Early and late outcomes after cord blood transplantation for pediatric patients with inherited leukodystrophies

Leukodystrophies (LD) are devastating inherited disorders leading to rapid neurological deterioration and premature death. Hematopoietic stem cell transplantation (HSCT) can halt disease progression for selected LD. Cord blood is a common donor source for transplantation of these patients because it is rapidly available and can be used without full HLA matching. However, precise recommendations allowing care providers to identify patients who benefit from HSCT are lacking. In this study, we define risk factors and describe the early and late outcomes of 169 patients with globoid cell leukodystrophy, X-linked adrenoleukodystrophy, and metachromatic leukodystrophy undergoing cord blood transplantation (CBT) at an European Society for Blood and Marrow Transplantation center or at Duke University Medical Center from 1996 to 2013. Factors associated with higher overall survival (OS) included presymptomatic status (77% vs 49%; P = .006), well-matched (80 preCBT, 50% remained stable, 20% declined to 60 to 80, and 30% to, 60. Overall, an encouraging OS was found for LD patients after CBT, especially for those who are presymptomatic before CBT and received adequately dosed grafts. Early identification and fast referral to a specialized center may lead to earlier treatment and, subsequently, to improved outcomes

In Vivo Analysis of the Role of O-Glycosylations of Von Willebrand Factor

The objective of this project was to study the function of O-glycosylations in von Willebrand factor (VWF) life cycle. In total, 14 different murine Vwf cDNAs mutated on one or several O-glycosylations sites were generated: 9 individual mutants, 2 doublets, 2 clusters and 1 mutant with all 9 murine glycosylation sites mutated (Del-O-Gly). We expressed each mutated cDNA in VWF deficient-mice by hydrodynamic injection. An immunosorbent assay with Peanut Agglutinin (PNA) was used to verify the O-glycosylation status. Wild-type (WT) VWF expressed by hepatocytes after hydrodynamic injection was able to bind PNA with slightly higher affinity than endothelial-derived VWF. In contrast, the Del-O-Gly VWF mutant did not bind PNA, demonstrating removal of O-linked glycans. All mutants displayed a normal multimeric pattern. Two mutants, Del-O-Gly and T1255A/T1256A, led to expression levels 50% lower than those induced by WT VWF and their half-life in vivo was significantly reduced. When testing the capacity of each mutant to correct the bleeding time of VWF-deficient mice, we found that S1486A, T1255A, T1256A and the doublet T1255A/T1256A were unable to do so. In conclusion we have shown that O-glycosylations are dispensable for normal VWF multimerization and biosynthesis. It also appears that some O-glycosylation sites, particularly the T1255 and T1256 residues, are involved in the maintenance of VWF plasma levels and are essential for normal haemostasis. As for the S1486 residue, it seems to be important for platelet binding as demonstrated in vitro using perfusion experiments

A Conformation-Sensitive Monoclonal Antibody against the A2 Domain of von Willebrand Factor Reduces Its Proteolysis by ADAMTS13

The size of von Willebrand factor (VWF), controlled by ADAMTS13-dependent proteolysis, is associated with its hemostatic activity. Many factors regulate ADAMTS13-dependent VWF proteolysis through their interaction with VWF. These include coagulation factor VIII, platelet glycoprotein 1bα, and heparin sulfate, which accelerate the cleavage of VWF. Conversely, thrombospondin-1 decreases the rate of VWF proteolysis by ADAMTS13 by competing with ADAMTS13 for the A3 domain of VWF. To investigate whether murine monoclonal antibodies (mAbs) against human VWF affect the susceptibility of VWF to proteolysis by ADAMTS13 in vitro, eight mAbs to different domains of human VWF were used to evaluate the effects on VWF cleavage by ADAMTS13 under fluid shear stress and static/denaturing conditions. Additionally, the epitope of anti-VWF mAb (SZ34) was mapped using recombinant proteins in combination with enzyme-linked immunosorbent assay and Western blot analysis. The results indicate that mAb SZ34 inhibited proteolytic cleavage of VWF by ADAMTS13 in a concentration-dependent manner under fluid shear stress, but not under static/denaturing conditions. The binding epitope of SZ34 mAb is located between A1555 and G1595 in the central A2 domain of VWF. These data show that an anti-VWF mAb against the VWF-A2 domain (A1555-G1595) reduces the proteolytic cleavage of VWF by ADAMTS13 under shear stress, suggesting the role of this region in interaction with ADAMTS13

