These evidence-based guidelines have been produced after a systematic literature review of a range of issues involving prevention, diagnosis and treatment of hospital-acquired pneumonia (HAP). Prevention is structured into sections addressing general issues, equipment, patient procedures and the environment, whereas in treatment, the structure addresses the use of antimicrobials in prevention and treatment, adjunctive therapies and the application of clinical protocols. The sections dealing with diagnosis are presented against the clinical, radiological and microbiological diagnosis of HAP. Recommendations are also made upon the role of invasive sampling and quantitative microbiology of respiratory secretions in directing antibiotic therapy in HAP/ventilator-associated pneumonia.

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h Mycoplasma amphoriforme is a recently described organism isolated from the respiratory tracts of patients with immunodeficiency and evidence of chronic infection. Novel assays for the molecular detection of the organism by real-time quantitative PCRs (qPCRs) targeting the uracil DNA glycosylase gene (udg) or the 23S rRNA gene are described here. The analytical sensitivities are similar to the existing conventional M. amphoriforme 16S rRNA gene PCR, with the advantage of being species specific, rapid, and quantitative. By using these techniques, we demonstrate the presence of this organism in 17 (19.3%) primary antibody-deficient (PAD) patients, 4 (5%) adults with lower respiratory tract infection, 1 (2.6%) sputum sample from a patient attending a chest clinic, and 23 (0.21%) samples submitted for viral diagnosis of respiratory infection, but not in normal adult control subjects. These data show the presence of this microorganism in respiratory patients and suggest that M. amphoriforme may infect both immunocompetent and immunocompromised people. Further studies to characterize this organism are required, and this report provides the tools that may be used by other research groups to investigate its pathogenic potential. Mycoplasma amphoriforme was first isolated in 1999 from a patient with primary antibody deficiency (PAD) with chronic bronchitis. It has also been isolated subsequently from both immunocompromised and immunocompetent patients with respiratory tract infections (RTI) in London, Denmark, France, and Tunisia (1-3). Based on 16S rRNA sequencing, M. amphoriforme belongs to the same phylogenetic group as other human pathogenic Mycoplasma species, the pneumoniae group (1, 2). The closest species phylogenetically for which there is a whole-genome sequence is Mycoplasma gallisepticum, a bird pathogen. Phenotypic studies have demonstrated that M. amphoriforme has features in common with this group, including gliding motility, a protruding polar tip resembling that of M. gallisepticum, and a cytoskeletal structure at its polar tip with homology to that of M. pneumoniae's attachment organelle (1, 4).To understand the role that this novel agent plays in human health, better laboratory tools are required. M. amphoriforme is fastidious, requiring specialized media for cultivation, and it takes approximately 2 weeks for colonies to appear on agar. The colonial morphology resembles granular droplets, making detection difficult, as the droplets can blend into the sample matrix and be overlooked. This paper reports the development and evaluation of two real-time quantitative PCR (qPCR) assays: one assay targeting M. amphoriforme's uracil DNA glycosylase gene (udg) and one assay targeting the variable region of the 23S rRNA gene that is unique to M. amphoriforme. The new qPCR assays were compared with a previously reported 16S rRNA gene assay (2) and used to test a range of human samples from the United Kingdom. MATERIALS AND METHODSPatients, samples, and ethical approval. Clinical samples from two hospitals were u...

We have demonstrated that the FRC of healthy term parturients increases significantly in the 30° head-up position in comparison with supine.

Our data, thus, provide objective evidence of a positive effect of aromatics inunction on mucus clearance in chronic airways obstruction.