Examination Of Bronchoalveolar Lavage Fluid Research Articles

Up-regulation of HSP90α in HDM-induced asthma causes pyroptosis of airway epithelial cells by activating the cGAS-STING-ER stress pathway

Heat Shock protein 90 α (HSP90α), an main subtype of chaperone protein HSP90, involves important biological functions such as DNA damage repair, protein modification, innate immunity. However, the potential role of HSP90α in asthma occurrence and development is still unclear. This study aimed to elucidate the underlying mechanism of HSP90α in asthma by focusing on the cGAS-STING-Endoplasmic Reticulum stress pathway in inflammatory airway epithelial cell death (i.e., pyroptosis; inflammatory cell death). To accomplish that, we modeled allergen exposure in C57/6BL mice and bronchial epithelial cells with house dust mite. Protein technologies and immunofluorescence utilized to study the expression of HSP90α, activation of cGAS-STING pathway and pyroptosis. The effect of inhibitors on HDM-exposed mice detected by histological techniques and examination of bronchoalveolar lavage fluid. Results showed that HSP90α promotes asthma inflammation via pyroptosis and activation of the cGAS-STING-ER stress pathway. Treatment with the HSP90 inhibitor tanespimycin (17-AAG) significantly relieved airway inflammation and abrogated the effect of HSP90α on pyroptosis and cGAS-STING-ER stress in vitro and in vivo models of HDM. Further data indicated that up-regulation of HSP90α stabilized STING through interaction, which increased localization of STING on the ER. Activation of STING triggered ER stress and leaded to pyroptosis-related airway inflammation. The finding showed the potential role of pyroptosis caused by dysregulation of HSP90α on airway epithelial cells in allergic inflammation, suggested that targeting HSP90α in airway epithelial cells might prove to be a potential additional treatment strategy for asthma.

Overview of canine eosinophilic lung and bronchial diseases

In dogs with chronic cough eosinophilic lung disease (ELD) may be present, especially in young dogs. A breed predisposition has been described in Alaskan Malamutes and Siberian Huskies. Chronic cough is the most common clinical sign. Other symptoms include nasal discharge, sneezing, poly- or dyspnea and exercise intolerance. The exact pathogenesis is unknown. Type 1 hypersensitivity reaction is suspected. Eosinophilic lung diseases may be classified into different groups (eosinophilic bronchitis, eosinophilic bronchopneumopathy and eosinophilic granuloma). Diagnostic work-up includes hematology, imaging, bronchoscopy and cytologic examination of bronchoalveolar lavage fluid. A wide spectrum is present in terms of the expression and severity of the changes. The current reported treatment is systemic or inhaled glucocorticoids, or a combination of both.Most patients respond well to therapy. Relapses after treatment discontinuation are common.

Correlation of clinical and radiographic variables in cats with lower airway disease.

Feline lower airway disease (FLAD) is frequently associated with radiographic abnormalities. To evaluate whether radiographic changes in cats with naturally occurring FLAD improve with treatment and if radiographic changes correlate with clinical signs. Twenty-four client-owned cats newly diagnosed with FLAD, based on medical history, typical clinical signs, radiographic findings, and examination of bronchoalveolar lavage fluid, were included in the prospective study. At 2 examination time points (days 0 and 60), an owner questionnaire, clinical examination, and thoracic radiography were carried out. Information from the questionnaire and clinical examination were evaluated on the basis of a 12-point clinical score. Radiographs were assessed using a 10-point radiographic score. Individual treatment was given to all cats over the study period, based on severity of the disease and compliance of the cat. Clinical and radiographic scores were compared statistically for both examination time points and evaluated for correlation. All cats showed radiographic abnormalities at initial presentation. In addition to significant improvement in clinical variables, the total radiographic score improved significantly (P = .01) during the study period, with significant improvement in the severity of bronchial (P = .01) and interstitial lung pattern (P = .04). Improvement of the clinical and radiographic score was not correlated. In addition to clinical signs, repeated radiographic examination can be used as a diagnostic tool to evaluate treatment response in cats with FLAD.

