The Solution Structures of Two Human IgG1 Antibodies Show Conformational Stability and Accommodate Their C1q and FcγR Ligands.

The human IgG1 antibody subclass shows distinct properties compared with the IgG2, IgG3, and IgG4 subclasses and is the most exploited subclass in therapeutic antibodies. It is the most abundant subclass, has a half-life as long as that of IgG2 and IgG4, binds the FcγR receptor, and activates complement. There is limited structural information on full-length human IgG1 because of the challenges of crystallization. To rectify this, we have studied the solution structures of two human IgG1 6a and 19a monoclonal antibodies in different buffers at different temperatures. Analytical ultracentrifugation showed that both antibodies were predominantly monomeric, with sedimentation coefficients s20,w (0) of 6.3-6.4 S. Only a minor dimer peak was observed, and the amount was not dependent on buffer conditions. Solution scattering showed that the x-ray radius of gyration Rg increased with salt concentration, whereas the neutron Rg values remained unchanged with temperature. The x-ray and neutron distance distribution curves P(r) revealed two peaks, M1 and M2, whose positions were unchanged in different buffers to indicate conformational stability. Constrained atomistic scattering modeling revealed predominantly asymmetric solution structures for both antibodies with extended hinge structures. Both structures were similar to the only known crystal structure of full-length human IgG1. The Fab conformations in both structures were suitably positioned to permit the Fc region to bind readily to its FcγR and C1q ligands without steric clashes, unlike human IgG4. Our molecular models for human IgG1 explain its immune activities, and we discuss its stability and function for therapeutic applications

Cosmoglobe DR1. III. First full-sky model of polarized synchrotron emission from all WMAP and Planck LFI data

We present the first model of full-sky polarized synchrotron emission that is derived from all WMAP and Planck LFI frequency maps. The basis of this analysis is the set of end-to-end reprocessed Cosmoglobe Data Release 1 sky maps presented in a companion paper, which have significantly lower instrumental systematics than the legacy products from each experiment. We find that the resulting polarized synchrotron amplitude map has an average noise rms of $3.2\,\mathrm{\mu K}$ at 30 GHz and $2^{\circ}$ FWHM, which is 30% lower than the recently released BeyondPlanck model that included only LFI+WMAP Ka-V data, and 29% lower than the WMAP K-band map alone. The mean $B$-to-$E$ power spectrum ratio is $0.40\pm0.02$, with amplitudes consistent with those measured previously by Planck and QUIJOTE. Assuming a power law model for the synchrotron spectral energy distribution, and using the $T$--$T$ plot method, we find a full-sky inverse noise-variance weighted mean of $\beta_{\mathrm{s}}=-3.07\pm0.07$ between Cosmoglobe DR1 K-band and 30 GHz, in good agreement with previous estimates. In summary, the novel Cosmoglobe DR1 synchrotron model is both more sensitive and systematically cleaner than similar previous models, and it has a more complete error description that is defined by a set of Monte Carlo posterior samples. We believe that these products are preferable over previous Planck and WMAP products for all synchrotron-related scientific applications, including simulation, forecasting and component separation.Comment: 15 pages, 15 figures, submitted to A&

Molecular and functional interactions between tumor necrosis factor-alpha receptors and the glutamatergic system in the mouse hippocampus : implications for seizure susceptibility

Tumor necrosis factor (TNF)-alpha is a proinflammatory cytokine acting on two distinct receptor subtypes, namely p55 and p75 receptors. TNF-alpha p55 and p75 receptor knockout mice were previously shown to display a decreased or enhanced susceptibility to seizures, respectively, suggesting intrinsic modifications in neuronal excitability. We investigated whether alterations in glutamate system function occur in these naive knockout mice with perturbed cytokine signaling that could explain their different propensity to develop seizures. Using Western blot analysis of hippocampal homogenates, we found that p55(-/-) mice have decreased levels of membrane GluR3 and NR1 glutamate receptor subunits while GluR1, GluR2, GluR6/7 and NR2A/B were unchanged as compared to wild-type mice. In p75(-/-) mice, GluR2, GluR3, GluR6/7 and NR2A/B glutamate receptor subunits were increased in the hippocampus while GluR1 and NR1 did not change. Extracellular single-cell recordings of the electrical activity of hippocampal neurons were carried out in anesthetized mice by standard electrophysiological techniques. Microiontophoretic application of glutamate increased the basal firing rate of hippocampal neurons in p75(-/-) mice versus wild-type mice, and this effect was blocked by 2-amino-5-phosphopentanoic acid and 6-nitro-7-sulfamoyl-benzo(f)quinoxaline-2,3-dione denoting the involvement of N-methyl-D-aspartic acid and AMPA receptors. In p55(-/-) mice, hippocampal neurons responses to glutamate were similar to wild-type mice. Spontaneous glutamate release measured by in vivo hippocampal microdialysis was significantly decreased only in p55(-/-) mice. No changes were observed in KCl-induced glutamate release in both receptor knockout mice strains versus wild-type mice. These findings highlight specific molecular and functional interactions between p55 and p75 receptor-mediated signaling and the glutamate system. These interactions may be relevant for controlling neuronal excitability in physiological and pathological conditions.peer-reviewe