Platelet clearance via shear-induced unfolding of a membrane mechanoreceptor

Mechanisms by which blood cells sense shear stress are poorly characterized. In platelets, glycoprotein (GP)Ib-IX receptor complex has been long suggested to be a shear sensor and receptor. Recently, a relatively unstable and mechanosensitive domain in the GPIba subunit of GPIb-IX was identified. Here we show that binding of its ligand, von Willebrand factor, under physiological shear stress induces unfolding of this mechanosensory domain (MSD) on the platelet surface. The unfolded MSD, particularly the juxtamembrane € Trigger' sequence therein, leads to intracellular signalling and rapid platelet clearance. These results illustrate the initial molecular event underlying platelet shear sensing and provide a mechanism linking GPIb-IX to platelet clearance. Our results have implications on the mechanism of platelet activation, and on the pathophysiology of von Willebrand disease and related thrombocytopenic disorders. The mechanosensation via receptor unfolding may be applicable for many other cell adhesion receptors

Measurement of cross sections for production of a Z boson in association with a flavor-inclusive or doubly b-tagged large-radius jet in proton-proton collisions at Formula Presented with the ATLAS experiment

We present measurements of cross sections for production of a leptonically decaying Z boson in association with a large-radius jet in 13 TeV proton-proton collisions at the LHC, using 36 fb - 1 of data from the ATLAS detector. Integrated and differential cross sections are measured at particle level in both a flavor inclusive and a doubly b -tagged fiducial phase space. The large-radius jet mass and transverse momentum, its kinematic relationship to the Z boson, and the angular separation of b -tagged small-radius track jets within the large-radius jet are measured. This measurement constitutes an important test of perturbative quantum chromodynamics in kinematic and flavor configurations relevant to several Higgs boson and beyond-Standard-Model physics analyses. The results highlight issues with modeling of additional hadronic activity in the flavor-inclusive selection, and a distinction between flavor-number schemes in the b -tagged phase space

Measurement of substructure-dependent jet suppression in Pb+Pb collisions at 5.02 TeV with the ATLAS detector

The ATLAS detector at the Large Hadron Collider has been used to measure jet substructure modification and suppression in Pb+Pb collisions at a nucleon–nucleon center-of-mass energy √sNN = 5.02 TeV in comparison with proton–proton (pp) collisions at √s = 5.02 TeV. The Pb+Pb data, collected in 2018, have an integrated luminosity of 1.72 nb−1, while the ppdata, collected in 2017, have an integrated luminosity of 260 pb−1. Jets used in this analysis are clustered using the anti-kt algorithm with a radius parameter R = 0.4. The jet constituents, defined by both tracking and calorimeter information, are used to determine the angular scale rg of the first hard splitting inside the jet by reclustering them using the Cambridge–Aachen algorithm and employing the soft-drop grooming technique. The nuclear modification factor, RAA, used to characterize jet suppression in Pb+Pb collisions, is presented differentially in rg, jet transverse momentum, and in intervals of collision centrality. The RAA value is observed to depend significantly on jet rg. Jets produced with the largest measured rg are found to be twice as suppressed as those with the smallest rg in central Pb+Pb collisions. The RAA values do not exhibit a strong variation with jet pT in any of the rg intervals. The rg and pT dependence of jet RAA is qualitatively consistent with a picture of jet quenching arising from coherence and provides the most direct evidence in support of this approach

Performance of the ATLAS forward proton Time-of-Flight detector in Run 2

We present performance studies of the Time-of-Flight (ToF) subdetector of the ATLAS Forward Proton (AFP) detector at the LHC. Efficiencies and resolutions are measured using high-statistics data samples collected at low and moderate pile-up in 2017, the first year when the detectors were installed on both sides of the interaction region. While low efficiencies are observed, of the order of a few percent, the resolutions of the two ToF detectors measured individually are 21 ps and 28 ps, yielding an expected resolution of the longitudinal position of the interaction, z vtx, in the central ATLAS detector of 5.3 ± 0.6 mm. This is in agreement with the observed width of the distribution of the difference between z vtx, measured independently by the central ATLAS tracker and by the ToF detector, of 6.0 ± 2.0 mm

Anomaly detection search for new resonances decaying into a Higgs boson and a generic new particle X in hadronic final states using Formula Presented pp collisions with the ATLAS detector

