Gene expression profiling of laser microdissected airway smooth muscle tissue in asthma and atopy.

BACKGROUND: Asthma and atopy share common characteristics including type 2 helper-T-cell-mediated inflammation. However, only asthma is associated with variable airways obstruction. The complex cellular and molecular pathways distinguishing asthma and atopy can now be captured by transcriptomic analysis (RNA-Seq). We hypothesized that the transcriptomic profile of airway smooth muscle (ASM) distinguishes atopic asthma from atopic healthy controls. First, we compared the ASM transcriptomic profiles of endobronchial biopsies between glucocorticoid-free, atopic asthma patients, and atopic and nonatopic healthy controls. Second, we investigated the association between ASM transcriptomic profiles and airway function. METHODS: Twelve asthma patients and 12 control subjects (six atopic, six nonatopic) underwent bronchoscopy. RNA of laser-dissected ASM from 96 bronchial biopsy specimens was sequenced with Roche GS FLX. Gene networks were identified using Ingenuity Pathway Analysis. RNA-Seq reads were assumed to follow a negative binomial distribution. With the current sample size, the estimated false discovery rate was approximately 1%. RESULTS: One hundred and seventy four ASM genes were differentially expressed between asthma patients and atopic controls, 108 between asthma patients and nonatopic controls, and 135 between atopic and nonatopic controls. A set of eight genes discriminated asthma patients from nonasthmatic controls, irrespective of atopy. Four of these genes (RPTOR, VANGL1, FAM129A, LEPREL1) were associated with airway hyper-responsiveness (P < 0.05). CONCLUSION: Airway smooth muscle from asthma patients can be distinguished from that of atopic and nonatopic control subjects by a specific gene expression profile, which is associated with airway hyper-responsiveness.

Extracellular matrix in airway smooth muscle is associated with dynamics of airway function in asthma.

BACKGROUND: Altered deposition of extracellular matrix (ECM) in the airway smooth muscle (ASM) layer as observed in asthma may influence ASM mechanical properties. We hypothesized that ECM in ASM is associated with airway function in asthma. First, we investigated the difference in ECM expression in ASM between asthma and controls. Second, we examined whether ECM expression is associated with bronchoconstriction and bronchodilation in vivo. METHODS: Our cross-sectional study comprised 19 atopic mild asthma patients, 15 atopic and 12 nonatopic healthy subjects. Spirometry, methacholine responsiveness, deep-breath-induced bronchodilation (ΔR(rs) ) and bronchoscopy with endobronchial biopsies were performed. Positive staining of elastin, collagen I, III and IV, decorin, versican, fibronectin, laminin and tenascin in ASM was quantified as fractional area and mean density. Data were analysed using Pearson's or Spearman's correlation coefficient. RESULTS: Extracellular matrix expression in ASM was not different between asthma and controls. In asthmatics, fractional area and mean density of collagen I and III were correlated with methacholine dose-response slope and ΔR(rs) , respectively (r = 0.71, P < 0.01; r = 0.60, P = 0.02). Furthermore, ASM collagen III and laminin in asthma were correlated with FEV(1) reversibility (r = -0.65, P = 0.01; r = -0.54, P = 0.04). CONCLUSION: In asthma, ECM in ASM is related to the dynamics of airway function in the absence of differences in ECM expression between asthma and controls. This indicates that the ASM layer in its full composition is a major structural component in determining variable airways obstruction in asthma.

Necessity of centralisation of EBUS.

In the study of Bauwens et al. endobronchial ultrasound (EBUS) was provocated as first step procedure in the staging of PET mediastinal hot spots in lung cancer patients. In case of negative findings a surgical procedure should be undertaken. We certainly agree that in case of a negative finding a surgical procedure should be performed, however, we disagree that the first step procedure should be EBUS. In our opinion the first step procedure in a standard clinical practice should be a standard transbronchial needle aspiration (TBNA).

