Bronchial Biopsy Samples Research Articles

Insight into the role of mitochondrion-related gene anchor signature in mitochondrial dysfunction of neutrophilic asthma

Objective At present, targeting molecular-pharmacological therapy is still difficult in neutrophilic asthma. The investigation aims to identify and validate mitochondrion-related gene signatures for diagnosis and specific targeting therapeutics in neutrophilic asthma. Methods Bronchial biopsy samples of neutrophilic asthma and healthy people were identified from the GSE143303 dataset and then matched with human mitochondrial gene data to obtain mitochondria-related differential genes (MitoDEGs). Signature mitochondria-related diagnostic markers were jointly screened by support vector machine (SVM) analysis, least absolute shrinkage, and selection operator (LASSO) regression. The expression of marker MitoDEGs was evaluated by validation datasets GSE147878 and GSE43696. The diagnostic value was evaluated by receiver operating characteristic (ROC) curve analysis. Meanwhile, the infiltrating immune cells were analyzed by the CIBERSORT. Finally, oxidative stress level and mitochondrial functional morphology for asthmatic mice and BEAS-2B cells were evaluated. The expression of signature MitoDEGs was verified by qPCR. Results 67 MitoDEGs were identified. Five signature MitoDEGs (SOD2, MTHFD2, PPTC7, NME6, and SLC25A18) were further screened out. The area under the curve (AUC) of signature MitoDEGs presented a good diagnostic performance (more than 0.9). There were significant differences in the expression of signature MitoDEGs between neutrophilic asthma and non-neutrophilic asthma. In addition, the basic features of mitochondrial dysfunction were demonstrated by in vitro and in vivo experiments. The expression of signature MitoDEGs in the neutrophilic asthma mice presented a significant difference from the control group. Conclusions These MitoDEGs signatures in neutrophilic asthma may hold potential as anchor diagnostic and therapeutic targets in neutrophilic asthma.

Correlation between allergic rhinitis and asthma

The concept of the „unified respiratory disease” emphasizes the close connection between the upper and lower respiratory pathways in allergic diseases. The structure and function of the upper and lower respiratory pathways are closely interconnected, forming a single morphofunctional entity. Studies show that allergic rhinitis and asthma share the same inflammatory cells and Th2-type cytokines in nasal and bronchial biopsy samples. Additionally, bronchial challenges can induce inflammation in the nasal area and vice versa, suggesting a close link between the two conditions. The epithelial barrier, through its junctions and constituent proteins, plays an essential role in maintaining homeostasis and protection against external factors, and the complex interactions between these two segments of the respiratory pathways are an important research topic. Tight junctions, adhesion junctions, gap junctions, as well as desmosomes contribute to maintaining epithelial integrity and regulating inflammation. The constituent proteins of these junctions and their interactions play crucial roles in the proper functioning of the epithelial barrier. Respiratory allergens such as dust mites, pollen, pets, and fungi are risk factors for both conditions. Recent studies have highlighted numerous environmental factors capable of compromising epithelial integrity and disrupting its barrier function, including allergens with protease activity. Allergenic proteases from various sources, such as dust mites, pollen, fungi, and kitchen cockroaches, have been identified and characterized. Studies have shown the significant role these allergens play in compromising the integrity of the respiratory epithelium and triggering allergic responses. They can activate specific receptors, inducing inflammation and disrupting the epithelium’s barrier function. For example, proteases from dust mites cleave junctional proteins, promoting allergen transport and triggering the release of proinflammatory cytokines. The same effect is observed with pollen, fungi, and kitchen cockroaches. The proteolytic activity of allergens contributes to sensitization and the progression of allergic diseases. Understanding these mechanisms can guide the development of effective therapeutic strategies for preventing and treating these conditions.

