Spatial Distribution of Macrophages During Callus Formation and Maturation Reveals Close Crosstalk Between Macrophages and Newly Forming Vessels

Macrophages are essential players in the process of fracture healing, acting by remodeling of the extracellular matrix and enabling vascularization. Whilst activated macrophages of M1-like phenotype are present in the initial pro-inflammatory phase of hours to days of fracture healing, an anti-inflammatory M2-like macrophage phenotype is supposed to be crucial for the induction of downstream cascades of healing, especially the initiation of vascularization. In a mouse-osteotomy model, we provide a comprehensive characterization of vessel (CD31+, Emcn+) and macrophage phenotypes (F4/80, CD206, CD80, Mac-2) during the process of fracture healing. To this end, we phenotype the phases of vascular regeneration-the expansion phase (d1-d7 after injury) and the remodeling phase of the endothelial network, until tissue integrity is restored (d14-d21 after injury). Vessels which appear during the bone formation process resemble type H endothelium (CD31hiEmcnhi), and are closely connected to osteoprogenitors (Runx2+, Osx+) and F4/80+ macrophages. M1-like macrophages are present in the initial phase of vascularization until day 3 post osteotomy, but they are rare during later regeneration phases. M2-like macrophages localize mainly extramedullary, and CD206+ macrophages are found to express Mac-2+ during the expansion phase. VEGFA expression is initiated by CD80+ cells, including F4/80+ macrophages, until day 3, while subsequently osteoblasts and chondrocytes are main contributors to VEGFA production at the fracture site. Using Longitudinal Intravital Microendoscopy of the Bone (LIMB) we observe changes in the motility and organization of CX3CR1+ cells, which infiltrate the injury site after an osteotomy. A transient accumulation, resulting in spatial polarization of both, endothelial cells and macrophages, in regions distal to the fracture site, is evident. Immunofluorescence histology followed by histocytometric analysis reveals that F4/80+CX3CR1+ myeloid cells precede vascularization

Limbostomy: Longitudinal Intravital Microendoscopy in Murine Osteotomies

Bone healing involves the interplay of immune cells, mesenchymal cells, and vasculature over the time course of regeneration. Approaches to quantify the spatiotemporal aspects of bone healing at cellular resolution during long bone healing do not yet exist. Here, a novel technique termed Limbostomy is presented, which combines intravital microendoscopy with an osteotomy. This design allows a modular combination of an internal fixator plate with a gradient refractive index (GRIN) lens at various depths in the bone marrow and can be combined with a surgical osteotomy procedure. The field of view (FOV) covers a significant area of the fracture gap and allows monitoring cellular processes in vivo. The GRIN lens causes intrinsic optical aberrations which have to be corrected. The optical system was characterized and a postprocessing algorithm was developed. It corrects for wave front aberration-induced image plane deformation and for background and noise signals, enabling us to observe subcellular processes. Exemplarily, we quantitatively and qualitatively analyze angiogenesis in bone regeneration. We make use of a transgenic reporter mouse strain with nucleargreen fluorescent protein and membrane-bound tdTomato under the Cadherin-5 promoter. We observe two phases of vascularization. First, rapid vessel sprouting pervades the FOV within 3-4 days after osteotomy. Second, the vessel network continues to be dynamically remodeled until the end of our observation time, 14 days after surgery. Limbostomy opens a unique set of opportunities and allows further insight on spatiotemporal aspects of bone marrow biology, for example, hematopoiesis, analysis of cellular niches, immunological memory, and vascularization in the bone marrow during health and disease

Ambulatory assessment for physical activity research. State of the science, best practices and future directions

