Bronchoalveolar Lavage Neutrophil Percentage Research Articles

Changes of breath volatile organic compounds in healthy volunteers following segmental and inhalation endotoxin challenge

It is still unclear how airway inflammation affects the breath volatile organic compounds (VOCs) profile in exhaled air. We therefore analyzed breath following well-defined pulmonary endotoxin (lipopolysaccharide, LPS) challenges. Breath was collected from ten healthy non-smoking subjects at eight time points before and after segmental and whole lung LPS inhalation challenge. Four Tenax-TA® adsorption tubes were simultaneously loaded from an aluminum reservoir cylinder and independently analyzed by two research groups using gas chromatography—mass spectrometry. Airway inflammation was assessed in bronchoalveolar lavage (BAL) and in sputum after segmental and inhaled LPS challenge, respectively. Segmental LPS challenge significantly increased the median (interquartile range, IQR) percentage of neutrophils in BAL from 3.0 (4.2) % to 64.0 (7.3) %. The inhalation challenge increased sputum neutrophils from 33.9 (26.8) % to 78.3 (13.5) %. We observed increases in breath aldehydes at both time points after segmental and inhaled LPS challenge. These results were confirmed by an independent laboratory. The longitudinal breath analysis also revealed distinct VOC patterns related to environmental exposures, clinical procedures, and to metabolic changes after food intake. Changes in breath aldehydes suggest a relationship to LPS induced inflammation compatible with lipid peroxidation processes within the lung. Findings from our longitudinal data highlight the need for future studies to better consider the potential impact of the multiple VOCs from detergents, hygiene or lifestyle products a subject is continuously exposed to. We suspect that this very individual ‘owncloud’ exposure is contributing to an increased variability of breath aldehydes, which might limit a use as inflammatory markers in daily clinical practice.

Clinical and radiological manifestations of lipoid pneumonia according to etiology: Squalene, omega-3-acid ethyl esters, and idiopathic.

Clinical manifestations of lipoid pneumonia (LP) vary depending on the causative agents or underlying causes. The aim of the present study was to investigate the clinical and radiological features of LP, classified according to etiologic agents. The clinico-radiological characteristics of LP patients were retrospectively compared among groups: exogenous versus idiopathic and squalene versus omega-3-acid ethyl esters (O-3-AEE) versus idiopathic. Idiopathic group was defined as LP with no proven or reported etiological evidence. Twenty-two patients met the diagnostic criteria for LP: squalene (9 [41%]), O-3-AEE (6 [27%]), olive oil (1 [5%]), and idiopathic (7 [32%]). Compared with the exogenous group, the idiopathic group showed a higher recurrence rate; higher frequencies of bronchial anthracofibrosis (BAF) and bronchoalveolar lavage (BAL) lymphocytosis; and a higher rate of crazy-paving pattern and lower rate of consolidation on computed tomography scan. In three-group tests, compared with the O-3-AEE group, the squalene group exhibited a significantly higher percentage of neutrophils and a higher rate of right middle lobe (RML) involvement. In comparison with the exogenous group, the idiopathic group demonstrated BAL lymphocytosis, higher rates of recurrence and BAF, and a higher rate of crazy-paving pattern. Compared with the O-3-AEE group, the squlaene group showed a higher percentage of BAL neutrophils and predominant RML involvement.

Calgranulin B and KL-6 in Bronchoalveolar Lavage of Patients with IPF and NSIP.

Idiopathic pulmonary fibrosis (IPF) and non-specific interstitial pneumonia (NSIP) are the most frequent idiopathic interstitial pneumonias. The aim of this study was to evaluate concentrations of calgranulin B and Krebs von den Lungen 6 (KL-6) in bronchoalveolar lavage (BAL) fluid of patients with IPF and idiopathic NSIP (i-NSIP) with fibrotic pattern. Thirty patients with IPF (68.73 ± 8.63years), 30 with i-NSIP (68.33 ± 7.45years), and healthy controls were included in the study. Calgranulin B and KL-6 both proved to be significantly higher in BAL of IPF and i-NSIP patients than in healthy controls (p < 0.05). Calgranulin B showed several significant correlations with functional parameters (oxygen demand at rest, 6-min walking test (6MWT), and PFTs); KL-6 was correlated with oxygen demand at rest and during 6MWT. Patients with higher concentrations of both biomarkers (> 75th percentile) had more advanced disease with lower values of FEV1%, FVC%, RV%, TLC%, DLCO% of predicted, distance walked in 6MWT, and BAL neutrophil percentage. Calgranulin B and KL-6 in BAL proved to be reliable biomarkers of IPF and i-NSIP and to have prognostic meaning, discriminating severe and advanced patients. The combination of the two biomarkers can facilitate the stratification of severity.