Allergic bronchopulmonary aspergillosis with marked peripheral blood eosinophilia and pulmonary eosinophilia: A case report

Allergic bronchopulmonary aspergillosis (ABPA) is an immune-related pulmonary disease caused by sensitization of airway by Aspergillus fumigatus. The disease manifests as bronchial asthma and recurring pulmonary shadows, which may be associated with bronchiectasis. The diagnosis of ABPA mainly depends on serological, immunological, and imaging findings. Pathological examination is not necessary but may be required in atypical cases to exclude pulmonary tuberculosis, tumor, and other diseases through lung biopsy. An 18-year-old man presented with recurrent wheezing, cough, and peripheral blood eosinophilia. Chest computed tomography showed pulmonary infiltration. There was a significant increase in eosinophils in bronchoalveolar lavage fluid. There was no history of residing in a parasite-endemic area or any evidence of parasitic infection. Pathologic examination of bronchoalveolar lavage fluid excluded fungal and mycobacterial infections. The patient was receiving medication for comorbid diseases, but there was no temporal correlation between medication use and clinical manifestations, which excluded drug-induced etiology. Histopathological examination of lung biopsy specimen showed no signs of eosinophilic granulomatosis with polyangiitis, IgG4-related diseases, or tumors. The diagnosis of ABPA was considered based on the history of asthma and the significant increase in serum Aspergillus fumigatus-specific immunoglobulin (Ig)E. Eosinophil-related diseases were excluded through pathological biopsy, which showed typical pathological manifestations of ABPA. The possibility of ABPA should be considered in patients with poorly controlled asthma, especially those with eosinophilia, lung infiltration shadows, or bronchiectasis. Screening for serum IgE, Aspergillus fumigatus-specific IgE and IgG, and alveolar lavage can help avoid misdiagnosis.

Cytologic scoring of equine exercise-induced pulmonary hemorrhage: Performance of human experts and a deep learning-based algorithm.

Exercise-induced pulmonary hemorrhage (EIPH) is a relevant respiratory disease in sport horses, which can be diagnosed by examination of bronchoalveolar lavage fluid (BALF) cells using the total hemosiderin score (THS). The aim of this study was to evaluate the diagnostic accuracy and reproducibility of annotators and to validate a deep learning-based algorithm for the THS. Digitized cytological specimens stained for iron were prepared from 52 equine BALF samples. Ten annotators produced a THS for each slide according to published methods. The reference methods for comparing annotator's and algorithmic performance included a ground truth dataset, the mean annotators' THSs, and chemical iron measurements. Results of the study showed that annotators had marked interobserver variability of the THS, which was mostly due to a systematic error between annotators in grading the intracytoplasmatic hemosiderin content of individual macrophages. Regarding overall measurement error between the annotators, 87.7% of the variance could be reduced by using standardized grades based on the ground truth. The algorithm was highly consistent with the ground truth in assigning hemosiderin grades. Compared with the ground truth THS, annotators had an accuracy of diagnosing EIPH (THS of < or ≥ 75) of 75.7%, whereas, the algorithm had an accuracy of 92.3% with no relevant differences in correlation with chemical iron measurements. The results show that deep learning-based algorithms are useful for improving reproducibility and routine applicability of the THS. For THS by experts, a diagnostic uncertainty interval of 40 to 110 is proposed. THSs within this interval have insufficient reproducibility regarding the EIPH diagnosis.

Comparison of Induced Sputum and Bronchoalveolar Lavage Fluid Examination in the Diagnosis of Sputum Negative Pulmonary Tuberculosis

Tuberculosis (TB) is one of the common infections in the world, especially in developing countries like India. Therefore, early diagnosis is important. This study was undertaken to compare the yield of sputum, induction with bronchoalveolar lavage (BAL) in smear-negative suspected pulmonary TB patients in a tertiary care hospital in Agra. Fifty patients were included in the study. In all patients, induced sputum, fibreoptic bronchoscopy and BAL fluid were subjected to diagnostic testing. On acid-fast smear examination, induced sputum and BAL fluid tested positive in 27/50 and 25/50 patients, respectively with a sensitivity of 83.3% and 90% respectively (p<0.0001). On comparing sputum induction versus BAL on culture, 30 patients were positive by sputum induction and 27 patients were positive on BAL fluid, with the sensitivity of 85.7% and 77.1%, respectively. The results showed that the sputum induction showed a significantly higher yield than that of BAL fluid (p=0.0013). Sputum induction offers an alternative approach in the diagnosis of smear-negative suspected pulmonary TB patients and would enhance sensitivity for the diagnosis of TB.

Prevalence of Antineutrophil Cytoplasmic Antibodies and Antinuclear Antibodies in Patients with Pulmonary Tuberculosis: A Tertiary Care Center Experience from North India.