Technological and digital progress benefits physical activity (PA) research. Here we compiled expert knowledge on how Ambulatory Assessment (AA) is utilized to advance PA research, i.e., we present results of the 2nd International CAPA Workshop 2019 "Physical Activity Assessment - State of the Science, Best Practices, Future Directions" where invited researchers with experience in PA assessment, evaluation, technology and application participated. First, we provide readers with the state of the AA science, then we give best practice recommendations on how to measure PA via AA and shed light on methodological frontiers, and we furthermore discuss future directions. AA encompasses a class of methods that allows the study of PA and its behavioral, biological and physiological correlates as they unfold in everyday life. AA includes monitoring of movement (e.g., via accelerometry), physiological function (e.g., via mobile electrocardiogram), contextual information (e.g., via geolocation-tracking), and ecological momentary assessment (EMA; e.g., electronic diaries) to capture self-reported information. The strengths of AA are data assessment that near real-time, which minimizes retrospective biases in real-world settings, consequentially enabling ecological valid findings. Importantly, AA enables multiple assessments across time within subjects resulting in intensive longitudinal data (ILD), which allows unraveling within-person determinants of PA in everyday life. In this paper, we show how AA methods such as triggered e-diaries and geolocation-tracking can be used to measure PA and its correlates, and furthermore how these findings may translate into real-life interventions. In sum, AA provides numerous possibilities for PA research, especially the opportunity to tackle within-subject antecedents, concomitants, and consequences of PA as they unfold in everyday life. In-depth insights on determinants of PA could help us design and deliver impactful interventions in real-world contexts, thus enabling us to solve critical health issues in the 21st century such as insufficient PA and high levels of sedentary behavior. (DIPF/Orig.

Measurement of the hadronic photon structure function F_{2}^{γ} at LEP2

The hadronic structure function of the photon F_{2}^{γ} (x, Q²) is measured as a function of Bjorken x and of the photon virtuality Q² using deep-inelastic scattering data taken by the OPAL detector at LEP at e⁺e⁻ centre-of-mass energies from 183 to 209 GeV. Previous OPAL measurements of the x dependence of F_{2}^{γ} are extended to an average Q² of 〈Q²〉=780 GeV² using data in the kinematic range 0.15<x<0.98. The Q² evolution of F_{2}^{γ} is studied for 12.1<〈Q²〉<780 GeV² using three ranges of x. As predicted by QCD, the data show positive scaling violations in F_{2}^{γ} with F_{2}^{γ} (Q²)/α = (0.08±0.02⁺⁰·⁰⁵_₀.₀₃) + (0.13±0.01⁺⁰·⁰¹_₀.₀₁) lnQ², where Q² is in GeV², for the central x region 0.10–0.60. Several parameterisations of F_{2}^{γ} are in qualitative agreement with the measurements whereas the quark-parton model prediction fails to describe the data

Measurement of the charm structure function F_{2,c)^{γ} of the photon at LEP

The production of charm quarks is studied in deep-inelastic electron–photon scattering using data recorded by the OPAL detector at LEP at nominal e⁺e⁻ centre-of-mass energies from 183 to 209 GeV. The charm quarks have been identified by full reconstruction of charged D* mesons using their decays into D⁰π with the D⁰ observed in two decay modes with charged particle final states, Kπ and Kπππ. The cross-section σ^{D*} for production of charged D* in the reaction e⁺e⁻→e⁺e⁻D*Χ is measured in a restricted kinematical region using two bins in Bjorken x, 0.00140.1 the perturbative QCD calculation at next-to-leading order agrees perfectly with the measured cross-section. For x<0.1 the measured cross-section is 43.8±14.3±6.3±2.8 pb with a next-to-leading order prediction of 17.0⁺²·⁹_₂.₃ pb

Measurement of triple gauge boson couplings from W⁺W⁻ production at LEP energies up to 189 GeV

A measurement of triple gauge boson couplings is presented, based on W-pair data recorded by the OPAL detector at LEP during 1998 at a centre-of-mass energy of 189 GeV with an integrated luminosity of 183 pb⁻¹. After combining with our previous measurements at centre-of-mass energies of 161–183 GeV we obtain κ = 0.97_{-0.16}^{+0.20}, g_{1}^{z} = 0.991_{-0.057}^{+0.060} and λ = -0.110_{-0.055}^{+0.058}, where the errors include both statistical and systematic uncertainties and each coupling is determined by setting the other two couplings to their Standard Model values. These results are consistent with the Standard Model expectations