Signal transduction pathways involved in proteolysis-inducing factor induced proteasome expression in murine myotubes

The proteolysis-inducing factor (PIF) is produced by cachexia-inducing tumours and initiates protein catabolism in skeletal muscle. The potential signalling pathways linking the release of arachidonic acid (AA) from membrane phospholipids with increased expression of the ubiquitin-proteasome proteolytic pathway by PIF has been studied using C2C12 murine myotubes as a surrogate model of skeletal muscle. The induction of proteasome activity and protein degradation by PIF was blocked by quinacrine, a nonspecific phospholipase A2 (PLA2) inhibitor and trifluroacetyl AA, an inhibitor of cytosolic PLA2. PIF was shown to increase the expression of calcium-independent cytosolic PLA2, determined by Western blotting, at the same concentrations as those inducing maximal expression of 20S proteasome α-subunits and protein degradation. In addition, both U-73122, which inhibits agonist-induced phospholipase C (PLC) activation and D609, a specific inhibitor of phosphatidylcholine-specific PLC also inhibited PIF-induced proteasome activity. This suggests that both PLA 2 and PLC are involved in the release of AA in response to PIF, and that this is important in the induction of proteasome expression. The two tyrosine kinase inhibitors genistein and tryphostin A23 also attenuated PIF-induced proteasome expression, implicating tyrosine kinase in this process. PIF induced phosphorylation of p44/42 mitogen-activated protein kinase (MAPK) at the same concentrations as that inducing proteasome expression, and the effect was blocked by PD98059, an inhibitor of MAPK kinase, as was also the induction of proteasome expression, suggesting a role for MAPK activation in PIF-induced proteasome expression. © 2003 Cancer Research UK

BeyondPlanck II. CMB map-making through Gibbs sampling

We present a Gibbs sampling solution to the map-making problem for CMB measurements, building on existing destriping methodology. Gibbs sampling breaks the computationally heavy destriping problem into two separate steps; noise filtering and map binning. Considered as two separate steps, both are computationally much cheaper than solving the combined problem. This provides a huge performance benefit as compared to traditional methods, and allows us for the first time to bring the destriping baseline length to a single sample. We apply the Gibbs procedure to simulated Planck 30 GHz data. We find that gaps in the time-ordered data are handled efficiently by filling them with simulated noise as part of the Gibbs process. The Gibbs procedure yields a chain of map samples, from which we may compute the posterior mean as a best-estimate map. The variation in the chain provides information on the correlated residual noise, without need to construct a full noise covariance matrix. However, if only a single maximum-likelihood frequency map estimate is required, we find that traditional conjugate gradient solvers converge much faster than a Gibbs sampler in terms of total number of iterations. The conceptual advantages of the Gibbs sampling approach lies in statistically well-defined error propagation and systematic error correction, and this methodology forms the conceptual basis for the map-making algorithm employed in the BeyondPlanck framework, which implements the first end-to-end Bayesian analysis pipeline for CMB observations.Comment: 11 pages, 10 figures. All BeyondPlanck products and software will be released publicly at http://beyondplanck.science during the online release conference (November 18-20, 2020). Connection details will be made available at the same website. Registration is mandatory for the online tutorial, but optional for the conferenc

BeyondPlanck VII. Bayesian estimation of gain and absolute calibration for CMB experiments