Tuberculosis (TB) can have manifestations closely mimicking autoimmune diseases. The prevalence of autoantibodies in TB varies among different populations. To study the prevalence of anti-neutrophilic cytoplasmic antibodies (ANCA) and antinuclear antibodies (ANA) in pulmonary tuberculosis (PTB). This was a cross-sectional, observational study. Subjects with microbiologically confirmed PTB, either via smear or culture positivity on sputum or bronchoalveolar lavage (BAL) fluid, or positive rapid diagnostic tests were included. ANCA against proteinase-3 (PR3), myeloperoxidase (MPO), lactoferrin, and elastase were tested using an enzyme-linked immunosorbent assay (ELISA). ANA was detected using indirect immunofluorescence (IIF). Eighty-nine subjects with a median [interquartile range (IQR)] age of 28 (20-46) years, 67.4% males, were recruited. Eighty-one subjects had microbiological confirmation on sputum examination, and eight required examination of BAL fluid. Sera were drawn from 62 treatment-naïve subjects, the rest (27) were on antitubercular therapy (ATT). Eighty-six (96.6%) subjects tested positive for anti-elastase antibody, seven of which were also positive for anti-PR3. None were positive for anti-MPO and anti-lactoferrin. Six (6.7%) subjects tested positive for ANA. None of the subjects had features of underlying connective tissue disease or vasculitis. PTB patients showed a high prevalence of anti-elastase and a low prevalence of ANA and anti-PR3 antibodies. ANCA positivity should be interpreted with caution in TB endemic areas. The role of anti-elastase antibodies in differentiating TB from ANCA-associated vasculitis (AAV) needs further research.

Effect of the Administration of a Nutraceutical Supplement in Racehorses with Lower Airway Inflammation.

Simple SummaryThe study of non-pharmacological products that could have a beneficial action on the respiratory system of the horse may have a great impact on the welfare and performance of equine athletes. As a matter of fact, young racehorses can be affected by a mild–moderate form of equine asthma (MEA), that can reach a prevalence of 80%, and pharmacological treatments during racing days are almost always forbidden. This study aimed to evaluate the efficacy of the administration of a feed supplement, composed of several nutraceutical herbs, in the treatment of lower airway inflammation. This product was administered to seven racehorses with MEA, while five horses were used as control. After 21 days, horses treated with the supplement showed a lower degree of clinical signs and lower mucus accumulation in the trachea, compared to controls. For this reason, the supplement was shown to be effective in controlling lower airway inflammation, and its use as an adjunctive treatment for MEA may deserve to be evaluated.Mild–moderate equine asthma (MEA) is a chronic inflammatory disorder of the lower airways of the horse, characterized by tracheal mucus accumulation, cough and poor performance. The therapeutic approach is based on pharmacological treatment and environmental management. Moreover, the efficacy of the administration of antioxidant molecules has been reported. The aim of the present study was to evaluate the effect of the administration of a commercial nutraceutical supplement, composed of several herbal extracts, on lower airway inflammation in racehorses. Twelve Thoroughbreds affected by MEA were selected. All horses underwent a clinical examination with assignment of a clinical score, airway endoscopy and cytological examination of bronchoalveolar lavage fluid. In seven horses, the supplement was administered for 21 days in association with environmental changes, while in five horses only environmental changes were performed. All procedures were repeated at the end of the study. Data concerning the clinical score, the endoscopic scores and the cytology at the beginning and at the end of the study were statistically compared. Data showed a significant reduction (p < 0.0156) of the clinical score and a significant reduction (p < 0.0156) of the tracheal mucus score. The results showed the beneficial effect of the supplement on mild–moderate lower airway inflammation, probably due to its antioxidant activity.

Bronchoscopy and Lung Fine-Needle Aspiration for Antemortem Evaluation of Pulmonary Involvement in Dogs with Naturally Occurring Canine Leishmaniosis.

Clinical manifestations from the lower respiratory tract are rare in canine leishmaniosis (CanL), making bronchoscopy and lung fine-needle aspiration (FNA) seldomly justified. The aim of this prospective study was to investigate the involvement of Leishmania infantum in the lungs of dogs with naturally occurring CanL by bronchoscopy and examination of bronchoalveolar lavage fluid (BALF), bronchial mucosa biopsies, and FNA, using immunodiagnostics. Dogs with relevant concurrent diseases and azotemia were excluded. Cough was detected in 5/31 (16.1%) dogs. Lesions (hyperemia, edema, mucosal granularity, secretions) were identified upon bronchoscopy in 19/31 (61.3%) dogs. The cytology of BALF revealed histiocytic inflammation in 14/31 (45.2%) dogs; the parasite was identified in one dog (3.2%). The immunofluorescence antibody test in BALF was positive in 15/31 (48.4%) dogs. Histopathology of bronchial mucosa and/or adjacent alveoli revealed lesions (mononuclear cell infiltration, fibrosis, edema, thickening of the inter-alveolar septa) in 24/31 (77.4%) dogs, with no Leishmania amastigotes. Positive antigen staining was observed within the cytoplasm of mononuclear cells in immunocytochemistry and immunohistochemistry. Μononuclear cells showed antigenic positivity in bronchial mucosa (27/31; 87.1%), BALF (30/31; 96.8%), and lung FNA (27/31; 87.1%). In conclusion, lungs seem to be affected from CanL more commonly than previously believed, and bronchoscopy allows obtaining valuable samples for antemortem diagnosis.