Pleural drainage and pleurodesis: implementation of guidelines in four hospitals.

The aim of the present study was to evaluate the implementation of the 2003 Dutch guideline on the diagnosis and treatment of malignant pleural effusions, and the potential effect of the implementation on the clinical outcome of pleurodesis. All patients with malignant pleural effusion who had a pleural drain placed with the intention of performing pleurodesis were registered prospectively in four centres. Details of the procedure and fluid recurrence and survival data were noted. Patients with a proven malignancy (n = 100) were entered into the registration database. Diagnostic guideline recommendations were followed in 60-70% of the patients. Surprisingly, pleurodesis was performed in only 75% of the patients, mainly due to the presence of a trapped lung. All pleurodeses were performed using talc, according to the guideline. Follow-up revealed fluid recurrence in 27 (36%) patients after a mean follow-up of 17 days (range 2-285 days); 14 patients with successful pleurodesis died with a median survival of 61 days (range 13-174 days). Systemic treatment following pleurodesis and good apposition of the pleural surfaces during drainage were good prognostic factors. Despite reasonable-to-good adherence to the guideline, the number of successful pleurodeses was low. Better predictors of a good pleurodesis outcome are needed.

A randomized controlled trial comparing indwelling pleural catheters with talc pleurodesis (NVALT-14).

BACKGROUND: Symptomatic malignant pleural effusion (MPE) occurs frequently in patients with metastatic cancer. The associated prognosis is poor and the success rate of talc pleurodesis (TP) is low. Indwelling pleural catheters (IPCs) are commonly inserted when TP has been unsuccessful. METHODS: We compared talc pleurodesis with the use of an indwelling pleural catheter in patients with recurrent MPE in a multicenter randomized controlled trial (superiority design). The primary endpoint was improvement from baseline in Modified Borg Score (MBS) 6weeks after randomized treatment. Secondary endpoints were hospitalization days, re-interventions, and adverse events. RESULTS: Dyspnea improved significantly (p<0.01) after either treatment, but the magnitude of this improvement did not differ significantly between arms (median 3 and 1 for TP:IPC respectively in rest, p=0.16, (TP 13:IPC 16) and 3 and 1 during exercise, p=0.72 (TP 13:IPC 17)). There was no difference in dyspnea during exercise between TP and IPC at week 6 following treatment, while at rest TP patients (n=13) reported less dyspnea than IPC patients (n=18) (median 0 vs 1, p=0.002). Compared to TP, patients with an IPC had significantly less hospital days during randomized treatment (median: 0 vs 5, p<0.0001), and total hospitalizations for all causes (median: 1.6 vs 1.0, p=0.0035). Fewer IPC patients underwent more than one re-intervention (7/45 vs 15/43, p=0.09). The mean number of re-interventions was lower following IPC (0.21 vs 0.53, p=0.05). Equal number of adverse events occurred. CONCLUSIONS: IPC was not superior in the primary endpoint, improvement of the modified Borg scale (MBS). However, IPC patients had lower hospital stay, fewer admissions and fewer re-interventions. The IPC is an effective treatment modality in patients with symptomatic malignant pleural effusion.

Evaluation of electrical impedance tomography in the measurement of PEEP-induced changes in lung volume.