We present a Bayesian calibration algorithm for CMB observations as implemented within the global end-to-end BeyondPlanck (BP) framework, and apply this to the Planck Low Frequency Instrument (LFI) data. Following the most recent Planck analysis, we decompose the full time-dependent gain into a sum of three orthogonal components: One absolute calibration term, common to all detectors; one time-independent term that can vary between detectors; and one time-dependent component that is allowed to vary between one-hour pointing periods. Each term is then sampled conditionally on all other parameters in the global signal model through Gibbs sampling. The absolute calibration is sampled using only the orbital dipole as a reference source, while the two relative gain components are sampled using the full sky signal, including the orbital and Solar CMB dipoles, CMB fluctuations, and foreground contributions. We discuss various aspects of the data that influence gain estimation, including the dipole/polarization quadrupole degeneracy and anomalous jumps in the instrumental gain. Comparing our solution to previous pipelines, we find good agreement in general, with relative deviations of -0.84% (-0.67%) for 30 GHz, -0.14% (0.02%) for 44 GHz and -0.69% (-0.08%) for 70 GHz, compared to Planck 2018 (NPIPE). The deviations we find are within expected error bounds, and we attribute them to differences in data usage and general approach between the pipelines. In particular, the BP calibration is performed globally, resulting in better inter-frequency consistency. Additionally, WMAP observations are used actively in the BP analysis, which breaks degeneracies in the Planck data set and results in better agreement with WMAP. Although our presentation and algorithm are currently oriented toward LFI processing, the procedure is fully generalizable to other experiments.Comment: 18 pages, 15 figures. All BeyondPlanck products and software will be released publicly at http://beyondplanck.science during the online release conference (November 18-20, 2020). Connection details will be made available at the same website. Registration is mandatory for the online tutorial, but optional for the conferenc

BeyondPlanck XII. Cosmological parameter constraints with end-to-end error propagation

We present cosmological parameter constraints as estimated using the Bayesian BeyondPlanck (BP) analysis framework. This method supports seamless end-to-end error propagation from raw time-ordered data to final cosmological parameters. As a first demonstration of the method, we analyze time-ordered Planck LFI observations, combined with selected external data (WMAP 33-61GHz, Planck HFI DR4 353 and 857GHz, and Haslam 408MHz) in the form of pixelized maps which are used to break critical astrophysical degeneracies. Overall, all results are generally in good agreement with previously reported values from Planck 2018 and WMAP, with the largest relative difference for any parameter of about 1 sigma when considering only temperature multipoles between 29<l<601. In cases where there are differences, we note that the BP results are generally slightly closer to the high-l HFI-dominated Planck 2018 results than previous analyses, suggesting slightly less tension between low and high multipoles. Using low-l polarization information from LFI and WMAP, we find a best-fit value of tau=0.066 +/- 0.013, which is higher than the low value of tau=0.051 +/- 0.006 derived from Planck 2018 and slightly lower than the value of 0.069 +/- 0.011 derived from joint analysis of official LFI and WMAP products. Most importantly, however, we find that the uncertainty derived in the BP processing is about 30% larger than when analyzing the official products, after taking into account the different sky coverage. We argue that this is due to marginalizing over a more complete model of instrumental and astrophysical parameters, and this results in both more reliable and more rigorously defined uncertainties. We find that about 2000 Monte Carlo samples are required to achieve robust convergence for low-resolution CMB covariance matrix with 225 independent modes.Comment: 13 pages, 10 figure

BeyondPlanck XIV. Polarized foreground emission between 30 and 70GHz

We constrain polarized foreground emission between 30 and 70GHz with the Planck Low Frequency Instrument (LFI) and WMAP data within the framework of BeyondPlanck global Bayesian analysis. We combine for the first time full-resolution Planck LFI time-ordered data with low-resolution WMAP sky maps at 33, 40 and 61GHz. Spectral parameters are fit with a likelihood defined at the native resolution of each frequency channel. This analysis represents the first implementation of true multi-resolution component separation applied to CMB observations for both amplitude and spectral energy distribution (SED) parameters. For synchrotron emission, we approximate the SED as a power-law in frequency and find that the low signal-to-noise ratio of the data set strongly limits the number of free parameters that may be robustly constrained. We partition the sky into four large disjoint regions (High Latitude; Galactic Spur; Galactic Plane; and Galactic Center), each associated with its own power-law index. We find that the High Latitude region is prior-dominated, while the Galactic Center region is contaminated by residual instrumental systematics. The two remaining regions appear to be both signal-dominated and clean of systematics, and for these we derive spectral indices of $\beta_{\mathrm s}^{\mathrm{Spur}}=-3.15\pm0.07$ and $\beta_{\mathrm s}^{\mathrm{Plane}}=-3.12\pm0.06$. This agrees qualitatively with the WMAP-only polarization constraints presented by Dunkley et al. (2009), but contrasts with several temperature-based analyses. For thermal dust emission we assume a modified blackbody model and we fit the power-law index across the full sky. We find $\beta_{\mathrm{d}}=1.62\pm0.04$, which is slightly steeper than that previously reported from Planck HFI data, but still statistically consistent at a 2$\sigma$ confidence level.Comment: 17 pages, 14 figures. All BeyondPlanck products and software will be released publicly at http://beyondplanck.science during the online release conference (November 18-20, 2020). Connection details will be made available at the same website. Registration is mandatory for the online tutorial, but optional for the conferenc

