Development of novel methods for non-canonical myeloma protein analysis with an innovative adaptation of immunofixation electrophoresis, native top-down mass spectrometry, and middle-down de novo sequencing

OBJECTIVES: Multiple myeloma (MM) is a malignant plasma cell neoplasm, requiring the integration of clinical examination, laboratory and radiological investigations for diagnosis. Detection and isotypic identification of the monoclonal protein(s) and measurement of other relevant biomarkers in serum and urine are pivotal analyses. However, occasionally this approach fails to characterize complex protein signatures. Here we describe the development and application of next generation mass spectrometry (MS) techniques, and a novel adaptation of immunofixation, to interrogate non-canonical monoclonal immunoproteins. METHODS: Immunoprecipitation immunofixation (IP-IFE) was performed on a Sebia Hydrasys Scan2. Middle-down de novo sequencing and native MS were performed with multiple instruments (21T FT-ICR, Q Exactive HF, Orbitrap Fusion Lumos, and Orbitrap Eclipse). Post-acquisition data analysis was performed using Xcalibur Qual Browser, ProSight Lite, and TDValidator. RESULTS: We adapted a novel variation of immunofixation electrophoresis (IFE) with an antibody-specific immunosubtraction step, providing insight into the clonal signature of gamma-zone monoclonal immunoglobulin (M-protein) species. We developed and applied advanced mass spectrometric techniques such as middle-down de novo sequencing to attain in-depth characterization of the primary sequence of an M-protein. Quaternary structures of M-proteins were elucidated by native MS, revealing a previously unprecedented non-covalently associated hetero-tetrameric immunoglobulin. CONCLUSIONS: Next generation proteomic solutions offer great potential for characterizing complex protein structures and may eventually replace current electrophoretic approaches for the identification and quantification of M-proteins. They can also contribute to greater understanding of MM pathogenesis, enabling classification of patients into new subtypes, improved risk stratification and the potential to inform decisions on future personalized treatment modalities

New Observations of Molecular Nitrogen by the Imaging Ultraviolet Spectrograph on MAVEN

International audienceThe Martian ultraviolet dayglow provides information on the basic state of the Martian upper atmosphere. The Imaging Ultraviolet Spectrograph (IUVS) on NASA’s Mars Atmosphere and Volatile Evolution (MAVEN) mission has observed Mars at mid and far-UV wavelengths since its arrival in September 2014. In this work, we describe a linear regression method used to extract components of UV spectra from IUVS limb observations and focus in particular on molecular nitrogen (N2) photoelectron excited emissions. We identify N2 Lyman-Birge-Hopfield (LBH) emissions for the first time at Mars and we also confirm the tentative identification of N2 Vegard-Kaplan (VK) emissions. We compare observed VK and LBH limb radiance profiles to model results between 90 and 210 km. Finally, we compare retrieved N2 density profiles to general circulation (GCM) model results. Contrary to earlier analyses using other satellite data that indicated N2 densities were a factor of three less than predictions, we find that N2 abundances exceed GCM results by about a factor of two at 130 km but are in agreement at 150 km

Mars’ seasonal mesospheric transport seen through nitric oxide nightglow

International audienceWe analyze the ultraviolet nightglow in the atmosphere of Mars through nitric oxide (NO) δ and γ band emissions as observed by the Imaging UltraViolet Spectrograph (IUVS) instrument onboard the Mars Atmosphere and Volatile EvolutioN (MAVEN) spacecraft when it is at apoapse and periapse.In the dayside thermosphere of Mars, solar extreme-ultraviolet radiation dissociates CO2 and N2 molecules. O(3P) and N(4S) atoms are carried from the dayside to the nightside by the day-night hemispheric transport process, where they descend through the nightside mesosphere and can radiatively recombine to form NO(C2Π). The excited molecules rapidly relax by emitting photons in the UV δ and γ bands. These emissions are indicators of the N and O atom fluxes from the dayside to Mars’ nightside and the descending circulation pattern from the nightside thermosphere to the mesosphere (e.g. Bertaux et al., 2005 ; Bougher et al., 1990 ; Cox et al., 2008 ; Gagné et al., 2013 ; Gérard et al., 2008 ; Stiepen et al., 2015, 2017).Observations of these emissions are gathered from a large dataset spanning different seasonal conditions.We present discussion on the variability in the brightness and altitude of the emission with season, geographical position (longitude), and local time, along with possible interpretation by local and global changes in the mesosphere dynamics. We show the possible impact of atmospheric waves forcing longitudinal variability and data-to-model comparisons indicating a wave-3 structure in Mars’ nightside mesosphere. Quantitative comparison with calculations of the Laboratoire de Météorologie Dynamique-Mars Global Climate Model (LMD-MGCM) suggests the model reproduces both the global trend of NO nightglow emission and its seasonal variation. However, it also indicates large discrepancies, with the emission up to a factor 50 times fainter in the model, suggesting that the predicted transport is too efficient toward the night winter pole in the thermosphere by ˜20° latitude to the north.These questions are now addressed through an extensive dataset of disk images, in complement to improved simulations of the LMD-MGCM and the Mars Global Ionosphere-Thermosphere Model (MGITM) models

