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Monocyte retention, activation phenotype and the effects of their depletion by intravenous clodronate-liposome treatment on lung inflammation and injury were determined. Results In mouse lungs following ischaemia-reperfusion, substantial numbers of lung-marginated monocytes remained within the pulmonary microvasculature, with reduced L-selectin and increased CD86 expression indicating their activation.

Monocyte depletion resulted in reductions in lung wet:dry ratios, bronchoalveolar lavage fluid protein, and perfusate levels of RAGE, MIP-2 and KC, while monocyte repletion resulted in a partial restoration of the injury. Pain is a common and distressing symptom experienced by intensive care patients.

Assessing pain in this environ-ment is challenging, and published guidelines have been inconsistently implemented. Data were collected from patients, reflecting the practice of physicians. Two of the 45 ICUs used validated behavioural pain assessment tools.

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The likelihood of receiving a physician painassessment was affected by the following factors: the number of nursing assessments performed; whether the patientwas admitted as a surgical patient; the presence of tracheal tube or tracheostomy; and the length of stay in ICU. Physician-documented pain assessments in the majority of participating ICUs were infrequent and did not utiliserecommended behavioural pain assessment tools.

Further research to identify factors influencing physician painassessment behaviour in ICU, such as human factors or cultural attitudes, is urgently needed. The lung has a unique structure consisting of three functionally different compartments alveolar, interstitial, and vascular situated in an extreme proximity.

The combined in vivo intravenous and intratracheal CD45 labelling enabled robust separation of the alveolar, interstitial, and vascular compartments of the lung.

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In naive mice, the alveolar compartment consisted predominantly of resident alveolar macrophages. The interstitial compartment, gated by events negative for both intratracheal and intravenous CD45 staining, showed two conventional dendritic cell populations, as well as a Ly6C lo monocyte population.

In mice exposed to acid-aspiration induced lung injury, this protocol also clearly distinguished the three lung compartments showing the dynamic trafficking of neutrophils and exudative monocytes across the lung compartments during inflammation and resolution. This simple in vivo dual labelling technique substantially increases the accuracy and depth of lung flow cytometric analysis, facilitates a more comprehensive examination of lung leukocyte pools, and enables the investigation of previously poorly defined 'interstitial' leukocyte populations during models of inflammatory lung diseases.

ShareModerators: G. Maksym, PhD, R. Harris, MD, J. Wilson, PhD, K.

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  • O'Dea, PhD, M. We present a novel method of in vivo antibody labelling to facilitate the accurate compartmental investigation of mouse lung leukocytes by flow cytometry. MethodsAnesthetized C57BL6 mice underwent tracheostomy and venous cannulation.

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    An anti-CD45 antibody PE-conjugated was injected intravenously and allowed to circulate for 5 minutes. Mice were then exsanguinated and lungs immediately flooded intratracheally with another anti-CD45 antibody PE-Cy5-conjugated. Lungs were harvested, and single cell suspensions prepared for flow cytometric analysis using a panel of myeloid markers.

    OSCEs for Medical and Surgical Finals

    This in vivo labelling protocol was also applied to mice exposed to acid-aspiration induced lung injury 3. To confirm the separation between the vascular and interstitial cell populations, a group of mice underwent intravenous labelling alone followed by lung perfusion for 15 minutes using an isolated-perfused lung apparatus 2. Whilst lung transplantation is a viable solution for end-stage lung disease, donor shortages, donor lung inflammation and perioperative lung injury remain major limitations. Ex vivo lung perfusion has emerged as the next frontier in lung transplantation to address and overcome these limitations, with multicentre clinical trials ongoing in the UK, rest of Europe and North America.

    Our research seeks to identify the poorly understood cellular and molecular mechanisms of primary graft dysfunction through the development of an isolated perfused lung model of transplantation and investigation of the role of pulmonary inflammation in this paradigm.

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    Margination and activation of monocytes within the pulmonary microcirculation contribute substantially to the development of acute lung injury in mice. The enhanced LPS-induced TNF expression exhibited by Gr-1 high compared with Gr-1 low monocytes within the lung microvasculature suggests differential roles for these subsets. We investigated the mechanisms responsible for such heterogeneity of lung-marginated monocyte proinflammatory response using a combined in vitro and in vivo approach.

    The monocyte subset inflammatory response was studied in vitro in mouse peripheral blood mononuclear cell-lung endothelial cell coculture and in vivo in a two-hit model of intravenous LPS-induced monocyte margination and lung inflammation in mice, by flow cytometry-based quantification of proinflammatory genes and intracellular phospho-kinases. Donec nec tortor at mi suscipit mollis. Phasellus fermentum venenatis dignissim. Etiam at enim nec lacus consectetur iaculis at quis arcu.

    Ut vitae est dui. Integer orci felis, vulputate sed maximus eget, vulputate eget nibh. Praesent scelerisque in sem quis vehicula. Vol 1 No 3 , Articles. Vol 1 No 3 Robin MacKenzie. Mary Johnson. Supplementary Files Data Files. Keywords OER open textbooks electronic textbooks open access. Cost-savings achieved in two semesters through the adoption of open educational resources. Demonstration Journal of the Health Sciences Theme , 1 3. Abstract Textbooks represent a significant portion of the overall cost of higher education in the United States.

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