A novel animal model of primary blast lung injury and its pathological changes in mice.

Primary blast lung injury (PBLI) is a major cause of death in military conflict and terrorist attacks on civilian populations. However, the mechanisms of PBLI are not well understood, and a standardized animal model is urgently needed. This study aimed to establish an animal model of PBLI for laboratory study. The animal model of PBLI was established using a self-made mini shock tube simulation device. In brief, mice were randomly divided into two groups: the control group and the model group, the model group were suffered 0.5 bar shock pressures. Mice were sacrificed at 2 hours, 4 hours, 6 hours, 12 hours, and 24 hours after injury. Lung tissue gross observation, hematoxylin and eosin staining and lung pathology scoring were performed to evaluated lung tissue damage. Evans blue dye leakage and bronchoalveolar lavage fluid examination were performed to evaluated pulmonary edema. The relative expression levels of inflammation factors were measured by real-time quantitative polymerase chain reaction and Western blotting analysis. The release of neutrophil extracellular traps was observed by immunofluorescence stain. In the model group, the gross observation and hematoxylin and eosin staining assay showed the inflammatory cell infiltration, intra-alveolar hemorrhage, and damaged lung tissue structure. The Evans blue dye and bronchoalveolar lavage fluid examination revealed that the lung tissue permeability and edema was significantly increased after injury. Real-time quantitative polymerase chain reaction and Western blotting assays showed that IL-1β, IL-6, TNF-α were upregulated in the model group. Immunofluorescence assay showed that the level of neutrophil extracellular traps in the lung tissue increased significantly in the model group. The self-made mini shock tube simulation device can be used to establish the animal model of PBLI successfully. Pathological changes of PBLI mice were characterized by mechanical damage and inflammatory response in lung tissue.

Neutrophils in Bronchoalveolar Lavage Fluid Indicating the Severity and Relapse of Pulmonary Sarcoidosis.

BackgroundPulmonary sarcoidosis is a highly heterogeneous granulomatous disease without any specific symptoms and manifestations. Neutrophils in bronchoalveolar lavage fluid (BALF) have been proposed to indicate the severity and prognosis of pulmonary sarcoidosis, but this needs confirmation in patients from different populations due to the heterogeneity of the disease. This study aimed to determine the characteristics of patients with pulmonary sarcoidosis in northeastern China and to explore the relationship between neutrophils in BALF and the severity of pulmonary sarcoidosis.MethodsWe enrolled 432 patients who were diagnosed with pulmonary sarcoidosis in this retrospective study. The symptoms, extrapulmonary involvement, forced vital capacity percentage predicted (FVC % pred), and diffusing capacity of the lung for carbon monoxide percentage predicted (DLco % pred) were recorded. BAL was performed in 319 patients, and the results of a cellular examination of BALF were collected. A total of 123 patients who received corticosteroid treatment were followed up for at least 12 months, and the outcomes were recorded.ResultsCough was the most common symptom, and cutaneous involvement was the most common extrapulmonary manifestation in 304 (70.4%) and 82 (19.0%) patients, respectively. The percentages of patients with high neutrophil counts in BALF (>3%) were higher at Stages 2 and 3 compared with Stages 0 and 1 (33.2 vs. 19.4%, p = 0.007), although the percentages of neutrophils in BALF showed no difference between patients at Stages 0, 1, 2, and 3. Patients with high neutrophil counts in BALF had lower FVC % pred compared with the other patients (79.5 ± 18.2 vs. 84.9 ± 14.5%, p = 0.025) and were prone to relapse after corticosteroids were tapered. High neutrophil counts in BALF were independently associated with relapse after corticosteroids were tapered in a binary logistic regression analysis (p = 0.027).ConclusionsPatients with pulmonary sarcoidosis lacked specific symptoms and manifestations. The neutrophil count in BALF could indicate the severity and outcomes of pulmonary sarcoidosis.

