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

Simultaneous Determination of Post-Translational Racemization and Isomerization of <i>N</i>‑Terminal Amyloid‑β in Alzheimer’s Brain Tissues by Covalent Chiral Derivatized Ultraperformance Liquid Chromatography Tandem Mass Spectrometry

Typical markers of protein aging are spontaneous post-translational modifications such as amino acid racemization (AAR) and amino acid isomerization (AAI) during the degradation of peptides. The post-translational AAR and AAI could significantly induce the density and localization of plaque deposition in brain tissues. Alzheimer’s disease (AD) is reliably related to the formation and aggregation of amyloid-β peptide (Aβ) plaques in the human brain. No current analytical methods can simultaneously determine AAR and AAI during the degradation of Aβ from AD patients. We now report a covalent chiral derivatized ultraperformance liquid chromatography tandem mass spectrometry (CCD-UPLC-MS/MS) method for the determination of post-translational AAR and AAI of <i>N</i>-terminal Aβ (<i>N</i>-Aβ_1–5) in human brain tissues. When subjected to tryptic <i>N</i>-Aβ_1–5 from post-translationally modified natural Aβ in focal brain tissues by the CCD procedure, it was monitored at <i>m</i>/<i>z</i> 989.6→637.0/678.9 during electrospray collision-induced dissociation. These <i>N</i>-Aβ_1–5 fragments with l-aspartic acid (l-Asp), d-Asp, l-isoAsp, and d-isoAsp could be separated using the UPLC system with a conventional reversed-phase column and mobile phase. The quantification of these peptides was determined using a stable isotope [¹⁵N]-labeled Aβ_1–40 internal standard. The CCD-UPLC-MS/MS assay of potential <i>N</i>-Aβ_1–5 allowed for the discovery of the present and ratio levels of these <i>N</i>-Aβ_1–5 sequences with l-Asp, d-Asp, l-isoAsp, and d-isoAsp

Demographic details of post-mortem brain specimens from controls and Creutzfeldt-Jakob disease patients.

<p>Demographic details of post-mortem brain specimens from controls and Creutzfeldt-Jakob disease patients.</p

Demographic details of post-mortem brain specimens from controls and Alzheimer’s disease patients.

<p>Demographic details of post-mortem brain specimens from controls and Alzheimer’s disease patients.</p

A decrease in GPI-PLD expression was observed in the caveolin-enriched membrane (CEM) fraction.

<p>Gradient fractions from control and from scrapie-infected brain lysates were assayed for GPI-PLD and PrP^Sc as described in the <i>Materials and Methods</i> section. Equal sucrose gradient fractions (1 ml each following centrifugation) were analyzed by Western blotting with the anti-GPI-PLD and anti-PrP 10E4 antibodies. Each experiment was repeated at least three times, and similar results were obtained in each experiment. An anti-caveolin-1 antibody was used as the positive control for the CEM fractions. Note lanes 5, 8 and 9, in which PrP^Sc and caveolin-1 are abundant. * Significance <i>p</i> < 0.05 compared with control levels; ** Significance <i>P</i> < 0.01 compared with control levels.</p

GPI-PLD is down-regulated in the scrapie-infected brains.

<p>(<b>A</b>) RT-PCR analysis of GPI-PLD expression levels using total RNA samples extracted from the whole brains of mice injected with either ME7 or normal brain homogenates at 160 dpi. The GPI-PLD mRNA levels were quantified using a densitometer analysis. Each value represents the mean ± SD of three samples. *Significance <i>P</i> < 0.05 compared with control levels. Actin was used as the control. (<b>B</b>) Western blot analysis of GPI-PLD protein levels in whole brain at 150 dpi from mice injected with either ME7 or normal brain homogenates. Total brain lysates of control and scrapie-infected mice were prepared as described in the <i>Materials and Methods</i> section and analyzed by western blotting with the GPI-PLD antibody. Each value represents the mean ± SD of three samples. **Significance <i>P</i> < 0.01 compared with control levels. Each experiment was repeated at least three times, and similar results were obtained in each experiment. β-actin was detected by anti-β-actin antibody and it served as the control. (<b>C-D</b>), GPI-PLD begins to be reduced at the time PrP^Sc is detected. Total lysates, which were prepared as described in the <i>Materials and Methods</i> section, were analyzed by Western blotting with the anti-GPI-PLD (C) and anti-PrP 10E4 antibodies; the latter experiment is done following PK treatment (D). Each experiment was repeated at least three times, and similar results were obtained in each experiment. β-actin was detected by anti-β-actin antibody and was used as the control. * Significance <i>p</i> < 0.05 compared to control 60 days after scrapie infection; ** Significance <i>p</i> < 0.01 compared to control 140 and 160 days after scrapie infection.</p

