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Experimental Lung Research 2019Although the isolation of rat and mouse mesothelial cells has previously been reported, most mesothelial cells used for experimental studies are obtained from...
Although the isolation of rat and mouse mesothelial cells has previously been reported, most mesothelial cells used for experimental studies are obtained from peritoneal cells. Here, we describe an optimized method for the isolation and propagation of rodent pleural mesothelial cells without the requirement for specialized surgical techniques. To harvest pleural mesothelial cells, the pleural space of 8-9-week-old rats or older mice was filled with 0.25% trypsin in ethylenediaminetetraacetic acid (EDTA) buffer for 20 min at 37 °C. Cells were then harvested, and incubated at 37 °C in a humidified atmosphere with 5% CO. Immunofluorescence analysis of plated pleural mesothelial cells was performed using Alexa 546 (calretinin). To investigate optimal proliferation conditions, medium enriched with various concentrations of fetal calf serum (FCS) was used for pleural mesothelial cell proliferation. By day 10, confluent cell cultures were established, and the cells displayed an obvious cobblestone morphology. Immunofluorescence analysis of the cells demonstrated that all stained positive for Alexa 546 (calretinin) expression. Mesothelial cells grew better in medium containing 20% FCS than with 10% FCS. This is a simple procedure for the efficient collection of primary pleural mesothelial cells, which were obtained in defined culture conditions from the euthanized rodent thoracic cavity using trypsin-EDTA treatment. The ability to easily culture and maintain identifiable pleural mesothelial cells from rodents will be helpful for future experiments using these cells.
Topics: Animals; Mice; Pleura; Primary Cell Culture; Rats
PubMed: 31250673
DOI: 10.1080/01902148.2018.1511002 -
Frontiers in Immunology 2022Filarial parasites are tissue dwelling worms transmitted by hematophagous vectors. Understanding the mechanisms regulating microfilariae (the parasite offspring)...
Filarial parasites are tissue dwelling worms transmitted by hematophagous vectors. Understanding the mechanisms regulating microfilariae (the parasite offspring) development is a prerequisite for controlling transmission in filarial infections. Th2 immune responses are key for building efficient anti-parasite responses but have been shown to also lead to detrimental tissue damage in the presence of microfilariae. , a rodent filaria residing in the pleural cavity was therefore used to characterize pleuropulmonary pathology and associated immune responses in wild-type and Th2 deficient mice. Wild-type and Th2-deficient mice ( ) were infected with and parasite outcome was analyzed during the patent phase (when microfilariae are in the general circulation). Pleuropulmonary manifestations were investigated and pleural and bronchoalveolar cells were characterized by RNA analysis, imaging and/or flow cytometry focusing on macrophages. mice were hypermicrofilaremic and showed an enhanced filarial survival but also displayed a drastic reduction of microfilaria-driven pleural cavity pathologies. In parallel, pleural macrophages from mice lacked expression of prototypical alternative activation markers RELMα and and showed an altered balance of some markers of the arginine metabolic pathway. In addition, monocytes-derived F4/80 macrophages from infected mice failed to mature into resident F4/80 large macrophages. Altogether these data emphasize that the presence of both microfilariae and IL-4R/IL-5 signaling are critical in the development of the pathology and in the phenotype of macrophages. In mice, the balance is in favor of parasite development while limiting the pathology associated with the host immune response.
Topics: Animals; Mice; Arginine; Filariasis; Filarioidea; Interleukin-5; Macrophages; Mice, Inbred BALB C; Microfilariae; Th2 Cells
PubMed: 36353644
DOI: 10.3389/fimmu.2022.866373 -
Respiratory Research Feb 2017Bacterial pleural infection requires prompt identification to enable appropriate investigation and treatment. In contrast to commonly used biomarkers such as C-reactive... (Comparative Study)
Comparative Study
BACKGROUND
Bacterial pleural infection requires prompt identification to enable appropriate investigation and treatment. In contrast to commonly used biomarkers such as C-reactive protein (CRP) and white cell count (WCC), which can be raised due to non-infective inflammatory processes, procalcitonin (PCT) has been proposed as a specific biomarker of bacterial infection. The utility of PCT in this role is yet to be validated in a large prospective trial. This study aimed to identify whether serum PCT is superior to CRP and WCC in establishing the diagnosis of bacterial pleural infection.