Seasonal Variation in Tracheal Mucous and Bronchoalveolar Lavage Cytology for Adult Clinically Healthy Stabled Horses

Seasonal changes affect the concentration of dust particles and aeroallergens in the equine stable environment and are hypothesized to change airway cytology in healthy stabled horses. The objective of the study was to evaluate if seasonal changes from May to November influenced the lower airway health cytological findings in clinically healthy stabled horses. Endoscopic examination, including tracheal aspiration (TA) and bronchoalveolar lavage (BAL) cytology was performed in November and May on 63 clinically healthy stabled Danish Warmblood geldings, aged 12.9 (±4.6) years. Tracheal aspiration and BAL cytology were evaluated via May-Grünwald-Giemsa stain after cytocentrifugation of samples. Five-hundred cells were identified microscopically as macrophages, lymphocytes, neutrophils, eosinophils, or mast cells. A significant difference in climatic conditions with lower temperatures (P < .001), fewer sunlight hours (P = .004), less precipitation (P = .006), and higher humidity (P < .001) was found in November compared to May. A significantly higher percentage of neutrophils (Median 6.10%, Quartiles 4.00–8.80, P < .001) and a higher tracheal mucus score (Median 3.50%, Quartiles 1.95–5.20, P = .010) were found during November compared to May period. Furthermore, a positive correlation between TA and BAL neutrophils was found for both sampling times. Mild winter weather conditions with lower temperatures, fewer minutes of sunlight, and a higher percent humidity were associated with significantly higher BAL neutrophil percentage and significantly higher mucus score in stabled clinical healthy horses. The clear seasonal influence on BAL neutrophilia and tracheal mucous findings in clinically normal horse needs to be included in the working diagnosis of asthma in horses.

The peroxisome proliferator-activated receptor agonist pioglitazone and 5-lipoxygenase inhibitor zileuton have no effect on lung inflammation in healthy volunteers by positron emission tomography in a single-blind placebo-controlled cohort study.

BackgroundAnti-inflammatory drug development efforts for lung disease have been hampered in part by the lack of noninvasive inflammation biomarkers and the limited ability of animal models to predict efficacy in humans. We used 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET) in a human model of lung inflammation to assess whether pioglitazone, a peroxisome proliferator-activated receptor-γ (PPAR-γ) agonist, and zileuton, a 5-lipoxygenase inhibitor, reduce lung inflammation.MethodsFor this single center, single-blind, placebo-controlled cohort study, we enrolled healthy volunteers sequentially into the following treatment cohorts (N = 6 per cohort): pioglitazone plus placebo, zileuton plus placebo, or dual placebo prior to bronchoscopic endotoxin instillation. 18F-FDG uptake pre- and post-endotoxin was quantified as the Patlak graphical analysis-determined Ki (primary outcome measure). Secondary outcome measures included the mean standard uptake value (SUVmean), post-endotoxin bronchoalveolar lavage (BAL) cell counts and differentials and blood adiponectin and urinary leukotriene E4 (LTE4) levels, determined by enzyme-linked immunosorbent assay, to verify treatment compliance. One- or two-way analysis of variance assessed for differences among cohorts in the outcome measures (expressed as mean ± standard deviation).ResultsTen females and eight males (29±6 years of age) completed all study procedures except for one volunteer who did not complete the post-endotoxin BAL. Ki and SUVmean increased in all cohorts after endotoxin instillation (Ki increased by 0.0021±0.0019, 0.0023±0.0017, and 0.0024±0.0020 and SUVmean by 0.47±0.14, 0.55±0.15, and 0.54±0.38 in placebo, pioglitazone, and zileuton cohorts, respectively, p<0.001) with no differences among treatment cohorts (p = 0.933). Adiponectin levels increased as expected with pioglitazone treatment but not urinary LTE4 levels as expected with zileuton treatment. BAL cell counts (p = 0.442) and neutrophil percentage (p = 0.773) were similar among the treatment cohorts.ConclusionsEndotoxin-induced lung inflammation in humans is not responsive to pioglitazone or zileuton, highlighting the challenge in translating anti-inflammatory drug efficacy results from murine models to humans.Trial registrationClinicalTrials.gov NCT01174056.

