Bronchial Aspirate Research Articles

Cytological Smears and Cell Blocks on Endobronchial Aspirates Complement each other and Allow a Higher Diagnostic Yield

The accurate diagnosis of lung carcinoma has become compulsory especially after the introduction of new target therapies. Most of the patients are non operable. This fact points out the diagnostic importance of the cytology specimen. Our target was to assess the diagnostic efficacy of bronchial smears and cell blocks by comparing their results with those of the final diagnoses made on biopsies. We conducted a prospective study of 55 bronchial aspirates. It showed a sensitivity of 100% and a specificity of 100% when coupling both techniques. These results put emphasis on the diagnostic efficacy of both bronchial smears and cell blocks in diagnosing bronchial carcinoma.

Surgical treatment of late‐diagnosed bronchial foreign body aspiration: a report of 23 cases

Late-diagnosed bronchial foreign bodies can lead to irreversible changes in the bronchi and the lungs. To date, few reports are available concerning surgical treatment for this condition. The present report summarizes clinicopathological features and surgical treatment options for late-diagnosed bronchial foreign body aspiration. Clinical data of 23 patients who underwent surgical treatment for late-diagnosed bronchial foreign body aspiration in our hospital from 1980 to 2010 were reviewed. The patients included 17 male and six female patients, aged 6 to 66 years. To remove the foreign body, bronchotomy was performed in six patients and pulmonary resection in 17 patients. Foreign bodies were located on the right side in 17 cases and on the left side in six cases. The majority of the foreign bodies were of food origin. Postoperative bronchopleural fistula occurred in two patients. One patient died of renal failure subsequent to lung infections. Detailed history, awareness of potential foreign body aspiration and thorough bronchoscopic examination are helpful to avoid pulmonary resection. However, the latter is necessary in patients with the associated substantial lung lesions to prevent these from progressing into serious lung infections such as pulmonary suppuration or empyema.

Performance Evaluation of Kits for Bisulfite-Conversion of DNA from Tissues, Cell Lines, FFPE Tissues, Aspirates, Lavages, Effusions, Plasma, Serum, and Urine

DNA methylation analyses usually require a preceding bisulfite conversion of the DNA. The choice of an appropriate kit for a specific application should be based on the specific performance requirements with regard to the respective sample material. In this study, the performance of nine kits was evaluated: EpiTect Fast FFPE Bisulfite Kit, EpiTect Bisulfite Kit, EpiTect Fast DNA Bisulfite Kit (Qiagen), EZ DNA Methylation-Gold Kit, EZ DNA Methylation-Direct Kit, EZ DNA Methylation-Lightning Kit (Zymo Research), innuCONVERT Bisulfite All-In-One Kit, innuCONVERT Bisulfite Basic Kit, innuCONVERT Bisulfite Body Fluids Kit (Analytik Jena). The kit performance was compared with regard to DNA yield, DNA degradation, DNA purity, conversion efficiency, stability and handling using qPCR, UV, clone sequencing, HPLC, and agarose gel electrophoresis. All kits yielded highly pure DNA suitable for PCR analyses without PCR inhibition. Significantly higher yields were obtained when using the EZ DNA Methylation-Gold Kit and the innuCONVERT Bisulfite kits. Conversion efficiency ranged from 98.7% (EpiTect Bisulfite Kit) to 99.9% (EZ DNA Methylation-Direct Kit). The inappropriate conversion of methylated cytosines to thymines varied between 0.9% (innuCONVERT Bisulfite kits) and 2.7% (EZ DNA Methylation-Direct Kit). Time-to-result ranged from 131 min (innuCONVERT kits) to 402 min (EpiTect Bisulfite Kit). Hands-on-time was between 66 min (EZ DNA Methylation-Lightning Kit) and 104 min (EpiTect Fast FFPE and Fast DNA Bisulfite kits). Highest yields from formalin-fixed and paraffin-embedded (FFPE) tissue sections without prior extraction were obtained using the innuCONVERT Bisulfite All-In-One Kit while the EZ DNA Methylation-Direct Kit yielded DNA with only low PCR-amplifiability. The innuCONVERT Bisulfite All-In-One Kit exhibited the highest versatility regarding different input sample materials (extracted DNA, tissue, FFPE tissue, cell lines, urine sediment, and cellular fractions of bronchial aspirates, pleural effusions, ascites). The innuCONVERT Bisulfite Body Fluids Kit allowed for the analysis of 3 ml plasma, serum, ascites, pleural effusions and urine.