STUDY OBJECTIVES: A new noninvasive practical technique called electrical impedance tomography (EIT) was examined for the measurement of alveolar recruitment. DESIGN: Prospective clinical study. SETTING: ICU of a general hospital. PATIENTS: Acute respiratory failure (ARF) patients. MEASUREMENTS: The ventilation-induced impedance changes (VICs) of the nondependent and the dependent part of the lung were determined by EIT as a measure of tidal volume distribution. By the use of an impedance ratio (IR), defined as the VIC of the nondependent part of the lung divided by the VIC of the dependent part of the lung, the ventilation performances in both parts of the lung were compared to each other. RESULTS: Between patients, the VIC of the nondependent part of the lung was significantly lower in the patients with a level of positive end-expiratory pressure (PEEP) of > 10 cm H2O than in patients with a PEEP of < 5 cm H2O (p < 0.05). A significantly lower IR (-/+ SD) was found in the group with PEEP of > 10 cm H2O than in the group with PEEP between 0 and 5 cm H2O (1.28+/-0.58 vs 2.99+/-1.24, respectively; p < 0.01). In individual patients, the VIC of the whole lung increased when the PEEP level was increased. The VICs of the nondependent part of the lung and of the dependent part of the lung showed significant increases at a PEEP of 10 cm H2O compared to a PEEP of 0 cm H2O (p < 0.05). Also the IR decreased in individual patients when the PEEP was increased; a significant decrease was found at 10 cm H2O compared to 0 cm H2O (1.67+/-1.24 vs 2.23+/-1.47, respectively; p < 0.05). CONCLUSIONS: The decrease in IR indicates an increase in VIC in the dependent part of the lung above the nondependent part of the lung. The increase in VIC can be regarded as an increase in lung volume, implying alveolar recruitment in the dependent part of the lung. The same results also have been shown in earlier reports by CT scan. Since EIT is far more practical than CT scanning and also is a bedside method, EIT might help in the adjustment of ventilator settings in ARF patients.

Electrical impedance tomography in the assessment of extravascular lung water in noncardiogenic acute respiratory failure.

STUDY OBJECTIVES: To establish the value of electrical impedance tomography (EIT) in assessing pulmonary edema in noncardiogenic acute respiratory failure (ARF), as compared to the thermal dye double indicator dilution technique (TDD). DESIGN: Prospective clinical study. SETTING: ICU of a general hospital. PATIENTS: Fourteen ARF patients. INTERVENTIONS: In order to use the TDD to determine the amount of extravascular lung water (EVLW), a fiberoptic catheter was placed in the femoral artery. MEASUREMENTS AND MAIN RESULTS: Fourteen consecutive ARF patients receiving mechanical ventilation were measured by EIT and TDD. EIT visualizes the impedance changes caused by the ventilation in two-dimensional image planes. An impedance ratio (IR) of the ventilation-induced impedance changes of a posterior and an anterior part of the lungs was used to indicate the amount of EVLW. For the 29 measurements in 14 patients, a significant correlation between EIT and TDD (r = 0. 85; p < 0.001) was found. The EIT reproducibility was good. The diagnostic value of the method was tested by receiver operator characteristic analysis, with 10 mL/kg of EVLW considered as the upper limit of normal. At a cutoff level of the IR of 0.64, the IR had a sensitivity of 93%, a specificity of 87%, and a positive predictive value of 87% for a supranormal amount of EVLW. Follow-up measurements were performed in 11 patients. A significant correlation was found between the changes in EVLW measured with EIT and TDD (r = 0.85; p < 0.005). CONCLUSION: We conclude that EIT is a noninvasive technique for reasonably estimating the amount of EVLW in noncardiogenic ARF.

Ventilation and perfusion imaging by electrical impedance tomography: a comparison with radionuclide scanning.

Electrical impedance tomography (EIT) is a technique that makes it possible to measure ventilation and pulmonary perfusion in a volume that approximates to a 2D plane. The possibility of using EIT for measuring the left-right division of ventilation and perfusion was compared with that of radionuclide imaging. Following routine ventilation (81mKr) and perfusion scanning (99mTc-MAA), EIT measurements were performed at the third and the sixth intercostal level in 14 patients with lung cancer. A correlation (r = 0.98, p < 0.005) between the left-right division for the ventilation measured with EIT and that with 81mKr was found. For the left-right division of pulmonary perfusion a correlation of 0.95 (p < 0.005) was found between the two methods. The reliability coefficient (RC) was calculated for estimating the left-right division with EIT. The RC for the ventilation measurements was 94% and 96% for the perfusion measurements. The correlation analysis for reproducibility of the EIT measurements was 0.95 (p < 0.001) for the ventilation and 0.93 (p < 0.001) for the perfusion measurements. In conclusion, EIT can be regarded as a promising technique to estimate the left-right division of pulmonary perfusion and ventilation.

