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Thirteen studies have been selected, showing a good safety. The data obtained were heterogeneous in terms of posology, administration scheme and patients' evaluation, ranging from a minimum of two to a maximum of three cycles. [https://www.selleckchem.com/products/cilofexor-gs-9674.html selleck chemicals] RTX led to a sustained clinical improvement with prolonged time to relapse, in parallel to a reduction or discontinuation of other immunosuppressive therapies. Treatment with RTX appears to work in some but not all patients with anti-AChR-Ab+MG, but randomised controlled trials are needed. Future studies should take into account the subtype of MG and employ reliable measures of outcome and severity focusing on how to identify patients who may benefit from the treatment. Trial registration number NCT02110706. © Author(s) (or their employer(s)) 2020. No commercial re-use. See rights and permissions. Published by BMJ.Apolipoprotein A-I (apoA-I) is the major protein constituent of high density lipoprotein (HDL) and a target of myeloperoxidase-dependent oxidation in the artery wall. In atherosclerotic lesions, apoA-I exhibits marked oxidative modifications at multiple sites including tryptophan 72 (Trp72). Site-specific mutagenesis studies have suggested, but not conclusively shown, that oxidative modification of Trp72 of apoA-I impairs many atheroprotective properties of this lipoprotein. Herein, we used genetic code expansion technology with an engineered Saccharomyces cerevisiae tryptophanyl tRNA-synthetase (Trp-RS) suppressor tRNA pair to insert the non-canonical amino acid 5-hydroxytryptophan (5-OHTrp) at position 72 in recombinant human apoA-I, and confirmed site-specific incorporation utilizing mass spectrometry. In functional characterization studies, 5-OHTrp72 apoA-I (compared with wild-type (WT) apoA-I) exhibited reduced ATP binding cassette subfamily A member 1 (ABCA1)-dependent cholesterol acceptor activity in vitro (41.73% ± 6.57% inhibition; P less then 0.01). Additionally, 5-OHTrp72 apoA-I displayed increased activation and stabilization of paraoxonase 1 (PON1) activity (µmoles/min/mg) when compared to WT apoA-I, and comparable PON1 activation/stabilization compared to reconstituted HDL (WT apoA-I, 1.92 ± 0.04; 5-OHTrp72 apoA-I, 2.35 ± 0.0; and HDL, 2.33 ± 0.1; P less then 0.001, P less then 0.001, and P less then 0.001, respectively). Following injection into apoA-I deficient mice, 5-OHTrp72 apoA-I reached plasma levels comparable to those of native apoA-I, yet exhibited significantly reduced (48%; P less then 0.01) lipidation and evidence of HDL biogenesis. Collectively, these findings unequivocally reveal that site-specific oxidative modification of apoA-I via 5-OHTrp at Trp72 impairs cholesterol efflux and the rate-limiting step of HDL biogenesis both in vitro and in vivo. Published under license by The American Society for Biochemistry and Molecular Biology, Inc.The E-protein transcription factors guide immune cell differentiation, with E12 and E47 (hereafter called E2A) being essential for B-cell specification and maturation. E2A and the oncogenic chimera E2A-PBX1 contain three transactivation domains (ADs), with AD1 and AD2 having redundant, independent, and cooperative functions in a cell-dependent manner. AD1 and AD2 both mediate their functions by binding to the KIX domain of the histone acetyltransferase paralogues CREB-binding protein (CBP) and E1A-binding protein P300 (p300). This interaction is necessary for B-cell maturation and oncogenesis by E2A-PBX1 and occurs through conserved ϕ-x-x-ϕ-ϕ motifs (with ϕ denoting a hydrophobic amino acid) in AD1 and AD2. However, disruption of this interaction via mutation of the KIX domain in CBP/p300 does not completely abrogate binding of E2A and E2APBX1. Here, we determined that E2A-AD1 and E2A-AD2 also interact with the TAZ2 domain of CBP/p300. Characterization of the TAZ2E2AAD1(1-37) complex indicated that E2A-AD1 adopts an α-helical structure and uses its ϕ-x-x-ϕ-ϕ motif to bind TAZ2. While this region overlapped with the KIX recognition region, key KIX-interacting E2A-AD1 residues were exposed, suggesting that E2A-AD1 could simultaneously bind both the KIX and TAZ2 domains. However, we did not detect a ternary complex involving E2A-AD1, KIX, and TAZ2 and found that E2A containing both intact AD1 and AD2 is required to bind to CBP/p300. Our findings highlight the structural plasticity and promiscuity of E2A-AD1 and suggest that E2A binds both the TAZ2 and KIX domains of CBP/p300 through AD1 and AD2. Published under license by The American Society for Biochemistry and Molecular Biology, Inc.The Escherichia coli outer membrane receptor FepA transports ferric enterobactin (FeEnt) by an energy- and TonB-dependent, but otherwise mechanistically undetermined process involving its internal, 150-residue N-terminal globular domain (N-domain). We genetically introduced pairs of Cys residues in different regions of FepA tertiary structure, with the potential to form disulfide bonds.&nbsp; These included Cys-pairs on adjacent β-strands of the N-domain (intra - N), and Cys-pairs that bridged the external surface of the N-domain to the interior of the C-terminal transmembrane β-barrel (inter - N-C).&nbsp;&nbsp; We characterized FeEnt uptake by these mutants with siderophore nutrition tests, [59Fe]Ent binding and uptake experiments, and fluorescence decoy (FD) sensor assays.&nbsp; The three methods consistently showed that the intra - N disulfide bonds, that restrict conformational motion within the N-domain, prevented FeEnt uptake, whereas most inter - N-C disulfide bonds did not prevent FeEnt uptake.&nbsp; These outcomes indicate that conformational rearrangements must occur in the N-terminus of FepA during FeEnt transport.&nbsp; They also argue against disengagement of the N-domain out of the channel as a rigid body, and suggest instead that it remains within the transmembrane pore as FeEnt enters the periplasm. Published under license by The American Society for Biochemistry and Molecular Biology, Inc.Exogenous and endogenous chemicals can react with DNA to produce DNA lesions that may block DNA replication. Not much is known about the roles of polymerase (Pol) ν and Pol θ in translesion synthesis (TLS) in cells. Here, we examined the functions of both polymerases in bypassing the major-groove O 6-alkyl-2'-deoxyguanosine (O 6-alkyl-dG) and the minor-groove N 2-alkyl-dG lesions in human cells, where the alkyl groups are ethyl (Et), n-butyl (nBu), and, for O 6-alkyl-dG, pyridyloxobutyl (POB). We found that Pol ν and Pol θ promote TLS across the major-groove O 6-alkyl-dG lesions. The O 6-alkyl-dG lesions mainly induced G-&gt;A mutations, which were modulated by the two TLS polymerases and the structures of the alkyl groups. Simultaneous ablation of Pol ν and Pol θ resulted in diminished mutation frequencies for all three O 6-alkyl-dG lesions, depletion of Pol ν alone reduced mutations only for O 6-nBu-dG, and sole loss of Pol θ attenuated the mutation rates for both O 6-nBu-dG and O 6-POB-dG. Replication across the two N 2-alkyl-dG lesions was error-free, and Pol ν and Pol θ were dispensable for their replicative bypass.