The Microbiome in Bronchial Biopsies from Smokers and Ex-Smokers with Stable COPD - A Metatranscriptomic Approach

Current knowledge about the respiratory microbiome is mainly based on 16S ribosomal RNA gene sequencing. Newer sequencing approaches, such as metatranscriptomics, offer the technical ability to measure the viable microbiome response to environmental conditions such as smoking as well as to explore its functional role by investigating host-microbiome interactions. However, knowledge about its feasibility in respiratory microbiome research, especially in lung biopsies, is still very limited. RNA sequencing was performed in bronchial biopsies from clinically stable smokers (n = 5) and ex-smokers (n = 6) with COPD not using (inhaled) steroids. The Trinity assembler was used to assemble non-human reads in order to allow unbiased taxonomical and microbial transcriptional analyses. Subsequently, host-microbiome interactions were analyzed based on associations with host transcriptomic data. Ultra-low levels of microbial mass (0.009%) were identified in the RNA-seq data. Overall, no differences were identified in microbiome diversity or transcriptional profiles of microbial communities or individual microbes between COPD smokers and ex-smokers in the initial test dataset as well as a larger replication dataset. We identified an upregulated host gene set, related to the simultaneous presence of Bradyrhizobium, Roseomonas, Brevibacterium.spp., which were related to PERK-mediated unfolded protein response (UPR) and expression of the microRNA-155-5p. Our results show that metatranscriptomic profiling in bronchial biopsy samples from stable COPD patients yields ultra-low levels of microbial mass. Further, this study illustrates the potential of using transcriptional profiling of the host and microbiome to gain more insight into their interaction in the airways.

Time-resolved proteomic profiling of cigarette smoke-induced experimental chronic obstructive pulmonary disease.

Chronic obstructive pulmonary disease (COPD) is the third leading cause of illness and death worldwide. Current treatments aim to control symptoms with none able to reverse disease or stop its progression. We explored the major molecular changes in COPD pathogenesis. We employed quantitative label-based proteomics to map the changes in the lung tissue proteome of cigarette smoke-induced experimental COPD that is induced over 8 weeks and progresses over 12 weeks. Quantification of 7324 proteins enabled the tracking of changes to the proteome. Alterations in protein expression profiles occurred in the induction phase, with 18 and 16 protein changes at 4- and 6-week time points, compared to age-matched controls, respectively. Strikingly, 269 proteins had altered expression after 8 weeks when the hallmark pathological features of human COPD emerge, but this dropped to 27 changes at 12 weeks with disease progression. Differentially expressed proteins were validated using other mouse and human COPD bronchial biopsy samples. Major changes in RNA biosynthesis (heterogeneous nuclear ribonucleoproteins C1/C2 [HNRNPC] and RNA-binding protein Musashi homologue 2 [MSI2]) and modulators of inflammatory responses (S100A1) were notable. Mitochondrial dysfunction and changes in oxidative stress proteins also occurred. We provide a detailed proteomic profile, identifying proteins associated with the pathogenesis and disease progression of COPD establishing a platform to develop effective new treatment strategies.

To evaluate the sensitivity of cytological examination of endobronchial biopsy, BAL, bronchial brushing and sputum in diagnosing lung carcinoma

All the cases of suspected bronchial malignancy were included in present study in which broncho-alveolar lavage (BAL) and bronchial biopsy samples were received in pathology department for evaluation. Respective bronchial brushings & sputum samples were also studied where ever available. Cross sectional study.In our study, correctly diagnosed adenocarcinoma of lung on BAL were 27.7%, sqaumous cell carcinoma were 26.6%, small cell carcinoma were 40% and BAL cytodiagnosis from miscellaneous tumor could only be made in 14.2% cases. Endobronchial biopsy and BAL, both were studied, sensitivity of cases increases and number of false negative cases decreases for diagnosis. Bronchial brushing has better sensitivity than BAL for diagnosis of lung carcinoma. Sputum is less sensitive than bronchial brushing for diagnosis of lung carcinoma.

Interplay among the Variants of One Carbon Metabolism, Methylenetetrahydrofolate Reductase Polymorphisms and Lung Cancer

