Coherent correlation imaging for resolving fluctuating states of matter.

Fluctuations and stochastic transitions are ubiquitous in nanometre-scale systems, especially in the presence of disorder. However, their direct observation has so far been impeded by a seemingly fundamental, signal-limited compromise between spatial and temporal resolution. Here we develop coherent correlation imaging (CCI) to overcome this dilemma. Our method begins by classifying recorded camera frames in Fourier space. Contrast and spatial resolution emerge by averaging selectively over same-state frames. Temporal resolution down to the acquisition time of a single frame arises independently from an exceptionally low misclassification rate, which we achieve by combining a correlation-based similarity metric1,2 with a modified, iterative hierarchical clustering algorithm3,4. We apply CCI to study previously inaccessible magnetic fluctuations in a highly degenerate magnetic stripe domain state with nanometre-scale resolution. We uncover an intricate network of transitions between more than 30 discrete states. Our spatiotemporal data enable us to reconstruct the pinning energy landscape and to thereby explain the dynamics observed on a microscopic level. CCI massively expands the potential of emerging high-coherence X-ray sources and paves the way for addressing large fundamental questions such as the contribution of pinning5-8 and topology9-12 in phase transitions and the role of spin and charge order fluctuations in high-temperature superconductivity13,14.

Structural deficits in key domains of Shank2 lead to alterations in postsynaptic nanoclusters and to a neurodevelopmental disorder in humans.

Postsynaptic scaffold proteins such as Shank, PSD-95, Homer and SAPAP/GKAP family members establish the postsynaptic density of glutamatergic synapses through a dense network of molecular interactions. Mutations in SHANK genes are associated with neurodevelopmental disorders including autism and intellectual disability. However, no SHANK missense mutations have been described which interfere with the key functions of Shank proteins believed to be central for synapse formation, such as GKAP binding via the PDZ domain, or Zn2+-dependent multimerization of the SAM domain. We identify two individuals with a neurodevelopmental disorder carrying de novo missense mutations in SHANK2. The p.G643R variant distorts the binding pocket for GKAP in the Shank2 PDZ domain and prevents interaction with Thr(-2) in the canonical PDZ ligand motif of GKAP. The p.L1800W variant severely delays the kinetics of Zn2+-dependent polymerization of the Shank2-SAM domain. Structural analysis shows that Trp1800 dislodges one histidine crucial for Zn2+ binding. The resulting conformational changes block the stacking of helical polymers of SAM domains into sheets through side-by-side contacts, which is a hallmark of Shank proteins, thereby disrupting the highly cooperative assembly process induced by Zn2+. Both variants reduce the postsynaptic targeting of Shank2 in primary cultured neurons and alter glutamatergic synaptic transmission. Super-resolution microscopy shows that both mutants interfere with the formation of postsynaptic nanoclusters. Our data indicate that both the PDZ- and the SAM-mediated interactions of Shank2 contribute to the compaction of postsynaptic protein complexes into nanoclusters, and that deficiencies in this process interfere with normal brain development in humans.

Deterministic Generation and Guided Motion of Magnetic Skyrmions by Focused He+-Ion Irradiation.

Magnetic skyrmions are quasiparticles with nontrivial topology, envisioned to play a key role in next-generation data technology while simultaneously attracting fundamental research interest due to their emerging topological charge. In chiral magnetic multilayers, current-generated spin-orbit torques or ultrafast laser excitation can be used to nucleate isolated skyrmions on a picosecond time scale. Both methods, however, produce randomly arranged skyrmions, which inherently limits the precision on the location at which the skyrmions are nucleated. Here, we show that nanopatterning of the anisotropy landscape with a He+-ion beam creates well-defined skyrmion nucleation sites, thereby transforming the skyrmion localization into a deterministic process. This approach allows control of individual skyrmion nucleation as well as guided skyrmion motion with nanometer-scale precision, which is pivotal for both future fundamental studies of skyrmion dynamics and applications.

Megahertz-rate ultrafast X-ray scattering and holographic imaging at the European XFEL.

The advent of X-ray free-electron lasers (XFELs) has revolutionized fundamental science, from atomic to condensed matter physics, from chemistry to biology, giving researchers access to X-rays with unprecedented brightness, coherence and pulse duration. All XFEL facilities built until recently provided X-ray pulses at a relatively low repetition rate, with limited data statistics. Here, results from the first megahertz-repetition-rate X-ray scattering experiments at the Spectroscopy and Coherent Scattering (SCS) instrument of the European XFEL are presented. The experimental capabilities that the SCS instrument offers, resulting from the operation at megahertz repetition rates and the availability of the novel DSSC 2D imaging detector, are illustrated. Time-resolved magnetic X-ray scattering and holographic imaging experiments in solid state samples were chosen as representative, providing an ideal test-bed for operation at megahertz rates. Our results are relevant and applicable to any other non-destructive XFEL experiments in the soft X-ray range.