Neutrophil extracellular traps in bronchial aspirates: a quantitative analysis

Neutrophil extracellular traps (NETs) are structures composed of DNA and granular proteins, which rapidly trap and kill pathogens. The formation of NETs has been detected during infection in animal experiments, but their role in humans is unclear. The purposes of this study were to quantitatively evaluate the production of NETs during acute respiratory infection and to study the relationship between the NET length and various inflammatory mediators. We examined bronchial aspirates collected from nine intubated patients in an intensive care unit. Samples were collected at the onset of acute respiratory infection (day 0) and on days 1, 3-5, and 6-8. The NET length was visualised by immunohistochemistry and quantified using computer tracing software. The NET length was measured and compared at each time point. The length differed significantly between time points (p<0.001). NETs were significantly longer on day 1 than on day 0 (p<0.001). Neutrophils released NETs abundantly in response to respiratory infection and regression analysis showed that NET length correlated with six clinical parameters (white blood cells, platelets, lactate, CXC ligand-2, interleukin-8, and procalcitonin) as the explanatory variables. NETs in bronchial aspirates may reflect disease progression of respiratory infections. Quantification of NETs in bronchial aspirates may provide a new indicator of inflammation.

Fulminant atypical Cryptococcus neoformans pneumonia confirmed by PLEX-ID

Use of the PLEX-ID system can lead to a rapid molecular diagnosis in microbiology. To illustrate the clinical implications of this new diagnostic tool, we present the case of a 46-year-old patient admitted with severe respiratory failure and septic shock. Cryptococcal pneumonia was diagnosed by Fungi-Fluor™ staining of the bronchoalveolar lavage (BAL) and the patient tested positive for HIV. Unfortunately, he died 12h after admission despite intensive care support and treatment with broad-spectrum antibiotics, amphotericin B, and flucytosine. Retrospective use of the PLEX-ID on the BAL, bronchial aspirate, and blood yielded Cryptococcus neoformans in all fluids tested. Rapid molecular diagnosis with PLEX-ID, especially when performed on the blood of septic patients, may reduce the time to adequate treatment and limit the number of diagnostic procedures needed.

Role of flexible fiberoptic bronchoscopy in suspected sputum smear negative pulmonary tuberculosis cases at microscopy centre under RNTCP

Background: Microbiological diagnosis is the main stay for the effective treatment of pulmonary tuberculosis. About 31%of the new cases may be smear-negative for AFB. Difficulties arise when a patient who is suspected of active tuberculosis, both clinically and radiologically, does not produce sputum or when it is available AFB may be negative. Fiberoptic bronchoscopy offers a mean of investigation whereby bronchial secretion and washing can be collected from the most likely abnormal site under direct vision. Aims & Objective: To study the role of flexible fiberoptic bronchoscopy in suspected sputum smear negative pulmonary tuberculosis cases at microscopy centre under RNTCP. Material and Methods: Thirty three patients aged Above 18 years old who were suspected of having pulmonary tuberculosis based on clinical and radiological appearances were prospectively studied. All subjects had at least 2 sputum smear examination which were negative for acid fast bacilli according to the revised national tuberculosis control program (RNTCP). The bronchoscopy was performed transnasally using fujinon bronchoscope by 2 bronchoscopists under local anaesthesia. A thorough examination of bronchial tree was carried out and bronchial aspirate (BA) bronchoalveolar lavage (BAL) bronchial brushing, transbronchial lung biopsy (TBLB) and post bronchoscopy sputum (PBS) were collected. The specimen obtained was placed on slides for Ziehl-Nielsen stain. Bronchial biopsy was performed on abnormal looking mucosa and stained with Eiosin-hematoxylin and Ziehl- Nielsen stains. Results: In total 33 sputum smear negative suspected pulmonary tuberculosis cases, at microscopy centre under RNTCP, early diagnosis of pulmonary tuberculosis was established in 10 (30.30%) cases and in 3 (9.09%) cases diagnosis of malignancy was established. Conclusion: Fiberoptic Bronchoscopy is a useful procedure to establish the diagnosis of pulmonary tuberculosis when sputum smear examination does not show acid fast bacilli. This allows appropriate treatment to be started with confidence.