Acute fibrinous and organizing pneumonia: two case reports and literature review

BackgroundAcute fibrinous and organizing pneumonia (AFOP) is a rare histologic interstitial pneumonia pattern characterized by the intra-alveolar fibrin deposition and organizing pneumonia. Its clinical characteristics are still not well known and there is no consensus on treatment yet.Case presentationWe report two female cases in their fifties diagnosed with AFOP confirmed by a second lung biopsy. Case 1 was idiopathic AFOP with manifestation of 6-week fever, dyspnea, and cough, while case 2 was secondary to systemic lupus erythematosus and fever was the major symptom. Their chest CT scans revealed bilateral multiple consolidations, predominantly in the lower lobes. Both cases were initially diagnosed with pneumonia, but did not improve after treatment with broad-spectrum antibiotics. In both cases, transbronchial biopsy and bronchoalveolar lavage fluid examination were inconclusive and the pathological diagnosis was confirmed by percutaneous lung biopsy. Both patients had a good clinical response to prednisone.ConclusionsWe report two rare AFOP cases to highlight the importance of awareness of this disease. We further perform the most comprehensive review to date in AFOP, including 150 patients since 2002. Consolidation was the most common imaging pattern, followed by ground-glass opacity and nodules. A lung biopsy is required for a definitive diagnosis. Corticosteroids is recommended as the most effective therapy, but treatment options should depend on the etiology and disease severity.

Krill oil and low-dose aspirin combination mitigates experimentally induced silicosis in rats: role of NF-κB/TGF-β1/MMP-9 pathway.

This study is an attempt to assess pulmonary protective and antifibrotic potentials of a combination of aspirin, a widely used anti-inflammatory and cardioprotective agent, and krill oil, a naturally occurring omega-3 fatty acid source, against silica-induced pulmonary injury. For silicosis induction, silica particles (50 mg/rat, 0.1 mL 0.9% NaCl) were instilled intranasally into rats. Aspirin (10 mg/kg/day), krill oil (40 mg/kg/day), or their combination was administered orally for 56 days following silica exposure. Results showed that oral aspirin and krill oil combination significantly mitigated silica-induced pulmonary injury. Bronchoalveolar lavage fluid examination showed a decreased lactate dehydrogenase activity, total protein content, and accumulation of total and differential inflammatory cells. Oral aspirin and krill oil combination significantly attenuated silica-induced oxidative stress through the restoration of reduced glutathione concentration and catalase activity in addition to alleviation of elevated malondialdehyde and total nitric oxide contents. Moreover, aspirin and krill oil combination revealed considerable mitigation of silica-induced upregulated expression of the inflammatory and fibrotic mediators: nuclear factor kappa-B, transforming growth factor-β1, and matrix metalloproteinase-9. The antifibrotic effect was also evidenced through the decreased hydroxyproline content and the obvious restoration of lung architecture, as demonstrated upon histopathological examination. In conclusion, oral aspirin and krill oil combination can confer pulmonary protective, anti-inflammatory, and antifibrotic potentials against silica-induced pulmonary injury. This impact could be credited to the ability of this combination to activate resolution mechanisms, which, in turn, suppress the expression of inflammatory and fibrotic biomarkers and replenish antioxidant stores.

Validity of flexible bronchoscopy in the diagnosis of chronic cough in children attending Assiut university children hospital

Context Chronic cough in children is a major health problem with an increased prevalence from 8 to 20.78% between 1999 and 2016. Flexible bronchoscope has become one of the most valuable tools in the diagnostic workup of unexplained chronic cough in children.Aims To assess the children with chronic cough using flexible bronchoscopy in order to diagnose the underlying causes that remain otherwise unexplained by noninvasive diagnostic tests.Settings and design Observational cohort study.Patients and methods Children with unexplained chronic cough underwent flexible bronchoscopy during the period from October 2017 to December 2019. Bronchoalveolar lavage (BAL) was done for cytologic examination and microbiologic studies.Results This study involved 60 children, aged from 2 months to 14 years. Flexible bronchoscopy demonstrated positive findings in 65% of cases. The most common finding was congenital airway anomalies in 25% of cases. The second most common finding was visible bronchitis in 21.67% of cases. Other bronchoscopic findings included endobronchial mucous plug (5%), foreign body aspiration (6.67%), extrinsic compression on the airways (10%), and adenoid hypertrophy (1.66%). Cytologic examination of BAL fluid showed that neutrophilia was the most common finding. Haemophilus influenzae was the most common isolated organism in the BAL fluid samples.Conclusions Flexible bronchoscopy is a valuable safe tool in the diagnosis of chronic cough in children. BAL helped in the diagnosis of the underlying etiology by providing samples for cytologic and microbiologic studies.