MUC18 regulates IL-13-mediated airway inflammatory response

To evaluate the effects of MUC18 on IL-13-mediated airway inflammatory responses in human airway epithelial cells and in mice. Primary normal human tracheobronchial epithelial (HTBE) cells, wild-type (WT) and Muc18 knockout (KO) mice, and mouse tracheal epithelial cells (mTECs) were utilized. Cultured HTBE cells treated with MUC18 siRNA or MUC18 expressing lentivirus were incubated with IL-13 (10ng/mL) for 24h. Mice were intranasally instilled with 500ng of IL-13 for 3days. mTECs were treated with IL-13 (10ng/mL) for 3days. PCR was used to measure mRNA expression. Western Blot and ELISAs were used to quantify protein expression. Cytospins of bronchoalveolar lavage (BAL) cells were used to obtain leukocyte differentials. MUC18 siRNA reduced IL-13-mediated eotaxin-3 (183±44 vs. 380±59pg/mL, p<0.05), while MUC18 overexpression increased IL-13-mediated eotaxin-3 (95±3 vs. 58±3pg/mL, p<0.05) in HTBE cells. IL-13-treated Muc18 KO mice had a lower percentage of neutrophils in BAL than WT mice (25±3 vs. 35±3%, p=0.0565). These results implicate MUC18 as a potential enhancer of airway inflammation in a type 2 cytokine (e.g., IL-13) milieu.

Persistence of respiratory and inflammatory responses after dermal sensitization to persulfate salts in a mouse model of non-atopic asthma.

BackgroundExposure to ammonium persulfate (AP) has been reported to be the main cause of occupational asthma in hairdressers. The aim of this study is to assess how long the asthmatic response to AP can be induced after dermal sensitization in a mouse model.MethodsBALB/c mice received dermal applications of AP or dimethylsulfoxide (DMSO) (control) on days 1 and 8. They then received a single nasal instillation (challenge) of AP or saline on days 15, 22, 29, 36, 45, 60 and 90. Respiratory responsiveness to methacholine was measured 24 h after the challenge using a non-specific methacholine provocation test. Pulmonary inflammation was analysed in bronchoalveolar lavage (BAL), and total serum immunoglobulin (Ig) E, IgG1 and IgG2a were measured in serum samples. Histological analysis of lung slides was performed.ResultsMice dermally sensitized and intranasally challenged with AP showed respiratory responsiveness to methacholine as long as 45 days after initial sensitization, as well as increased percentage of neutrophils in BAL compared with the control group. At day 60, dermally sensitized mice still presented bronchial hyperresponsiveness, while the percentage of neutrophils returned to baseline levels similar to those of controls. Total serum IgE increased significantly on day 22 after dermal sensitization. Total serum IgG1 and IgG2a increased from 45 days after dermal sensitization and remained high at 90 days.ConclusionsBoth respiratory responsiveness to methacholine and airway inflammation responses decrease with increasing time between sensitization and challenge. Respiratory responsiveness to methacholine tends to persist longer than inflammation.

Clinical features of usual interstitial pneumonia with anti-neutrophil cytoplasmic antibody in comparison with idiopathic pulmonary fibrosis.