A 15.13 kDaEnterobacter gergoviae Biofilm Predictive Protein is Recognized bysIgA in Bronchial Aspirate from Patients with VAP

Ventilator-associated pneumonia (VAP) is difficult to diagnose, expensive to treat, and is associated with significant morbidity and mortality. The major part of VAP pathogenesis is the formation of a biofilm structure which also plays role in its resistance to treatment. A strategy to preventVAP isby removing the biofilm structure. The aim of this study is looking up the biofilm predictive protein of Enterobacter gergoviae (E. gergoviae) which is recognized by Secretory Immunoglobulin A (sIgA) as a mucosal immune response in VAP patient. This study used a tube adherence method for the detection of biofilms, protein profiling techniques with SDS PAGE and blotting for the evaluation of the antigen antibody response. The sIgAwere isolated from bronchial aspirate of VAP patient at ICU RSSA Malang. From two E. gergoviae samples, one sample biofilm positive (sample 1), and theother was biofilm negative (sample 2). The biofilm predictive protein was detected using SDS PAGE, and is discovered to be 15.13 kDa. The sIgA from sample 1 (p 1) reacted with protein band 15.13 kDa, 38.02 kDa, 45.71 kDa and 47.21 kDa whilesIgA from sample 2 (p 2) reacted with 38.02 kDa, 45.71 kDa, 47.21 kDa, and 50 kDa. This finding was predicted that protein MW 15.13 kDa played role in biofilm formation.

Hemoptysis secondary to actinomycosis: A rare presentation.

We present a 70-year-old female patient who had the history of hypertension and presented with massive haemoptysis. She had been complaining of cough with expectoration and mild streaking of blood in sputum for about 3 days with only crepts in right infrascapular and infra-axillary regions as positive clinical findings. Bronchoscopy revealed a cauliflower-like lesion in the upper- right lobe bronchus; bronchial aspirate showed occasional colonies of gram positive filamentous bacteria surrounded by neutrophils. The Trucut biopsy showed sheets of neutrophils with colonies of filamentous bacteria consistent with actinomycotic infection. She was started on intravenous benzyl penicillin 20 million units 6 hourly. She recovered with no further bouts of hemoptysis and was discharged on amoxicillin + clavulanic acid in a stable condition and she remained under similar condition for more than a year on follow up. Actinomycosis is a rare disease caused by a harmless commensal species, Actinomyces. Diagnosis of actinomycosis is a challenging situation, and more so, very few cases causing hemoptysis have come to light so far.

Natural surfactant combined with superoxide dismutase and catalase decreases oxidative lung injury in the preterm lamb

We performed a randomized study in preterm lambs to assess the hypothesis that treatment with a natural surfactant combined with superoxide dismutase (SOD) and catalase (CAT) might decrease pulmonary oxidative stress in an animal model of respiratory distress syndrome (RDS). Animals received 200 mg/kg of porcine natural surfactant or 200 mg/kg of natural surfactant combined with 2 mg/ml of SOD and 3,000 U/ml of CAT. Lung tissue oxidation was studied by measuring total hydroperoxide (TH), advanced oxidation protein products (AOPP), and non-protein bound iron (NPBI) in bronchial aspirate samples. In addition, the animal's lung mechanics were evaluated. TH, AOPP, and NPBI were lower in the groups treated with surfactant plus SOD and CAT than in the surfactant group, while lung mechanics did not vary. We concluded that natural surfactant combined with SOD and CAT is effective in reducing the oxidative lung stress in an animal model of RDS.

Simultaneous analysis of antibiotics in biological samples by ultra high performance liquid chromatography–tandem mass spectrometry

