Effect of External Positive and Negative Pressure on Venous Flow in an Experimental Model.

OBJECTIVE: Positive external pressure is said to decrease transmural pressure; negative pressure in the pleural cavity is widely believed to result in negative pressure in systemic chest veins. The discrepancy between erect column height and foot venous pressure has been explained on this basis. METHODS: These core concepts rest on static closed models that may not be appropriate. This study examined the effects of external pressures in a dynamic open model that may better reflect in vivo conditions. Flow in a Penrose drain enclosed in a chamber that could be positively or negatively pressurized was used. Input and output reservoirs with pressures in the physiological range provided flow. Flow and pressure were monitored in horizontal and erect models with modifications to suit particular experiments. RESULTS: The discrepancy between foot venous pressure and erect venous column height was shown in this experimental model to be a result of two flows in opposite directions (superior and inferior vena cavae) meeting at the zero reference level at the heart; the upper column pressure therefore does not register at the foot. Positive external pressure results in slowing of velocity with conversion to pressure. Internal and transmural pressures therefore do not decrease. Negative external pressure has only a marginal effect on flow; importantly, internal pressure does not become negative. In an experimental set-up it was shown that negative pressure in chest veins was not necessary for air embolism to occur. CONCLUSION: Persistent negative pressure in systemic chest veins probably does not occur. The reason for the discrepant foot venous pressure is likely to be a result of dynamic flow and not negative pressure in chest veins. External positive pressure results in slowing of velocity but the transmural pressure remains largely unchanged.

Airway smooth muscle proliferation and survival is not modulated by mast cells.

BACKGROUND: Airway smooth muscle (ASM) hyperplasia and mast cell localization within the ASM bundle are important features of asthma. The cause of this increased ASM mass is uncertain and whether it is a consequence of ASM-mast cell interactions is unknown. OBJECTIVE: We sought to investigate ASM proliferation and survival in asthma and the effects of co-culture with mast cells. METHODS: Primary ASM cultures were derived from 11 subjects with asthma and 12 non-asthmatic controls. ASM cells were cultured for up to 10 days in the presence or absence of serum either alone or in co-culture with the human mast cell line-1, unstimulated human lung mast cells (HLMC) or IgE/anti-IgE-activated HLMC. Proliferation was assessed by cell counts, CFSE assay and thymidine incorporation. Apoptosis and necrosis were analysed by Annexin V/propidium iodide staining using flow cytometry and by assessment of nuclear morphology using immunofluorescence. Mast cell activation was confirmed by the measurement of histamine release. RESULTS: Using a number of techniques, we found that ASM proliferation and survival was not significantly different between cells derived from subjects with or without asthma. Co-culture with mast cells did not affect the rate of proliferation or survival of ASM cells. CONCLUSION: Our findings do not support a role for increased airway smooth proliferation and survival as the major mechanism driving ASM hyperplasia in asthma.

Airway wall geometry in asthma and nonasthmatic eosinophilic bronchitis.

BACKGROUND: Variable airflow obstruction and airway hyperresponsiveness (AHR) are features of asthma, which are absent in nonasthmatic eosinophilic bronchitis (EB). Airway remodelling is characteristic of both conditions suggesting that remodelling and airway dysfunction are disassociated, but whether the airway geometry differs between asthma and nonasthmatic EB is uncertain. METHODS: We assessed airway geometry by computed tomography (CT) imaging in asthma vs EB. A total of 12 subjects with mild-moderate asthma, 14 subjects with refractory asthma, 10 subjects with EB and 11 healthy volunteers were recruited. Subjects had a narrow collimation (0.75 mm) CT scan from the aortic arch to the carina to capture the right upper lobe apical segmental bronchus (RB1). In subjects with asthma and EB, CT scans were performed before and after a 2-week course of oral prednisolone (0.5 mg/kg). RESULTS: Mild-moderate and refractory asthma were associated with RB1 wall thickening in contrast to subjects with nonasthmatic EB who had maintained RB1 patency without wall thickening [mean (SD) % wall area and luminal area mild-t0-moderate asthma 67.7 (7.3)% and 6.6 (2.8) mm(2)/m(2), refractory asthma 67.3 (5.6)% and 6.7 (3.4) mm(2)/m(2), healthy control group 59.7 (6.3)% and 8.7 (3.8) mm(2)/m(2), EB 61.4 (7.8)% and 11.1 (4.6) mm(2)/m(2) respectively; P < 0.05]. Airway wall thickening of non-RB1 airways generation three to six was a feature of asthma only. There was no change in airway geometry of RB1 after prednisolone. Proximal airway wall thickening was associated with AHR in asthma (r = -0.56; P = 0.02). CONCLUSIONS: Maintained airway patency in EB may protect against the development of AHR, whereas airway wall thickening may promote AHR in asthma.

