Acute-phase protein alpha-1-acid glycoprotein mediates neutrophil migration failure in sepsis by a nitric oxide-dependent mechanism.

The reduction of circulating neutrophil migration to infection sites is associated with a poor outcome of severe sepsis. alpha-1-Acid glycoprotein (AGP) was isolated from the sera of severely septic patients by HPLC and acrylamide gel electrophoresis and identified by mass spectrometry. Both the isolated protein and commercial AGP inhibited carrageenin-induced neutrophil migration into the rat peritoneal cavity when administered i.v. at a dose of 4.0 microg per rat (95 pmol per rat). Analysis by intravital microscopy demonstrated that both proteins inhibited the rolling and adhesion of leukocytes in the mesenteric microcirculation. The inhibitory activity was blocked by 50 mg/kg aminoguanidine, s.c., and was not demonstrable in inducible nitric oxide synthase (iNOS) knockout mice. Incubation of AGP with neutrophils from healthy subjects induced the production of NO and inhibited the neutrophil chemotaxis by an iNOS/NO/cyclic guanosine 3,5-monophosphate-dependent pathway. In addition, AGP induced the l-selectin shedding by neutrophils. The administration of AGP to rats with mild cecal ligation puncture sepsis inhibited neutrophil migration and reduced 7-day survival from approximately 80% to 20%. These data demonstrate that AGP, an acute-phase protein, inhibits neutrophil migration by an NO-dependent process and suggest that AGP also participates in human sepsis.

Peroxynitrite mediates the failure of neutrophil migration in severe polymicrobial sepsis in mice.

BACKGROUND AND PURPOSE: Sepsis is a systemic inflammatory response resulting from the inability of the host to restrict local infection. The failure of neutrophil migration to the infection site is one of the mechanisms involved in this process. Recently, it was demonstrated that this event is mediated by nitric oxide (NO). The present study addresses the possibility that peroxynitrite (ONOO(-)), a NO-derived powerful oxidizing and nitrating compound, could also be involved in neutrophil migration failure. EXPERIMENTAL APPROACH: Male C57Bl/6 mice were subjected to moderate (MSI) or severe (SSI) septic injury, both induced by cecal ligation and puncture (CLP). The leukocyte rolling and adhesion in the mesentery was evaluated by intravital microscopy. Cytokines (TNF-alpha and MIP-1alpha) were measured by ELISA and 3-nitrotyrosine (3-NT) by immunofluorescence. KEY RESULTS: Compared with saline pretreatment of SSI mice, pre-treatment with uric acid, a ONOO(-) scavenger, partially restored the failure of neutrophil rolling, adhesion and migration to the site of infection. These mice also presented low circulating bacterial counts and diminished systemic inflammatory response. Pretreatment with uric acid reduced 3-NT labelling in leukocytes in mesenteric tissues and in neutrophils obtained from peritoneal exudates. Finally, uric acid pretreatment enhanced significantly the survival rate in the SSI mice. Similarly, treatment with FeTPPs, a more specific ONOO(-) scavenger, re-established neutrophil migration and increased mice survival rate. CONCLUSIONS AND IMPLICATIONS: These results indicate that ONOO(-) contributed to the reduction of neutrophil/endothelium interaction and the consequent failure of neutrophil migration into infection foci and hence susceptibility to severe sepsis.

[Geometric changes of the left ventricle during the isovolumic contraction in healthy people and patients with coronary disease]. / Geometrische Anderungen des linken Ventrikels während der isovolumetrischen Kontraktion bei Gesunden und Koronarkranken

Biplane cineventriculograms of 5 normal and 12 patients with coronary artery disease were analysed in order to determine the dimensional changes of the left ventricle during the isovolumic phase of contraction. In postero-arterior projection the ventricular diameters in both groups decreased by an average of 3-6%, the wall thickness increased by 5-10%, In lateral projection there were no significant changes of geometry. The changes in regional diameters showed significantly greater maximal differences in diseased than in normal ventricles. Using ventriculographic data of enddiastole instead of aortic valve opening for determination of ejection phase parameters resulted in an overestimation of the mean velocity of circumferential fiber shortening by an average of 10-20%, of the stroke volume of 5-15% and of the ejection fraction of 5%. These results indicate that ejection phase indices can only be determined exactly from cineventriculograms when aortic valve opening and closure is considered.