable area of the kidney.     

The Role of Heparan Sulfate in CCL26-Induced Eosinophil Chemotaxis

Proinflammatory chemokine ligand 26 (CCL26, eotaxin-3) mediates transendothelial cell migration of eosinophils by binding and activating the G-protein-coupled (GPC) chemokine receptor 3 on the surface of eosinophilic cells. Here we have investigated the role of glycosaminoglycans (GAGs) as potential co-receptors in the process of CCL26-induced eosinophil chemotaxis. For this purpose, we have first identified the GAG-binding site of CCL26 by a site-directed mutagenesis approach in the form of an alanine screening. A panel of GAG-binding-deficient mutants has been designed, generated, and analyzed with respect to their binding affinities to heparan sulphate (HS) by isothermal fluorescence titration studies. This showed that basic amino acids in the α-helical part of CCL26 are strongly involved in GAG-binding. In chemotaxis experiments, we found that decreased GAG-binding affinity correlated with decreased chemotactic activity, which indicates an involvement of GAGs in eosinophil migration. This was further proven by the negative impact of heparinase III treatment and, independently, by the incubation of eosinophils with an anti heparan sulfate antibody. We finally investigated eosinophils’ proteoglycan (PG) expression patterns by real-time PCR, which revealed the highest expression level for serglycin. Including an anti-serglycin antibody in CCL26-induced eosinophil migration experiments reduced the chemotaxis of these immune cells, thereby proving the dependence of eosinophil mobilization on the proteoglycan serglycin.     

Design and Development of a Bimodal Optical Instrument for Simultaneous Vibrational Spectroscopy Measurements

Vibrational spectroscopy techniques are widely used in analytical chemistry, physics and biology. The most prominent techniques are Raman and Fourier-transform infrared spectroscopy (FTIR). Combining both techniques delivers complementary information of the test sample. We present the design, construction, and calibration of a novel bimodal spectroscopy system featuring both Raman and infrared measurements simultaneously on the same sample without mutual interference. The optomechanical design provides a modular flexible system for solid and liquid samples and different configurations for Raman. As a novel feature, the Raman module can be operated off-axis for optical sectioning. The calibrated system demonstrates high sensitivity, precision, and resolution for simultaneous operation of both techniques and shows excellent calibration curves with coefficients of determination greater than 0.96. We demonstrate the ability to simultaneously measure Raman and infrared spectra of complex biological material using bovine serum albumin. The performance competes with commercial systems; moreover, it presents the additional advantage of simultaneously operating Raman and infrared techniques. To the best of our knowledge, it is the first demonstration of a combined Raman-infrared system that can analyze the same sample volume and obtain optically sectioned Raman signals. Additionally, quantitative comparison of confocality of backscattering micro-Raman and off-axis Raman was performed for the first time.     

The Functional Characterization of GCaMP3.0 Variants Specifically Targeted to Subcellular Domains

Calcium (Ca²⁺) ions play a pivotal role in physiology and cellular signaling. The intracellular Ca²⁺ concentration ([Ca²⁺]_i) is about three orders of magnitude lower than the extracellular concentration, resulting in a steep transmembrane concentration gradient. Thus, the spatial and the temporal dynamics of [Ca²⁺]_i are ideally suited to modulate Ca²⁺-mediated cellular responses to external signals. A variety of highly sophisticated methods have been developed to gain insight into cellular Ca²⁺ dynamics. In addition to electrophysiological measurements and the application of synthetic dyes that change their fluorescent properties upon interaction with Ca²⁺, the introduction and the ongoing development of genetically encoded Ca²⁺ indicators (GECI) opened a new era to study Ca²⁺-driven processes in living cells and organisms. Here, we have focused on one well-established GECI, i.e., GCaMP3.0. We have systematically modified the protein with sequence motifs, allowing localization of the sensor in the nucleus, in the mitochondrial matrix, at the mitochondrial outer membrane, and at the plasma membrane. The individual variants and a cytosolic version of GCaMP3.0 were overexpressed and purified from E. coli cells to study their biophysical properties in solution. All versions were examined to monitor Ca²⁺ signaling in stably transfected cell lines and in primary cortical neurons transduced with recombinant Adeno-associated viruses (rAAV). In this comparative study, we provide evidence for a robust approach to reliably trace Ca²⁺ signals at the (sub)-cellular level with pronounced temporal resolution.     

