Research-based flow cytometry assays for pathogenic assessment in the human B-cell biology of gene variants revealed in the diagnosis of inborn errors of immunity: a Bruton’s tyrosine kinase case-study

IntroductionInborn errors of immunity (IEI) are an expanding group of rare diseases whose field has been boosted by next-generation sequencing (NGS), revealing several new entities, accelerating routine diagnoses, expanding the number of atypical presentations and generating uncertainties regarding the pathogenic relevance of several novel variants.MethodsResearch laboratories that diagnose and provide support for IEI require accurate, reproducible and sustainable phenotypic, cellular and molecular functional assays to explore the pathogenic consequences of human leukocyte gene variants and contribute to their assessment. We have implemented a set of advanced flow cytometry-based assays to better dissect human B-cell biology in a translational research laboratory. We illustrate the utility of these techniques for the in-depth characterization of a novel (c.1685G>A, p.R562Q) de novo gene variant predicted as probably pathogenic but with no previous insights into the protein and cellular effects, located in the tyrosine kinase domain of the Bruton’s tyrosine kinase (BTK) gene, in an apparently healthy 14-year-old male patient referred to our clinic for an incidental finding of low immunoglobulin (Ig) M levels with no history of recurrent infections.Results and discussionA phenotypic analysis of bone marrow (BM) revealed a slightly high percentage of pre-B-I subset in BM, with no blockage at this stage, as typically observed in classical X-linked agammaglobulinemia (XLA) patients. The phenotypic analysis in peripheral blood also revealed reduced absolute numbers of B cells, all pre-germinal center maturation stages, together with reduced but detectable numbers of different memory and plasma cell isotypes. The R562Q variant allows Btk expression and normal activation of anti-IgM-induced phosphorylation of Y551 but diminished autophosphorylation at Y223 after anti IgM and CXCL12 stimulation. Lastly, we explored the potential impact of the variant protein for downstream Btk signaling in B cells. Within the canonical nuclear factor kappa B (NF-κB) activation pathway, normal IκBα degradation occurs after CD40L stimulation in patient and control cells. In contrast, disturbed IκBα degradation and reduced calcium ion (Ca2+) influx occurs on anti-IgM stimulation in the patient’s B cells, suggesting an enzymatic impairment of the mutated tyrosine kinase domain

International Consensus Statement on Rhinology and Allergy: Rhinosinusitis

Background: The 5 years since the publication of the first International Consensus Statement on Allergy and Rhinology: Rhinosinusitis (ICAR‐RS) has witnessed foundational progress in our understanding and treatment of rhinologic disease. These advances are reflected within the more than 40 new topics covered within the ICAR‐RS‐2021 as well as updates to the original 140 topics. This executive summary consolidates the evidence‐based findings of the document. Methods: ICAR‐RS presents over 180 topics in the forms of evidence‐based reviews with recommendations (EBRRs), evidence‐based reviews, and literature reviews. The highest grade structured recommendations of the EBRR sections are summarized in this executive summary. Results: ICAR‐RS‐2021 covers 22 topics regarding the medical management of RS, which are grade A/B and are presented in the executive summary. Additionally, 4 topics regarding the surgical management of RS are grade A/B and are presented in the executive summary. Finally, a comprehensive evidence‐based management algorithm is provided. Conclusion: This ICAR‐RS‐2021 executive summary provides a compilation of the evidence‐based recommendations for medical and surgical treatment of the most common forms of RS

Effect of poloxamer 188 vs placebo on painful vaso-occlusive episodes in children and adults with sickle cell disease : a randomized clinical trial

Key PointsQuestionCan poloxamer 188, an agent that is reported to reduce blood viscosity and cell-cell interactions, effectively reduce the duration of vaso-occlusive episodes (painful crises) in hospitalized patients with sickle cell disease? FindingsIn this randomized clinical trial that included 388 children and adults with sickle cell disease, treatment with poloxamer 188 vs placebo resulted in mean time to last dose of parenteral opioids during vaso-occlusive episodes of 81.8 vs 77.8 hours, a difference that was not statistically significant. MeaningAmong patients with sickle cell disease, poloxamer 188 did not significantly shorten the duration of painful vaso-occlusive episodes

Effect of Poloxamer 188 vs Placebo on Painful Vaso-Occlusive Episodes in Children and Adults With Sickle Cell Disease: A Randomized Clinical Trial.

Key PointsQuestionCan poloxamer 188, an agent that is reported to reduce blood viscosity and cell-cell interactions, effectively reduce the duration of vaso-occlusive episodes (painful crises) in hospitalized patients with sickle cell disease? FindingsIn this randomized clinical trial that included 388 children and adults with sickle cell disease, treatment with poloxamer 188 vs placebo resulted in mean time to last dose of parenteral opioids during vaso-occlusive episodes of 81.8 vs 77.8 hours, a difference that was not statistically significant. MeaningAmong patients with sickle cell disease, poloxamer 188 did not significantly shorten the duration of painful vaso-occlusive episodes

CTA – the World’s largest ground-based gamma-ray observatory

International audienc

Monte Carlo Simulations and Validation of NectarCAM, a Medium Sized Telescope Camera for CTA

The upcoming Cherenkov Telescope Array (CTA) ground-based gamma-ray observatory will open up our view of the very high energy Universe, offering an improvement in sensitivity of 5-10 times that of previous experiments. NectarCAM is one of the proposed cameras for the Medium-Sized Telescopes (MST) which have been designed to cover the core energy range of CTA, from 100 GeV to 10 TeV. The final camera will be capable of GHz sampling and provide a field of view of 8 degrees with its 265 modules of 7 photomultiplier each (for a total of 1855 pixels). In order to validate the performance of NectarCAM, a partially-equipped prototype has been constructed consisting of only the inner 61-modules. It has so far undergone testing at the integration test-bench facility in CEA Paris-Saclay (France) and on a prototype of the MST structure in Adlershof (Germany). To characterize the performance of the prototype, Monte Carlo simulations were conducted using a detailed model of the 61 module camera in the CORSIKA/sim_telarray framework. This contribution provides an overview of this work including the comparison of trigger and readout performance on test-bench data and trigger and image parameterization performance during on-sky measurements

Sensitivity of CTA to gamma-ray emission from the Perseus galaxy cluster

In these proceedings we summarize the current status of the study of the sensitivity of the Cherenkov Telescope Array (CTA) to detect diffuse gamma-ray emission from the Perseus galaxy cluster. Gamma-ray emission is expected in galaxy clusters both from interactions of cosmic rays (CR) with the intra-cluster medium, or as a product of annihilation or decay of dark matter (DM) particles in case they are weakly interactive massive particles (WIMPs). The observation of Perseus constitutes one of the Key Science Projects to be carried out by the CTA Consortium. In this contribution, we focus on the DM-induced component of the flux. Our DM modelling includes the substructures we expect in the main halo which will boost the annihilation signal significantly. We adopt an ON/OFF observation strategy and simulate the expected gamma-ray signals. Finally we compute the expected CTA sensitivity using a likelihood maximization analysis including the most recent CTA instrument response functions. In absence of signal, we show that CTA will allow us to provide stringent and competitive constraints on TeV DM, especially for the case of DM decay