Data_Sheet_1_HMGN1 and R848 Synergistically Activate Dendritic Cells Using Multiple Signaling Pathways.pdf

High mobility group nucleosome-binding protein 1 (HMGN1 or N1) is a Th1-polarizing alarmin, but alone is insufficient to induce antitumor immunity. We previously showed that combination of N1 and R848, a synthetic TLR7/8 agonist, synergistically activates dendritic cells (DCs) and induces therapeutic antitumor immunity, however, it remained unclear how N1 and R848 synergistically activate DCs. Here, we show that co-stimulation with N1 and R848 of human monocyte-derived DCs (MoDCs) markedly upregulated DC's surface expression of CD80, CD83, CD86, and HLA-DR, as well as synergistic production of pro-inflammatory cytokines including IL-12p70, IL-1β, and TNF-α. This combination also synergistically activated NF-κB and multiple MAPKs that are involved in DC maturation. Moreover, N1 and R848 synergistically increased nuclear translocation of interferon (IFN) regulatory transcription factors (e.g., IRF3 and IRF7) and promoted the expression of type 1 IFNs such as IFN-α2, IFN-α4, and IFN-β1. Similar signaling pathways were also induced in mouse bone marrow-derived DCs (BMDCs). RNA-seq analysis in human MoDCs revealed that N1 plus R848 synergistically upregulated the expression of genes predominantly involved in DC maturation pathway, particularly genes critical for the polarization of Th1 immune responses (e.g., IL12A, IL12B, and IFNB1, etc.). Overall, our findings show that (1) N1 synergizes with R848 in activating human and mouse DCs and (2) the synergistic effect based on various intracellular signaling events culminated in the activation of multiple transcriptional factors. These findings have important implications for future clinical trials since N1 and R848 synergistically promoted optimal Th1 lineage immune responses resulting in tumor rejection in mice.</p

Stabilization of Ruthenium(II) Polypyridyl Chromophores on Mesoporous TiO₂ Electrodes: Surface Reductive Electropolymerization and Silane Chemistry

Stabilization is a critical issue in the long term operation of dye-sensitized photoelectrosynthesis cells (DSPECs) for water splitting or CO2 reduction. The cells require a stable binding of the robust molecular chromophores, catalysts, and chromophore/catalyst assemblies on metal oxide semiconductor electrodes under the corresponding (photoelectro)­chemical conditions. Here, an efficient stabilization strategy is presented based on functionalization of FTO|nanoTiO2 (mesoporous, nanostructured TiO2 deposited on fluorine-doped tin oxide (FTO) glass) electrodes with a vinylsilane followed by surface reductive electropolymerization of a vinyl-derivatized Ru­(II) polypyridyl chromophore. The surface electropolymerization was dominated by a grafting-through mechanism, and rapidly completed within minutes. Chromophore surface coverages were controlled up to three equivalent monolayers by the number of electropolymerization cycles. The silane immobilization and cross-linked polymer network produced highly (photo)­stabilized chromophore-grafted FTO|nanoTiO2 electrodes. The electrodes showed significant improvements over structures based on atomic layer deposition and polymer dip-coating stabilization methods in a wide pH range from pH ≈ 1 to pH ≈ 12.5 under both dark and light conditions. Under illumination, with hydroquinone added as a sacrificial electron transfer donor, a photoresponse for sustained electron transfer mediation occurred for at least ∼20 h in a pH ≈ 7.5 phosphate buffer (0.1 M NaH2PO4/Na2HPO4, with 0.5 M NaClO4). The overall procedure provides an efficient way to fabricate highly stabilized molecular assemblies on electrode surfaces with potential applications for DSPECs in solar fuels

Simultaneous Electrosynthesis of Syngas and an Aldehyde from CO₂ and an Alcohol by Molecular Electrocatalysis

A tandem cell for artificial photosynthesis with CO2 and water as the oxidants and an organic alcohol as the reductant is described. The use of molecular catalysts with high activity and selectivity, in an appropriate cell configuration, leads to electrochemical reduction of CO2 and water to CO and H2 (syngas) in tandem with benzyl alcohol oxidation to benzaldehyde. A faradaic efficiency (FE) of ∼70% for the formation of benzaldehyde was obtained with simultaneous syngas generation with varying ratios of H2 and CO at the cathode. The maximum energy efficiency obtained for the electrochemical cell was 17.6%

