Researching peers and disaster vulnerable communities: an insider perspective

Conducting research among peers and communities that a researcher also serves may be both daunting and rewarding. Researching peers may make the researcher feel uncomfortable raising certain questions that are sensitive or that could be construed to be testing their competencies. This paper is inclined more towards showing that it is advantageous to be an insider, whose position can facilitate collection of information that could not have been accessed, or revealed to an outsider. The paper reports on fieldwork conducted in a low-income country in Sub-Sahara Africa as part of a doctoral study with communities affected by disasters and those that work with such communities. The paper demonstrates the complexities of conducting such research and provides some insights that may be useful to insiders, outsiders or “in-betweeners” embarking on fieldwork in low-income countries and among vulnerable population struggling with manifold stresses and shocks

Additional file 1 of Attenuation of OX40 signaling suppression by age disrupts peripheral deletion of CD4+ T cells specific for the epidermal autoantigen desmoglein 3

Additional file 1: Table S1. A comprehensive list of antibodies used in the experiments as well as information on the target antigen, clone, type of fluorescence, catalog number, and the respective supplying companies. Fig. S1. a,Outline of the adoptive transfer of Dsg3H1-Rag2−/− T cells and WT CD4+ T cells to Dsg3−/− or WT mice. b, FCM plots of Dsg3−/−and WT mice after the transfer of CFSE-labeled Ly5.1+ Dsg3H1-Rag2−/− T cells and Ly5.2+ WT T cells. The gating strategy used to identify 7AAD− CD4+ cells is illustrated. Ly5.1+Dsg3H1-Rag2−/− T cells and CFSE+Ly5.1− co-transferred WT T cells were gated (red and black squares, respectively). Proliferation indicated by CFSE intensity reduction of Ly5.1+Dsg3H1-Rag2−/− T cells are observed only in WT mice. Fig. S2. a, FCM plots demonstrate the gating strategy employed to identify proliferating Ly5.1+ Dsg3H1-Rag2−/− Tcells in SLN. Cutoff values for the expression of IFN-γ, Birc5, and OX40 in Dsg3H1-Rag2−/− T cells were based on their expression levels in recipient T cells. Fluorescence minus one (FMO) controls of CD4, Ly5.1,CFSE, IFN-γ, Birc5, and OX40 are also shown. b, FCM plots show the gating strategy employed to identify four subsets of migrating dendritic cells in SLN. FMO controls of CD45, CD11c, IA/IE, CD11b, Langerin and CD103 are also shown. c, FCM plots demonstrate the gating strategy employed to identify Tregs in SLN. FMO controls of Foxp3 and OX40 are also shown

Additional file 3: of A GRIA2 and PAX8-positive renal solitary fibrous tumor with NAB2-STAT6 gene fusion

Results of reverse transcription-polymerase chain reaction (RT-PCR). Fusion of the NAB2-STAT6 gene was detected by RT-PCR with several sets of specifically designed forward and reverse PCR primers. Sequences of the PCR primers are given in Additional file 2. SFT, solitary fibrous tumor. (PNG 73 kb

Differential glycan profiles of PNGase F-treated IgA1 of mIgA-MIDD.

<p>IgA1 purified from mIgA-MIDD serum was incubated with (left) or without (right) PNGase F. After digestion, the reaction mixture was labeled with Cy3-SE and subjected to the lectin microarray. The relative intensity of each lectin was normalized to the maximum fluorescence intensity. mIgA-MIDD, monoclonal immunoglobulin deposition disease associated with monoclonal IgA; PNGase F, peptide N-glycosidase F.</p

Lectin blot analysis of IgA1 purified from mIgA-MIDD serum under reducing and non-reducing conditions.

<p>IgA1 purified from mIgA-MIDD serum was pretreated in SDS buffer with or without 2-ME, separated by SDS-PAGE under non-reducing (lane 1) or reducing (lane 2) conditions, and then subjected to western blotting with an anti-IgA1 mAb (A), ConA (B), and WFA (C). Cross-reacting bands were detected using Konica immunostaining kit (Konica, Tokyo, Japan) for anti-IgA1 mAb and ConA and Western Lightning Chemiluminescence Plus (Perkin-Elmer, Boston, MA) for WFA. A Gal deficient IgA (lane 3), enzymatically deglycosylated with neuraminidase and galactosidase, was used as a positive control for WFA lectin. mIgA-MIDD, monoclonal immunoglobulin deposition disease associated with monoclonal IgA; 2-ME, 2-mercaptoethanol; ConA, jack bean lectin concanavalin A; WFA, Wisteria floribunda agglutinin.</p

IgA Nephropathy Caused by Unusual Polymerization of IgA1 with Aberrant N-Glycosylation in a Patient with Monoclonal Immunoglobulin Deposition Disease

<div><p>Immunoglobulin A nephropathy (IgAN) is a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region. The cause of IgAN is unknown, but multiple mechanisms have been suggested. We previously reported a rare case of mesangioproliferative glomerulonephritis in a patient with monoclonal immunoglobulin deposition disease associated with monoclonal IgA1. In this study, we performed the detailed analyses of serum IgA1 from this patient in comparison with those from patients with mIgA plasma cell disorder without renal involvement and healthy volunteers. We found unusual polymerization of IgA1 with additional <i>N</i>-glycosylation distinctive in this patient, which was different from known etiologies. Glycan profiling of IgA1 by the lectin microarray revealed an intense signal for <i>Wisteria floribunda</i> agglutinin (WFA). This signal was reduced by disrupting the native conformation of IgA1, suggesting that the distinct glycan profile was reflecting the conformational alteration of IgA1, including the glycan conformation detected as additional <i>N</i>-glycans on both the heavy and light chains. This unusually polymerized state of IgA1 would cause an increase of the binding avidity for lectins. WFA specifically recognized highly polymerized and glycosylated IgA1. Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation.</p></div

Sandwich lectin ELISA of sequential deglycosylated IgA1 in mIgA-MIDD.

<p>IgA1 purified from mIgA-MIDD serum was digested with neuraminidase, and then β-galactosidase or β1,4-galactosidase. Digested and undigested samples were subjected to a sandwich lectin ELISA with HPA (A), VVA (B), PNA (C), and WFA (D) as described in the Methods. The relative intensity of each lectin was normalized to the IgA1 concentration. HPA, <i>Helix pomatia</i> agglutinin; VVA, <i>Vicia villosa</i> lectin; PNA, peanut agglutinin.</p

SDS-polyacrylamide gel electrophoresis analyses of purified IgA1.

<p>IgA1 purified from HV, MPCD, and mIgA-MIDD was boiled in SDS buffer with (A) or without (B) 2-ME and subjected to SDS-PAGE.</p

SDS-agarose gel electrophoresis of purified IgA1.

<p>IgA1 purified from sera of two HV, two MPCD patients, and an mIgA-MIDD patient was incubated in SDS buffer and then subjected to agarose gel electrophoresis analysis.</p

Differential glycan profiles of purified IgA1 after trypsin digestion.

<p>IgA1 purified from sera of three HVs, two MPCD patients, and one mIgA-MIDD patient was incubated with trypsin. Each tryptic digest was labeled with Cy3-SE and subjected to the lectin microarray. The relative intensities of lectins were normalized to the maximum fluorescence intensity.</p