Additional file 1 of Polygenic effects on the risk of Alzheimer’s disease in the Japanese population

Additional file 1: Figure S1. The excluded region around the APOE gene. We removed the APOE region, consisting of ±500 kb, from around the top-hit SNP rs1160985 (chr19:45403412) in our data. Each data point indicates GWAS p values from Jansen et al. [32] used as SNP weights in the PRS calculation. Figure S2. Associations between the PRS and covariates. Age at baseline examination and years of education were examined by Spearman correlation. Sex and doses of APOE ε4 and ε2 alleles were analysed by t tests or ANOVAs. CN = cognitively normal; MCI = mild cognitive impairment; ADD = Alzheimer's disease dementia. Figure S3. Associations between the PRS.adjLD and covariates. Age at examination and years of education were examined by Spearman correlations. Sex and dose of APOE ε4 and ε2 alleles were analysed by t tests or ANOVAs. Figure S4. Comparison of AD conversion between the APOE ε4 carriers and the APOE ε4 noncarriers with high PRS. Kaplan–Meier survival curves for conversion rates of MCI to AD in the APOE ε4 carriers and the APOE ε4 noncarriers with high PRS values. p-values were calculated by log-rank test. Figure S5. Age differences between the low- and high-PRS groups and between the nonconverters and converters. Baseline ages were compared between groups using the Wilcoxon rank-sum test. Each violin plot includes the kernel probability density of the data at different values and the box plots with the median value and the interquartile range

Additional file 1: Figure S1. of A diagnostic marker for superficial urothelial bladder carcinoma: lack of nuclear ATBF1 (ZFHX3) by immunohistochemistry suggests malignant progression

Specificity and sensitivity of the seven anti-ATBF1 antibodies. Western blot analysis of ATBF1 in HEK293T cells using the seven anti-ATBF1 antibodies (Fig. 5a MB33, MB34, MB39, D1-120, MB44, MB47 and MB49). Lanes 1, 3, 5, 7, 9, 11, and 13 represent HEK293T cells with an HA-tag expression vector (pCI-HA). Lanes 2, 4, 6, 8, 10, 12, and 14 represent HEK293T cells containing an HA-tagged ATBF1 expression vector (pCI-HA-ATBF1). HEK293T cells were grown in DMEM supplemented with 10 % fetal bovine serum at 37 °C and 5 % CO2. HEK293T cells were transfected with the HA-tagged expression vector or the HA-tagged ATBF1 expression vector (HA-ATBF1) using transIT-293 reagent. (PPTX 215 kb

Meta-analysis of top-ranked association results with <i>SORL1</i> in Japanese, Korean, and Caucasian datasets.

<p>CH:MB, chromosome:position (in megabase pairs, build 19); MA, minor allele; MAF, minor allele frequenc; OR, odds ratio; <i>P</i> P-value.</p

Top-ranked genome-wide association results in the Japanese discovery (Stage 1) sample (P<2.5×10⁻⁵) and their replication in Japanese (Stage 2).

<p>CH:MB, chromosome:position (in megabasepairs, build 19); MA, minor allele; MAF, minor allele frequency; # SNPs, the number of SNPs for which P≤1×10⁻⁴ in the discovery (Stage 1) sample; OR, odds ratio; <i>P</i> P-value;</p><p>Selected SNPs represent the strongest association within each locus.</p