Observation of fluctuation-mediated picosecond nucleation of a topological phase.

Topological states of matter exhibit fascinating physics combined with an intrinsic stability. A key challenge is the fast creation of topological phases, which requires massive reorientation of charge or spin degrees of freedom. Here we report the picosecond emergence of an extended topological phase that comprises many magnetic skyrmions. The nucleation of this phase, followed in real time via single-shot soft X-ray scattering after infrared laser excitation, is mediated by a transient topological fluctuation state. This state is enabled by the presence of a time-reversal symmetry-breaking perpendicular magnetic field and exists for less than 300 ps. Atomistic simulations indicate that the fluctuation state largely reduces the topological energy barrier and thereby enables the observed rapid and homogeneous nucleation of the skyrmion phase. These observations provide fundamental insights into the nature of topological phase transitions, and suggest a path towards ultrafast topological switching in a wide variety of materials through intermediate fluctuating states.

Reference shape effects on Fourier transform holography.

Soft-x-ray holography which utilizes an optics mask fabricated in direct contact with the sample, is a widely applied x-ray microscopy method, in particular, for investigating magnetic samples. The optics mask splits the x-ray beam into a reference wave and a wave to illuminate the sample. The reconstruction quality in such a Fourier-transform holography experiment depends primarily on the characteristics of the reference wave, typically emerging from a small, high-aspect-ratio pinhole in the mask. In this paper, we study two commonly used reference geometries and investigate how their 3D structure affects the reconstruction within an x-ray Fourier holography experiment. Insight into these effects is obtained by imaging the exit waves from reference pinholes via high-resolution coherent diffraction imaging combined with three-dimensional multislice simulations of the x-ray propagation through the reference pinhole. The results were used to simulate Fourier-transform holography experiments to determine the spatial resolution and precise location of the reconstruction plane for different reference geometries. Based on our findings, we discuss the properties of the reference pinholes with view on application in soft-x-ray holography experiments.

Legal vs. Extra-Legal Responses to Public Health Emergencies.

There is a long-established claim that emergency action through the law is impossible, or bound to be ineffective. This article seeks to challenge this position by reference to the response of many European states to the Coronavirus pandemic and by drawing on Lon Fuller's theory of law. It argues that there are a number of reasons why a fragmentation of governance between ordinary, legal action and emergency, extra-legal action is neither necessary nor desirable in this specific context. In societies that are generally rule of law compliant governance according to formal legal principles is not only constraining, it also possesses the quality of a 'liberating limitation', creating the room for effective, sustainable action. Too little has been made of this positive dimension of the legal form as an instrument for emergency action.

Vaccination as an Equaliser? Evaluating COVID-19 Vaccine Prioritisation and Compensation.

This article assesses the equity of COVID-19 vaccination programmes in three jurisdictions that have historically taken different approaches to the institutionalisation of equity considerations. The Sars-Cov-2 pandemic has thrown into sharp relief persistent societal inequalities and has added novel dimensions to these problems. Certain groups have proved particularly vulnerable, both in terms of infection risk and severity as well as the accompanying social fallout. Against this background the implementation of 'objective' vaccination programmes may seem like a great leveller, addressing the disparate risks that are tied to social determinants of health and the pandemic behemoth. However, implementing vaccination programmes in an equitable manner is itself essential for the realisation of such a vision. This article undertakes a comparative analysis of the English, Italian, and American jurisdictions and critically assesses two aspects of their vaccination frameworks: (i) the prioritisation of groups for vaccination and (ii) the nature of public compensation schemes for those who have suffered vaccine-related injuries. It examines whether and to what extent these measures address the inequalities raised by COVID-19 and the role of the law in this pursuit.

Ultrafast Demagnetization Dominates Fluence Dependence of Magnetic Scattering at Co M Edges.

