Psychometric validity of the distress thermometer and problem check list in ART-na\uefve HIV infected patients in Northern Nigeria

Background: HIV diagnosis comes with a lot of worry and distress. Ability to objectively estimate this distress by non-psychiatrist will enhance early detection of psychological distress for intervention. Objectives: To investigate the validity of the Distress Thermometer (DT) and its problem checklist in achieving early detection of mental distress among ART-na\uefve HIV infected patient. Materials and Methods: A total of 90 ART-na\uefve HIV infected patients completed the DT and its problem check list, Hospital Anxiety Depression Scale (HADS), Oslo Social Support Scale and the 14-item Resilience Scale. Results: The DT was positively correlated with all the measures of distress and reversely correlated with all the positive wellness in this study. The correlations were only significant for the negative measures of psychological wellness. The internal consistency of the DT\u2019s problem list overall and sub-categories were within acceptable range (i.e. \u3b1 > 0.50). The Receiver Operating Characteristic (ROC) curves and Area Under the Curves (AUC) analysis were significant and found the DT and Problem List to respectively differentiate between cases of distress, anxiety and depression. The DT\u2019s cut-off was >5.0 with AUC range (0.754 \u2013 0.709); sensitivity range (81.0% \u2013 70.4%); specificity range (68.3% \u2013 65.2%) for distress, anxiety and depression as determined by HADS. And the Problem List cut-off was >6.0 with AUC range (0.854 \u2013 0.821); sensitivity range (90.5% - 85.7%); specificity range (68.3% - 65.2%) for distress, anxiety and depression as measured by HADS. Conclusion: The DT and Problem List were found to be valid measures of distress in ART-na\uefve HIV infected patients

Protein Engineering of the N‑Terminus of NEMO: Structure Stabilization and Rescue of IKKβ Binding

NEMO is a scaffolding protein that, together with the catalytic subunits IKKα and IKKβ, plays an essential role in the formation of the IKK complex and in the activation of the canonical NF-κB pathway. Rational drug design targeting the IKK-binding site on NEMO would benefit from structural insight, but to date, the determination of the structure of unliganded NEMO has been hindered by protein size and conformational heterogeneity. Here we show how the utilization of a homodimeric coiled-coil adaptor sequence stabilizes the minimal IKK-binding domain NEMO(44–111) and furthers our understanding of the structural requirements for IKK binding. The engineered constructs incorporating the coiled coil at the N-terminus, C-terminus, or both ends of NEMO(44–111) present high thermal stability and cooperative melting and, most importantly, restore IKKβ binding affinity. We examined the consequences of structural content and stability by circular dichoism and nuclear magnetic resonance (NMR) and measured the binding affinity of each construct for IKKβ­(701–745) in a fluorescence anisotropy binding assay, allowing us to correlate structural characteristics and stability to binding affinity. Our results provide a method for engineering short stable NEMO constructs to be suitable for structural characterization by NMR or X-ray crystallography. Meanwhile, the rescuing of the binding affinity implies that a preordered IKK-binding region of NEMO is compatible with IKK binding, and the conformational heterogeneity observed in NEMO(44–111) may be an artifact of the truncation

Reining in Polyoma Virus Associated Nephropathy: Design and Characterization of a Template Mimicking BK Viral Coat Protein Cellular Binding

The BK polyoma virus is a leading cause of chronic post kidney transplantation rejection. One target for therapeutic intervention is the initial association of the BK virus with the host cell. We hypothesize that the rate of BKV infection can be curbed by competitively preventing viral binding to cells. The X-ray structures of homologous viruses complexed with N-terminal glycoproteins suggest that the BC and HI loops of the viral coat are determinant for binding and thereby infection of the host cell. The large size of the viral coat precludes it from common biophysical and small molecule screening studies. Hence, we sought to develop a smaller protein template incorporating the identified binding loops of the BK viral coat in a manner that adequately mimics the binding characteristics of the BK virus coat protein to cells. Such a mimic may serve as a tool for the identification of inhibitors of BK viral progression. Herein, we report the design and characterization of a reduced-size and soluble template derived from a four-helix proteinTM1526 of <i>Thermatoga maritima</i> archaea bacteriawhich maintains the topological display of the BC and HI loops as found in the viral coat protein, VP1, of BKV. We demonstrate that the GT1b and GD1b sialogangliosides, which bind to the VP1 of BKV, also associate with our BKV template. Employing a GFP-tagged template, we show host cell association that is dose dependent and that can be reduced by neuraminidase treatment. These data demonstrate that the BKV template mimics the host cell binding observed for the wild-type virus coat protein VP1