Myeloperoxidase anti-neutrophil cytoplasmic antibody (MPO-ANCA) is occasionally positive in patients with usual interstitial pneumonia (UIP). However, the differences from idiopathic pulmonary fibrosis (IPF/UIP) have not been well documented. We aimed to clarify the clinical, radiological and pathological features of UIP associated with MPO-ANCA (ANCA/UIP). We retrospectively reviewed the medical records of 12 consecutive ANCA/UIP patients not manifesting microscopic polyangiitis and 108 IPF/UIP patients with no autoantibodies, both diagnosed by surgical lung biopsy. There was no significant difference in clinical background, laboratory results and pulmonary function tests between ANCA/UIP patients and IPF/UIP patients except for the percentage of bronchoalveolar lavage neutrophils. HRCT showed subpleural reticulation in both groups. Increased attenuation around honeycombing and cysts was significantly observed in ANCA/UIP. Pathologically, ANCA/UIP had more prominent inflammatory cell infiltration, lymphoid follicles with germinal centres and cellular bronchiolitis. During the disease course, three of 12 patients (25%) developed microscopic polyangiitis. Immunosuppressive treatment tended to be more effective in ANCA/UIP patients, and the survival time in ANCA/UIP patients tended to be longer than those with IPF/UIP. ANCA/UIP may be distinguishable from IPF/UIP with a combination of HRCT findings of increased attenuation around honeycombing and cysts and some of the characteristic pathological findings. In contrast to IPF/UIP, immunosuppressive treatment could be a therapeutic option for ANCA/UIP.

Expression microarray as a tool to identify differentially expressed genes in horses suffering from inflammatory airway disease.

Inflammatory airway disease (IAD) affects performance and well-being of horses. Diagnosis is primarily reached by bronchoalveolar lavage (BAL) cytology which is invasive and requires sedation. The purpose of this study was to identify differential gene expression in peripheral blood of horses with IAD using species-specific expression microarrays. Equine gene expression microarrays were used to investigate global mRNA expression in circulating leukocytes from healthy, IAD-affected, and low-performing Standardbred and endurance horses. Nine genes in Standardbred and 61 genes in endurance horses were significantly differentially regulated (P<.001). These genes were related to inflammation (eg, ALOX15B, PLA2G12B, and PENK), oxidant/antioxidant balance (eg, DUOXA2 and GSTO1-1), and stress (eg, V1aR, GRLF1, Homer-2, and MAOB). All these genes were up-regulated, except down-regulated Homer-2 and MAOB. DUOXA2, ALOX15B, PLA2G12B, MAOB, and GRLF1 expression was further validated by RT-qPCR. An increase in glutathione peroxidase (GPx) activity in heparinized whole blood of IAD-affected Standardbred (P=.0025) and endurance horses (P=.0028) also suggests a deregulation of the oxidant/antioxidant balance. There was good correlation (r=.7354) between BAL neutrophil percentage and whole blood GPx activity in all horses. This study showed that circulating blood cell gene expression reflects inflammatory responses in tissues. Whether any of the genes have potential for diagnostic applications in the future remains to be investigated. Although not specific for IAD, whole blood GPx activity appears to be correlated with BAL neutrophil percentage. This finding should be further assessed by testing a larger number of horses.

Comparison of Bronchoalveolar Lavage and Tracheal Aspirate Results in 64 Non‐Thoroughbred Horses Presenting with Respiratory Signs or Exercise Intolerance

IntroductionIt has been debated which airway sampling method, bronchoalveolar lavage (BAL) or tracheal aspirate (TA), is superior in evaluating equine respiratory disease. Most studies suggest BAL is the best sampling method due to better correlation with lung histopathology. In this study we compared BAL and TA results of athletic non‐Thoroughbred horses with chronic respiratory signs or exercise intolerance during 2009–2013.Methods64 horses, 2–25 years of age (16 Coldbloods, 30 Warmbloods, 11 Standardbreds, 7 ponies) with recurrent or chronic (duration &gt;1 month) respiratory signs or exercise intolerance that were subject to both BAL and TA sampling at rest were included. Neutrophil counts &gt;5% in BAL and &gt;20% in TA were considered abnormal. Statistical analysis was performed with Spearman's correlation coefficient.ResultsBoth BAL and TA were abnormal in 29 horses and normal in 26 horses. 7 horses had only abnormal TA and 2 horses only abnormal BAL. Correlation was found between the amount of mucus in the trachea and both BAL (P = 0.002) and TA (P&lt;0.001) neutrophil percentage. Neutrophil (P&lt;0.001), eosinophil (P&lt;0.001), lymphocyte (P = 0.008) and macrophage (P = 0.004) percentages correlated between BAL and TA.ConclusionsIn our university hospital population, mucus in the trachea was a good indicator of airway neutrophilia. Unlike previous studies, in only 14% of the horses the interpretation of BAL and TA result was different with TA being slightly more sensitive at detecting neutrophilia. However, a gold standard for detecting inflammation in the equine airways is still lacking and the type and severity of the disease may greatly influence the best choice of sampling method.Ethical Animal ResearchRetrospective analysis of case records. Sources of funding: none. Competing interests: none.