A rapid and reliable multiclass method was developed for the simultaneous analysis of 21 antibiotics (beta-lactams, aminoglycosides, penicillins, cephalosporins, carbapenems or quinolones) in urine, serum, cerebrospinal fluid (CSF) and bronchial aspirations by ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC–MS/MS). Prior to chromatographic determination, the analytes were extracted from human biological fluids by simple sample treatments, which imply dilution, liquefaction, or protein precipitation. Several chromatographic conditions were optimized in order to obtain a fast separation (<6min for each chromatographic run). MS/MS conditions were evaluated in order to increase selectivity and sensitivity and all compounds were detected in electrospray (ESI) positive ion mode, except clavulanic acid and sulbactam, which were monitored in negative ion mode. The developed method was validated in terms of linearity, selectivity, limits of detection (LODs) and quantification (LOQs), trueness, repeatability and interday precision. The LOQs ranged from 0.01 to 1.00mg/L for urine, serum and CSF. In case of bronchial aspirations, the LOQs were between 0.02 and 0.67mg/kg. In all matrices the recovery results were in the range 70–120% and interday precision was lower than 25%. Finally, the optimized method was applied to the analysis of biological samples from 10 patients in the intensive care unit (ICU) of a hospital located in Almeria (Spain). Several antibiotics (e.g., amoxicillin, tobramycin, levofloxacin, or linezolid) were found in the studied samples, observing that the highest concentrations were obtained in urine samples.

Pathological findings and diagnostic implications of a rhesus macaque (Macaca mulatta) model of aerosol-exposure melioidosis (Burkholderia pseudomallei)

Aerosolized Burkholderia pseudomallei, the causative agent of melioidosis, can infect many species of mammals (including humans), causing rapid, severe pneumonia with high mortality. Diagnosis in humans is challenging, as few organisms can be detected in blood or other non-invasive samples. Although it cannot be said that the model is established, studies to date indicate that rhesus macaques may represent a good model of human melioidosis. This is supported by the results of this study. The early progression of meliodosis in the rhesus macaque was studied in an effort to better understand the disease and the application of rapid diagnostic methods. Results indicate that a PCR analysis of key diagnostic samples such as nasal swabs, throat swabs, tracheo bronchial lymph node aspirates and broncho-alveolar lavage may be a useful component of a rapid diagnostic algorithm in case of aerosol exposure.

Abstract A36: Quantitative determination of methylation patterns in FHIT and APC genes in NSCLC: comparative analysis in tumor and normal lung tissue, peripheral blood, sputum and bronchial aspirate

Abstract Introduction and objective. Quantifying and comparing the degree of methylation in the promoter region of APC gene and coding region of FHIT gene in patients (p) with non small cell lung cancer (NSCLC) in different samples. Correlate these methylation patterns with clinical variables. Methods. DNA was extracted from peripheral blood simples (B), sputum (S) and bronchial aspirate (BA), obtained from patients with NSCLC prior to the completion of surgery, as well as resected lung tumor tissue (TT) and normal lung tissue (TN). Methylation patterns were analyzed by bisulfito conversion and subsequent pyrosequencing (QIagen PyroMark System).We analyzed 5 CpG islands in the promoter region of the APC gene and 5 CpG islands in the coding region of the FHIT gene. Results: We analyzed 20p, with a median age of 64 years (range 48-70), 16 men and 4 women. Smoking status: 2p never smokers, 11p former smokers, 7p current smokers. Histology: 10p adenocarcinoma, 9p squamous, 1p large cell. Stage: I 8p, II 8p, III 4p. No statistically significant differences were observed between the samples studied to any of the islands analyzed. The degree of methylation in TT CpG1 was higher in smokers and former smokers &amp;lt;5 years, compared to never smokers and ex-smokers &amp;gt;5 years, mean 4 (0-10) vs 0, p=0.022. There was no other difference when analyzing the degree of methylation as a function of the variables age, sex, smoking status, cumulative tobacco consumption, histological type and clinical stage. Respect FHIT gene, no statistically significant differences were found between the tissues studied respect to any of the CpG islands analyzed and likewise, no differences were observed when analyzed for degree of methylation depending on the clinical variables studied. Conclusions. The area of the APC gen promoter and FHIT coding region analyzed showed a low degree of methylation, with no significant difference observed between the samples studied. The degree of methylation in the TT CpG1 island of the APC gene was higher in current smokers and former smokers &amp;lt;5 years. However, these findings have to be confirmed in a larger sample. *This study was funded by the Carlos III Health Institute (PS09/00308), Spain. Note: This abstract was not presented at the conference. Citation Format: Rocio Lavado-Valenzuela, Sr., Carlos Pagés, Sr., Roberto Mongil, Sr., Francisco Páez, Sr., Enrique Bermejo, Sr., Miguel Angel Berciano, Sr., Dolores Bautista, Sr., Cynthia Robles, Jr., Ricardo Arrabal, Sr., Jose Luis De la Cruz, Sr., Manuel Benavides, Sr., Manuel Cobo, Sr.. Quantitative determination of methylation patterns in FHIT and APC genes in NSCLC: comparative analysis in tumor and normal lung tissue, peripheral blood, sputum and bronchial aspirate. [abstract]. In: Proceedings of the Third AACR International Conference on Frontiers in Basic Cancer Research; Sep 18-22, 2013; National Harbor, MD. Philadelphia (PA): AACR; Cancer Res 2013;73(19 Suppl):Abstract nr A36.

