Expression levels of TLR2 and TLR4 on monocytes (A) and granulocytes (B) FFR-positive and FFR-negative patients and healthy control subjects.

<p>*Comparison by Kruskal-Wallis test for all groups.** Between subgroups with Mann Whitney U test. TLR = toll-like receptor.</p

Concentrations of cytokines and surface markers on monocytes and granulocytes according to FSS tertiles and FFR-negative patients.

<p>Concentrations of cytokines (A) and surface markers on monocytes (B) and granulocytes (C) according to FSS tertiles (of FFR-positive patients) and FFR-negative patients. Comparison of all subgroups was performed with the Jonckheere-Terpstra test. FSS = functional syntax score.</p

Comparison of levels of cytokines for FFR-positive and FFR-negative patients and healthy controls.

<p>*Comparison by Kruskal-Wallis test.** Between subgroup differences compared with Mann Whitney U test.</p

Spearman rank correlation between NO and MMP9 levels in BM of Wistar rats.

<p>A positive and statistically significant (0.016 cells, total MMP9 amount was quantified in the supernatants from the same BM samples. Errors in MNIC yields and MMP9 (not indicated) are ca 10% and 5% respectively.</p

Baseline Characteristics of patients with stable angina.

<p> <i>Continuous values are presented as means ± standard deviation (SD). Categorical values are presented as number (percentages).</i></p>*<p> <i>Significance level 0.05. FFR = fractional flow reserve BMI = body mass index ;DM = diabetes mellitus; MI = myocardial infarction; PCI = percutaneous coronary intervention; CABG = coronary artery bypass grafting; LVEF = left ventricular ejection fraction.</i></p>**<p> <i>Defined as a serum creatinin >150 µmol/l.</i></p

Medication use of patients with stable angina at inclusion.

<p> <i>Proportions were compared using Chi-square testing.</i></p>*<p> <i>Significance level 0.05. FFR = fractional flow reserve; ASA = Acetylsalicylic acid; ACE = angiotensin-converting enzyme.</i></p

Relative contribution of NOS isoforms to basal NO production in BM of Lewis rats.

<p>The basal NO level that was obtained in absence of a stimulus was taken as 100%. Contribution of the NOS isotypes was determined using two concentrations of the NOS isoform-specific inhibitor 1400 W. 1400 W was used at final concentrations of 0.1 µM for iNOS inhibition and 10 µM for inhibition of both iNOS and nNOS. With L-NAME (500 µM, final column), the signal intensity was below the detection threshold. Values represent biological triplicates.</p

MAP and BM-derived MNIC yields in control and hypertensive Wistar rats.

<p>b.d.: below detection</p>*<p>Two BM samples, where MNIC remained below the detection threshold are excluded from this average.</p

X-band EPR spectra of MNIC at 77°K.

<p>(A). The EPR spectrum from BM cell suspension of <i>ca</i> 100 mln cells. These cells were collected from a DOCA-treated Wistar male rat and stimulated for NO production with Ca-ionophore A23187. The intensity of the spectrum was not changed and was quantified as 60±7 pmol MNIC. (B) The reference spectrum of a strong calibration sample with 4 nmol MNIC, showing the characteristic triplet structure centered around g = 2.035 with a hyperfine splitting of 25.2 G = 2A_N¹⁴. The intensity of the reference spectrum was scaled down by a factor 25.</p

The effect of NOS inhibitors and/or activators on NO level in BM of Wistar rats.

<p>X-band EPR spectra at 77°K of frozen BM cell suspensions from Wistar rats after 30 min of NO trapping with Fe²⁺-DETC. The samples (53±2×10⁶ BM cells) were reduced by 50 mM dithionite to remove the overlapping signal from paramagnetic Cu²⁺-DETC. A basal yield of 21±2 pmol MNIC is obtained in absence of stimulus (top), clearly recognizable by its characteristic hyperfine triplet structure at g = 2.035. Stimulation with ionomycin or A23187 calcium ionophores raises the MNIC yields to 45±4 and 49±5 pmol. In presence of the NOS inhibitor L-NAME, the MNIC yields remained below the detection threshold of <i>ca</i> 10 pmol. The EPR spectra show significant background signals not related to MNIC, as visible in the bottom spectra. Such background signals derive from paramagnetic iron centers, and some residual Cu²⁺-DETC as commonly found in biological samples <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057761#pone.0057761-vanFaassen1" target="_blank">[18]</a>.</p