Palmitate‐TLR4 signaling regulates the histone demethylase, JMJD3, in macrophages and impairs diabetic wound healing

Chronic macrophage inflammation is a hallmark of type 2 diabetes (T2D) and linked to the development of secondary diabetic complications. T2D is characterized by excess concentrations of saturated fatty acids (SFA) that activate innate immune inflammatory responses, however, mechanism(s) by which SFAs control inflammation is unknown. Using monocyte‐macrophages isolated from human blood and murine models, we demonstrate that palmitate (C16:0), the most abundant circulating SFA in T2D, increases expression of the histone demethylase, Jmjd3. Upregulation of Jmjd3 results in removal of the repressive histone methylation (H3K27me3) mark on NFκB‐mediated inflammatory gene promoters driving macrophage‐mediated inflammation. We identify that the effects of palmitate are fatty acid specific, as laurate (C12:0) does not regulate Jmjd3 and the associated inflammatory profile. Further, palmitate‐induced Jmjd3 expression is controlled via TLR4/MyD88‐dependent signaling mechanism, where genetic depletion of TLR4 (Tlr4−/−) or MyD88 (MyD88−/−) negated the palmitate‐induced changes in Jmjd3 and downstream NFκB‐induced inflammation. Pharmacological inhibition of Jmjd3 using a small molecule inhibitor (GSK‐J4) reduced macrophage inflammation and improved diabetic wound healing. Together, we conclude that palmitate contributes to the chronic Jmjd3‐mediated activation of macrophages in diabetic peripheral tissue and a histone demethylase inhibitor‐based therapy may represent a novel treatment for nonhealing diabetic wounds.Palmitate drives chronic macrophage mediated inflammation in diabetic tissue via upregulation of the histone demethylase, Jmjd3, and a histone demethylase inhibitor‐based therapy may represent a novel treatment for non∖healing diabetic wounds.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/163876/1/eji4870.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/163876/2/eji4870-sup-0001-SuppMat.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/163876/3/eji4870_am.pd

Epigenetic regulation of the PGE2 pathway modulates macrophage phenotype in normal and pathologic wound repair

Macrophages are a primary immune cell involved in inflammation, and their cell plasticity allows for transition from an inflammatory to a reparative phenotype and is critical for normal tissue repair following injury. Evidence suggests that epigenetic alterations play a critical role in establishing macrophage phenotype and function during normal and pathologic wound repair. Here, we find in human and murine wound macrophages that cyclooxygenase 2/prostaglandin E2 (COX-2/PGE2) is elevated in diabetes and regulates downstream macrophage-mediated inflammation and host defense. Using single-cell RNA sequencing of human wound tissue, we identify increased NF-κB–mediated inflammation in diabetic wounds and show increased COX-2/PGE2 in diabetic macrophages. Further, we identify that COX-2/PGE2 production in wound macrophages requires epigenetic regulation of 2 key enzymes in the cytosolic phospholipase A2/COX-2/PGE2 (cPLA2/COX-2/PGE2) pathway. We demonstrate that TGF-β–induced miRNA29b increases COX-2/PGE2 production via inhibition of DNA methyltransferase 3b–mediated hypermethylation of the Cox-2 promoter. Further, we find mixed-lineage leukemia 1 (MLL1) upregulates cPLA2 expression and drives COX-2/PGE2. Inhibition of the COX-2/PGE2 pathway genetically (Cox2fl/fl Lyz2Cre+) or with a macrophage-specific nanotherapy targeting COX-2 in tissue macrophages reverses the inflammatory macrophage phenotype and improves diabetic tissue repair. Our results indicate the epigenetically regulated PGE2 pathway controls wound macrophage function, and cell-targeted manipulation of this pathway is feasible to improve diabetic wound repair