Exogenous Surfactant Attenuates Lung Injury From Gastric-Acid Aspiration During Ex Vivo Reconditioning in Pigs

Lung injury (LI) due to gastric-acid aspiration is associated with poor posttransplantation outcomes. We investigated the effects of ex vivo lung perfusion (EVLP) reconditioning and surfactant administration on LI due to gastric-acid aspiration. Thirty piglets were allocated at random to five groups: the lungs were studied 24 hr after gastric juice-induced LI of the left lower lobe (LLL), LI followed by EVLP (4 hr), or LI followed by LLL surfactant lavage immediately before EVLP; sham animals were studied 24 hr after saline infusion alone or followed by EVLP. Gross anatomy, hemodynamics, and aerodynamics were evaluated; neutrophil and bacterial counts were determined in bronchoalveolar lavage (BAL) fluid and blood. LLLs were evaluated based on a semi-quantitative histologic score, apoptotic cell death (TUNEL), and inflammatory cytokine levels. The sham and sham-EVLP groups were not significantly different. Compared with sham, LI animals had irreversible atelectasis, higher lung infection rates (P<0.0001) and BAL neutrophil percentages (P<0.0001), lower PaO2 (P=0.0006), higher IL-1 (P=0.022) and IL-8 (P=0.006), higher apoptotic cell percentages (P=0.007), and worse histologic severity scores (P<0.0001). EVLP alone did not improve these findings. Adding surfactant before EVLP returned PaO2, pulmonary vascular resistance, and apoptotic-cell percentage to sham-EVLP values but only partially improved the histologic severity score. Local surfactant infusion immediately before EVLP improved the function of donor lungs injured by gastric juice aspiration. This strategy may hold promise for decreasing the shortage of donor lungs.

Bronchial collapse during bronchoalveolar lavage in horses is an indicator of lung inflammation

The bronchoalveolar lavage (BAL) procedure can return variable volumes of fluid, possibly depending on the presence of bronchial collapse during fluid aspiration and on the severity of lung inflammation. We tested the hypothesis that horses with bronchial collapse during BAL are at higher risk of having severe lung inflammation. Prospective field study. Bronchial collapse was graded using a new simple scoring method (0, 1 or 2) during a standardised BAL procedure in the field on 131 horses with normal, mild/moderate or severe lower airway inflammation on cytology of BAL fluid. Of the 131 horses, 37 (28%), 55 (42%) and 39 (30%) horses had bronchial collapse scores of 0, 1 and 2, respectively. There was a difference in collapse scores between all the BAL inflammation categories (P<0.001). Severe collapse had a positive predictive value of 0.95 for both mild/moderate and severe BAL inflammation, with a prevalence of 63% and 20%, respectively. The BAL fluid return volume in the horses with severe collapse scores was lower than volumes in the partial (score 1/2) and no collapse (score 0/2) groups (P<0.001). The BAL fluid volume was negatively correlated with BAL neutrophil percentage (P<0.001). Airway collapse during BAL is associated with airway inflammation and neutrophilia. During a standardised BAL procedure, clinicians can expect lung inflammation in horses that have bronchial collapse and bronchial collapse in horses with lung inflammation. Lung inflammation may be a contributing factor in the mechanism of bronchial collapse during BAL in horses.