Introduction: Folates perform an integral task in DNA synthesis, methylationand repair. Methylenetetrahydrofolate Reductase (MTHFR) potrays a key part in the metabolism of folate and regulates the equilibrium between the various forms of folate for DNA synthesis and DNA methylation. MTHFR irrevocably transforms 5,10 methylenetetrahydrofolate to 5-methyltetrahydrofolate, the principal circulating folate and the carbon donor for remethylation processes. MTHFR is vastly polymorphic in the general population. Materials and Methods: It was a case-control study conducted during March 2010 to September 2011 in the Department of Pulmonary Medicine in collaboration with the Department of Biochemistry at Government Medical College and Hospital, Chandigarh, to see whether any association exists between the variants of one carbon metabolism, MTHFR polymorphisms (C677T and A1298C), and lung cancer. Twenty biopsy proven lung cancer patients and 20 age and sex matched cancer-free controls were selected. Results: The mean serum folate in cases was higher (12.84 ng/mL±7.527 ng/mL) as compared to controls (4.46 ng/mL±1.346 ng/mL), suggesting that high levels of serum folate are associated with lung cancer. There was no significant variance in the levels of vitamin B12 and plasma homocysteine between cases and controls. No MTHFR polymorphism C677T was seen in the blood and the bronchial biopsy samples of all cases as well as blood samples of all the controls. The MTHFR polymorphism A1298C was present in the blood as well as bronchial biopsy samples of cases as well as blood of controls. Thus, in the present study, there was no relation of this polymorphism with lung cancer. Conclusion: Polymorphisms in MTHFR may contribute to lung cancer. More research on the basis of cellular and molecular mechanisms of lung cancer is urgently needed to aid in understanding of pathogenesis of the disease.

Bronchial brushing cytology is comparable to bronchial biopsy for epidermal growth factor receptor mutation test in non-small cell lung cancer.

Objectives:Bronchial brushing (BB) is often used to obtain supplementary samples for diagnosing lung cancer. We examined the possibility of epidermal growth factor receptor (EGFR) testing on BB samples and compared them with bronchial biopsy samples.Material and Methods:We used 150 BB samples with non-small cell carcinoma submitted to our department within 2 years. Biopsy samples were concurrently submitted for histologic diagnosis. We used the peptide nucleic acid clamping method for EFGR mutation test. Histologic diagnosis identified 137 cases of adenocarcinomas and 13 cases of non-small cell lung carcinoma, not otherwise specified. Each sample was assessed for adequacy and DNA content for EGFR mutation test.Results:Among BB samples, 28 had exon 19 deletion, 21 had mutations in exon 21, 99 were wild type, and analysis of two failed. The EGFR mutation rate in BB samples was 33.1% (49/148). Among bronchial biopsy samples, 26 had exon 19 deletion, 20 had mutations in exon 21, 92 were wild type, and analysis of 12 failed. The EGFR mutation rate using biopsy sample was 33.8% (46/136). The mutation detection results were nearly identical in both groups of samples (131/138, 94.9%). However, in two cases, an exon 21 mZutation was detected in biopsy samples but not in BB samples. In five cases, exon 19 deletion (two cases) and exon 21 mutation (three cases) were detected in BB but not in biopsy samples. The median DNA content was 58.83 ng for BB samples and 48.47 ng for biopsy samples. The failure rate for BB samples was lower than for biopsy samples. Overall, the BB samples were comparable to bronchial biopsy samples in terms of DNA quantity and mutation detection results.Conclusion:We conclude that in case of inadequate biopsy samples, BB samples can be used as a substitute material for EGFR mutation test.

Clinical Significance of Psoriasiform Sarcoidosis

Sarcoidosis is a chronic granulomatous inflammatory disease characterized by the presence of noncaseating granulomas in various organs. Psoriasis is a persistent and recurrent autoimmune disorder mainly affecting the skin. Cutaneous involvement is observed in one fourth of the sarcoidosis patients and psoriatic plaques may be a manifestation of cutaneous sarcoidosis. The pathogenic mechanism of psoriasis is relevant to the overstimulation of CD4 TH1 and TH17 lymphocytes that are also involved in the granuloma formation of sarcoidosis. We present seven cases of sarcoidosis patients presenting with psoriasiform sarcoidosis. All the patients had cutaneous plaques indistinguishable from psoriasis. Diagnosis of sarcoidosis was confirmed by biopsy in at least two organs. Psoriasiform sarcoidosis was identified by skin biopsy of the involved cutaneous lesions. All patients had stage III sarcoidosis and presented with one or more clinical or laboratory manifestations of sarcoidosis. BAL was negative for infectious agents. Histopathologic examination of the bronchial and transbronchial biopsy samples revealed noncaseiting granulomatous inflammation. Differentiation of sarcoidosis and psoriasis may constitute a diagnostic dilemma for the clinician in terms of the identical skin lesions. The presence of psoriatic plaques may suggest an overlap between the underlying pathogenetic mechanisms of both disorders. Despite the great clinical similarity, the only way to differentiate the two syndromes is pathological examination. The common pathogenetic mechanism involving the TH1 and TH17 pathways for both sarcoidosis and psoriasis may be the fundamental linkage for this occurrence. We present seven cases of sarcoidosis patients presenting with psoriasiform lesions to define the clinical profile of sarcoidosis in these patients including presentation, laboratory findings and prognosis. The coexistence of sarcoidosis and psoriasis, on the other hand may shed light on the common mechanisms that may play a role in the development of both disorders.