We systematically study the fluence dependence of the resonant scattering cross-section from magnetic domains in Co/Pd-based multilayers. Samples are probed with single extreme ultraviolet (XUV) pulses of femtosecond duration tuned to the Co M_{3,2} absorption resonances using the FERMI@Elettra free-electron laser. We report quantitative data over 3 orders of magnitude in fluence, covering 16 mJ/cm^{2}/pulse to 10 000 mJ/cm^{2}/pulse with pulse lengths of 70 fs and 120 fs. A progressive quenching of the diffraction cross-section with fluence is observed. Compression of the same pulse energy into a shorter pulse-implying an increased XUV peak electric field-results in a reduced quenching of the resonant diffraction at the Co M_{3,2} edge. We conclude that the quenching effect observed for resonant scattering involving the short-lived Co 3p core vacancies is noncoherent in nature. This finding is in contrast to previous reports investigating resonant scattering involving the longer-lived Co 2p states, where stimulated emission has been found to be important. A phenomenological model based on XUV-induced ultrafast demagnetization is able to reproduce our entire set of experimental data and is found to be consistent with independent magneto-optical measurements of the demagnetization dynamics on the same samples.

Imaging Nanometer Phase Coexistence at Defects During the Insulator-Metal Phase Transformation in VO2 Thin Films by Resonant Soft X-ray Holography.

We use resonant soft X-ray holography to image the insulator-metal phase transition in vanadium dioxide with element and polarization specificity and nanometer spatial resolution. We observe that nanoscale inhomogeneity in the film results in spatial-dependent transition pathways between the insulating and metallic states. Additional nanoscale phases form in the vicinity of defects which are not apparent in the initial or final states of the system, which would be missed in area-integrated X-ray absorption measurements. These intermediate phases are vital to understand the phase transition in VO2, and our results demonstrate how resonant imaging can be used to understand the electronic properties of phase-separated correlated materials obtained by X-ray absorption.

Quantification of silver nanoparticle uptake and distribution within individual human macrophages by FIB/SEM slice and view.

BACKGROUND: Quantification of nanoparticle (NP) uptake in cells or tissues is very important for safety assessment. Often, electron microscopy based approaches are used for this purpose, which allow imaging at very high resolution. However, precise quantification of NP numbers in cells and tissues remains challenging. The aim of this study was to present a novel approach, that combines precise quantification of NPs in individual cells together with high resolution imaging of their intracellular distribution based on focused ion beam/ scanning electron microscopy (FIB/SEM) slice and view approaches. RESULTS: We quantified cellular uptake of 75 nm diameter citrate stabilized silver NPs (Ag 75 Cit) into an individual human macrophage derived from monocytic THP-1 cells using a FIB/SEM slice and view approach. Cells were treated with 10 µg/ml for 24 h. We investigated a single cell and found in total 3138 ± 722 silver NPs inside this cell. Most of the silver NPs were located in large agglomerates, only a few were found in clusters of fewer than five NPs. Furthermore, we cross-checked our results by using inductively coupled plasma mass spectrometry and could confirm the FIB/SEM results. CONCLUSIONS: Our approach based on FIB/SEM slice and view is currently the only one that allows the quantification of the absolute dose of silver NPs in individual cells and at the same time to assess their intracellular distribution at high resolution. We therefore propose to use FIB/SEM slice and view to systematically analyse the cellular uptake of various NPs as a function of size, concentration and incubation time.

Curved gratings as an integrated photon fluence monitor in x-ray transmission scattering experiments.

A concept to obtain a measure of the photon flux accepted by a solid sample in single-shot transmission experiments with extreme ultraviolet (XUV) or soft x-ray radiation is demonstrated. Shallow, continuously distorted gratings are used to diffract a constant fraction of the incident photons onto an extended area of a CCD detector. The signal can be tailored to fit the dynamic range of the detector, i.e. matching the scattered intensity of the studied structure of interest. Furthermore, composite grating designs that also allow for the measurement of the spatial photon distribution on the sample are demonstrated. The gratings are directly fabricated by focused ion-beam (FIB) lithography into a Si3N4 membrane that supports the actual sample layer. This allows for rapid fabrication of hundreds of samples, making the concept suitable for systematic studies in destructive single-shot measurements at free-electron laser (FEL) sources. We demonstrate relative photon flux measurements in magnetic scattering experiments with synchrotron and FEL radiation at 59.6 eV photon energy.

Significance of endothelial progenitor cells (EPC) for tumorigenesis of head and neck squamous cell carcinoma (HNSCC): possible marker of tumor progression and neovascularization?