Abstract C20: Quantitative determination of EGFR gene aberrant methylation in NSCLC: a comparative analysis in tumor tissue, normal lung tissue, peripheral blood, sputum and bronchial aspirate

Abstract Introduction and objective. To evaluate and compare quantitatively aberrant methylation in the promoter area of EGFR gene in patients (p) with in non small cell lung cancer (NSCLC) lung tumor tissue (TT), lung normal tissue (NT), blood (B), bronchial aspirate (BA) and sputum (S). Correlate these methylation patterns with clinical variables. Methods. DNA was extracted from samples of B, S and BA from patients with NSCLC prior to surgery and resected TT and NT. Methylation patterns were analyzed by the bisulfite conversion and subsequent pyrosequencing (QIagen PyroMark System). We analyzed three CpG islands in the promoter region of EGFR gene. Results. 43p were included, median age 66 years (range 46-79), 35 men and 8 women. Histology: 26p adenocarcinoma, 14 squamous and 3 others. 2p had EGFR mutation. Smoking status: 4p never smokers, 21p former smokers, 18p smokers. Significant differences in methylation percentage (%Mth) were observed between TT and S in the following islands: CpG2 (11.8vs7.1, p = 0.008), CpG3 (10vs7.4, p = 0.037) and in overall promoter area (10.6vs7. 6, p = 0.034). The %Mth was higher in women vs men in TT CpG1 island (16.2vs24.9, p = 0.042) and TT CpG2 island (15.8vs26.5, p = 0.013). The %Mth was higher in squamous histology compared to adenocarcinoma in NT CpG2 island (22.6vs14.1, p = 0.017) and S CpG1 island (13.7vs8.2, p = 0.047). However, the %Mth was higher in the overall promoter area of adenocarcinoma respect to squamous histology (19.7vs12.8 TT, p = 0.042). The %Mth was higher in overall promoter area of NT respect Stage I vs II vs III (22.3 vs 13 vs 17.5, p = 0.03). The %Mth was higher in former smokers compared to current smokers and non smokers in NT CpG3 (22.8 vs 13 vs 14.1, p = 0.032). The %Mth in NT CpG3 was higher in never smokers and former smokers &amp;gt; 5 years, respect to current smokers and former smokers &amp;lt;5 years, (23.3 vs 15.8, p = 0.05). Conclusion. EGFR aberrant hypermethylation was higher in tumor tissue respect to sputum, with no correlation with EGFR mutation. The %Mth in normal tissue was higher in never smoker and former smokers &amp;gt; 5 years patients. * This study was funded by the Carlos III Health Institute (PS09/00308), Spain Note: This abstract was not presented at the conference. Citation Format: Rocio Lavado-Valenzuela, Enrique Bermejo, Roberto Mongil, Francisco Páez, Carlos Pagés, Miguel Angel Berciano, Ricardo Arrabal, Dolores Bautista, Jose Luis De la Cruz, Cynthia Robles, Manuel Benavides, Manuel Cobo. Quantitative determination of EGFR gene aberrant methylation in NSCLC: a comparative analysis in tumor tissue, normal lung tissue, peripheral blood, sputum and bronchial aspirate. [abstract]. In: Proceedings of the Third AACR International Conference on Frontiers in Basic Cancer Research; Sep 18-22, 2013; National Harbor, MD. Philadelphia (PA): AACR; Cancer Res 2013;73(19 Suppl):Abstract nr C20.