Levels of CD4+ CD25+ T regulatory cells in bronchial mucosa and peripheral blood of chronic obstructive pulmonary disease indicate involvement of autoimmunity mechanisms.

Many theories have been proposed to explain pathogenesis of COPD; however, remains unclear why the majority of smokers (~80%) do not develop COPD, or only develop a mild disease. To explore if COPD has an autoimmune component, the role of T regulatory lymphocytes (Tregs) in the lung tissue of COPD patients is of crucial importance. Bronchial tissue biopsy samples were prospectively collected from 64 patients (39 COPD and 25 controls - 15 smokers and 10 non-smokers). The patients with COPD were subdivided into mild/moderate (GOLD stage I-II) and severe/very severe (GOLD stage III-IV) groups. Digital image analysis was performed to estimate densities of CD4+ CD25+ cell infiltrates in double immunohistochemistry slides of the biopsy samples. Blood samples were collected from 42 patients (23 COPD and 19 controls) and tested for CD3+ CD4+ CD25+ bright lymphocytes by flow cytometry. The number of intraepithelial CD4+ CD25+ lymphocytes mm-2 epithelium was significantly lower in the severe/very severe COPD (GOLD III-IV) group as well as in the control non-smokers (NS) group (p < 0,0001). Likewise, the absolute number of Treg (CD3+ CD4+ CD25+ bright) cells in the peripheral blood samples was significantly different between the four groups (p = 0.032). The lowest quantity of Treg cells was detected in the severe/very severe COPD and healthy non-smokers groups. Our findings suggest that severe COPD is associated with lower levels of Tregs in the blood and bronchial mucosa, while higher Tregs levels in the smokers without COPD indicate potential protective effect of Tregs against developing COPD.

High frequency of infection of lung cancerpatients with the parasite Toxoplasma gondii.

BackgroundToxoplasma gondii is an intracellular protozoan parasite that can cause a wide range of clinical conditions, including miscarriage and pneumonia. The global prevalence is 30% in humans, but varies by locality (e.g. in the UK it is typically 10%). The association between lung cancer and T. gondii infection was investigated by direct detection in lung tissue samples.MethodsLung tissue samples were taken from patients undergoing lung resection surgery (n=72) for suspected lung cancer (infection prevalence 100% (95% CI: 93.9–100%)). All 72 participants were confirmed as having lung cancer following subsequent diagnostic tests. In addition, bronchial biopsy samples were collected from non-lung cancer healthy control subjects (n=10). Samples were tested for T. gondii using PCR amplification of T. gondii specific gene markers and T. gondii specific immunohistochemistry.ResultsAll 72 lung cancer patients were infected with T. gondii (prevalence 100% (95% CI: 93.9–100%)). Of which, 95.8% (n=69) of patients showed evidence of active parasite stages. Infection prevalence in the controls (10%) was significantly lower (p<0.0001).ConclusionsClinicians treating lung cancer patients should be aware of the potential presence of the parasite, the potential for induction of symptomatic complications and interference with treatment success.

Anti-inflammatory effects of roflumilast in chronic obstructive pulmonary disease (ROBERT): a 16-week, randomised, placebo-controlled trial