418 Reconditioning of Gastric Acid Injured Lung Grafts with Surfactant in Lung Transplantation

Purpose: The number of available donor lungs is still the limiting factor in lung transplantation. We have recently shown during ex-vivo lung evaluation that diluted surfactant lavage improved the graft function after gastric acid aspiration. In the present study we hypothesized that the same treatment in lungs injured by gastric acid aspiration may improve graft function in transplanted porcine lungs. Methods and Materials: Lung injury was induced by intratracheal instillation of 1 ml/kg Oroacid® by a flexible fiberoptic bronchoscope under direct vision into the left main bronchus and the animals were ventilated for 24 hours thereafter. Following harvest the donor lungs were stored at 4°C for 4 hours. In control group (n 4) left lung transplantation was performed without any treatment. In surfactant treated group (n 6) the recipients received intratracheal diluted surfactant (1 ml/kg) lavage just before reperfusion and ventilation. During 7 hours of reperfusion hemodynamic and ventilatory variables were recorded hourly. Bronchoalveolar lavage (BAL) was performed at the end of the experiment for bubble surfactometry and further analysis. Results: Surfactant lavage resulted in lower mean pulmonary artery pressure, lower airway pressure and better oxygenation compared to control group (p 0.001). BAL IL-6 level and neutrophil percentage at the end of the experiment was significantly higher in control compared to surfactant group (p 0.05). Minimal surface tension was significantly lower in surfactant group compared to controls (p 0.05). Conclusions: In this model diluted surfactant administration before reperfusion attenuates the injury and improves the graft function in lung transplantation.

Detection of Mycoplasma pneumoniae in different respiratory specimens

Mycoplasma pneumoniae (M. pneumoniae) is an important community-acquired pneumonia pathogen. Serological test and polymerase chain reaction (PCR) assay are the two main laboratory tests to detect M. pneumoniae now. Little information was compared about the sensitivity and specificity of PCR using different specimens including bronchoalveolar lavage (BAL) and nasopharyngeal aspirate (NPA). The aim of the present study was to evaluate diagnostic values of different specimens by fluorescence quantitative real-time PCR and to find clinical features helpful to diagnose M. pneumoniae pneumonia (MPP). Four hundred and six hospitalized pneumonia children were studied. M. pneumoniae DNA in NPA and BAL samples were detected by fluorescence quantitative real-time PCR. M. pneumoniae-specific IgM was tested by ELISA. MPP were diagnosed based on positive M. pneumoniae-specific IgM in 101 (24.9%) children. The median ages of MPP and non-MPP children were 4.1 and 2.4 years, respectively, with significant difference between them (p < 0.001). Laboratory results including leukocyte count, neutrophil percentage, immunoglobulins, except serum IgM, subgroups of T lymphocyte, and BAL cell count had no significant differences in MPP and non-MPP. BAL macrophage cell percentage was lower in BAL-PCR positive children (p = 0.003), while BAL neutrophil percentage was higher in BAL-PCR positive children (p = 0.007). PCR from NPA and BAL were similar in diagnostic parameters, including sensitivity, specificity, PPV, and NPV (78.6%, 63.4%, 39.8%, and 90.6% for NPA-PCR, respectively; 70.3%, 58.7%, 36.0%, and 85.6% for BAL-PCR, respectively). NPA is better than BAL as PCR sample in MPP diagnosis for similar performance in PCR assay, cheap, and less invasive. BAL is useful in defining local inflammatory condition. Age is the only prefigurative factor in MPP.

Murine mechanical ventilation stimulates alveolar epithelial cell proliferation

ABSTRACTHigh tidal volume mechanical ventilation can cause inflammation and lung damage. Mechanical strain is also necessary for normal lung growth. The current work was performed to determine if mechanical ventilation with clinically utilized tidal volumes stimulates a proliferative response in the lung. Six- to 8-week-old C57/Bl6 mice, anesthetized with ketamine/xylozine, were ventilated for 6 hours with 10 mL/kg tidal volume, positive end-expiratory pressure (PEEP) 3cm H2O. Pulmonary function testing demonstrated decreased compliance within 3 hours of ventilation. Assessment of bronchoalveolar lavage (BAL) demonstrated no significant increase in lactate dehydrogenase, total lavagable cell number, or total protein after ventilation. There was evidence of inflammation in the lungs of ventilated mice, with an increased percentage of lymphocytes and neutrophils in BAL, and an increase in macrophage inflammatory protein (MIP)-2 and interleukin (IL)-1β message in lung tissue. Immunohistochemistry of inflation-fixed lungs demonstrated increased alveolar cell proliferation, as measured by both proliferating cell nuclear antigen and Ki67 staining. Dual staining confirmed that proliferating cells labeled with proSP-B, demonstrating that ventilation induces proliferation of alveolar type II cells. Ventilation did not increase apoptosis in alveolar type II cells, as measured by TUNEL staining. Ventilation at low tidal volumes leads to a mild inflammatory response and alveolar epithelial cell proliferation.

