Phase II testing of Xenon International on Mount Schauinsland, Germany.

Station RN33 on Mount Schauinsland near Freiburg, Germany, is part of the International Monitoring System monitoring radioxenon in air (131mXe, 133Xe, 133mXe, and 135Xe) for verification of the Comprehensive Nuclear Test Ban Treaty. Here, we present data from phase II testing of a new system, Xenon International at RN33, July 14th, 2021 to Jan 22nd, 2022, together with SPALAX data from the same time period. Radioxenon could be detected in 473 of 719 samples, among them many multiple isotope detections. Activity concentrations of spiked and selected environmental samples were verified by laboratory reanalysis. The sensitivity of Xenon International for radioxenons is up to one order of magnitude better for the metastable isotopes than that of the SPALAX, with a shorter sampling duration of 6 h.

The articular cartilage surface is impaired by a loss of thick collagen fibers and formation of type I collagen in early osteoarthritis.

Osteoarthritis (OA) is a joint disease affecting millions of patients worldwide. During OA onset and progression, the articular cartilage is destroyed, but the underlying complex mechanisms remain unclear. Here, we uncover changes in the thickness of collagen fibers and their composition at the onset of OA. For articular cartilage explants from knee joints of OA patients, we find that type I collagen-rich fibrocartilage-like tissue was formed in macroscopically intact cartilage, distant from OA lesions. Importantly, the number of thick fibers (>100 nm) has decreased early in the disease, followed by complete absence of thick fibers in advanced OA. We have obtained these results by a combination of high-resolution atomic force microscopy imaging under near-native conditions, immunofluorescence, scanning electron microscopy and a fluorescence-based classification of the superficial chondrocyte spatial organization. Taken together, our data suggests that the loss of tissue functionality in early OA cartilage is caused by a reduction of thick type II collagen fibers, likely due to the formation of type I collagen-rich fibrocartilage, followed by the development of focal defects in later OA stages. We anticipate that such an integrative characterization will be very beneficial for an in-depth understanding of other native biological tissues and the development of sustainable biomaterials. STATEMENT OF SIGNIFICANCE: In early osteoarthritis (OA) the cartilage appears macroscopically intact. However, this study demonstrates that the collagen network already changes in early OA by collagen fiber thinning and the formation of fibrocartilage-like tissue. Both nanoscopic deficiencies already occur in macroscopically intact regions of the human knee joint and are likely connected to processes that result in a weakened extracellular matrix. This study enhances the understanding of earliest progressive cartilage degeneration in the absence of external damage. The results suggest a determination of the mean collagen fiber thickness as a new target for the detection of early OA and a regulation of type I collagen synthesis as a new path for OA treatment.

Modelling 85Kr datasets with python for applications in tracer hydrology and to investigate atmospheric circulation.

We present a model written in python to evaluate data from comprehensive 85Kr collection schemes comprising 11 datasets from different monitoring stations around the globe. The model is designed to (1) calculate atmospheric input functions for the application of 85Kr as a dating tracer and (2) to investigate atmospheric circulation based on a two-box model of the atmosphere. Different functions were implemented, to (1) filter the data, (2) fit polynomials and running means, (3) extrapolate fits from the northern to the southern hemisphere, (4) calculate interhemispheric exchange times and 85Kr emission rates and (5) export data to a csv file. Although the model is designed to evaluate atmospheric 85Kr datasets, some functionality and basic concepts can be applied to other dating tracers, like tritium and SF6.•Standardized method to systematically analyse atmospheric 85Kr activity concentration time series for dating water and ice and to investigate atmospheric circulation.•Easily modifiable python script to adapt functions for similar data analysis procedures.

Hybrid fluorescence-AFM explores articular surface degeneration in early osteoarthritis across length scales.

Atomic force microscopy (AFM) has become a powerful tool for the characterization of materials at the nanoscale. Nevertheless, its application to hierarchical biological tissue like cartilage is still limited. One reason is that such samples are usually millimeters in size, while the AFM delivers much more localized information. Here a combination of AFM and fluorescence microscopy is presented where features on a millimeter sized tissue sample are selected by fluorescence microscopy on the micrometer scale and then mapped down to nanometer precision by AFM under native conditions. This served us to show that local changes in the organization of fluorescent stained cells, a marker for early osteoarthritis, correlate with a significant local reduction of the elastic modulus, local thinning of the collagen fibers, and a roughening of the articular surface. This approach is not only relevant for cartilage, but in general for the characterization of native biological tissue from the macro- to the nanoscale. STATEMENT OF SIGNIFICANCE: Different length scales have to be studied to understand the function and dysfunction of hierarchically organized biomaterials or tissues. Here we combine a highly stable AFM with fluorescence microscopy and precisely motorized movement to correlate micro- and nanoscopic properties of articular cartilage on a millimeter sized sample under native conditions. This is necessary for unraveling the relationship between microscale organization of chondrocytes, micrometer scale changes in articular cartilage properties and nanoscale organization of collagen (including D-banding). We anticipate that such studies pave the way for a guided design of hierarchical biomaterials.

AFM Imaging Suggests Receptor-Free Penetration of Lipid Bilayers by Toxins.

A crucial step of exotoxin action is the attack on the membrane. Many exotoxins show an architecture following the AB model, where a binding subunit translocates an "action" subunit across a cell membrane. Atomic force microscopy is an ideal technique to study these systems because of its ability to provide structural as well as dynamic information at the same time. We report first images of toxins Photorhabdus luminescens TcdA1 and Clostridium difficile TcdB on a supported lipid bilayer. A significant amount of toxin binds to the bilayer at neutral pH in the absence of receptors. Lack of diffusion indicates that toxin particles penetrate the membrane. This observation is supported by fluorescence recovery after photobleaching measurements. We mimic endocytosis by acidification while imaging the particles over time; however, we see no large conformational change. We therefore conclude that the toxin particles we imaged in neutral conditions had already formed a pore and speculate that there is no "pre-pore" state in our imaging conditions (i.e., in the absence of receptor).

Cysteine scanning reveals minor local rearrangements of the horizontal helix of respiratory complex I.

The NADH:ubiquinone oxidoreductase, respiratory complex I, couples electron transfer from NADH to ubiquinone with the translocation of protons across the membrane. The complex consists of a peripheral arm catalyzing the redox reaction and a membrane arm catalyzing proton translocation. The membrane arm is almost completely aligned by a 110 Å unique horizontal helix that is discussed to transmit conformational changes induced by the redox reaction in a piston-like movement to the membrane arm driving proton translocation. Here, we analyzed such a proposed movement by cysteine-scanning of the helix of the Escherichia coli complex I. The accessibility of engineered cysteine residues and the flexibility of individual positions were determined by labeling the preparations with a fluorescent marker and a spin-probe, respectively, in the oxidized and reduced states. The differences in fluorescence labeling and the rotational flexibility of the spin probe between both redox states indicate only slight conformational changes at distinct positions of the helix but not a large movement.