Time-course comparison of pulmonary inflammation induced by intratracheal instillation of four different nickel oxide nanoparticles in male Fischer rats

Occupational exposure to nickel oxide (NiO) is an important cause of respiratory tract cancer. Toxicity is known to be associated with the dissociated component, i.e. nickel (II) ions. To address the relationship between physicochemical properties, including solubility in artificial lysosomal fluid, of NiO and time-course changes in the pulmonary response, we conducted an intratracheal instillation study in male Fischer rats using four different well-characterized NiO products, US3352 (NiO A), NovaWireNi01 (NiO B), I small particle (NiO C), and 637130 (NiO D). The NiOs were suspended in purified water and instilled once intratracheally into male F344 rats (12 weeks old) at 0 (vehicle control), 0.67, 2, and 6 mg/kg body weight. The animals were euthanized on days 3, 28, or 91 after instillation, and blood analysis, bronchoalveolar lavage fluid (BALF) testing, and histopathological examination were performed. The most soluble product, NiO B, caused the most severe systemic toxicity, leading to a high mortality rate, but the response was transient and surviving animals recovered. The second-most-soluble material, NiO D, and the third, NiO A, caused evident pulmonary inflammation, and the responses persisted for at least 91 days with collagen proliferation. In contrast, NiO C induced barely detectable inflammation in the BALF examination, and no marked changes were noted on histopathology. These results indicate that the early phase toxic potential of NiO products, but not the persistence of pulmonary inflammation, is associated with their solubility.

The Relationship between Lung Inflammation and Aerobic Threshold in Standardbred Racehorses with Mild-Moderate Equine Asthma.

Simple SummaryHorses can be affected by equine asthma, a disease that shares several similarities with human asthma. Young racehorses can be affected by a mild form of equine asthma, which can have a negative impact on racing performance. In this study, we evaluated the relation between the increase in inflammatory cells in the bronchoalveolar lavage fluid of horses affected by poor performance due to equine asthma and the speed at which the horse blood lactate reaches the value of 4 mmol/L, which is a parameter of athletic capacity. With this aim, we evaluated the results of a treadmill test with blood lactate analysis of 30 horses that had a bronchoalveolar lavage fluid cytology consistent with a mild form of equine asthma. The results showed a significant relation between the increase in the differential count of neutrophils in bronchoalveolar lavage and the decrease in the speed at 4 mmol/L of lactate. This confirm the negative impact of neutrophilic lung inflammation in the presence of equine asthma on athletic capacity and, consequently, on racing performance.Mild–moderate equine asthma (MEA) is a common disease of young racehorses characterized by inflammation of the lower airways diagnosed by cytological examination of bronchoalveolar lavage fluid (BALF), and it is considered an important cause of poor performance. The most common parameter of athletic capacity associated with horse performance is the speed at 4 mmol/L of lactate (VLA4). The aim of the present work was to evaluate the relation between the different BALF inflammatory cells and VLA4 in racehorses affected by MEA. A population of Standardbred racehorses that underwent an accurate protocol for poor performance evaluation was collected for a retrospective study. Horses with any other alteration potentially influencing performance were excluded, thus considering only subjects with BALF cytology consistent with MEA. A sample of 30 horses (3.4 ± 1.0 years old) was selected. The relationship between BALF inflammatory cell differential count and VLA4 was evaluated by means of linear regression. Statistical analysis showed a significant relation (p = 0.015, r2 = 0.19) between the increase in the differential count of neutrophils in BALF and the decrease in VLA4. The results obtained suggested that the accumulation of neutrophils in the airways of MEA horses may have a direct impact on athletic capacity, possibly due to impaired alveolar blood-gas exchanges during strenuous exercise.

The mechanisms of antibiotic resistance in major pathogens of purulent-inflammatory complications in cancer patients