Stable Molecular Surface Modification of Nanostructured, Mesoporous Metal Oxide Photoanodes by Silane and Click Chemistry

Binding functional molecules to nanostructured mesoporous metal oxide surfaces provides a way to derivatize metal oxide semiconductors for applications in dye-sensitized photoelectrosynthesis cells (DSPECs). The commonly used anchoring groups, phosphonates and carboxylates, are unstable as surface links to oxide surfaces at neutral and high pH, leading to rapid desorption of appended molecules. A synthetically versatile molecular attachment strategy based on initial surface modification with a silyl azide followed by click chemistry is described here. It has been used for the stable installation of surface-bound metal complexes. The resulting surfaces are highly stabilized toward complex loss with excellent thermal, photochemical, and electrochemical stabilities. The procedure involves binding 3-azidopropyltrimethoxysilane (APTMS) to nanostructured mesoporous TiO2 or tin-doped indium oxide (ITO) electrodes by silane attachment followed by azide-terminated, Cu­(I)-catalyzed azide–alkyne cycloaddition (CuAAC) reactions with an alkyne-derivatized ruthenium­(II) polypyridyl complex. The chromophore-modified electrodes display enhanced photochemical and electrochemical stabilities compared to phosphonate surface binding with extended photoelectrochemical oxidation of hydroquinone for more than ∼6 h with no significant decay

Completing a Charge Transport Chain for Artificial Photosynthesis

A ruthenium polypyridyl chromophore with electronically isolated triarylamine substituents has been synthesized that models the role of tyrosine in the electron transport chain in photosystem II. When bound to the surface of a TiO₂ electrode, electron injection from a Ru­(II) Metal-to-Ligand Charge Transfer (MLCT) excited state occurs from the complex to the electrode to give Ru­(III). Subsequent rapid electron transfer from the pendant triarylamine to Ru­(III) occurs with an observed rate constant of ∼10¹⁰ s^–1, which is limited by the rate of electron injection into the semiconductor. Transfer of the oxidative equivalent away from the semiconductor surface results in dramatically reduced rates of back electron transfer, and a long-lived (τ = ∼165 μs) triarylamine radical cation that has been used to oxidize hydroquinone to quinone in solution

The most parsimonious tree generated from analysis of 138 <i>Alu</i> insertions in Lemuriformes.

<p>The amplification patterns of the <i>Alu</i> insertions were used to construct a Dollo parsimony tree of phylogenetic relationships with <i>G. senegalensis</i> and <i>H. sapiens</i> as outgroups using the MESQUITE and PAUP* programs. Numbers above branches are bootstrap values. The significance level of each node supported by insertions as determined by likelihood testing is indicated by either *(p<0.05) or **(p<0.01). Numbers below arrows indicate the number of unambiguous loci supporting that node. Numbers in brackets below arrows indicate the number of loci at a given informative node identified by McLain et al. (2012). Numbers in parentheses represent insertions that are only present in one species or group. These insertions are not parsimony-informative. Consistency index (CI): 1.000; Homoplasy index (HI): 0.000; Retention index (RI): 1.000.</p

PCR amplification of polymorphic <i>Alu</i> insertions in Lemuriformes.

<p>Gel photographs displaying the methodology for establishing evolutionary relationships using <i>Alu</i> elements. The presence and absence of elements, supplemented by sequencing to eliminate the possibility of confounding events, is used to determine which species are more closely related. A total of 5 gel electrophoresis results on a 24-species primate panel are shown with <i>H. sapiens</i> and <i>G. senegalensis</i> as outgroups. <b>A:</b> Amplification of locus Str71B, an <i>Alu</i> insertion shared by the infraorder Lemuriformes. <b>B:</b> Amplification of locus MmA39, an <i>Alu</i> insertion shared by the family Cheirogaleidae. <b>C:</b> Amplification of locus MmA27, an <i>Alu</i> insertion shared by the sister genera <i>Microcebus</i> and <i>Mirza</i>. <b>D:</b> Amplification of locus Str59, an <i>Alu</i> insertion specific to the genus <i>Microcebus</i>. <b>E:</b> Amplification of locus Em6, an <i>Alu</i> insertion affirming the monophyly of the family Lemuridae to the exclusion of other lemur species and outgroups.</p