Unadjusted simple Pearson correlation pattern between plasma and RBC analytes.

*<p>P<0.05,</p>†<p>P<0.01,</p>‡<p>P<0.001.</p

RBC levels of analytes involved in Arginine/NO pathway.

<p>(A) L-arginine (Arg), L-citrulline (Cit), L-ornithine (Orn) were simultaneously determined by LC-MS/MS. (B) Arg bioavailability and the relative activity of arginase and RBC-NOS enzymes are expressed as Arg/(Orn+Cit) and Orn/Cit ratios, respectively. (C) The endogenous inhibitors asymmetric dimethylarginine (ADMA) and symmetric dimethylarginine (SDMA) were determined by LC-MS/MS. (D) Tetrahydrobiopterin (BH₄) and oxidized biopterins (Box) were detected by HPLC after selective oxidation with iodine. Data are presented as age and sex adjusted geometric means and 95% C.I. Comparisons between groups (CAD, n = 22; Ctrl, n = 20) were performed by covariance analysis, adjusting for age and sex.</p

Partial Pearson correlation pattern between plasma and RBC analytes, adjusted for the effects of all other analytes.

*<p>P<0.05,</p>†<p>P<0.01,</p>‡<p>P<0.001.</p

Demographic and clinical characteristics of CAD patients and Ctrl subjects.

<p>Quantitative variables were expressed as mean ± SD and categorical variables as n (%).</p><p>P value: Wilcoxon test for continous variables and Chi Square test for categorical variables.</p><p>P value adjusted for sex and age after log-transformation of the data.</p

Immunostaining of NO synthase (NOS) protein in human red blood cells (RBCs).

<p>NOS was detected in RBCs from CAD patients (A) and from Ctrl (B). RBCs were incubated with a monoclonal anti eNOS antibody (2.5 µg/ml) and with an anti-mouse conjugated secondary antibody (40 µg/ml; Alexa Fluor488). The samples were mounted with fluorescent mounting medium and examined by laser scanning confocal microscope (LSM710, Carl Zeiss) using a 63×/1.3 oil immersion objective lens. Fluorescent images were captured with a digital camera using the image processor Zen (Carl Zeiss). (C) Fluorescence intensity (densitometric sum of grey) was quantified. Data are expressed as the log median of total fluorescence intensity per field ± interquartile range subtracted of the negative control value. Means derive from n = 10 CAD and n = 10 Ctrl.</p