Concentration of surfactant protein D, Clara cell protein CC-16 and IL-10 in bronchoalveolar lavage (BAL) in patients with sarcoidosis, hypersensivity pneumonitis and idiopathic pulmonary fibrosis.

The process of interstitial inflammation, often chronic, goes fluently from alveolitis through granuloma formation to irreversible fibrosis and lung remodeling. Eventually, the loss of functional alveolar units leads to chronic respiratory failure. The pneumoproteins (e.g. SP-D, CC-16) are considered to be markers of interstitial inflammation. We measured BAL concentration of SP-D, CC-16 and IL-10 in patients with sarcoidosis (27), IPF (7) and HP (9). The level of each marker was determined by ELISA specific kit. We found the highest SP-D and CC-16 BAL concentration in patients with the III stage of sarcoidosis (96,67 ng/ml and 31,78 ng/ml, respectively). The lowest SP-D concentration was observed in patients with IPF (76,49 ng/ml), and the lowest CC-16 concentration in patients with HP (21,39 ng/ml). The differences were not statistically significant. In the group of the III stage of sarcoidosis higher SP-D levels were related to higher BAL cytosis and higher percentage of BAL neutrophils, just the opposite as in the IPF and HP group. In the III stage of sarcoidosis and HP, the lower SP-D levels, the lower FEV1 and VC values. The results show, that in acute interstitial inflammation with larger parenchyma engagement (III stage of sarcoidosis) the levels of SP-D were higher then in chronic interstitial inflammation (IPF).

Non-acid gastro-oesophageal reflux in children with suspected pulmonary aspiration

In a group of children with suspected pulmonary aspiration, we aimed to describe the type and physical characteristics of gastro-oesophageal reflux (GOR) episodes, and to determine their correlation with the lipid-laden macrophage (LLM) content in bronchoalveolar lavage (BAL). Twenty-one children with a diagnosis of bronchial asthma, recurrent lung consolidations and recurrent laryngotracheitis underwent 24-h multichannel intraluminal impedance and pH (MII-pH) monitoring, fibreoptic bronchoscopy and BAL. The following parameters were evaluated: total number of reflux episodes, number of acid reflux [AR; pH<4] and non-acid reflux [NAR] episodes [pH>4], height of reflux episodes, LLM content and percentage of neutrophils in the BAL. The number of NAR episodes and the number of those reaching the proximal oesophagus were significantly higher in patients with recurrent lung consolidations than in those with bronchial asthma and laryngotracheitis (p<0.01 and p<0.01). BAL studies showed a significantly higher LLM content in children with recurrent lung consolidations than in those with bronchial asthma and laryngotracheitis (p<0.01). The LLM content correlated significantly with the total number of reflux episodes (r=0.73; p<0.001) and with those reaching the proximal oesophagus (r=0.67; p<0.001). Finally, the LLM content correlated with the number of NAR episodes (r=0.61; p<0.01), with those reaching the proximal oesophagus (r=0.64; p<0.01) and with the percentage of BAL neutrophils (r=0.7; p<0.01). NAR episodes reaching the proximal oesophagus correlate with diagnostic marker for pulmonary micro-aspiration. MII-pH monitoring increases the yield in identifying types and proximal extension of reflux episodes, that discriminate between patients with and without pulmonary aspiration.