Seasonal Transport in Mars’ Mesosphere-Thermosphere revealed by Nitric Oxide nightglow

International audienceWe analyze the ultraviolet nightglow in the atmosphere of Mars through the Nitric Oxide (NO) δ and γ band emissions observed by the Imaging Ultraviolet Spectrograph (IUVS, McClintock et al., 2015) when the Mars Atmosphere and Volatile EvolutioN (MAVEN) spacecraft is at apoapsis and periapsis.On the dayside thermosphere of Mars, solar extreme ultraviolet radiation dissociates CO2 and N2 molecules. O(3P) and N(4S) atoms are carried by the day-to-night hemispheric transport. They descend in the nightside mesosphere, where they can radiatively recombine to form NO(C2Π). The excited molecules rapidly relax by emitting UV photons in the δ and γ bands. These emissions are thus indicators of the N and O atom fluxes transported from the dayside to Mars’ nightside and the descending circulation pattern from the nightside thermosphere to the mesosphere (e.g. Bertaux et al., 2005 ; Bougher et al., 1990 ; Cox et al., 2008 ; Gagné et al., 2013 ; Gérard et al., 2008 ; Stiepen et al., 2015, 2017).A large dataset of nightside disk images and vertical limb scans during southern winter, fall equinox and southern summer conditions have been accumulated since the beginning of the mission.We will present a discussion regarding the variability of the brightness and altitude of the emission with season, geographical position (longitude) and local time and possible interpretation for local and global changes in the mesosphere dynamics. We show the possible impact of atmospheric waves structuring the emission longitudinally and indicating a wave-3 structure in Mars’ nightside mesosphere. Quantitative comparison with calculations from the LMD-MGCM (Laboratoire de Météorologie Dynamique-Mars Global Climate Model) show that the model globally reproduces the trends of the NO nightglow emission and its seasonal variation but also indicates large discrepancies (up to a factor 50 fainter in the model) suggesting that the predicted transport is too efficient toward the night winter pole in the thermosphere by ∼20° latitude north.These questions are now addressed with an extensive dataset of disk images, complemented with improved simulations from the LMD-MGCM and new M-GITM (Mars Global Ionosphere-Thermosphere Model) simulations of emissions for selected sampling periods

First Retrieval of Thermospheric Carbon Monoxide From Mars Dayglow Observations

International audienceAs a minor species in the Martian thermosphere, Carbon Monoxide (CO) is a tracer that can be used to constrain changing circulation patterns between the lower thermosphere and upper mesosphere of Mars. By linking CO density distributions to dynamical wind patterns, the structure and variability of the atmosphere will be better understood. Direct measurements of CO can therefore provide insight into the magnitude and pattern of winds and provide a metric for studying the response of the atmosphere to solar forcing. In addition, CO measurements can help solve outstanding photochemical modeling problems in explaining the abundance of CO at Mars. CO is directly observable by electron impact excitation and solar resonance fluorescence emissions in the far-ultraviolet (FUV). The retrieval of CO from solar fluorescence was first proposed over 40 years ago, but has been elusive at Mars due to significant spectral blending. However, by simulating the spectral shape of each contributing emission feature, electron impact excitation and solar fluorescence brightnesses can be extracted from the composite spectrum using a multiple linear regression approach. We use CO Fourth Positive Group (4PG) molecular band emission observed on the limb (130 - 200 km) by the Imaging Ultraviolet Spectrograph (IUVS) on NASA’s Mars Atmosphere and Volatile Evolution (MAVEN) spacecraft over both northern and southern hemispheres from October 2014 to December 2016. We present the first direct retrieval of CO densities by FUV remote sensing in the upper atmosphere of Mars. Atmospheric composition is inferred using the terrestrial Atmospheric Ultraviolet Radiance Integrated Code adapted to the Martian atmosphere. We investigate the sensitivity of CO density retrievals to variability in solar irradiance, solar longitude, and local time. We compare our results to predictions from the Mars Global Ionosphere-Thermosphere Model as well as in situ measurements by the Neutral Gas and Ion Mass Spectrometer on MAVEN and quantify any differences

Loss of the Martian atmosphere to space: Present-day loss rates determined from MAVEN observations and integrated loss through time

International audienceObservations of the Mars upper atmosphere made from the Mars Atmosphere and Volatile Evolution (MAVEN) spacecraft have been used to determine the loss rates of gas from the upper atmosphere to space for a complete Mars year (16 Nov 2014 – 3 Oct 2016). Loss rates for H and O are sufficient to remove ∼2-3 kg/s to space. By itself, this loss would be significant over the history of the planet. In addition, loss rates would have been greater early in history due to the enhanced solar EUV and more-active Sun. Integrated loss, based on current processes whose escape rates in the past are adjusted according to expected solar evolution, would have been as much as 0.8 bar CO2 or 23 m global equivalent layer of H2O; these losses are likely to be lower limits due to the nature of the extrapolation of loss rates to the earliest times. Combined with the lack of surface or subsurface reservoirs for CO2 that could hold remnants of an early, thick atmosphere, these results suggest that loss of gas to space has been the dominant process responsible for changing the climate of Mars from an early, warmer environment to the cold, dry one that we see today