Pulmonary perfusion measured by means of electrical impedance tomography.

Electrical impedance tomography (EIT) is a recent imaging technique based on electrical impedance, offering the possibility of measuring pulmonary perfusion. In the present study the influence of several pulmonary haemodynamical parameters on the EIT signal were investigated. First, the influence on the systolic wave of the EIT signal (delta Zsys) of stroke volume, large pulmonary artery distensibility (both assessed by means of MRI) and the extent of the pulmonary peripheral vascular bed in 11 emphysematous patients (reduced peripheral vascular bed) and 9 controls (normal peripheral vascular bed) was investigated. Second, the influence of hypoxic pulmonary vasoconstriction on delta Zsys was examined in 14 healthy subjects. Finally, the origin of the diastolic wave was examined in three patients with atrioventricular dissociation. Multiple regression analysis showed that delta Zsys was only dependent on the variable emphysema (p < 0.02), but not dependent on stroke volume (p < 0.3) or pulmonary artery distensibility (p > 0.9). The mean value of delta Zsys for emphysematous patients (131 +/- 32 arbitrary units (AU)) was significantly lower (p < 0.001) than in the control group (200 +/- 39). In the group of healthy subjects delta Zsys decreased significantly (p < 0.001) during hypoxia (193 +/- 38 AU) compared with rest measurements (260 +/- 62 AU). The absence of the diastolic wave in the cardiological patients suggests the influence of reverse venous blood flow on the EIT signal. It is concluded that volume changes in the small pulmonary vessels contribute significantly to the EIT signal. Moreover, the hypoxia induced decrease in delta Zsys indicates the potential of EIT for measuring pulmonary vascular responses to external stimuli.

Influences of lung parenchyma density and thoracic fluid on ventilatory EIT measurements.

Ventilatory impedance changes can be measured by electrical impedance tomography (EIT). Several studies have pointed out that the ventilatory-induced impedance change measured over the lungs shows a linear relationship with tidal volume. However, EIT measures the ventilatory impedance changes relative to a reference. Therefore, changes in the reference due to lung parenchyma destruction (increase of thoracic impedance) or lung water (decrease of thoracic impedance) might influence ventilatory EIT measurements. A study was designed to evaluate the influence of the density of lung parenchyma and the thoracic fluid content on ventilatory EIT measurements. Eleven emphysema patients with a variable degree of lung parenchyma destruction, nine haemodialysis patients with general fluid overload and ten healthy subjects were measured. The impedance changes were measured with the subject in the supine position breathing a constant tidal volume of 1 litre starting at the maximum end-expiratory level. In the emphysema group a significantly lower impedance change between ins- and expiration was found in comparison with the healthy subjects (11.6 +/- 6.4 AU l-1 versus 18.6 +/- 4.2 AU l-1, p < 0.05), whereas the haemodialysis group showed a significantly larger impedance change between ins- and expiration before haemodialysis (30.5 +/- 13.1 AU l-1, p < 0.05). A significant decrease in ventilation-induced impedance change during dialysis was found (30.5 +/- 13.1 AU l-1 versus 21.4 +/- 8.6 AU l-1, p < 0.01). Furthermore, a significant correlation between lung function parameters, which indicate the severity of lung parenchyma destruction, and the measured impedance change was found in emphysema patients. From these results it can be concluded that the density of lung parenchyma and the thoracic fluid content have a serious impact on the ventilation-induced impedance change.

The influence of extravascular lung water on cardiac output measurements using thoracic impedance cardiography.

