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Poster Sessions ALI and Inflammation 0035-0045 S14 19th ESICM Annual Congress – Barcelona, Spain – 24–27 September 2006 Poster Sessions 0037 CRITICAL ROLE OF NEUTROPHIL P21-ACTIVATED KINASE IN LPS-INDUCED ACUTE ALI and inflammation 0035-0045 LUNG INJURY 0035 Reutershan J1, Stockton R2, Zarbock A2, Sullivan G W2, Chang D2,ScottD2,SchwartzMA2,LeyK2 1 PENTRAXIN 3 (PTX3) AND C REACTIVE PROTEIN IN ALI/ARDS: PRELIMINARY University of Tuebingen, Dept. of Anaesthesiology and Intensive Care Medicine, Tuebingen, 2 RESULTS FROM A PROSPECTIVE STUDY Germany, University of Virginia, Robert M. Berne Cardiovascular Research Center, Charlottesville, United States Mauri T1, Coppadoro A1, Bellani G1, Bombino M1, Valentino S2, Patroniti N1, Mantovani A2, Pesenti A1 INTRODUCTION. Excessive recruitment of polymorphonuclear leukocytes (PMNs) into the lung 1Perioperative Medicine and Intensive Care, St Gerardo Hospital, Monza, 2Laboratory of Immunology is critical in the early phase of acute lung injury (ALI/ARDS). Although chemokines and adhesion and Cell Biology, Humanitas Clinical Institute, Rozzano, Italy molecules have been demonstrated to initiate leukocyte-endothelial interaction, pathways which regu- late PMN migration through the alveolo-capillary membrane remain to be identified. Here, we studied INTRODUCTION. PTX3 has recently been described as an acute phase mediator of innate immunity. the role of the small GTPases effector p21-activated kinase (PAK) for PMN migration in LPS-induced Together with C Reactive Protein (CRP), it belongs to the pentraxin superfamily. PTX3 is produced ALI. by a variety of cells, located almost everywhere (e.g. dendritic cells), while CRP is produced only by the liver. PTX3 plays a key role in the activation and the amplification of inflammation. ALI/ARDS is METHODS. LPS-induced PMN migration into the different compartments of the lung was investi- characterized by an important inflammatory reaction, often both local and systemic. gated in wild type male C57Bl/6 mice. PAK activity was inhibited by injecting a specific blocking peptide. A fluorescent blocking peptide was injected to investigate its cellular targets in vivo. PMN METHODS. We enrolled 9 patients affected by ALI/ARDS. We measured PTX3 and CRP levels migration through a layer of pulmonary endothelial cells was investigated to determine the role of neu- on the first 2 days from ICU admission, then every three days for the first month and then once a trophil PAK. To test whether this translates to humans, F-actin polymerization, oxidative burst, and week, until ICU discharge. We also measured PTX3 levels in the bronco alveolar lavage (BAL) fluid. adhesion were investigated in human PMNs. 102 plasma samples and 14 BAL fluids were collected and tested. We registered data on etiology, pulmonary mechanics, gas exchange, systemic inflammation, organs function, blood cells counts and RESULTS. PMN migration into the lung interstitium and alveolar airspace, but not accumulation in outcome at hospital discharge. the pulmonary vasculature was reduced when PAK activity was blocked. Injection of a fluorescently labeled peptide identified leukocytes as a major cellular target for its blocking activity. The majority of RESULTS. PTX3 plasma levels were high and within a very wide range (2.02-3872.64 ng/ml; 111.47 fluorescent PMNs were found in the blood, while no positive PMNs were found in the BAL, indicating ± 467.21 ng/ml), while CRP levels were relatively low and less variable (0.16-49 mg/dl; 15.94 ± 11.14 that PAK-activation was required for PMNs to emigrate from the circulation. In addition, migration mg/dl). The levels of the two mediators showed no correlation. PTX3 was present in BAL fluids in vari- towards a chemotactic gradient was reduced when PAK activity was blocked in PMNs, highlighting a able concentrations (0-19.4 ng/ml; 3.28 ± 5.63 ng/ml). We found a significant correlation (p<0.05; r critical role for PAK in PMNs. In human PMNs, blocking PAK activity disrupted F-actin polymeriza- 0.673) between the levels of PTX3 in BAL fluid and plasma. PTX3 levels on the first day were sig- tion, static adhesion, and oxidative burst of adherent PMNs. nificantly different between survivors and not-survivors at hospital discharge (p<0.05; 1.310±0.457 log(ng/ml) vs 2.401±0.825 log(ng/ml)). Moreover PTX3 values were significantly higher in patients CONCLUSION. PAK is a critical mediator of LPS-induced PMN migration into the lung and may be with septic shock than in non shock patients (p<0.05; 1.532±0.794 vs 1.223±0.478 log(ng/ml)). PTX3 an attractive target for the treatment of acute lung injury. values were significantly (p<0.05) related to SvO (r -0.316), arterial blood pH (r -0.371) and SOFA 2 REFERENCE(S). 1. Reutershan J, Sequential recruitment of neutrophils into lung and bronchoalve- values (r 0.591). olar lavage fluid in LPS-induced acute lung injury. AJP Lung 2005 289:L807-L815. 