Prognostic Value of Bronchoalveolar Lavage Neutrophilia in Stable Lung Transplant Recipients

Bronchoalveolar lavage (BAL) neutrophilia may identify patients prone to develop bronchiolitis obliterans syndrome (BOS) after lung transplantation (LTx). This study assessed the predictive value of BAL neutrophilia in stable recipients. Evaluated were 63 consecutive recipients 3 to 12 months after LTx demonstrating no acute rejection (AR) and lymphocytic bronchitis (LB; B < or = 1 without infection; BOS, 0). Recipients were subdivided into never-BOS (follow-up > or = 12 months) and ever-BOS groups (i.e., BOS development > or = 1 after bronchoscopy). The groups were statistically indistinguishable for demographic data and preceding AR and LB episodes. Onset of BOS was at a median of 232 days (range, 87-962) after bronchoscopy. The ever-BOS group (16 patients) demonstrated a significantly higher percentage of neutrophils compared with the never-BOS group (47 patients) at the time of bronchoscopy (33.6% +/- 2.1% vs 9.9% +/- 1.1%, p < 0.05). By Cox regression analysis, a BAL neutrophil percentage of > or = 20% remained a significant predictor for BOS > or = 1 (hazard ratio, 3.57; 95% confidence interval, 1.71-8.40, p < 0.05) distinct from known potential BOS predictor variables. The positive and negative predictive value of BAL neutrophilia of > or = 20% for future BOS was 0.72 and 0.93, respectively (p < 0.05). BAL neutrophilia in stable recipients is of predictive value to identify recipients at risk for BOS. These data warrant prospective confirmation and further studies to evaluate the benefit of preemptive therapy for potential BOS patients.

Investigation of children with chronic nonspecific cough: Any clinical benefit of bronchoscopy and bronchoalveolar lavage?

Chronic cough can be a complicated and frustrating diagnostic dilemma. The aim of this study was to identify the possible causes of chronic nonspecific cough in seemingly healthy children using fiberoptic bronchoscopy (FOB) and bronchoalveolar lavage (BAL). Eighteen children responded to criteria of selection for chronic cough. The average age was 5.8 years (range, 1.7-10.7 years) and BAL findings were compared with those of 16 nonatopic controls. Children with chronic cough had an increased percentage of BAL neutrophils in comparison with the control group (p = 0.098). Using a BAL neutrophil percent cutoff of 17%, 6 children had high BAL neutrophils (HBNs; median, 77%; range, 27-96%) and 12 children had normal BAL neutrophils (NBNs; median, 3%; range, 0-13%). In the HBN group, FOB showed endoscopic abnormalities in four patients, BAL culture was positive in three patients, and chest x-ray (CXRs) showed minimal densities in four. The IL-8 levels showed a significant increase with respect to the NBN group (p = 0.005). The combination of endoscopic anomalies, BAL culture, BAL IL-8 levels, and minor CXR changes can support the diagnosis of subclincal infection in seemingly healthy children with chronic nonspecific cough and HBN.

Microaspiration in infants with laryngomalacia.

Laryngomalacia is the most common congenital laryngeal anomaly and the most frequent cause of stridor in infants. Infants with laryngomalacia may have associated respiratory symptoms other than stridor. In this study, we evaluated whether infants with laryngomalacia have episodes of microaspiration by calculating the amount of lipids present in the alveolar macrophages. We reviewed bronchoalveolar lavage (BAL) findings obtained in 14 infants with laryngomalacia and in 11 control children. BAL was performed during flexible fiberoptic bronchoscopy (FFB) to diagnose persistent stridor. The presence of microaspiration was determined by evaluating the amount of lipid per single macrophage, assigning each cell a score from 0 to 4 according to the amount of lipid in the cytoplasm. The lipid-laden macrophage index was similar in patients and controls (31.5, range 12-177 vs. 45, range 35-106). In 5 of the 14 (36%) patients the index exceeded 100, and more than 25% of the cells were graded 3 and 4, indicating the presence of microaspiration. The lipid-laden macrophage index correlated significantly with the number and percentage of BAL neutrophils (Pearson correlation test, r = 0.83, P < .001 and r = 0.925, P < .001) as a marker of lung inflammation. Infants with laryngomalacia may have episodes of microaspiration. BAL performed during FFB to evaluate stridor may add helpful therapeutic information without increasing the risks of the procedure.