OBJECTIVES: Angiogenesis and neovascularisation plays a crucial role for tumorigenesis and tumor progression in head and neck squamous cell carcinoma (HNSCC). The aim of our study was to investigate the neovascularization capacity by endothelial progenitor cells (EPC) in tumor patient as a possible predictor for tumor progression and tumor stage. MATERIALS AND METHODS: Therefore, we investigated the cell number and biologic activity by cell migration and colony-forming ability of EPC. Cells were isolated from the peripheral venous blood of 79 patients who suffer HNSCC in different stages of disease. Thirty-three healthy individuals served as the control group. RESULTS: Significantly increased biological activities were reflected by expression of the migration rate (1027 ± 1510) in comparison to the control group (632 ± 269) and the clonal potency measured by colony-forming unit (CFU) (tumor patients (19.7 ± 12.3) vs. control group (10.84 ± 4.8)). To determine whether or not EPC number can be used as a valid prognostic marker for clinical outcome of tumor patients, we furthermore compared a "high EPC-number-subgroup" (HI) with a "low EPC-number-subgroup" (LO) in a Kaplan-Meier survival curve. The HI-subgroup shows herein clearly a worse outcome. CONCLUSIONS: Our findings indicate a possible pathway for EPC to play a critical role in the vasculogenesis and consequently in the progression of HNSCC. CLINICAL RELEVANCE: Our findings could serve as possible predictors for the neovascularisation potential in HNSCC tumor patients.

Restoring implant fixation strength in osteoporotic bone with a hydrogel locally delivering zoledronic acid and bone morphogenetic protein 2. A longitudinal in vivo microCT study in rats.

Osteoporosis poses a major public health challenge, and it is characterized by low bone mass, deterioration of the microarchitecture of bone tissue, causing a consequent increase in bone fragility and susceptibility to fractures and complicating bone fixation, particularly screw implantation. In the present study, our aim was to improve implant stability in osteoporotic bone using a thermoresponsive hyaluronan hydrogel (HA-pNIPAM) to locally deliver the bisphosphonate zoledronic acid (ZOL) to prevent bone resorption and bone morphogenetic protein 2 (BMP2) to induce bone formation. Adult female Wistar rats (n = 36) were divided into 2 treatment groups: one group of SHAM-operated animals and another group that received an ovariectomy (OVX) to induce an osteoporotic state. All animals received a polyetheretherketone (PEEK) screw in the proximal tibia. In addition, subgroups of SHAM or OVX animals received either the HA-pNIPAM hydrogel without or with ZOL/BMP2, placed into the defect site prior to screw implantation. Periprosthetic bone and implant fixation were monitored using longitudinal in vivo microCT scanning post-operatively and at 3, 6, 9, 14, 20 and 28 days. Histological assessment was performed post-mortem. Our data showed that pure hydrogel has no impact of implant fixation The ZOL/BMP2-hydrogel significantly increased bone-implant contact and peri-implant bone fraction, primarily through reduced resorption. STATEMENT OF CLINICAL SIGNIFICANCE: Local delivery of ZOL and BMP2 using a biocompatible hydrogel improved implant stability in osteoporotic bone. This approach could constitute a potent alternative to systemic drug administration and may be useful in avoiding implant loosening in clinical settings.

Automatable sample fabrication process for pump-probe X-ray holographic imaging.

Soft X-ray holography is a recently developed imaging technique with sub-50 nm spatial resolution. Key advantages of this technique are magnetic and elemental sensitivity, compatibility with imaging at free electron laser facilities, and immunity to in-situ sample excitations and sample drift, which enables the reliable detection of relative changes between two images with a precision of a few nanometers. In X-ray holography, the main part of the experimental setup is integrated in the sample, which consequently requires a large number of fabrication steps. Here we present a generic design and an automatable fabrication process for samples suitable, and optimized for, efficient high resolution X-ray holographic dynamic imaging. The high efficiency of the design facilitates the acquisition of magnetic images in a few minutes and makes fully automatic image reconstruction possible.

Midterm results after subtrochanteric end-to-side valgization osteotomy in severe infantile coxa vara.

