Patient characteristics during ICU stay.

<p>Data are n (%) or median (interquartile range).</p><p>LOD: logistic organ dysfunction; Pcuff: cuff pressure; ACV: assist control ventilation, PSV: pressure support ventilation.</p

Study flowchart.

<p>Study flowchart.</p

Mean α-amylase levels in patients with microaspiration.

<p>Mean α-amylase levels in patients with microaspiration.</p

Alpha amylase results.

#<p>p<0.05 versus microaspiration.</p>§<p>p<0.05 versus abundant microaspiration.</p><p>*Youden’s index.</p

Accuracy of all α-amylase levels, coming from all tracheal aspirates, in diagnosing microaspiration.

<p>Area under the receiver operator curve 0.56±0.05 [95% CI 0.53–0.60].</p

Patient characteristics at ICU admission.

<p>Data are n (%) or median (interquartile range).</p><p>SAPS: simplified acute physiology score; LOD: logistic organ dysfunction; COPD: chronic obstructive pulmonary disease; ICU: intensive care unit, ARDS: acute respiratory distress syndrome.</p>#<p>p<0.05 versus microaspiration,</p>§<p>p<0.05 versus abundant microaspiration.</p><p>*Several patients had more than one cause for ICU admission.</p

Correlation between α-amylase and pepsin.

<p>r² =  O.305, P = 0.001.</p

Isolation and Characterization of a Primary Proximal Tubular Epithelial Cell Model from Human Kidney by CD10/CD13 Double Labeling

<div><p>Renal proximal tubular epithelial cells play a central role in renal physiology and are among the cell types most sensitive to ischemia and xenobiotic nephrotoxicity. In order to investigate the molecular and cellular mechanisms underlying the pathophysiology of kidney injuries, a stable and well-characterized primary culture model of proximal tubular cells is required. An existing model of proximal tubular cells is hampered by the cellular heterogeneity of kidney; a method based on cell sorting for specific markers must therefore be developed. In this study, we present a primary culture model based on the mechanical and enzymatic dissociation of healthy tissue obtained from nephrectomy specimens. Renal epithelial cells were sorted using co-labeling for CD10 and CD13, two renal proximal tubular epithelial markers, by flow cytometry. Their purity, phenotypic stability and functional properties were evaluated over several passages. Our results demonstrate that CD10/CD13 double-positive cells constitute a pure, functional and stable proximal tubular epithelial cell population that displays proximal tubule markers and epithelial characteristics over the long term, whereas cells positive for either CD10 or CD13 alone appear to be heterogeneous. In conclusion, this study describes a method for establishing a robust renal proximal tubular epithelial cell model suitable for further experimentation.</p></div

Sorting proximal tubular cells using specific antibodies.

<p>(A) Fluorescence plot showing cells labeled with antibodies against CD10 (APC: allophycocyanin) and CD13 (PE: phycoerythrin). FACS analysis revealed about 4% double-positive cells. (B) Fluorescence plot showing cells treated with isotypes to both antibodies to determine the upper threshold for non-specific fluorescence.</p

Evaluation of PT cells and CD10/CD13 double-negative cells phenotypic stability.

<p>(A) Fluorescence plot showing PT cells labeled with antibodies against CD10 (APC: allophycocyanin) and CD13 (PE: phycoerythrin) after four passages. Flow cytometry revealed about 94% double-positive cells. (B) Relative percentage of CD10/CD13 double-positive cells at passages 2, 3, 4 and 5 in the PT cells populations (n = 4). NS: non-significant (p>0.05). (C) Representative western blots for PT cells over 5 passages. Blots were incubated with antibodies against aquaporin-1, N-cadherin, MUC1. The β-actin protein was used as an internal control (D) Fluorescence plot showing the CD10/CD13 double-negative cell population labeled with antibodies against CD10 and CD13 after two passages. Flow cytometry revealed about 15% double-negative cells.</p