The clinical effects of roflumilast, a selective phosphodiesterase-4 inhibitor, are well established, but little is known about the anti-inflammatory mechanisms underlying the drug's efficacy. The aim of the ROflumilast Biopsy European Research Trial (ROBERT) was to assess the anti-inflammatory effects of roflumilast on bronchial mucosal inflammation in patients with moderate-to-severe chronic obstructive pulmonary disease (COPD) and chronic bronchitis. ROBERT was a randomised, double-blind, placebo-controlled trial done at 18 sites in five countries. Eligible patients were aged 40-80 years, had COPD, and had had a chronic productive cough for 3 months in each of the two previous years. Patients also had to have a post-bronchodilator predicted FEV1 30-80% and a post-bronchodilator FEV1/forced vital capacity ratio of 70% or less. Patients entered a 6-week run-in period before being randomly assigned (1:1) via a computerised central randomisation system to roflumilast 500 μg once daily or placebo for 16 weeks, in addition to bronchodilator therapy (inhaled corticosteroids were not permitted). Randomisation was stratified by concomitant use of long-acting β agonist. Both participants and investigators were masked to group assignment. Roflumilast and placebo were supplied as identical yellow, triangular tablets. Airway inflammation was assessed by quantification of inflammatory cells in bronchial biopsy samples and induced sputum samples. The primary endpoint was the change in the number of CD8 inflammatory cells in bronchial biopsy submucosa from randomisation to week 16 in the intention-to-treat population. Changes in cell counts of additional inflammatory markers, including eosinophils, were assessed as secondary endpoints. This trial is registered with ClinicalTrials.gov, number NCT01509677, and is closed to new participants, with follow-up completed. Between Jan 4, 2012, and Feb 11, 2016, 158 patients were randomly assigned: 79 to the roflumilast group, and 79 to the placebo group. At week 16, the change in the number of CD8 cells in the bronchial submucosa did not differ significantly between the roflumilast and placebo groups (treatment ratio 1·03 [95% CI 0·82-1·30]; p=0·79). However, compared with placebo, roflumilast was associated with a significant reduction in eosinophils in bronchial biopsy samples at week 16 (treatment ratio 0·53 [95% CI 0·34-0·82]; p=0·0046). Significant reductions in both absolute (p=0·0042) and differential (p=0·0086) eosinophil cell counts in induced sputum were also noted with roflumilast compared with placebo, but peripheral blood eosinophil counts were not significantly affected. We noted no other significant effects of roflumilast on bronchial mucosal inflammatory cells. The most common (ie, occurring in >5% patients) moderate adverse events were worsening of COPD (three [4%] patients in the roflumilast group vs seven [9%] in the placebo group), cough (six [8%] vs four [5%]), diarrhoea (four [5%] vs three [4%]), and nasopharyngitis (three [4%] vs five [6%]). Severe adverse events included worsening of COPD, which occurred in four (5%) patients in the roflumilast group and two (3%) in the placebo group. No deaths occurred during the study. Serious adverse events occurred in eight (10%) patients in the roflumilast group and five (6%) in the placebo group. 16 weeks of treatment with roflumilast did not affect the number of CD8 cells in bronchial submucosa compared with placebo. However, we noted significant reductions in eosinophil cell counts in bronchial biopsy samples and induced sputum, generating the hypothesis that the effect of roflumilast in COPD could be mediated by an effect on lung eosinophils. Takeda and AstraZeneca.

Airway pathological heterogeneity in asthma: Visualization of disease microclusters using topological data analysis

Asthma is a complex chronic disease underpinned by pathological changes within the airway wall. How variations in structural airway pathology and cellular inflammation contribute to the expression and severity of asthma are poorly understood. Therefore we evaluated pathological heterogeneity using topological data analysis (TDA) with the aim of visualizing disease clusters and microclusters. A discovery population of 202 adult patients (142 asthmatic patients and 60 healthy subjects) and an external replication population (59 patients with severe asthma) were evaluated. Pathology and gene expression were examined in bronchial biopsy samples. TDA was applied by using pathological variables alone to create pathology-driven visual networks. In the discovery cohort TDA identified 4 groups/networks with multiple microclusters/regions of interest that were masked by group-level statistics. Specifically, TDA group 1 consisted of a high proportion of healthy subjects, with a microcluster representing a topological continuum connecting healthy subjects to patients with mild-to-moderate asthma. Three additional TDA groups with moderate-to-severe asthma (Airway Smooth MuscleHigh, Reticular Basement MembraneHigh, and RemodelingLow groups) were identified and contained numerous microclusters with varying pathological and clinical features. Mutually exclusive TH2 and TH17 tissue gene expression signatures were identified in all pathological groups. Discovery and external replication applied to the severe asthma subgroup identified only highly similar "pathological data shapes" through analyses of persistent homology. We have identified and replicated novel pathological phenotypes of asthma using TDA. Our methodology is applicable to other complex chronic diseases.

