[Show abstract][Hide abstract] ABSTRACT: In large series of nonresponding community-acquired pneumonia (CAP) patients, chronic obstructive pulmonary disease (COPD) was observed to be a protective factor for nonresponse to initial antibiotics. This intriguing fact may be linked to changes in the phenotype of inflammatory cells and, in particular, to the induction of classical-M1 or alternative-M2 activation of macrophages, which result in different inflammatory profiles. We evaluated the effect of sputum obtained from patients with acute exacerbation of COPD (AECOPD), CAP and COPD+CAP on the phenotypic changes in macrophages. Human THP1 cells differentiated to macrophages were incubated with sputum from patients with AECOPD, CAP or COPD+CAP, and expression of tumour necrosis factor-alpha, interleukin-6, mannose receptor and arginase was measured to evaluate the phenotype acquired by macrophages. We found that sputum from CAP patients induced the M1 phenotype and that from AECOPD patients induced an M2-like phenotype. Sputum from CAP+COPD patients did not present a clear M1 or M2 phenotype. These results indicate that the microenvironment in the lung modulates the activation of macrophages, resulting in different phenotypes in AECOPD, CAP and COPD+CAP patients. This different type of activation induces different inflammatory responses and may be involved in the different outcome observed when COPD and CAP present simultaneously.
Preview · Article · Aug 2010 · European Respiratory Journal
[Show abstract][Hide abstract] ABSTRACT: Linezolid is the first synthetic compound of a new group of antimicrobials, the oxazolidinones, which inhibit protein synthesis. It shows a broad spectrum of activity against Gram positive organisms. With respect to its pharmacokinetics, linezolid shows a relatively high volume of distribution and good penetration into inflammatory fluids, bone, fat and muscle.
A reversed-phase isocratic high-performance liquid chromatographic method for linezolid analysis in piglet pulmonary tissue is described. Tissue samples and controls were prepared in 1 x TBE (1 M Tris, 0.9 M boric acid, 0.01 M EDTA). The mobile phase consisted of 20% ultrafiltered water and 80% of (A) 15 mM potassium monohydrogen phosphate buffer (pH = 5) with (B) acetonitrile (80%/20%; v/v). Samples were homogenized and precipitated with HClO(4) 3% (1/1, v/v). The injection volume was 100 microL. Ofloxacin was used as an internal standard.
The assay was linear over a linezolid concentration range: 1.6-100 microg/mL. The method provided good validation data (n = 15): inaccuracy (3.6%), intra and inter-day variability (4.2% and 5.2%, respectively), recovery (91.8%), limit of detection (0.8 microg/mL) and quantitation (1.6 microg/mL) and acceptable stability within 24 h in the auto-sampler.
The method offers a fast and simple approach to determine linezolid in pulmonary tissue which could be of use in pharmacokinetic studies.
No preview · Article · Mar 2010 · Clinical Chemistry and Laboratory Medicine
[Show abstract][Hide abstract] ABSTRACT: The objectives of the study were to validate a model of methicillin-resistant Staphylococcus aureus (MRSA) pneumonia in ventilated piglets and to study the time-course of biological markers and histopathological changes. 12 piglets were intubated and inoculated with 15 mL of a suspension of 10(6) colony forming units of MRSA in every lobe through the bronchoscope channel. The piglets were ventilated for 12 h (n = 6) and 24 h (n = 6). Clinical parameters were assessed every 6 h and pro-inflammatory cytokines were measured in serum and in bronchoalveolar lavage (BAL) at baseline and sacrifice. Histopathology of each lobe and cultures from blood, lungs and BAL were performed. Animals developed histopathological evidence of pneumonia at necropsy. At 12 h, pneumonia was present in all animals and was severe pneumonia at 24 h. Microbiological studies confirmed the presence of MRSA. A significant increase in interleukin (IL)-6, IL-8 and tumour necrosis factor-α values was seen in BAL at 24 h and IL-6 at 12 h. In serum, only IL-6 levels had increased significantly at 24 h. In ventilated piglets, bronchoscopic inoculation of MRSA induces pneumonia at 12 h and severe pneumonia at 24 h. This severity was associated with a corresponding increase in systemic and local inflammatory response.
Preview · Article · Mar 2010 · European Respiratory Journal
[Show abstract][Hide abstract] ABSTRACT: The mannose-binding lectin (MBL) pathway of complement system is activated when carbohydrate-bound MBL forms complexes with different serine proteases (MASP-1, MASP-2 and MASP-3), among which MASP-2 has a predominant functional role. Polymorphisms impairing the quantity and/or the functional activity of proteins encoded by the MBL2 and MASP2 genes have been reported in all human populations showing different allelic frequency and distribution. This likely reflects the existence of environmental influences on MBL2 and MASP2 genetic evolution. Herewith, we conducted a study in a children population from Mozambique to analyse the genetic diversity of sequences corresponding to the promoter and collagen-like region (exon 1) of MBL2 and to the CUB-1 and epidermal growth factor domain (exon 3) of MASP2, which are critical regions for the formation of functional MBL/MASP-2 complexes. Our results show a high prevalence of MBL-intermediate/low genotypes (43.5%); the description of new alleles and a high level of sequence polymorphism at both MBL2 and MASP2, with no statistical evidence for positive or balancing selection. Furthermore, Biacore analyses performed to explore the functional relevance of the MASP2 variants found [T73M (2.9%), R84Q (12.7%) and P111L (25.4%)] were compared with those of two previously reported variants (R103C and D105G). None of the analysed MASP2 variants, with the exception of D105G, interfered with interactions with either MBL or ficolins (H and L).
