1. Chronic carbon monoxide exposure is associated with the increases in carotid intima-media thickness and C-reactive protein level
Suat Zengin, Cuma Yildirim, Vedat Davutoglu, Ibrahim Sari, Behcet Al, Suleyman Ercan, Murat Yuce Tohoku J Exp Med . 2009 Nov;219(3):201-6. doi: 10.1620/tjem.219.201.
Being the most common cause of death from poisoning worldwide, cardiovascular manifestations of acute carbon monoxide (CO) poisoning have been subject of various studies but current evidence about effects of chronic CO exposure on atherosclerosis is limited which is very common. We aimed to investigate association of chronic CO exposure with atherosclerosis by measuring carotid intima-media thickness (CIMT) and high-sensitivity C-reactive protein (hs-CRP). Forty healthy male non-smoker indoor barbecue workers (mean age; 33.0 +/- 9.0 years) working in different restaurants for at least three years and 48 age-matched healthy men (mean age; 34.3 +/- 6.6 years) enrolled in the study. Clinical characteristics of indoor barbecue workers and control group were comparable in terms of body mass index, blood pressure, and lipid profile. However, carboxyhemoglobin (COHb) (6.4 +/- 1.5% vs. 2.0 +/- 1.1%), hs-CRP (2.7 +/- 2.0 mg/L vs. 1.1 +/- 0.8 mg/L) and CIMT (1.1 +/- 0.3 mm vs. 0.9 +/- 0.1 mm) were higher in indoor barbecue workers (p < 0.001 for each). In Pearson correlation analysis, CIMT was correlated with COHb concentration (r = 0.635, p < 0.001) and hs-CRP level (r = 0.466, p < 0.001). Among indoor barbecue workers, the years worked (years exposed to CO) are correlated with COHb, hs-CRP and CIMT. In multivariate analysis, COHb concentration is the only independent predictor of CIMT (beta = 0.571, p < 0.001). The increased CIMT and hs-CRP in indoor barbecue workers suggest that chronic CO exposure may increase the risk of atherosclerotic cardiovascular events.
2. Aminopeptidase P from human leukocytes
A Yaron, I Rusu Eur J Biochem . 1992 Nov 15;210(1):93-100. doi: 10.1111/j.1432-1033.1992.tb17395.x.
Cytosolic aminopeptidase P was obtained in highly purified form from human leukocytes by a four-step procedure. Buffy coats were the starting material. A M(r) of 140,000 was obtained by size-exclusion HPLC for the native enzyme. As shown by SDS/PAGE under reducing and denaturing conditions, the enzyme consisted of likely identical subunits with M(r) of 71,000. Purified aminopeptidase P cleaved off, specifically and efficiently, the N-terminal residues from peptides with N-terminal Xaa-Pro sequences. The penultimate proline was not replaceable by hydroxyproline, alanine and glycine in di-, tri- and tetrapeptides. Polyproline was not hydrolyzed. Dipeptides were cleaved (Arg-Pro, Phe-Pro > Trp-Pro > Pro-Pro) although slower than longer peptides. Cleavage was observed of several biologically active peptides; C-terminal fragment (residues 201-206) of C-reactive protein, oxytocin fragment Tyr-Pro-Leu-Gly, morphiceptin, peptide Gly-Pro-Arg-Pro (inhibitor of fibrin polymerization) and kentsin. In addition, cleavage of a protein, interleukin-6, was also demonstrated. Aminopeptidase P was maximally activated by Mn2+, and to a lesser extent by Co2+. The activity was optimal at pH 8. Ni2+, Zn2+ and especially Cd2+ caused marked inhibition. EDTA, 1,10-phenantroline and dithiothreitol were also inhibitory. Carbobenzoxy-phenylalanine, as well as several N-carbobenzoxy-proline-containing peptides, caused partial inhibition. The observed resistance of Gly-Pro, Pro-Gly, Pro-Phe and Pro-Ile to hydrolysis by the purified enzyme strongly indicates absence of known proline-specific dipeptidases in the aminopeptidase-P preparation.
3. C-reactive protein: a physiological activator of interleukin 6 receptor shedding
A J Szalai, G M Fuller, S Horiuchi, N Yamamoto, S A Jones, D Novick J Exp Med . 1999 Feb 1;189(3):599-604. doi: 10.1084/jem.189.3.599.
The soluble interleukin 6 receptor (sIL-6R) circulates at elevated levels in various diseases. This suggests that inflammatory mediators control sIL-6R release. Through examination of human neutrophils, it was found that the acute phase reactant C-reactive protein (CRP) activates a threefold increase in sIL-6R production. Maximal release occurred after 30-60 min exposure to CRP (50 micrograms/ml), and was mimicked by peptides corresponding to amino acid residues 174- 185 and 201-206 of native CRP. A third peptide fragment (77-82) had no effect. Differential mRNA splicing did not account for the CRP-mediated release of sIL-6R, since this isoform was not detected in conditioned media. Furthermore, stimulation of neutrophils with CRP or with peptides 174-185 or 201-206 promoted a loss of membrane-bound IL-6R, suggesting release by proteolytic shedding. The metalloprotease inhibitor TAPI had only a marginal effect on CRP-mediated sIL-6R release, suggesting that shedding occurs via a mechanism distinct from that previously reported. It well established that IL-6 stimulates the acute phase expression of CRP. Our current findings demonstrate a novel relationship between these two mediators, since CRP may affect IL-6-mediated inflammatory events by enabling formation of the sIL-6R/IL-6 complex.