The purpose of this study was to investigate the influence of pulmonary oedema as measured with the double indicator dilution technique on the accuracy of cardiac output (CO) measurement using thoracic impedance cardiography (TIC) compared with thermodilution in thirteen sepsis patients. Differences in the Kubicek and Sramek-Bernstein equation with respect to pulmonary oedema were explored theoretically and experimentally. From a parallel two cylinder model a hypothesis can be derived that CO determined with the Kubicek equation is oedema independent, whereas CO determined using the Sramek-Bernstein equation is oedema dependent. Experimentally, CO determined using Kubicek's equation correlated better with thermodilution CO (r = 0.75) than CO determined with the Sramek-Bernstein equation (r = 0.25). The effect of oedema on the accuracy of TIC was investigated by comparing the differences in the CO of impedance and thermodilution to the extravascular lung water index. For the Kubicek equation the difference was not influenced by oedema (r = 0.04, p = 0.84), whereas for the Sramek-Bernstein equation the difference was affected by oedema (r = 0.39, p = 0.05). Thus, the effects of pulmonary oedema on the accuracy of TIC measurements can better be understood with the parallel cylinder model. Moreover, the Kubicek equation still holds when pulmonary oedema is present, in contrast to the Sramek-Bernstein equation.

Validity and reproducibility of electrical impedance tomography for measurement of calf blood flow in healthy subjects.

The Sheffield electrical impedance tomography; (EIT) system produces images of changes in the distribution of resistivity within tissue. The paper reports on the application of electrical impedance tomography in monitoring volume changes in the limb during venous occlusion. The aim of the study is to assess the feasibility, reproducibility and validity of calf blood flow measurements by EIT. In 14 healthy volunteers calf blood flow is compared, as determined in a calf segment by strain-gauge plethysmography (SGP), with the impedance changes measured by EIT during rest and post-ischaemic hyperaemia. The measurements are repeated to assess reproducibility. The reproducibility for the EIT, assessed from the repeated measurements and expressed as a reproducibility coefficient, is 0.88 during rest and 0.89 during hyperaemia. The reproducibility coefficient for SGP data is 0.83 at rest and 0.67 during hyperaemia. Flow measurements, assessed by means of two methods, correlate well at rest (r = 0.89), but only moderately during hyperaemia (r = 0.51). The correlation coefficient for the pooled flow measurements is 0.98. It is concluded that EIT is a valid and reliable method for assessing blood flow in the limb. Possible applications of EIT in localising fluid changes are discussed.

Estimation of non-cardiogenic pulmonary oedema using dual-frequency electrical impedance.

The study investigates the effects of non-cardiogenic oedema, especially the accumulation of protein in extracellular fluid, on thoracic impedance and proposes a new method of oedema measurement based on an impedance ratio from a dual-frequency measurement. In vitro measurements in a cell containing an albumin-in-saline solution yield a resistance increase when the albumin concentration increases. Subsequently, 13 patients having acute respiratory failure are measured. The single-frequency Z0 measurements and the proposed impedance ratio are compared with extravascular lung water (EVLW) determined by the double indicator dilution method. The single-frequency measurement correlates poorly with EVLW (r = -0.24, p = 0.56). In some patients, a total thoracic impedance increase is found with increasing EVLW. The correlation between the impedance ratio and EVLW is r = -0.79 (p < 0.0005). The ratio decreases as EVLW increases. Thus, when oedema is measured using bio-impedance, cardiogenic and non-cardiogenic oedema yield different results. It is well recognised that cardiogenic oedema decreases total thoracic impedance. In non-cardiogenic oedema, however, protein accumulation causes an impedance increase. The decrease in the impedance ratio as EVLW increases can be explained by the accumulation of albumin in the extracellular compartment.

[Patient with a lung abscess, primarily treated with drainage followed by antibiotics]. / Een patiënt met een longabces, primair behandeld met drainage en aanvullend met antibiotica.