Sonochemical Activation of Al/Ni Hydrogenation Catalyst

This paper proposes a sonochemical approach to the nanostructuring of Al/Ni catalyst with high content of accessible Ni centers and a high reusability. The surface and bulk composition as well as pore size distribution of this catalyst are controlled synergistically by adjusting the ultrasound intensity in aqueous solution. Sonochemical activation of Al/Ni alloy leads to formation of mesoporous Al/Ni metallic based frameworks with surface area up to 125 m² g^?1, and regular distribution of nickel active center in the porous matrix. One of the opportunities of porous Al/Ni catalyst is that due to a time‐resolved controllable formation of protective oxide layer it can be stored and handled under air in comparison to traditional Raney catalysts which need inert conditions. The Al/Ni catalyst is characterized by scanning electron microscopy (SEM), electron diffraction spectroscopy (EDS), X‐ray photoelectron spectroscopy (XPS), confocal scanning fluorescence microscopy (CSFM), solid‐state NMR experiments, and powder X‐ray diffraction analysis (PXRD). The catalytic activity was investigated for the hydrogenation of acetophenone.     

Structural Analysis of Mitochondrial Dynamics—From Cardiomyocytes to Osteoblasts: A Critical Review

Mitochondria play a crucial role in cell physiology and pathophysiology. In this context, mitochondrial dynamics and, subsequently, mitochondrial ultrastructure have increasingly become hot topics in modern research, with a focus on mitochondrial fission and fusion. Thus, the dynamics of mitochondria in several diseases have been intensively investigated, especially with a view to developing new promising treatment options. However, the majority of recent studies are performed in highly energy-dependent tissues, such as cardiac, hepatic, and neuronal tissues. In contrast, publications on mitochondrial dynamics from the orthopedic or trauma fields are quite rare, even if there are common cellular mechanisms in cardiovascular and bone tissue, especially regarding bone infection. The present report summarizes the spectrum of mitochondrial alterations in the cardiovascular system and compares it to the state of knowledge in the musculoskeletal system. The present paper summarizes recent knowledge regarding mitochondrial dynamics and gives a short, but not exhaustive, overview of its regulation via fission and fusion. Furthermore, the article highlights hypoxia and its accompanying increased mitochondrial fission as a possible link between cardiac ischemia and inflammatory diseases of the bone, such as osteomyelitis. This opens new innovative perspectives not only for the understanding of cellular pathomechanisms in osteomyelitis but also for potential new treatment options.     

A note on the time travel approach for handling time windows in vehicle routing problems

In this paper, we identify two cases in which the proposition for calculating time window penalties presented in Nagata, Y., Bräysy, O. and Dullaert, W. A penalty-based edge assembly memetic algorithm for the vehicle routing problem with time windows, Computers & Operations Research 2010;37(4): 724–37 yields incorrect results. We derive the corrected proposition and use numerical studies to show that a significant proportion of the evaluations performed by a Tabu Search for VRPTW falls under the two incorrect cases. Moreover, we demonstrate that the incorrect time window handling has a significant negative impact on the solution quality of the Tabu Search.     

Solid state transformations of Au-Cu-Pt alloys studied by in-situ X-ray synchrotron radiation and DSC

Seven alloys of the ternary Au-Cu-Pt system, containing 75 or 76.5 wt.% Au with 0–15 wt.% Pt and heat-treated to a fully ordered condition, are examined using in-situ synchrotron radiation to determine transformation temperatures and phases formed, and to investigate mechanisms of ordering and disordering. These experiments are complemented by differential scanning calorimetry. Diffraction data show that the alloys can be separated into three groups according to their stable low-temperature ordered phase(s), namely L1₀, L1₂, or the two combined. Platinum increases transformation temperatures in comparison with binary AuCu, two-phase ordered alloys showing the highest transformation temperatures. Details of the evolution of the X-ray diffraction peak structures upon heating and cooling point to significant differences between mechanisms of disordering and ordering: whereas ordering visibly proceeds at significant undercooling by nucleation and growth, disordering appears, from the visible shifts in peak position, to progress in more homogeneous fashion within the alloy.     

Investigation of energy performance of newly built low-energy buildings in Sweden

Energy use in the built environment represents a large part of total energy use in Sweden and is one important sector where energy conservation needs to be significantly improved in order to meet the national implementation of the European goals. One key question that needs to be investigated in relation to these goals is the performance and implementation of passive or low-energy houses. This paper presents results and an evaluation of a newly built house in an area with passive houses in Linköping, Sweden. Nine passive houses were built with the aim to be energy efficient, with an annual space heating demand of 21 kWh/m², and at the same time to have the same visual appearance as any other building in the surrounding area.This study evaluates the energy performance of a residential area with low-energy buildings based on Building Energy Simulation (BES) (IDA ICE 4), and measurements from the real object. Both annual and hourly validation is performed using room by room modeling and internal heat gains. A novel approach to internal heat gain modeling is presented using time-use data (TUD). The results show possible improvements in the design, the building envelope and in the heating control.