Bronchial foreign body misdiagnosed as pulmonary tuberculosis

Retained and neglected foreign bodies in the respiratory tract in children are a common occurrence. Undiagnosed foreign bodies can cause mechanical effects or chemical reactions and may present as chronic pulmonary infection, bronchiectasis, asthma, and lung collapse or lung abscess. Delay in diagnosis, appears to result from a failure to give serious consideration to the diagnosis, normal chest roentgenographic findings, and negative bronchoscopic findings. A case of left bronchial foreign body aspiration is herein reported to illustrate the causes and complications of the delayed diagnosis. It is hoped that this will help sensitize the medical practitioner of the need for a high index of suspicion in children presenting with symptoms suggestive of foreign body aspiration. This study will also serve as advocacy for the provision of adequate bronchoscopic equipments for paediatric practice in our health facilities.

Draft Genome Sequence of Mycobacterium massiliense Strain M159, Showing Phenotypic Resistance to β-Lactam and Tetracycline Antibiotics

Mycobacterium massiliense is a nontuberculous mycobacterium associated with human infections. We report here the draft genome sequence of M. massiliense strain M159, isolated from the bronchial aspirate of a patient who had a pulmonary infection. This strain showed genotypic and in vitro resistance to a number of tetracyclines and beta-lactam antibiotics.

Comparison of Performances of MTD Gene-Probe® Test, BACTEC 960™ System and Löwenstein-Jensen Culture Methods in the Diagnosis of Smear-Negative Tuberculosis Cases

The aim of this study was to compare the results of nucleic acid amplification-based MTD (Mycobacterium tuberculosis direct test) Gene-Probe® method in samples obtained from acid-fast bacilli (ARB) smear-negative patients with suspected tuberculosis (TB), with the culture results obtained from automated BACTEC 960™ (MGIT) system and Löwenstein-Jensen (LJ) medium. In addition, the contribution of molecular methods in early diagnosis of pulmonary TB and the effect of radiological prevalence of the disease associated with or without cavity to the molecular diagnosis and/or growth time in culture media have been evaluated. A total of 107 patients (86 male, 21 female; mean age: 49.89 ± 17.1 years, age range: 18-81 years) who were clinically and radiologically suspected of having pulmonary TB and/or TB pleurisy, were included in the study. Of the samples 65 (60.7%) were sputum, 32 (29.9%) were bronchial aspiration, 5 (4.7%) were pleural fluid, and 5 (4.7%) were transthoracic fine needle aspiration biopsy materials. Patient samples were cultured in solid LJ media and liquid-based BACTEC 960 system (Becton Dickinson Co., USA) in the same working day. Meanwhile, MTD Gen-Probe test (Gen-Probe Inc., USA) was studied in two separate working days of the week as specified by the laboratory. The samples were incubated until positivity was determined in BACTEC 960 system and/or growth was detected in LJ medium. Negative cultures were incubated for 42 days and were finalized. When mycobacterial growth was determined in the culture, identification of M.tuberculosis complex (MTBC) and differentiation from nontuberculous mycobacteria were performed by conventional methods and BACTEC 460 NAP test. Forty five (42%) patients were diagnosed as pulmonary paranchimal TB (40 were active pulmonary TB, 1 was miliary TB and 4 were culture-negative pulmonary TB), while 4 (3.7%) patients diagnosed as extrapulmonary TB and 58 (57.9%) patients were diagnosed as other pulmonary diseases unrelated with TB. LJ cultures yielded positive results in 32 of 45 (71%) pulmonary TB patients, and BACTEC 960 were found positive in 84.4% (38/45) of those patients. On the other hand the positivity rate of MTD Gen-Probe test was detected as 37.4% (40/107). The sensitivity, specificity, positive and negative predictive values for MTD Gen-Probe test were estimated as 89%, 100%, 100% and 93%, respectively. Those values for BACTEC 960 system were found as 82%, 98%, 97% and 88%, and for LJ culture method as 71%, 100%, 100% and 83%, respectively. Average periods to make a decision for diagnosis of TB by MTD Gen-Probe, BACTEC 960 (MGIT) and LJ culture methods were calculated as 2.36 days, 20.11 days and 32.49 days, respectively. In comparison of the methods in terms of turnaround times, MTD Gen-Probe test was found superior to LJ culture method, however the turnaround times for BACTEC 960 and LJ culture methods were similar. When the clinical data were evaluated, no effect of radiological density of lesion was identified on the diagnosis time of molecular test and time of growth in liquid based automated BACTEC system and/or LJ culture method. However, LJ culture demonstrated earlier reactivity in patients with cavitary lesions. As a result, MTD Gene-Probe test was observed as a reliable and rapid method for the early diagnosis of pulmonary TB patients, early initiation of therapy, prevention of disease progression and transmission.