Functional KCa3.1 K+ channels are required for human lung mast cell migration.

BACKGROUND: Mast cell recruitment and activation are critical for the initiation and progression of inflammation and fibrosis. Mast cells infiltrate specific structures in many diseased tissues such as the airway smooth muscle (ASM) in asthma. This microlocalisation of mast cells is likely to be key to disease pathogenesis. Human lung mast cells (HLMC) express the Ca2+ activated K+ channel K(Ca)3.1 which modulates mediator release, and is proposed to facilitate the retraction of the cell body during migration of several cell types. A study was undertaken to test the hypothesis that blockade of K(Ca)3.1 would attenuate HLMC proliferation and migration. METHODS: HLMC were isolated and purified from lung material resected for bronchial carcinoma. HLMC proliferation was assessed by cell counts at various time points following drug exposure. HLMC chemotaxis was assayed using standard Transwell chambers (8 microm pore size). Ion currents were measured using the single cell patch clamp technique. RESULTS: K(Ca)3.1 blockade with triarylmethane-34 (TRAM-34) did not inhibit HLMC proliferation and clotrimazole had cytotoxic effects. In contrast, HLMC migration towards the chemokine CXCL10, the chemoattractant stem cell factor, and the supernatants from tumour necrosis factor alpha stimulated asthmatic ASM was markedly inhibited with both the non-selective K(Ca)3.1 blocker charybdotoxin and the highly specific K(Ca)3.1 blocker TRAM-34 in a dose dependent manner. Although K(Ca)3.1 blockade inhibits HLMC migration, K(Ca)3.1 is not opened by the chemotactic stimulus, suggesting that it must be involved downstream of the initial receptor-ligand interactions. CONCLUSIONS: Since modulation of K(Ca)3.1 can inhibit HLMC chemotaxis to diverse chemoattractants, the use of K(Ca)3.1 blockers such as TRAM-34 could provide new therapeutic strategies for mast cell mediated diseases such as asthma.

Activation of human lung mast cells by monomeric immunoglobulin E.

The mechanism of chronic mast cell activation in asthma is unclear. Monomeric immunoglobulin (Ig)E in the absence of allergen induces mediator release from rodent mast cells, indicating a possible role for IgE in the continued activation of mast cells within the asthmatic bronchial mucosa. In this study it was investigated whether monomeric IgE induces Ca2+ influx and mediator release from human lung mast cells (HLMC). Purified HLMC were cultured for 4 weeks and then exposed to monomeric human myeloma IgE. Ratiometric Ca2+ imaging was performed on single fura-2-loaded cells. Histamine release was measured by radioenzymatic assay; leukotriene C4 (LTC4) and interleukin (IL)-8 were measured by ELISA. At concentrations experienced in vivo, monomeric IgE induced dose-dependent histamine release, LTC4 production and IL-8 synthesis. This was associated with a rise in cytosolic free Ca2+. Enhanced histamine release was still evident 1 week after initial exposure to IgE suggesting that continued exposure maintains enhanced secretion. Monomeric immunoglobulin E alone activates cultured human lung mast cells initiating Ca2+ influx, degranulation, arachidonic acid metabolism and cytokine synthesis. These findings support the hypothesis that immunoglobulin E loading of mast cells within the asthmatic airway contributes to the disordered airway physiology of this disease.

Venous flow restriction: the role of vein wall motion in venous admixture.

OBJECTIVE: There are wide differences in flow between vascular beds at rest, even more during stress. The hydrodynamic energy (Energy grade line or EGL) of venous outflows must also vary considerably between vascular beds. We explored the mechanism of venous admixture of differing energy flows using a mechanical model. MATERIALS AND METHODS: The model simulated two venous flows coalescing at a venous junction and then flowing through collapsible venous pumps. Flow rates and pressures were monitored when the venous pumps were full (steady state) and when they were compressed and allowed to refill inducing wall motion (pump flow). RESULTS: With increasing EGL differences between two coalescing venous flows, reduction or cessation (venous flow restriction) of the weaker flow occurred during steady state; higher base EGL of both flows ameliorated venous flow restriction and lower base EGL the opposite. Outflow obstruction favoured venous flow restriction. Pump action in the vicinity of the venous junction abolished venous flow restriction and maximized both venous flows. CONCLUSION: The model suggests a pivotal role for vein wall motion in venous admixture and regional perfusion. Observations in the model are explained on the basis of network flow principles and collapsible tube mechanics.