Shared immunological targets in the lungs and joints of patients with rheumatoid arthritis: identification and validation

ObjectivesImmunological events in the lungs might trigger production of anti-citrullinated protein antibodies during early rheumatoid arthritis (RA). We investigated the presence of shared immunological citrullinated targets in joints and lungs...

Asthma heterogeneity and severity—why is comprehensive phenotyping important?

Asthma heterogeneity and severity—why is comprehensive phenotyping important?

A microdose study of 14C-AR-709 in healthy men: pharmacokinetics, absolute bioavailability and concentrations in key compartments of the lung

To explore, in a microdose (phase-0) study, the pharmacokinetics, bioavailability and concentrations in key compartments of the lung, of AR-709, a novel diaminopyrimidine antibiotic for the treatment of respiratory infection. Four healthy men each received two single, 100 μg microdoses of ¹⁴C-AR-709, 7 days apart: the first was administered intravenously (IV), the second orally. Plasma pharmacokinetics of ¹⁴C and unchanged AR-709 were obtained by high-performance liquid chromatography and accelerator mass spectrometry (AMS). Next, 15 healthy men received a single, 100 μg microdose of ¹⁴C-AR-709 IV. Plasma, bronchoalveolar lavage fluid, alveolar macrophages and bronchial mucosal biopsy samples were analysed by AMS. After IV administration, clearance of AR-709 was 496 mL/min, volume of distribution was 1,700 L and the absolute oral bioavailability was 2.5 %. Excretion in urine was negligible. At 8-12 h after IV dosing, ¹⁴C concentrations in lung samples were 15- (bronchial mucosa) to 200- (alveolar macrophages) fold higher than in plasma. In alveolar macrophages, ¹⁴C was still mostly associated with AR-709 at 12 h after dosing. The results of this microdose study indicate that AR-709 attains concentrations appreciably higher within the lung than in plasma. Its low oral bioavailability however, precludes oral administration. Although IV administration would appear to be an effective route of administration, this would limit the use of AR-709 to a clinical setting and would therefore be economically unsustainable. If further clinical development were to be undertaken, therefore, an alternative route of administration would be necessary.

Regulation of Wnt4 in chronic obstructive pulmonary disease

Chronic obstructive pulmonary disease (COPD) is associated with persistent inflammation and oxidative stress in susceptible individuals. Using microarray analysis of bronchial biopsy samples from patients with COPD and controls, we identified Wnt4 as being up-regulated in COPD. Analysis of bronchial biopsy samples showed a very strong correlation between Wnt4 and IL8 gene expression, suggesting that Wnt4 plays a role in chronic lung inflammation. In vitro, Wnt4 induced proliferation and inflammation in human epithelial cells (BEAS-2B) and normal primary human bronchial epithelial cells in a concentration-dependent manner. This effect was enhanced in the presence of interleukin-1β (IL-1β) as a result of activation of the p38 and c-Jun NH2-terminal kinase mitogen-activated protein kinase pathways. Hydrogen peroxide, but not proinflammatory stimuli, up-regulated Wnt4 expression in epithelial cells. In monocytic THP-1 and primary airway smooth muscle cells, Wnt4 induced inflammation and enhanced the inflammatory response to lipopolysaccharide and IL-1β but did not induce proliferation. In addition, these other cell types did not have enhanced Wnt4 expression in response to hydrogen peroxide. Our results indicate that airway epithelial activation, due to oxidative stress, may lead to Wnt4 induction. Wnt4, in turn, acts through the noncanonical pathway to activate epithelial cell remodeling and IL8 gene expression, leading to neutrophil infiltration and inflammation.

Comparison of the expression levels of napsin A, thyroid transcription factor-1, and p63 in nonsmall cell lung cancer using cytocentrifuged bronchial brushings