BACKGROUND: For the treatment of the severe infantile coxa vara it is mandatory for the orthopaedic surgeon to observe the mechanobiology of the growing hip before and after the surgical intervention. We hereby would like to present our experiences with the subtrochanteric end-to-side valgization osteotomy and to compare the procedure with the alternatively used Y-shaped osteotomy as described by Pauwels. METHODS: Thirteen patients (20 hips) who had undergone subtrochanteric end-to-side valgization were followed for a mean 6.2 years (range, 0.8 to 12.8 y). At the time of surgery the mean age was 7.1 years (range, 2.0 to 13.3 y), last follow-up examination was performed at a mean of 13.4 years of age (range, 5.1 to 18.3 y). The deformities were etiologically based on 5 entities: congenital coxa vara (n=1), osteochondrodysplasias (n=12), postosteomyelitic coxa vara (n=5), and avascular femoral head necrosis in the course of congenital dysplasia of the hip (n=2). The follow-up rate was 100%.In addition, we analyzed a total of 93 pelvic radiographies with a total of 139 hip joints. Thirty angles and distances were assessed according to parameters described in the literature. RESULTS: Although preoperatively 12 patients presented with a positive Trendelenburg's sign, it was only present postoperatively in 2 patients. Duchenne's limp reduced from 10 to 1. All of the 15 preoperatively apparent nonunions could be healed by means of surgery. Two hips redeveloped pathologically lowered collodiaphyseal angles postoperatively, one of which had to undergo revision surgery. Preoperatively 15 out of 20 patients (75%) showed nonunions all of which healed after surgery. No recurrence could be seen at the time of the last follow-up.The following angles were assessed on plain radiographies of the pelvis preoperatively and directly postoperatively as well as on the last follow-up at a mean of 85 months: CCD-angle 98 degrees/156 degrees/144 degrees, EY-angle 55 degrees/5 degrees/15.7 degrees, AY-angle 32 degrees/75 degrees/66 degrees, CE-angle 20 degrees/25 degrees/18 degrees, AC-angle 20 degrees/18 degrees/20 degrees. The articulotrochanteric distance was 5 mm/26 mm/14 mm. CONCLUSIONS: The subtrochanteric end-to-side valgization osteotomy showed to be highly effective in the management of the infantile coxa vara, improving the clinical impairment of the patients postoperatively. All of the preoperatively present nonunions showed osseous consolidation at follow-up examination. Only minor revarization tendencies could be found. The procedure is technically less demanding, safer and more efficient regarding the lengthening of the affected limb in comparison to the Y-shaped intertrochanteric osteotomy as described by Pauwels. LEVEL OF EVIDENCE: Case-control study (EBM-level III).

Manifestations of hypertensive encephalopathy in cats.

OBJECTIVES: Hypertensive encephalopathy in cats is an important entity but is underestimated in clinical practice. This could be explained, in part, by non-specific clinical signs. The objective of this study was to characterise the clinical manifestations of hypertensive encephalopathy in cats. METHODS: Cats with systemic hypertension (SHT) recognised by routine screening, associated with underlying predisposing disease or a clinical presentation suggestive of SHT (neurological or non-neurological), were prospectively enrolled over a 2-year period. Confirmation of SHT was based on at least two sets of measurements of systolic blood pressure >160 mmHg by Doppler sphygmomanometry. RESULTS: Fifty-six hypertensive cats with a median age of 16.5 years were identified; 31 had neurological signs. In 16/31 cats, neurological abnormalities were the primary complaint. The other 15 cats were first presented to the medicine or ophthalmology service, and neurological disease was recognised based on the cat's history. The most common neurological signs were ataxia, various manifestations of seizures and altered behaviour. Individual cats also showed paresis, pleurothotonus, cervical ventroflexion, stupor and facial nerve paralysis. In 28/30 cats, retinal lesions were detected. Of these 28 cats, six presented with a primary complaint of visual deficits, and neurological signs were not the primary complaint; nine presented with non-specific medical issues, without suspicion of SHT-induced organ damage; in 13 cats, neurological issues were the primary complaint and fundic abnormalities were detected subsequently. CONCLUSIONS AND RELEVANCE: SHT is common in older cats and the brain is an important target organ; however, neurological deficits are commonly ignored in cats with SHT. Gait abnormalities, (partial) seizures and even mild behavioural changes should prompt clinicians to consider the presence of SHT. A fundic examination in cats with suspected hypertensive encephalopathy is a sensitive test to support the diagnosis.

Pump-probe x-ray microscopy of photo-induced magnetization dynamics at MHz repetition rates.

We present time-resolved scanning x-ray microscopy measurements with picosecond photo-excitation via a tailored infrared pump laser at a scanning transmission x-ray microscope. Specifically, we image the laser-induced demagnetization and remagnetization of thin ferrimagnetic GdFe films proceeding on a few nanoseconds timescale. Controlling the heat load on the sample via additional reflector and heatsink layers allows us to conduct destruction-free measurements at a repetition rate of 50 MHz. Near-field enhancement of the photo-excitation and controlled annealing effects lead to laterally heterogeneous magnetization dynamics which we trace with 30 nm spatial resolution. Our work opens new opportunities to study photo-induced dynamics on the nanometer scale, with access to picosecond to nanosecond time scales, which is of technological relevance, especially in the field of magnetism.