Diabetic Peripheral Neuropathy as a Predictor of Asymptomatic Myocardial Ischemia in Type 2 Diabetes Mellitus: A Cross-Sectional Study

<p><b>Article full text</b></p> <p><br></p> <p>The full text of this article can be found here<b>. </b><a href="https://link.springer.com/article/10.1007/s12325-016-0399-1">https://link.springer.com/article/10.1007/s12325-016-0399-1</a></p> <p><br></p> <p><b>Provide enhanced content for this article</b></p> <p><br></p> <p>If you are an author of this publication and would like to provide additional enhanced content for your article then please contact <a href="http://www.medengine.com/Redeem/”mailto:[email protected]”"><b>[email protected]</b></a>.</p> <p><br></p> <p>The journal offers a range of additional features designed to increase visibility and readership. All features will be thoroughly peer reviewed to ensure the content is of the highest scientific standard and all features are marked as ‘peer reviewed’ to ensure readers are aware that the content has been reviewed to the same level as the articles they are being presented alongside. Moreover, all sponsorship and disclosure information is included to provide complete transparency and adherence to good publication practices. This ensures that however the content is reached the reader has a full understanding of its origin. No fees are charged for hosting additional open access content.</p> <p><br></p> <p>Other enhanced features include, but are not limited to:</p> <p><br></p> <p>• Slide decks</p> <p>• Videos and animations</p> <p>• Audio abstracts</p> <p>• Audio slides</p

Characteristics of the subjects included in the study.

<p>Gender data are presented as number (%) in each group; all other data are presented as median (interquartile range). AHI  =  apnea hypopnea index, BMI  =  body mass index, BP  =  blood pressure, HDL  =  high density lipoprotein, LDL  =  low density lipoprotein, SaO₂  =  oxygen saturation.</p>*<p><i>p</i>≤0.05 OSA severe hypoxemia versus controls; ^#<i>p</i>≤0.05 OSA mild hypoxemia versus controls; ^†<i>p</i>≤0.05 OSA severe hypoxemia versus OSA mild hypoxemia.</p

Expression of select genes in skin of OSA patients is differently regulated in severely hypoxemic (<75% blood oxygen saturation) and mildly hypoxemic (≥75% blood oxygen saturation) OSA groups.

<p>Gene expression in skin biopsies obtained from OSA patients and control subjects was analyzed by qRT-PCR using specific primers, and is presented as relative mRNA expression versus a control group. Results for each sample were normalized versus 28S. n = 10–12. Data are presented as mean +/− SDEV. * p<0.05; ** p<0.005. There is a global statistical significance for all genes in three groups of studied subjects. ≥75% – OSA group with mild hypoxemia; <75% – OSA group with severe hypoxemia.</p

Expression of select genes in mouse aortas is affected by IH.

<p>Mice were exposed to intermittent hypoxia (IH) or intermittent air (IA) for 4 weeks. RNA was isolated from mouse aortas followed by cDNA generation and qPCR analysis. The gene expression is presented as relative mRNA expression versus a control (IA) group. Results for each sample were normalized versus 28S. Data are presented as mean +/− SDEV. n = 7–12. * p<0.05.</p

Expression of select genes in HMVEC exposed to IH.

<p>RNA was isolated from HMVEC exposed to IH, followed by cDNA generation and qRT-PCR analysis. The gene expression is presented as relative mRNA expression versus a control group. Results were obtained from at least 3 experiments and each sample was normalized versus 28S. Data are presented as mean +/− SDEV. * p<0.05; *** p<0.001.</p

Endothelium-dependent microvascular reactivity is decreased in severely hypoxemic OSA patients.

<p>Changes in skin blood flow were measured at the forearm by scanning LASER Doppler following administration of acetylcholine (ACh) that stimulates the endothelium-dependent release of nitric oxide (<b>A</b>) or SNP that releases nitric oxide in an endothelium-independent manner (<b>B</b>). Data are presented as box-plots with medians, quartiles and minimum and maximum values; 12 subjects per group. C – control group; ≥75% – OSA group with mild hypoxemia; <75% – OSA group with severe hypoxemia.</p

Sequences of primers used in qRT-PCR experiments.

<p>H – human, M – mouse.</p

A and B: Forearm skin biopsy staining for SDF-1in a diabetic patient (Figure 4A) and a healthy control subject (Figure 4B), (frozen sections, x100).

<div><p>SDF-1 was expressed by stromal cells (black arrows) and endothelial cells (red arrows) and the staining pattern was mostly cytoplasmic and occasionally nuclear in cases of increased expression. The number of stained stromal cells and the intensity of staining were increased in in diabetic patients while no difference was found in the number of stained endothelial cells.</p> <p><i>C</i> and <i>D</i>: Foot skin staining for CXCR4 in a diabetic patient (Figure 4C) and a healthy control subject (Figure 4D) (frozen sections, x200). CXCR4 was expressed by stromal cells (black arrows), endothelial cells (red arrows) and epithelial cells (blue arrows) and the staining pattern was mostly membranar and cytoplasmic. The intensity of staining was higher in in the diabetic group (p<0.05) but no differences were observed between the two groups in the number of positive stromal and endothelial cells (p=NS).</p></div

Changes in EPCs measurements during the four study visits between the patients who did not heal their ulcers (NH) and those who did (H).

<p>There were no differences in all EPC measurements at baseline but patients who healed their ulcers had lower CD34⁺KDR⁺ counts at visits 3 and 4 and CD34⁺CD133⁺ at visit 4. Data are presented as the median and interquartile range box.</p

An example of flow cytometric analysis of human peripheral blood sorted on CD45^dim cells in a patient with DFU (A) and a healthy control subject (B).

<p>The triple positive phenotype (CD34⁺/KDR⁺/CD133⁺) was determined by gating the CD133⁺ cells on the CD34⁺/KDR⁺. 1.000.000 events per sample were acquired and the counts for each phenotype are shown in the picture. Smaller counts in all phenotypew were observed in the diabetic patient (A) when compared to the healthy subject (B) in the double and triple measurements. </p