Obstructive sleep apnea and rhonchopathy are associated with downregulation of trefoil factor family peptide 3 (TFF3)—Implications of changes in oral mucus composition - Fig 2

<p><b>TFF analysis by RT-PCR (A) and Western blot (B).</b> (A), Detection of TFF3 at ∼300 bp and ß-actin as loading control at ∼240 bp. Different samples of uvula tissue from mild OSA (lanes 1 and 2), moderate OSA (lanes 3 and 4), severe OSA (lane 5) and rhonchopathy (lane 6) are shown. DEPC-H₂O served as negative control (lane 8), RNA-extract from lung tissue was used as a positive control (lane 7). (B) Detection of TFF3 after SDS gel electrophoresis under reducing conditions in protein isolates from the uvula tissue of patients suffering from mild OSA (lane 1), moderate OSA (lane 2), severe OSA (lane 3) and rhonchopathy (lane 4). Lung (6) served as positive control. Bands are visible at 20 kDa and 9 kDa for TFF3 and at 42 kDa for the loading control GAPDH.</p

ELISA quantification of TFF3 and -2 protein concentration with regard to gender in saliva samples from patients with rhonchopathy and mild, moderate or severe OSA.

<p>No significant changes can be observed between the different groups evaluated for TFF3 (A) or TFF2 (B).</p

ELISA quantification of TFF3 and -2 in terms of BMI correlation in saliva samples from patients with rhonchopathy and mild, moderate or severe OSA.

<p>(A) The TFF3 protein concentration reveals a tendency (red line) to decline from rhonchopathy to severe OSA. However, this concentration decrease is not significant. (B) The TFF2 protein concentration reveals no significant changes between the different groups evaluated. In order to match patients in groups, we used the BMI classification of Diet and Health depending on age: </p><p></p><p></p><p>Age                BMI</p><p></p><p></p><p>19–24 years            19-24</p><p></p><p></p><p>23–34 years            20-25</p><p></p><p></p><p>35–44 years            21-26</p><p></p><p></p><p>45–54 years            22-27</p><p></p><p></p><p>55–64 years            23-28</p><p></p><p></p><p>>65 years            24-29 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0185200#pone.0185200.ref028" target="_blank">28</a>]</p><p></p><p></p><p></p> <p>Age                BMI</p> <p>19–24 years            19-24</p> <p>23–34 years            20-25</p> <p>35–44 years            21-26</p> <p>45–54 years            22-27</p> <p>55–64 years            23-28</p> <p>>65 years            24-29 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0185200#pone.0185200.ref028" target="_blank">28</a>]</p

ELISA quantification of TFF3 and -2 protein concentration with regard to the Epworth Sleepiness Scale (ESS) in saliva samples from patients with rhonchopathy and mild, moderate or severe OSA.

<p>No significant changes are observed between the different groups evaluated for TFF3 (A) or TFF2 (B). The Epworth Sleepiness Scale assessment is based on a standardized questionnaire that retrospectively measures the probability of falling asleep in typical daily situations. The subjective assessment is done by patients who can choose between a scale of 4 probabilities (0 = never falling asleep to 3 = high probability of falling asleep). The sum results in a code distinguishing between low ESS <11/24 and high ESS ≥11/24.</p

Data of AHI, age, BMI, ESS, gender and smoking from healthy controls, rhonchopathy patients, mild, moderate and severe OSA.

<p>Data of AHI, age, BMI, ESS, gender and smoking from healthy controls, rhonchopathy patients, mild, moderate and severe OSA.</p

ELISA quantification of TFF3 and -2 protein concentration with regard to age in saliva samples from patients with rhonchopathy and mild, moderate or severe OSA.

<p>No significant changes are observed between the different groups evaluated for TFF3 (A) or TFF2 (B). We defined six age groups: </p><p></p><p></p><p>Group 1 = 19–24 years</p><p></p><p></p><p>Group 2 = 25–34 years</p><p></p><p></p><p>Group 3 = 35–44 years</p><p></p><p></p><p>Group 4 = 45–54 years</p><p></p><p></p><p>Group 5 = 55–64 years</p><p></p><p></p><p>Group 6 = >65 years</p><p></p><p></p><p></p> <p>Group 1 = 19–24 years</p> <p>Group 2 = 25–34 years</p> <p>Group 3 = 35–44 years</p> <p>Group 4 = 45–54 years</p> <p>Group 5 = 55–64 years</p> <p>Group 6 = >65 years</p

ELISA quantification of TFF3 and -2 concentrations with regard to smoking in saliva samples from patients with rhonchopathy and mild, moderate or severe OSA.

<p>(A) The TFF3 protein concentration reveals a tendency to decline between non-smokers and smokers from rhonchopathy to severe OSA (red line). However, this concentration decrease is not significant. (B) The TFF2 protein concentration does not reveal any significant changes.</p

ELISA protein quantification of TFF3 and -2 in saliva samples from healthy volunteers and from patients with rhonchopathy and mild, moderate or severe OSA.

<p>(A) A significant decline in the TFF3 protein concentration is visible in all samples from patients in relation to the healthy control group without rhonchopathy. A slight but not significant fall in the protein concentrations between rhonchopathy and the different forms of OSA is visible (red line). (B) No significant differences are detectable between saliva from healthy volunteers and patients (independent of the disease or disease severity) with regard to TFF2 protein concentration.</p

RMSD plots of SP-G models during 50 ns MD simulation.

<p>A representative simulation for the unmodified and posttranslationally modified SP-G model is shown in black and red, respectively. Both structure model variants are stable with respect to the RMSD plots of the backbone atoms.</p

Predicted posttranslational modifications and their sequence positions in the SP-G model.

<p>Present modification types: phosphorylation (PHOS), palmitoylation (PALM), <i>O</i>-glycosylation with GlcNAc (<i>O</i>-GLYC), and <i>N-</i>glycosylation with a pentasaccaride core consisting of 2 GlcNAc and 3 mannose moieties (<i>N</i>-GLYC).</p