A search is presented for a heavy resonance Formula Presented decaying into a Standard Model Higgs boson Formula Presented and a new particle Formula Presented in a fully hadronic final state. The full Large Hadron Collider run 2 dataset of proton-proton collisions at Formula Presented collected by the ATLAS detector from 2015 to 2018 is used and corresponds to an integrated luminosity of Formula Presented. The search targets the high Formula Presented-mass region, where the Formula Presented and Formula Presented have a significant Lorentz boost in the laboratory frame. A novel application of anomaly detection is used to define a general signal region, where events are selected solely because of their incompatibility with a learned background-only model. It is constructed using a jet-level tagger for signal-model-independent selection of the boosted Formula Presented particle, representing the first application of fully unsupervised machine learning to an ATLAS analysis. Two additional signal regions are implemented to target a benchmark Formula Presented decay into two quarks, covering topologies where the Formula Presented is reconstructed as either a single large-radius jet or two small-radius jets. The analysis selects Higgs boson decays into Formula Presented, and a dedicated neural-network-based tagger provides sensitivity to the boosted heavy-flavor topology. No significant excess of data over the expected background is observed, and the results are presented as upper limits on the production cross section Formula Presented) for signals with Formula Presented between 1.5 and 6 TeV and Formula Presented between 65 and 3000 GeV. A search is presented for a heavy resonance Y decaying into a Standard Model Higgs boson H and a new particle X in a fully hadronic final state. The full Large Hadron Collider run 2 dataset of proton-proton collisions at √ s = 13     TeV collected by the ATLAS detector from 2015 to 2018 is used and corresponds to an integrated luminosity of 139     fb − 1 . The search targets the high Y -mass region, where the H and X have a significant Lorentz boost in the laboratory frame. A novel application of anomaly detection is used to define a general signal region, where events are selected solely because of their incompatibility with a learned background-only model. It is constructed using a jet-level tagger for signal-model-independent selection of the boosted X particle, representing the first application of fully unsupervised machine learning to an ATLAS analysis. Two additional signal regions are implemented to target a benchmark X decay into two quarks, covering topologies where the X is reconstructed as either a single large-radius jet or two small-radius jets. The analysis selects Higgs boson decays into b ¯ b , and a dedicated neural-network-based tagger provides sensitivity to the boosted heavy-flavor topology. No significant excess of data over the expected background is observed, and the results are presented as upper limits on the production cross section σ ( p p → Y → X H → q ¯ q b ¯ b ) for signals with m Y between 1.5 and 6 TeV and m X between 65 and 3000 GeV

The Hepatitis B Virus Ribonuclease H Is Sensitive to Inhibitors of the Human Immunodeficiency Virus Ribonuclease H and Integrase Enzymes

Nucleos(t)ide analog therapy blocks DNA synthesis by the hepatitis B virus (HBV) reverse transcriptase and can control the infection, but treatment is life-long and has high costs and unpredictable long-term side effects. The profound suppression of HBV by the nucleos(t)ide analogs and their ability to cure some patients indicates that they can push HBV to the brink of extinction. Consequently, more patients could be cured by suppressing HBV replication further using a new drug in combination with the nucleos(t)ide analogs. The HBV ribonuclease H (RNAseH) is a logical drug target because it is the second of only two viral enzymes that are essential for viral replication, but it has not been exploited, primarily because it is very difficult to produce active enzyme. To address this difficulty, we expressed HBV genotype D and H RNAseHs in E. coli and enriched the enzymes by nickel-affinity chromatography. HBV RNAseH activity in the enriched lysates was characterized in preparation for drug screening. Twenty-one candidate HBV RNAseH inhibitors were identified using chemical structure-activity analyses based on inhibitors of the HIV RNAseH and integrase. Twelve anti-RNAseH and anti-integrase compounds inhibited the HBV RNAseH at 10 μM, the best compounds had low micromolar IC50 values against the RNAseH, and one compound inhibited HBV replication in tissue culture at 10 μM. Recombinant HBV genotype D RNAseH was more sensitive to inhibition than genotype H. This study demonstrates that recombinant HBV RNAseH suitable for low-throughput antiviral drug screening has been produced. The high percentage of compounds developed against the HIV RNAseH and integrase that were active against the HBV RNAseH indicates that the extensive drug design efforts against these HIV enzymes can guide anti-HBV RNAseH drug discovery. Finally, differential inhibition of HBV genotype D and H RNAseHs indicates that viral genetic variability will be a factor during drug development. © 2013 Tavis et al