A 45-year-old woman presented herself with coughing, nocturnal sweating, weight loss, and chest pain, left laterally. In the previous 5 months she had been treated twice with antibiotics due to a suspected pneumonia. With the help of a CT scan, 2 subpleural lung abscesses were diagnosed. The primary treatment was CT-guided drainage, as a result of which the largest abscess was emptied and a microbiological diagnosis could be established. Subsequently, the patient made a quick recovery with the help of specific antibiotics. It might be better to drain lung abscesses, especially subpleural ones, at an early stage rather than wait for the results of a trial treatment with antibiotics.

Regional pressure volume curves by electrical impedance tomography in a model of acute lung injury.

OBJECTIVE: A new noninvasive method, electrical impedance tomography (EIT), was used to make pressure-impedance (PI) curves in a lung lavage model of acute lung injury in pigs. The lower inflection point (LIP) and the upper deflection point (UDP) were determined from these curves and from the traditional pressure-volume (PV) curves to determine whether the PI curves resemble the traditional PV curves. Furthermore, regional differences in the mentioned determinants were investigated. DESIGN: Prospective, experimental study. SETTING: Animal research laboratory. INTERVENTIONS: In nine anesthetized pigs, repeated lung lavage was performed until a Pao2 <80 torr was reached. Thereafter, an inspiratory PV curve was made using a constant flow of oxygen. During the intervention, EIT measurements were performed. MEASUREMENTS AND MAIN RESULTS: In this study, the LIP(EIT) was within 2 cm H2O of the LIP(PV). Furthermore, it was possible to visualize regional PI curves by EIT. No significant difference was found between the LIP(PV) (21.3+/-3.0 cm H2O) and the LIP(EIT) of the total lung (21.5+/-3.0 cm H2O) or the anterior parts of the lung (21.5+/-2.9 cm H2O). A significantly higher LIP (29.5+/-4.9 cm H2O) was found in the posterior parts of the lung. A UDP(PV) could be found in three animals only, whereas in all animals a UDP(EIT) could be determined from the anterior part of the lung. CONCLUSIONS: Using EIT, determination of LIP and UDP from the regional PI curves is possible. The obtained information from the regional PI curves may help in understanding alveolar recruitment. The use of this new bedside technique for clinical decision making remains to be examined.

Monitoring of recruitment and derecruitment by electrical impedance tomography in a model of acute lung injury.

OBJECTIVE: To evaluate a noninvasive system for obtaining information about alveolar recruitment and derecruitment in a model of acute lung injury. DESIGN: Prospective experimental study. SETTING: Animal research laboratory. SUBJECTS: Nine anesthetized pigs. INTERVENTIONS: Electrical impedance tomography measurements were performed. Electrical impedance tomography is an imaging technique that can register the ventilation-induced impedance changes in different parts of the lung. In nine anesthetized pigs, repeated lung lavages were performed until a PaO2 of <80 mm Hg was reached. Thereafter, the lungs were recruited according to two different recruitment protocols: the open lung approach and the open lung concept. Five time points for measurements were chosen: healthy (reference), lavage (atelectasis), recruitment, derecruitment, and maintain recruited (final). MEASUREMENTS AND MAIN RESULTS: After lavage, there was a significant increase in the impedance ratio, defined as the ventilation-induced impedance changes of the anterior part of the lung divided by that of the posterior part (from 1.75 +/- 0.63 to 4.51 +/- 2.22; p < .05). The impedance ratio decreased significantly after performing the recruitment protocol (from 4.51 +/- 2.22 to 1.18 +/- 0.51). During both recruitment procedures, a steep increase in baseline impedance change was seen. Furthermore, during derecruitment, a decrease in the slope in baseline impedance change was seen in the posterior part of the lung, whereas the anterior part showed no change. CONCLUSION: Electrical impedance tomography is a technique that can show impedance changes resembling recruitment and derecruitment of alveoli in the anterior and posterior parts of the lung. Therefore, electrical impedance tomography may help in determining the optimal mechanical ventilation in a patient with acute lung injury.