17 research outputs found
Confocal microscopy images of double immunohistochemical stainings for insulin (green), the acinar enzyme carboxypeptidase A (red) or both the markers (yellow in merge panels).
<p>Blue staining is DNA (DAPI, Hoechst dye. Representative cells are shown from a control non-diabetic subject (left panels) and from a subject with type 2 diabetes (right panel).</p
Representative image (left panel) of a pancreatic cell in T2D samples containing both amylase (black arrows) and insulin (white arrows) granules, with adjacent cells of the immune system.
<p>The image has been reconstructed from several pictures at 10,000x final magnification. Enlargments are provided in the right panels. Mc: macrophage, Lymph: lymphocyte, Ms: mast cell.</p
Electron microscopy images (magnification 10,000x) of pancreatic cells containing both zymogen-like (black arrows) and insulin-like (white arrows) granules in the four cases with T2D.
<p>Electron microscopy images (magnification 10,000x) of pancreatic cells containing both zymogen-like (black arrows) and insulin-like (white arrows) granules in the four cases with T2D.</p
Immunogold labeling of amylase (particle size: 5 nm) and/or insulin (particle size: 15 nm) in an acinar cell (A, square enlarged in a, showing only amylase particles), a beta cell (B, square enlarged in b, showing only insulin particles), and a hybrid cell (C, square enlarged in c, showing both amylase and insulin particles) in a T2D sample.
<p>A, B and C: magnification 38,000x; the cell membrane is indicated by the dotted black arrows. Lymph indicates a lymphocyte close to the hybrid cell (see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0179398#pone.0179398.g007" target="_blank">Fig 7</a> for additional images of cells of the immune system).</p
<i>AKT1</i> and <i>SELP</i> Polymorphisms Predict the Risk of Developing Cachexia in Pancreatic Cancer Patients
<div><p>Pancreatic ductal adenocarcinoma (PDAC) patients have the highest risk of developing cachexia, which is a direct cause of reduced quality of life and shorter survival. Novel biomarkers to identify patients at risk of cachexia are needed and might have a substantial impact on clinical management. Here we investigated the prognostic value and association of <i>SELP-rs6136</i>, <i>IL6-rs1800796</i> and <i>AKT1-rs1130233</i> polymorphisms with cachexia in PDAC. Genotyping was performed in DNA from blood samples of a test and validation cohorts of 151 and 152 chemo-naive locally-advanced/metastatic PDAC patients, respectively. The association of <i>SELP-rs6136</i>, <i>IL6-rs1800796</i> and <i>AKT1-rs1130233</i> polymorphisms with cachexia as well as the correlation between cachexia and the candidate polymorphisms and overall survival were analyzed. Akt expression and phosphorylation in muscle biopsies were evaluated by specific ELISA assays. <i>SELP-rs6136-AA</i> and <i>AKT1-rs1130233-AA/GA</i> genotypes were associated with increased risk of developing cachexia in both cohorts (<i>SELP: p</i> = 0.011 and <i>p</i> = 0.045; <i>AKT1: p</i> = 0.004 and <i>p</i> = 0.019 for the first and second cohorts, respectively), while patients carrying <i>AKT1-rs1130233-GG</i> survived significantly longer (<i>p</i> = 0.002 and <i>p</i> = 0.004 for the first and second cohorts, respectively). In the multivariate analysis <i>AKT1-rs1130233-AA/GA</i> genotypes were significant predictors for shorter survival, with an increased risk of death of 1.7 (<i>p</i> = 0.002) and 1.6 (<i>p</i> = 0.004), in the first and second cohorts, respectively. This might be explained by the reduced phosphorylation of Akt1 in muscle biopsies from patients harboring <i>AKT1-rs1130233-AA/GA</i> (<i>p</i> = 0.003), favoring apoptosis induction. In conclusion, <i>SELP</i> and <i>AKT1</i> polymorphisms may play a role in the risk of cachexia and death in PDAC patients, and should be further evaluated in larger prospective studies.</p></div
Representative semithin and electron microscopy images of pancreatic cells in T2D slides.
<p>A: Semithin section (stained with a 1:1 mixture of toluidine blue, 1% in bidistilled water, and methylen blue, 1% in bidistilled water) showing a cluster of 4 cells with islet-like appearance (magnification: 1,000x), enlarged in B (N: nucleus); C and D: Electron microscopy of cells containing both zymogen-like (black arrows) and insulin-like (white arrows) granules (magnification: 10,000x). ER in D indicates the endoplasmic reticulum, looking expanded and convoluted in this specific T2D beta cell.</p
Immunogold labeling of amylase (particle size: 5 nm) and insulin (particle size: 15 nm) of an hybrid cell (magnification: 38,000x) in T2D samples, showing different cellular areas (numbered squares are enlarged) with both positivities in the cytoplasm.
<p>Immunogold labeling of amylase (particle size: 5 nm) and insulin (particle size: 15 nm) of an hybrid cell (magnification: 38,000x) in T2D samples, showing different cellular areas (numbered squares are enlarged) with both positivities in the cytoplasm.</p
Akt1 expression in muscle samples according to the <i>AKT1-rs1130233</i> polymorphism.
<p>Bar graphs illustrating the mean±SD expression of total Akt1 (<b>A</b>) and phospho-Akt1 (<b>B</b>) in muscle samples from patients with differential <i>AKT1-rs1130233</i> genotypes (N = 9 samples in each group) *<i>p</i><0.05.</p
Correlation between cachexia and candidate <i>SELP</i>, <i>AKT1</i> and <i>IL-6</i> SNPs.
<p><i>*p-values were calculated with Fisher's exact test.</i></p><p><i>SNPs, single nucleotide polymorphisms.</i></p><p>Note: <i>SELP</i> and <i>IL6</i> SNPs was detectable in all the samples, while the <i>AKT1</i> genotype could not be determined in 2 of the patients of the second cohort.</p><p>Correlation between cachexia and candidate <i>SELP</i>, <i>AKT1</i> and <i>IL-6</i> SNPs.</p
Clinical outcome according to cachexia and <i>AKT1-rs1130233</i> polymorphism.
<p>(<b>A</b> and <b>B</b>) Kaplan–Meier survival curves according to cachexia in the first and second cohort of patients. (<b>C</b> and <b>D</b>) Kaplan–Meier survival curves according to the <i>AKT1-rs1130233</i> polymorphism in the first and second cohort of patients. <i>p</i>-values were calculated with the log-rank test. OS: Overall survival.</p