Telomerase as a potential marker for inflammation and cancer detection in bronchial washing: A prospective study

ObjectivesThe diagnosis of lung cancer remains difficult especially in peripheral tumors, given the absence of relevant markers and of sensitive imaging techniques. Telomerase is a ribonucleotide enzyme responsible for the immortalization of cancerous cells and seems to increase in bronchial aspirates of lung cancer patients. The purpose of our study is to further investigate the value of telomerase measurement in bronchial aspirates as a diagnostic tool for lung cancer. Design and methodsRandom 82 bronchial aspirates were obtained from patients undergoing bronchoscopy to diagnose any lung illness including inflammation and cancer. Cytology examination, quantification of proteins by Bradford method, and telomerase activity measurement by quantitative Real-time PCR were performed. Out of 82 specimens, 11 were excluded because of hemolysis, absence of elements or lack of final diagnosis. ROC curve analysis was done. ResultsA significant difference in telomerase activity average was noted between normal patients and those with inflammation and cancer. Discriminatory capacity of telomerase activity was: for cancer vs. non cancer, AUC =0.74 (95% CI: 0.62–0.84), sensitivity=78%, specificity=72%, Negative Predictive Value=87%, at cut-off >0.46 atmol/mg protein/20min; for cancer vs. normal, AUC=0.87 (95% CI: 0.72–0.96), se=78%, sp=92%, NPV=71%, at cut-off >0.46; for cancer vs. inflammation, AUC=0.69 (95% CI: 0.55–0.80), se=74%, sp=70%, NPV=79%, at cut-off >1.03, and for inflammation vs. normal, AUC=0.76 (95% CI: 0.62–0.88), se=79%, sp=77%, NPV=59%, at cut-off >0. ConclusionTelomerase activity in bronchial aspirates is a promising diagnostic marker for lung cancer and inflammation detection.

Volume dependency for culture of fungi from respiratory secretions and increased sensitivity of Aspergillus quantitative PCR

Diagnosis of aspergillosis is often difficult. We compared fungal yields from respiratory specimens using the Health Protection Agency standard culture method (BSOP57), a higher volume undiluted culture method Mycology Reference Centre Manchester (MRCM) and Aspergillus quantitative real time polymerase chain reaction (qPCR). Sputum, bronchial aspirate and bronchoalveolar lavage (BAL) samples (total 23) were collected from aspergillosis patients. One fraction of all samples was cultured using the MRCM method, one BSOP57 and one was used for qPCR. The recovery rate for fungi was significantly higher by MRCM (87%) than by BSOP57 (8.7%) from all 23 specimens. Sputum samples were 44% positive by MRCM compared to no fungi isolated (0%) by BSOP57. Bronchial aspirates were 75% positive by MRCM and 0% by BSOP57. BAL samples were positive in 20% by MRCM and 10% by BSOP57. qPCR was always more sensitive than culture (95.6%) from all samples. In general, over 100 mould colonies (81 Aspergillus fumigatus) were grown using the MRCM method compared with only one colony from BSOP57. This study provides a reference point for standardisation of respiratory sample processing in diagnostic laboratories. Culture from higher volume undiluted respiratory specimens has a much higher yield for Aspergillus than BSOP57. qPCR is much more sensitive than culture and the current UK method requires revision.

Accidental severe bronchial aspiration of barium uncovers diagnosis of bronchiectasis