Bone Mass and Strength in Older Men With Type 2 Diabetes: The Osteoporotic Fractures in Men Study

The effects of type 2 diabetes mellitus (T2DM) on bone volumetric density, bone geometry, and estimates of bone strength are not well established. We used peripheral quantitative computed tomography (pQCT) to compare tibial and radial bone volumetric density (vBMD, mg/cm3), total (ToA, mm2) and cortical (CoA, mm2) bone area and estimates of bone compressive and bending strength in a subset (n = 1171) of men (≥65 years of age) who participated in the multisite Osteoporotic Fractures in Men (MrOS) study. Analysis of covariance–adjusted bone data for clinic site, age, and limb length (model 1) and further adjusted for body weight (model 2) were used to compare data between participants with (n = 190) and without (n = 981) T2DM. At both the distal tibia and radius, patients with T2DM had greater bone vBMD (+2% to +4%, model 1, p < .05) and a smaller bone area (ToA −1% to −4%, model 2, p < .05). The higher vBMD compensated for lower bone area, resulting in no differences in estimated compressive bone strength at the distal trabecular bone regions. At the mostly cortical bone midshaft sites of the radius and tibia, men with T2DM had lower ToA (−1% to −3%, p < .05), resulting in lower bone bending strength at both sites after adjusting for body weight (−2% to −5%, p < .05) despite the lack of difference in cortical vBMD at these sites. These data demonstrate that older men with T2DM have bone strength that is low relative to body weight at the cortical-rich midshaft of the radius despite no difference in cortical vBMD. © 2010 American Society for Bone and Mineral Researc

Cosmoglobe DR1 results. II. Constraints on isotropic cosmic birefringence from reprocessed WMAP and Planck LFI data

Cosmic birefringence is a parity-violating effect that might have rotated the plane of linearly polarized light of the cosmic microwave background (CMB) by an angle $\beta$ since its emission. This has recently been measured to be non-zero at a statistical significance of $3.6\sigma$ in the official Planck PR4 and 9-year WMAP data. In this work, we constrain $\beta$ using the reprocessed BeyondPlanck LFI and Cosmoglobe DR1 WMAP polarization maps. These novel maps have both lower systematic residuals and a more complete error description than the corresponding official products. Foreground $EB$ correlations could bias measurements of $\beta$, and while thermal dust $EB$ emission has been argued to be statistically non-zero, no evidence for synchrotron $EB$ power has been reported. Unlike the dust-dominated Planck HFI maps, the majority of the LFI and WMAP polarization maps are instead dominated by synchrotron emission. Simultaneously constraining $\beta$ and the polarization miscalibration angle, $\alpha$, of each channel, we find a best-fit value of $\beta=0.35^{\circ}\pm0.70^{\circ}$ with LFI and WMAP data only. When including the Planck HFI PR4 maps, but fitting $\beta$ separately for dust-dominated, $\beta_{>70\,\mathrm{GHz}}$, and synchrotron-dominated channels, $\beta_{\leq 70\,\mathrm{GHz}}$, we find $\beta_{\leq 70\,\mathrm{GHz}}=0.53^{\circ}\pm0.28^\circ$. This differs from zero with a statistical significance of $1.9\sigma$, and the main contribution to this value comes from the LFI 70 GHz channel. While the statistical significances of these results are low on their own, the measurement derived from the LFI and WMAP synchrotron-dominated maps agrees with the previously reported HFI-dominated constraints, despite the very different astrophysical and instrumental systematics involved in all these experiments.Comment: 10 pages, 7 figures, 2 tables. Submitted to A&