[Show abstract][Hide abstract] ABSTRACT: Inflammatory markers have been assessed for the diagnosis and follow-up of ventilator-associated pneumonia (VAP), but their potential role in predicting the risk for VAP is unknown. We prospectively assessed the evolution of cytokines in mechanically ventilated patients and their predictive and diagnostic role for VAP.
Prospective observational study.
Medical intensive care unit.
Mechanically ventilated patients. Exclusion criteria were active infection at admission and subsequent extrapulmonary infection.
Sequential measurements of interleukin (IL)-1, IL-6, IL-8, IL-10, and tumor necrosis factor-alpha were done in 44 ventilated patients. VAP was suspected in 20 cases and microbiologically confirmed in nine. At admission, demographics, severity scores, and clinical and standard laboratory values did not discriminate patients with and without VAP, but the median (interquartile range) serum levels of IL-6 were higher in patients who subsequently developed VAP, compared with those without VAP (235 [141-803] vs. 113 [60-170] pg/mL, p = 0.015). The sensitivity and specificity of IL-6 to predict VAP was 71% and 78%, respectively, using 198 pg/mL as optimal cutoff, with relative risk (95% confidence interval) 8.9 (1.4-56.3). When VAP was suspected, serum levels of IL-6 were higher in patients with confirmed compared with nonconfirmed VAP (1131 [496-1987] vs. 236 [115-357] pg/mL, p = 0.016). The sensitivity and specificity to discriminate between confirmed and nonconfirmed VAP was 71% and 89%, respectively, using 620 pg/mL as optimal cutoff, with relative risk (95% confidence interval) 15.0 (1.2-185.2).
IL-6 at admission is an early and accurate indicator of patients at increased risk for VAP. IL-6 is also accurate in discriminating patients with VAP from other causes of pulmonary infiltrates. Extrapolation of these results to the overall population of critically ill patients is limited by the small number of patients.
No preview · Article · Apr 2009 · Critical care medicine
[Show abstract][Hide abstract] ABSTRACT: The recent Infectious Disease Society of America/American Thoracic Society guidelines for the management of community-acquired pneumonia (CAP) in adults defined a predictive rule to identify patients with severe CAP to determine the need for intensive care unit (ICU) admission. We clinically validated this rule.
We analyzed 2102 episodes of CAP in consecutively hospitalized patients over a 7-year period. The predictive rule consists of at least 1 of 2 major severity criteria (septic shock and invasive mechanical ventilation) or at least 3 of 9 minor severity criteria. We assessed the association of the predictive rule with ICU admission and mortality.
A total of 235 episodes of CAP (11%) occurred in patients who were admitted to the ICU, whereas the predictive rule identified 397 (19%) of 2102 episodes as severe CAP. The predictive rule and the decision for ICU admission agreed in 1804 (86%) of the episodes (kappa coefficient, 0.45), with a sensitivity of 71% and a specificity of 88%, similar to the 2001 American Thoracic Society guidelines (sensitivity, 66%; specificity, 90%) in predicting ICU admission. Severe CAP criteria had higher sensitivity (58% vs. 46%) and similar specificity (88% vs. 90%), compared with the 2001 American Thoracic Society guidelines in predicting hospital mortality. Invasive mechanical ventilation was the main determinant for ICU admission, followed by septic shock. In the absence of major criteria, ICU admission was not related to survival of patients with minor severity criteria.
The predictive rule to identify severe CAP is accurate for ICU admission and improved the prediction of mortality, compared with the previous American Thoracic Society guidelines. The need for ICU admission derived from minor severity criteria alone is uncertain and deserves further investigation.
Full-text · Article · Feb 2009 · Clinical Infectious Diseases
[Show abstract][Hide abstract] ABSTRACT: To determine the incidence and trends of pneumococcal community-acquired pneumonia (CAP) resistant to antibiotics, to describe clinical and microbiological features of pneumococcal CAP, and to ascertain prognostic risk factors in a third-level hospital. Design and setting: We performed a prospective study of all well-defined pneumococcal CAP hospitalizations in the Hospital Clínic de Barcelona (Spain) over 2 years of follow-up, and results were compared with a previous study.
One hundred twenty-five patients were included (mean age, 59.6 years; 71.2% male and 28.8% female). Mortality was 7% (n = 9). Twenty-four percent were HIV-1 seropositive (n = 30), and 53% had at least one comorbidity (n = 65). Nonsusceptibility to penicillin, ceftriaxone, and erythromycin accounted for 34%, 9%, and 33%, respectively. A decrease in penicillin (p = 0.01) and cephalosporin (p < 0.001) resistance was observed on comparison with a previous study, while macrolide resistance remained unchanged. Serotype 1 infection was overrepresented (8%, n = 10). A bad outcome was related to female gender (relative risk [RR], 9.1; confidence interval [CI], 1.3 to 61.3), pleural effusion (RR, 13.35; CI, 1.9 to 93.1), and prior oral corticoid intake (RR, 10.59; CI, 1.2 to 91.2), whereas drug-resistant strains were not.
We found a decrease in drug resistance compared with a previous report and a relatively high incidence of serotype 1 pneumococcal CAP. We also observed a high prevalence of HIV-1 infection among individuals with pneumococcal pneumonia. We confirm the lack of association of drug resistance with mortality and length of hospitalization. Mortality was associated with female gender, pleural effusion, and previous oral corticoid treatment. These results should be better ascertained in further studies.