The problem of microbial antibiotic resistance and investigation of its underlying mechanisms is of paramount importance for all fields of clinical medicine, including oncology. The aim of the study was to examine the mechanisms of antibiotic resistance for major pathogens causing purulent-inflammatory complications in cancer patients. Materials and methods. In 2012—2015 there was conducted a prospective examination of 184 cancer patients, including 67 patients at the Department of Surgery no. 1 and 117 patients at the Intensive Care Unit of the Krasnoyarsk Regional Clinical Oncology Center named after A.I. Kryzhanovsky. For this, we collected bronchoalveolar lavage fluid, wound discharge and investigated the material by using bacteriological method, as well as MALDI-TOF. Antibiotic sensitivity was studied as follows: disco-diffusion, double disc method, carbapenem inactivation method, staphylococcal sensitivity — by screening method, PCR, E-test method, and serial dilutions in Muller-Hinton broth. Genotyping and antibiotic resistance mechanisms study were performed by using PCR, M-PCR, and sequencing. The WHONET program (WHO) was used, with significance level set at p 0.05. Results. Microbiological examination of bronchoalveolar lavage fluid and wound discharge samples allowed to uncover prevalent associations of multi-resistant (MDR) and extremely resistant pathogens (XDR). The microflora of the lower respiratory tract and in the wound secretion in cancer patients were found to be dominated by non-fermenting Gram-negative bacteria reaching up to 44.5% and 48%, respectively; as well as order Enterobacteriales found in 24% and 34.9%, respectively; Gram-positive bacteria — 24% and 17.1%, respectively. Imipenem- and/or meropenem-resistant P. aeruginosa and A. baumannii, K. pneumoniae strains, were assessed for MBL production phenotypically, as well as the genes of the most common VIM, IMP types, whereas A. baumannii — for OXA-23, OXA-40, and OXA-58; and K. pneumoniae — for OXA-48. 20 strains and 16 strains of P. aeruginosa and A. baumannii, respectively, were studied by PCR. It was found that A. baumannii strains formed no MBL, but 56.3% of A. baumannii isolates (9 strains) produced OXA-23 and OXA-40 carbapenemases. Among P. aeruginosa strains there were three of them which possessed VIM (15.0%), whereas the remaining strains formed no MBL, but were resistant to carbapenems being associated with other resistance mechanisms, e.g. efflux, decreased permeability of cell wall etc. Among 6 isolates of K. pneumoniae, 1 strain produced OXA-48. In cancer patients, the percentage of methicillin-resistant strains among all members of the genus Staphylococcus was 48.9% (4 strains belonged to MRSA). PVL- MRSA strains belonged to the clones ST239/spa3(t037)/SCCmecIIIA/tst,sek,seq+ (75%) and ST8/ spa1(t008)/SCCmecIVc/sea+ (25%). MRSA ST239 showed multiple antibiotic resistance: to aminoglycosides (aacA-aphD, aadD genes were detected), linkcosamides/macrolides (the ermA gene was detected), fluoroquinolones (mutations in the GyrA gene — Ser84Leu; in GrlA- Ser80Phe), rifampicin (MIC more than 128 gg/ml; mutations in the rpoB gene are His481Asn, Ile527Met), sulfamethoxazole, tetracycline (tetM gene), and chloramphenicol (66.7% of isolates, the cat gene encoding chloramphenicol acetyl transferase was detected); but sensitive to vancomycin (MIC 1.0 gg/ ml), linezolid in 100% of cases. MRSA ST8 are resistant to aminoglycosides (aacA-aphD, aadD genes), lincosamides/macrolides (ermC gene), tetracyclines (tetK gene), chloramphenicol (cat gene); and 100% sensitive to fluoroquinolones, rifampicin (MIC 0.006 gg/ml), sulfamethaxazole, vancomycin (MIC 1.0 gg/ml), daptomycin (MIC 0.094 gg/ml), linezolid (MIC 0.75 gg/ml). Conclusion. Thus, it was found that members of the order Enterobacteriales, A. baumannii, P. aeruginosa and MRSA retain high resistance to a large number of antibacterial drugs of almost all classes. These data should be taken into account while choosing proper antibiotic therapy, as well as controlling spread of nosocomial infections caused by multiresistant microorganisms.

Bronchoalveolar Lavage Fluid Caseating Granulomas in A Case of Pulmonary Tuberculosis

Pulmonary tuberculosis (TB) is a common infectious disease that is considered one of the leading causes of morbidity and mortality worldwide. Special diagnostic studies for TB can be used including smear microscopy and cultures for acid fast bacilli (AFB) and Mycobacterium tuberculosis (MTB) nucleic acid amplification test (NAAT). The specimen is usually taken from the sputum but might also include pleural and bronchoalveolar lavage (BAL) fluids. In addition, BAL cytology and lung biopsy are commonly done if bronchoscopy is performed. Here, we present a case of suspected pulmonary TB in a patient who was complaining of cough and hemoptysis for several months. Chest x-ray showed right upper lung opacity with cystic changes that was confirmed by CT chest. Work up done for this case included three samples of sputum and one BAL fluid specimen for AFB smear microscopy, culture and NAAT in addition to transbronchial lung biopsies and they were all negative. However, Cytologic examination of BAL fluid using Papanicolaou stain surprisingly showed evidence of intact caseating granulomas characteristic for tuberculous infection. Ultimately, the patient was started on the standard anti-TB regimen for 6 months with significant clinical improvement confirming TB diagnosis. To the best of our knowledge, this unusual cytologic finding in culture-negative BAL fluid coupled with unremarkable lung biopsy and persistently negative sputum samples for TB was never reported in the English literature.