Colour reconnection in e+e- -> W+W- at sqrt(s) = 189 - 209 GeV

The effects of the final state interaction phenomenon known as colour reconnection are investigated at centre-of-mass energies in the range sqrt(s) ~ 189-209 GeV using the OPAL detector at LEP. Colour reconnection is expected to affect observables based on charged particles in hadronic decays of W+W-. Measurements of inclusive charged particle multiplicities, and of their angular distribution with respect to the four jet axes of the events, are used to test models of colour reconnection. The data are found to exclude extreme scenarios of the Sjostrand-Khoze Type I (SK-I) model and are compatible with other models, both with and without colour reconnection effects. In the context of the SK-I model, the best agreement with data is obtained for a reconnection probability of 37%. Assuming no colour reconnection, the charged particle multiplicity in hadronically decaying W bosons is measured to be (nqqch) = 19.38+-0.05(stat.)+-0.08 (syst.).Comment: 30 pages, 9 figures, Submitted to Euro. Phys. J.

Determination of alpha_s using Jet Rates at LEP with the OPAL detector

Hadronic events produced in e+e- collisions by the LEP collider and recorded by the OPAL detector were used to form distributions based on the number of reconstructed jets. The data were collected between 1995 and 2000 and correspond to energies of 91 GeV, 130-136 GeV and 161-209 GeV. The jet rates were determined using four different jet-finding algorithms (Cone, JADE, Durham and Cambridge). The differential two-jet rate and the average jet rate with the Durham and Cambridge algorithms were used to measure alpha(s) in the LEP energy range by fitting an expression in which order alpah_2s calculations were matched to a NLLA prediction and fitted to the data. Combining the measurements at different centre-of-mass energies, the value of alpha_s (Mz) was determined to be alpha(s)(Mz)=0.1177+-0.0006(stat.)+-0.0012$(expt.)+-0.0010(had.)+-0.0032(theo.) \.Comment: 40 pages, 17 figures, Submitted to Euro. Phys. J.

Measurement of Rb in e+e- Collisions at 182 - 209 GeV

Measurements of Rb, the ratio of the bbbar cross-section to the qqbar cross- section in e+e- collisions, are presented. The data were collected by the OPAL experiment at LEP at centre-of-mass energies between 182 GeV and 209 GeV. Lepton, lifetime and event shape information is used to tag events containing b quarks with high efficiency. The data are compatible with the Standard Model expectation. The mean ratio of the eight measurements reported here to the Standard Model prediction is 1.055+-0.031+-0.037, where the first error is statistical and the second systematic.Comment: 21 pages, 5 figures, Submitted to Phys. Letts

Scaling violations of quark and gluon jet fragmentation functions in e+e- annihilations at sqrt(s) = 91.2 and 183-209 GeV

Flavour inclusive, udsc and b fragmentation functions in unbiased jets, and flavour inclusive, udsc, b and gluon fragmentation functions in biased jets are measured in e+e- annihilations from data collected at centre-of-mass energies of 91.2, and 183-209 GeV with the OPAL detector at LEP. The unbiased jets are defined by hemispheres of inclusive hadronic events, while the biased jet measurements are based on three-jet events selected with jet algorithms. Several methods are employed to extract the fragmentation functions over a wide range of scales. Possible biases are studied in the results are obtained. The fragmentation functions are compared to results from lower energy e+e- experiments and with earlier LEP measurements and are found to be consistent. Scaling violations are observed and are found to be stronger for the fragmentation functions of gluon jets than for those of quarks. The measured fragmentation functions are compared to three recent theoretical next-to-leading order calculations and to the predictions of three Monte Carlo event generators. While the Monte Carlo models are in good agreement with the data, the theoretical predictions fail to describe the full set of results, in particular the b and gluon jet measurements.Comment: 46 pages, 17 figures, Submitted to Eur. Phys J.