A 77-year-old man, with a history of chronic obstructive pulmonary disease (COPD) and radiotherapy-treated laryngeal malignancy diagnosed 15 years prior, presented with a 1-year history of intermittent dysphagia. As a part of his investigations, a barium swallow was obtained; but during the initial phases of the procedure, the patient aspirated the contrast agent of barium sulphate. A chest radiograph (Fig. 1a), taken few minutes after the aspiration, showed barium within the different segments and subsegments of the bronchial tree bilaterally, and uncovered features, such as irregular dilatation of the bronchial tree, non-tapering of the bronchi, luminal filling defects and lack of bronchial side branches, characteristic of bronchiectasis. This incident precipitated an exacerbation of COPD with type II respiratory failure, requiring bronchodilators, intravenous antibiotics and non-invasive ventilation using a bilevel positive airway pressure machine. On recovery, direct laryngoscopy revealed post-radiotherapy changes but no evidence of tumour recurrence. A speech and language pathology assessment using a few millilitres of water revealed weak laryngeal elevation, and immediate aspiration of water on swallowing. Given the high risk of further aspirations, advice regarding the benefits and risks of nasogastric tube insertion or percutaneous endoscopic gastrostomy tube insertion were given to the patient, but he declined both procedures. Six months following discharge, he was readmitted with an episode of food aspiration. A chest radiograph (Fig. 1b) revealed residual barium in both lung bases. This event precipitated another exacerbation of the COPD and bronchiectasis, which caused the patient to succumb to the disease. As early as the 1920s, the concept of utilizing contrast agents to aid in the diagnosis of pulmonary diseases, in particular bronchiectasis, had transpired. This involved instilling the contrast medium within the bronchial tree, by means of a catheter introduced in the trachea, and followed by the acquisition of various radiographs; a procedure known as bronchography. The initial contrast agents that were used in this regard were mainly waterand oil-based solutions. Subsequently in 1950s, barium sulphate, an inert and non-toxic agent, became a popular bronchographic medium that produced similar images as the earlier contrast agents, until the mid-1970s, when computed tomography became the gold standard imaging study for the investigation of pulmonary diseases. Although the earlier contrast agents, as well as barium sulphate, were considered safe bronchographic media, reports of pneumonitis and even death had been described with both [1]. The instilled contrast agent was found to cause obstruction of the small bronchi, followed by atelectasis of lung tissue and pneumonitis [2]. COPD patients often developed exacerbations after such procedures, as seen in our case. To minimize these risks, after the acquisition of the radiographs, patients were asked to cough and expectorate as much contrast dye as possible in an attempt to clear the airways. However, this mechanism would have been ineffective in cases of extensive small airway filling or ‘alveolarization’ with the contrast medium, as the alveoli lies outside the range of the normal mechanisms (cough and ciliary action) for cleansing the bronchial tree. The case we present demonstrates bronchographic features of bronchiectasis following accidental aspiration of E. Gerada (&) J. Gerada Department of Medicine, Mater Dei Hospital, Msida MSD 2090, Malta e-mail: egerada@gmail.com

Bronchopulmonary complications associated to enteral nutrition devices in patients admitted to Internal Medicine Departments

Background and aimsEnteral nutrition using feeding devices such as nasogastric (NG) tube or percutaneous endoscopic gastrostomy (PEG) is an effective feeding method subject that may give rise to complications. We have studied the relationship between enteral nutrition feeding devices in patients admitted to the Internal Medicine Departments and the development of pulmonary complications (bronchial aspiration and aspiration pneumonia). Patients and methodsAll of the patients discharge between 2005 and 2009 from the Internal Medicine (IM) Departments of the public hospitals of the National Health System in Spain were analyzed. The data of patients with bronchial aspiration or aspiration pneumonia who also were carriers of NG tubes or PEG, were obtained from the Minimum Basic Data Set (MBDS). ResultsFrom a total of 2,767,259 discharges, 26,066 (0.92%) patients with nasogastric tube (NG tube) or percutaneous gastrostomy (PEG) were identified. A total of 21.5% of patients with NG tube and 25.9% of patients with PEG had coding for a bronchopulmonary aspiration on their discharge report versus 1.2% of patients without an enteral feeding tube. In the multivariate analysis, the likelihood of suffering bronchoaspiration was 9 times greater in patients with SNG (OR 9.1; 95% CI 8.7–9.4) and 15 greater in subjects with PEG (OR 15.2; 95% CI 14.5–15.9) than in subjects without SNG or PEG. Mean stay (9.2 and 12.7 more days), diagnostic complexity and costs were much higher in patients with SNG or PEG compared to patients in hospital who did not require these devices. ConclusionsAn association was found between SNG and PEG for enteral feeding and pulmonary complications. Mean stay, diagnostic complexity and cost per admission of these patients were higher in patients who did not require enteral nutrition.