Appropriate treatment of nonsmall cell lung cancer (NSCLC) depended on histologic type. The aim of this study was to evaluate the expression levels of Napsin A, thyroid transcription factor-1 (TTF-1), and p63 by immunostaining using CytoRich Red preserved cytocentrifuged bronchial brushings. The expression levels of Napsin A, TTF-1, and p63 were semiquantitatively evaluated using proportion and intensity scores in 12 patients from whom both bronchial brushing cytology and bronchial biopsy samples had been obtained, as well as in a selection of 64 resected NSCLC tissue samples, and 8 normal or benign bronchial brushing cytology samples. The nuclear expressions of TTF-1 and p63 allowed for good visualization of cancer cells, whereas the cytoplasmic expression of Napsin A in cancer cells was often more difficult to evaluate because medium-sized and large macrophages also expressed Napsin A. Napsin A and TTF-1 were reliable adenocarcinoma markers, and p63 was a reliable squamous cell carcinoma marker in cytology, and histology samples. The average scores of Napsin A, TTF-1, and p63 were highly correlated in bronchial brushing cytology and bronchial biopsy samples, whereas TTF-1 expression was significantly lower than Napsin A expression in resected NSCLC tissue samples (P < .001). Immunostaining of bronchial brushings fixed with CytoRich Red was useful in determining histologic types in NSCLC. It was suggested that the panels of Napsin A, TTF-1, and p63 were effective in identifying histologic type in NSCLC, and that a combination of either Napsin A and p63 or TTF-1 and p63 should be chosen, depending on morphology.

Effect of Inhaled Corticosteroid Therapy on CT Scan-Estimated Airway Dimensions in a Patient With Chronic Bronchitis Related to Ulcerative Colitis

CT scanning allows assessment of airway-wall thickness and is useful for diagnosing and assessing various lung diseases, including asthma and COPD. Several studies have reported that CT scan-estimated dimensions are positively correlated with pulmonary function test results and a positive response to treatment. However, to our knowledge, airway disease complicated by inflammatory bowel disease has not yet been assessed using CT scan-estimated dimensions. A 61-year-old woman with ulcerative colitis (UC) complained of cough, sputum, and fever 4 months after undergoing a total colectomy and rectal mucosectomy. Examination of bronchial biopsy samples demonstrated lymphocytic and plasma cell infiltration of the epithelium and submucosa. CT scan-estimated dimensions indicated thickening of the airways. Chronic bronchitis complicated with UC was diagnosed, and high-dose inhaled corticosteroid therapy was prescribed. Three months later, the CT scan-estimated dimensions of the patient's airways, her symptoms, and pulmonary function test results were dramatically improved.

Exhaled Nitric Oxide is Associated with Allergic Inflammation in Children

Exhaled nitric oxide (eNO) has been proposed as a noninvasive marker of airway inflammation in asthma. In asthmatic patients, exhaled NO levels have been shown to relate with other markers of eosinophilic recruitment, which are detected in blood, sputum, bronchoalveolar lavage fluid and bronchial biopsy samples. The purpose of this study was to assess the possible relationship between eNO and allergic inflammation or sensitization in childhood asthma and allergic rhinitis. Subjects consisted of 118 asthmatic children, 79 patients with allergic rhinitis, and 74 controls. Their age ranged from 6 to 15 yr old. eNO level, peripheral blood eosinophil count, eosinophil cationic protein (ECP), serum total IgE level and specific IgE levels were measured. Methacholine challenge test and allergic skin prick test for common allergens were performed in all subjects. Atopic group (n = 206, 44.48 ± 30.45 ppb) had higher eNO values than non-atopic group (n = 65, 20.54 ± 16.57 ppb, P < 0.001). eNO level was significantly higher in patients with asthma (42.84 ± 31.92 ppb) and in those with allergic rhinitis (43.59 ± 29.84 ppb) than in healthy controls (27.01 ± 21.34 ppb, P < 0.001) but there was no difference between asthma and allergic rhinitis group. eNO also had significant positive correlations with Dermatophagoides pteronyssinus IgE level (r = 0.348, P < 0.001), Dermatophagoides farinae IgE level (r = 0.376, P < 0.001), and the number of positive allergens in skin prick test (r = 0.329, P = 0.001). eNO had significant positive correlations with peripheral blood eosinophil count (r = 0.356, P < 0.001), serum total IgE level (r = 0.221, P < 0.001), and ECP (r = 0.436, P < 0.001). This study reveals that eNO level is associated with allergic inflammation and the degree of allergic sensitization.

S39 Sub typing of non small cell carcinoma in EBUS-TBNA samples

IntroductionDifferentiation and accurate classification of NSCLC (Squamous, Adenocarcinoma, Large cell carcinoma) is crucial in determining the prognosis and selecting targeted chemotherapy regimens. However, it is not always possible to subtype...