Epicardial/Intrathoracic Fat Volume with Sample Weight Adjustment by Gender.

<p>The box whisker plot displays min and max (lower and upper whiskers), first (Q1) and third (Q3) quartile (central rectangle), median (the segment inside the rectangle), and mean (○/+ inside the rectangle).</p

Pericardial Tracing to Define Epicardial Fat Volume.

<p>Pericardial Tracing to Define Epicardial Fat Volume.</p

Pearson Correlation^* and Minimally Adjusted Model^@: Epicardial Fat Volume and Intrathoracic Fat Volume Associated with Selected Covariates.

<p>Pearson Correlation^{<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0159958#t002fn002" target="_blank">*</a>} and Minimally Adjusted Model<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0159958#t002fn003" target="_blank">^@</a>: Epicardial Fat Volume and Intrathoracic Fat Volume Associated with Selected Covariates.</p

Lymphoblastoid Cell Lines as a Tool to Study Inter-Individual Differences in the Response to Glucose

<div><p>Background</p><p>White blood cells have been shown in animal studies to play a central role in the pathogenesis of diabetic retinopathy. Lymphoblastoid cells are immortalized EBV-transformed primary B-cell leukocytes that have been extensively used as a model for conditions in which white blood cells play a primary role. The purpose of this study was to investigate whether lymphoblastoid cell lines, by retaining many of the key features of primary leukocytes, can be induced with glucose to demonstrate relevant biological responses to those found in diabetic retinopathy.</p><p>Methods</p><p>Lymphoblastoid cell lines were obtained from twenty-three human subjects. Differences between high and standard glucose conditions were assessed for expression, endothelial adhesion, and reactive oxygen species.</p><p>Results</p><p>Collectively, stimulation of the lymphoblastoid cell lines with high glucose demonstrated corresponding changes on molecular, cellular and functional levels. Lymphoblastoid cell lines up-regulated expression of a panel of genes associated with the leukocyte-mediated inflammation found in diabetic retinopathy that include: a cytokine (<i>IL-1B</i> fold change = 2.11, p-value = 0.02), an enzyme (<i>PKCB</i> fold change = 2.30, p-value = 0.01), transcription factors <i>(NFKB-p50</i> fold change = 2.05, p-value = 0.01), <i>(NFKB-p65</i> fold change = 2.82, p-value = 0.003), and an adhesion molecule (<i>CD18</i> fold change = 2.59, 0.02). Protein expression of CD18 was also increased (p-value = 2.14x10^-5). The lymphoblastoid cell lines demonstrated increased adhesiveness to endothelial cells (p = 1.28x10^-5). Reactive oxygen species were increased (p = 2.56x10^-6). Significant inter-individual variation among the lymphoblastoid cell lines in these responses was evident (F = 18.70, p < 0.0001).</p><p>Conclusions</p><p>Exposure of lymphoblastoid cell lines derived from different human subjects to high glucose demonstrated differential and heterogeneous gene expression, adhesion, and cellular effects that recapitulated features found in the diabetic state. Lymphoblastoid cells may represent a useful tool to guide an individualized understanding of the development and potential treatment of diabetic complications like retinopathy.</p></div

High glucose induces gene expression up-regulation in lymphoblastoid cell lines.

<p>Diabetes associated genes are increased in lymphoblastoid cell lines (n = 23) exposed to high glucose (HG) (30 mM). Figure is a bar graph of the fold change in gene expression in response to high glucose. Expression was normalized to <i>GAPDH</i>. Change in gene expression is based on the difference in expression of each gene under high and standard glucose conditions (11 mM). Error bars represent 95% confidence intervals of fold change. * p-value < 0.05. NS Not significant p > 0.05.</p

Methods in the DCCT/EDIC Study.

<p>Methods in the DCCT/EDIC Study.</p

Expression of CD18 by flow cytometry is increased in lymphoblastoid cell lines exposed to high glucose.

<p>CD18 expression was compared between standard glucose (11 mM) and high glucose (HG) (30 mM) conditions. CD18 is increased in high glucose (p = 2.14x10^-5) consistent with the mRNA expression findings. Figure is a univariate scatter plot showing the distribution of change in relative fluorescence units (RFU) for CD18 expression in high glucose compared to standard glucose conditions in twenty-two lymphoblastoid cell lines. Most but not all cell lines demonstrate an increase in CD18 under high glucose conditions. Mean and 95% confidence intervals shown.</p

Certification and Standardized Processes in the DCCT/EDIC Study.

<p>Certification and Standardized Processes in the DCCT/EDIC Study.</p

Quality Control Monitoring in the DCCT/EDIC Study.

<p>^a Data analyzed at the central reading units (e.g. CBL, CORU, CERC);</p><p>^b EDIC-specific quality assessments</p><p>^c Height: two measurements within 1 cm; if not, measure twice more</p><p>^d Weight: two measurements within 0.2 kg; if not, measure twice more</p><p>Quality Control Monitoring in the DCCT/EDIC Study.</p

Reactive oxygen species are increased in lymphoblastoid cell lines exposed to high glucose.

<p>Measurements performed in twenty-three subject lymphoblastoid cell lines. Reactive oxygen species were measured under both standard (11 mM) and high glucose (HG) (30 mM) cell culture conditions for each lymphoblastoid cell line. Reactive oxygen species assayed by mean CM-H2DCFDA fluorescence and reported in relative fluorescence units (RFU). Univariate scatter plot demonstrates differential response for each of 23 subject lymphoblastoid cell lines to high glucose for the formation of reactive oxygen species. Significant increases in reactive oxygen species production in high glucose are evident (p = 2.56x10^–6). Scatter plot reveals increased reactive oxygen species formation for most but not all lymphoblastoid cell lines in high glucose compared to standard glucose conditions. Mean and 95% confidence intervals shown.</p