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Klug, C.

Publications and source records attributed to Klug, C..

3 recordsLinked to original sources

Ultra-precision 3D Printed Sample Tubes of Complex Cross Section for Enhanced Electron Paramagnetic Resonance Sensitivity of AqueousSamples

This paper discusses the applications of ultra-precision 3D printing in developing EPR sample tubes for X-band measurements of aqueous samples. The seed idea of this work started from the flat cell idea discussed in Mett and Hyde et al.(2003), Sidabras et al. (2005), and Sidabras et al. (2017), where placing multiple aqueous flat cells perpendicular to the electric field in the microwave cavity reduce RF losses and improve EPR signal sensitivity. In this work, we present the 3D printed aquastar sample holders for dielectric resonators and the aquasun sample holders for cavity resonators. Even though the ultra-precision 3D printing enables the fabrication of complex cross-sections at 25um resolution, the photopolymer material has comparatively higher dielectric losses compared to the conventional materials like glass and PTFE. To reduce the RF losses due to the inherent dielectric properties, we introduced the latticing approach in reducing the amount of plastic without compromising the structural stability. In addition to this, we present the slicing and dielectric approximation approach in reducing the computation cost for these complex cross-sections. The novel sample tubes discussed here present signal improvements through both simulation and experiments, which were validated through power saturation experiments. The sample tube development methodology presented here provide a general framework for designing and implementing 3D printed EPR sample holders, based on resonator electromagnetic field distribution and dielectric properties of the photopolymer material.

biophysics↗

Distinct Membrane Binding Properties of the Two Non-visual Arrestins

Membrane interactions play a crucial role in regulating arrestin activation and its binding to phosphorylated G protein-coupled receptors (GPCRs). Here, we utilize in vitro biophysical approaches and cell-based fluorescence intensity fluctuation analysis to systematically compare the membrane-binding properties of the two highly conserved arrestin subtypes, arrestin-2 and arrestin-3, under basal and stimulated conditions. Our findings reveal that arrestin-2 selectively engages the PI(4,5)P2-containing nanodiscs via its C-edge, whereas arrestin-3 primarily utilizes its finger loop to interact with negatively charged lipids. Notably, while the lipid bilayer alone does not activate arrestin, it synergistically enhances arrestin-2/3 activation in conjunction with a phosphorylated GPCR C-tail. Additionally, the spacing between receptor phosphorylation sites and the lipid bilayer modulates arrestin-membrane assembly. Live cell tracking further demonstrates that arrestin-2 and arrestin-3 exhibit distinct plasma membrane dissociation dynamics. These findings provide novel insights into the mechanisms governing arrestin activation and its functional interplay with membranes.

biochemistry↗

Chronic administration of XBD173 ameliorates cognitive deficits and neuropathology via 18 kDa translocator protein (TSPO) in a mouse model of Alzheimer disease

Alzheimers disease (AD) is characterized by the accumulation of {beta}-amyloid peptide (A{beta}). There is increasing evidence that depression may precede AD and may be an early manifestation of dementia, suggesting common mechanisms underlying both diseases. Ligands targeting the mitochondrial translocator protein (18 kDa) (TSPO), promote neurosteroidogenesis and may be neuroprotective. Moreover, TSPO is upregulated in AD. To study whether the TSPO ligand XBD173 may exert early neuroprotective effects in AD pathology we investigated the impact of XBD173 on amyloid toxicity and neuroplasticity in mouse models. We show that XBD173 (emapunil), via neurosteroid-mediated signaling via delta subunit-containing GABAA receptors, prevents the neurotoxic effect of A{beta} on long-term potentiation (CA1-LTP) in the hippocampus and prevents the loss of spines. Chronic but not acute administration of XBD173 ameliorates spatial learning deficits in transgenic AD mice with arctic mutation (ArcA{beta}) mice. The heterozygous TSPO-knockout crossed with the transgenic arctic mutation model of AD mice (het TSPOKO X ArcA{beta}) treated with XBD173 does not show this improvement in spatial learning suggesting TSPO is needed for procognitive effects of XBD173. The neuroprotective profile of XBD173 in AD pathology is further supported by a reduction in plaques and soluble A{beta} levels in the cortex, increased synthesis of neurosteroids, rescued spine density, reduction of complement protein C1q deposits, and reduced astrocytic phagocytosis of functional synapses both in the hippocampus and cortex. Our findings suggest that XBD173 may exert therapeutic effects via TSPO in a mouse model of AD.

neuroscience↗