2. Stockton RA, CONCLUSION. PTX3 seems to be a key factor for the development, the amplification and the out- p21-activated kinase regulates endothelial permeability through modulation of contractility. JBC 2004 come of this syndrome; the levels of the two mediators appear not to be related, suggesting a possible 279:46621-46630. individual role for each one. Grant acknowledgement. German Research Foundation (grant RE 1683/2-1 to JR) and NIH grant Grant acknowledgement. MIUR, Roma, Italy HL73361 to KL. 0036 0038 NATRIURETIC PEPTIDES FOR THE ASSESSMENT OF SMOKE INHALATION INJURY IMPACT OF TRANSFORMING GROWTH FACTOR BETAˆ 1 AND PROCOLLAGEN III ON THE MORTALITY OF ACUTE RESPIRATORY DISTRESS SYNDROME Reper P A1,VanLoeyKE1 1Critical Care Department, Queen Astrid Hospital, Brussels, Belgium Forel J1,DemoryD1, Guervilly C1,RochA1,MarinV2,ArnalJ3, Donati Y3, Embriaco N1, Gainnier M1, Papazian L1 INTRODUCTION. Smoke intoxication remains a major cause of mortality after smoke inhalation 1Medical Intensive Care Unit, 2Immunology Laboratory, Hˆopitaux Sud Assistance Publique - Hˆopitaux and remains difficult to evaluate by biological markers. de Marseille, Marseille, 3Intensive Care Unit, Hˆopital Font-Pr´e, Toulon, France Increased plasma levels of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) have been identified as predictors of cardiac dysfunction and prognosis in congestive heart failure and is- INTRODUCTION. To determine whether blood and bronchoalveolar lavage fluid levels of the TGFβ- chemic heart disease. 1 and the Procollagen III (PIIINP) are associated with the mortality of patients with ARDS. In severe sepsis patients, some data are now available about the possible prognostic value of natriuretic peptides. METHODS. We included 56 patients with ARDS, after patient’s next of kin consent, in a prospec- The aim of the present study was to determine the role of the N-terminal prohormone forms of ANP tive study. Patients were ventilated with a protective mechanical ventilation according to NIH. Bron- (NT-proANP) and BNP (NT-proBNP) after smoke inhalation injury. choalveolar lavages (BAL) and blood samples were performed on the first 48 hrs of ARDS for dosing TGFβ-1 and PIIINP. Survival was registered at day 28. BAL and blood levels of TGFβ-1 and PIIINP METHODS. Retrospective, clinical study in a specialised burn intensive care unit. Population: 9 pa- were compared by Mann-Whitney test. We determined cut-off values of these mediators using ROC tients with severe inhalation injury confirmed by fiberoptic examination and clinical findings (close curves. Multivariate logistic regression was performed to determine the effect of TGFβ-1 and PIIINP space fire, soot on the face and in the mouth; HbCO >10%) and 10 control subjects with no smoke on the risk of death while controlling the effect of age, sexe, SOFA, Lung injury score, PaO2/FiO2, exposure. All the patients had burn wounds < 5 ± 2% TBSA. polymorphonuclear count in BAL. Collection of clinical and demographic data in relation to ANP, BNP, and IL-6 in plasma over a period of 4 days. RESULTS. Table 1 shows the levels of PIIINP in survivors and non survivors. BAL fluid and blood levels of TGFβ-1 were not statistically differents in survivors and nonsurvivors. Table 2 summarizes RESULTS. There was no significant difference between the two groups for age, burn surface area, and the results of the multivariate analysis. medical history. We found a significant increase in ANP (113.7+/-9.9 vs. 13.8+/-1.3 pg/ml) and BNP (81.4+/-3.6 vs. TABLE 1. 6.5+/-0.8 pg/ml) after inhalation injury. Levels of PIIINP (µg/L) in BAL fluid and blood in survivors and non survivors Plasma ANP peaked together with IL-6 and peaks of ANP and IL-6 were significantly correlated Survivors (n=36) Nonsurvivors (n=20) p value (p<0.01). BAL levels of PIIINP 1.7 [0.7-4.1] 5.9 [1.7-24.1] 0.024 CONCLUSION. ANP and BNP increase significantly in patients with severe inhalation injury. Blood levels of PIIINP 7.7 [5.0-10.7] 14.9 [9.3-18.6] 0.001 BNP reflects left ventricular dysfunction and could be related to the potential cardiac depression fol- lowing smoke intoxication and particularly cyanide and carbon monoxyde exposition associated with TABLE 2. hypoxaemic conditions. Results of multivariate analysis for the risk of death determined on inclusion ANP is related to IL-6 production rather than to cardiovascular dysfunction. Odds Ratio (95% CI) p value Further studies are necessary to confirm these preliminary results. Blood levels of PIIINP>=8.95µg/L 4.82 (1.16-20.01) 0.03 BAL levels of PIIINP >=4.0µg/L 4.27 (1.07-16.99) 0.04 CONCLUSION. Early increased levels of Procollagen III Peptide in blood and in BAL fluid, but not levels of Transforming Growth Factor β-1, are associated with an increased risk of death in patient with acute respiratory distress syndrome.
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