BAL fluid analysis in the identification of infectious agents in patients with hematological malignancies and pulmonary infiltrates.

The present study aims to evaluate the diagnostic yield of bronchoalveolar lavage (BAL) fluid in patients with hematological malignancies and describe the most common pathogens detected in BAL fluid (BALF.) An analysis of 480 BALF samples was performed in patients with hematological malignancies over a period of 7 years. The results of culture methods, PCR, and immunoenzymatic sandwich microplate assays for Aspergillus galactomannan (GM) in BALF were analyzed. Further, the diagnostic thresholds for Aspergillus GM and Pneumocystis jiroveci were also calculated. Microbiological findings were present in 87% of BALF samples. Possible infectious pathogens were detected in 55% of cases; 32% were classified as colonizing. No significant difference in diagnostic yield or pathogen spectrum was found between non-neutropenic and neutropenic patients. There was one significant difference in BALF findings among intensive care units (ICU) versus non-ICU patients for Aspergillus spp. (22% versus 9%, p = 0.03). The most common pathogens were Aspergillus spp. (n = 86, 33% of BAL with causative pathogens) and Streptococcus pneumoniae (n = 46, 18%); polymicrobial etiology was documented in 20% of cases. A quantitative PCR value of > 1860 cp/mL for Pneumocystis jirovecii was set as a diagnostic threshold for pneumocystis pneumonia. The absorbance index of GM in BALF of 0.5 was set as a diagnostic threshold for aspergillosis. The examination of BAL fluid revealed the presence of pathogen in more than 50% of cases and is, therefore, highly useful in this regard when concerning pulmonary infiltrates.Electronic supplementary materialThe online version of this article (10.1007/s12223-019-00712-4) contains supplementary material, which is available to authorized users.

31P - Do two lungs form an integrated immune system? Learning from BALF examination in lung cancer

Background: Bronchoalveolar lavage fluid (BALF) examination was found to be useful in the evaluation of local immune status in the site of lung cancer development. This procedure may be performed during lung cancer diagnosis and repeated during therapy. It especially concerns the majority of lung cancer cases, which are beyond the possibility of resection at recognition. However, the procedure of BAL in the lung affected by cancer is often complicated. The aim of this study was an analysis of differences in the character of immune response between the lung affected by cancer versus the opposite, ’healthy‘ lung. May BALF from the lung free of tumor serve for the characterization of local immune status? Methods: BALF of 36 patients was harvested from the lung affected by cancer and opposite site: the lung without tumor. BALF cells were analyzed by means of the following sets of antibodies anti: the first probe CD4, CD127, CD25, Foxp3, CTLA4, the second probe: CD3, CD16/CD56, CD45, CD19, CD8. For macrophage phenotyping the immunofluorescent method with antibodies anti: CCR7 and CD163 was used. The concentrations of 27 cytokines were analyzed in the duplicate using Bio-Plex Pro Human Cytokine Assay. The next part of the study was performed in other 32 patients by the same method with a panel of antibodies anti: CD3, CD4, CD8, CD127, CD45RA, CD69, CTLA-4, PD-1. Results: By the analysis of the BALF profile we found that only the cells and cytokines involved in regulatory pathways showed significant differences between both the ’healthy‘ and the cancerous lung, being elevated in the latter. The proportion of Tregs, CTLA4+Tregs, activated and memory PD-1+ lymphocytes, CCR7lowCD163high macrophages were significantly higher in the lung with cancer than in the healthy lung. On the other hand, the proportion of effectory cells and inflammatory cytokines concentration did not differ between both lungs, which may indicate that they form an integrated immune system and individual homeostasis. Conclusions: Regulatory mechanisms which are up-regulated in the tumourbearing lung affect BALF profile in the lung with cancer, while the examination of BALF from the “healthy” lung may serve for the evaluation of individual basic profile of the anti-cancer response. Legal entity responsible for the study: The authors. Funding: Has not received any funding. Disclosure: J. Domagala-Kulawik: Travel grant, consultant: BMS, MSD. All other authors have declared no conflicts of interest.