METHODS
Consecutive patients presenting to a tertiary pleural service between 2008 and 2013 were recruited to a well-established pleural disease study. Consent was obtained to store pleural fluid and relevant clinical information. Serum CRP, WCC and PCT were measured. A diagnosis was agreed upon by two independent consultants after a minimum of 12 months. The study was performed and reported according to the STARD reporting guidelines.
RESULTS
80/425 patients enrolled in the trial had a unilateral pleural effusion secondary to infection. 10/80 (12.5%) patients had positive pleural fluid microbiology. Investigations for viral causes of effusion were not performed. ROC curve analysis of 425 adult patients with unilateral undiagnosed pleural effusions showed no statistically significant difference in the diagnostic utility of PCT (AUC 0.77), WCC (AUC 0.77) or CRP (AUC 0.85) for the identification of bacterial pleural infection. Serum procalcitonin >0.085 μg/l has a sensitivity, specificity, negative predictive value and positive predictive value of 0.69, 0.80, 0.46 and 0.91 respectively for the identification of pleural infection. The diagnostic utility of procalcitonin was not affected by prior antibiotic use (p = 0.80).
CONCLUSIONS
The study presents evidence that serum procalcitonin is not superior to CRP and WCC for the diagnosis of bacterial pleural infection. The study suggests routine procalcitonin testing in all patients with unilateral pleural effusion is not beneficial however further investigation may identify specific patient subsets that may benefit.
TRIAL REGISTRATION
The trial was registered with the UK Clinical Research Network ( UKCRN ID 8960 ). The trial was approved by the South West Regional Ethics Committee (Ethical approval number 08/H0102/11).
Topics: Aged; Aged, 80 and over; Bacterial Infections; Biomarkers; C-Reactive Protein; Calcitonin; Female; Humans; Incidence; Leukocyte Count; Male; Middle Aged; Pleural Effusion; Prognosis; Risk Factors; Survival Rate; United Kingdom
PubMed: 28158976
DOI: 10.1186/s12931-017-0501-5 -
BMC Infectious Diseases Aug 2019Pulmonary Cryptococcosis (PC) is diagnosed with increasing incidence in recent years, but it does not commonly involve the pleural space. Here, we report a HIV-negative... (Review)
Review
BACKGROUND
Pulmonary Cryptococcosis (PC) is diagnosed with increasing incidence in recent years, but it does not commonly involve the pleural space. Here, we report a HIV-negative case with advanced stage IIIB non-small cell lung cancer (NSCLC) treated with radiation therapy presented with dyspnea, a new PET-positive lung mass and bilateral pleural effusion suspecting progressive cancer. However, the patient has been diagnosed as pulmonary cryptococcal infection and successfully treated with oral fluconazole therapy.
CASE PRESENTATION
A 77-year-old male with advanced stage non-small cell lung cancer treated with combined chemo-radiation therapy who presented with progressive dyspnea, a new PET-positive left lower lobe lung mass and bilateral pleural effusions. Initial diagnostic thoracentesis and bronchoscopy yielded no cancer, but instead found yeast forms consistent with cryptococcal organisms in the transbronchial biopsies of the left lower lobe lung mass. Subsequent to this, the previously collected pleural fluid culture showed growth of Cryptococcus neoformans. The same sample of pleural effusion was tested and was found to be positive for crytococcal antigen (CrAg) by a lateral flow assay (LFA). The patient has been treated with oral fluconazole therapy resulting in gradual resolution of the nodular infiltrates.
CONCLUSION
PC should be considered in immunosuppressed cancer patients. Additionally, concomitant pleural involvement in pulmonary cryptococcal infections may occur. The incidence of false positive FDG-PET scans in granulomatous infections and the use of CrAg testing in pleural fluid to aid in diagnosis are reviewed.
Topics: Administration, Oral; Aged; Antifungal Agents; Carcinoma, Non-Small-Cell Lung; Cryptococcosis; Cryptococcus neoformans; Fluconazole; Humans; Immunocompromised Host; Lung Diseases, Fungal; Lung Neoplasms; Male; Pleural Effusion; Positron-Emission Tomography
PubMed: 31405376
DOI: 10.1186/s12879-019-4343-2 -
The Clinical Respiratory Journal Feb 2019Thoracoscopy in the endoscopy suite, has a high diagnostic yield of undiagnosed pleural effusions with minimal and mild complications. Whereas relatively minimal... (Review)
Review
Thoracoscopy in the endoscopy suite, has a high diagnostic yield of undiagnosed pleural effusions with minimal and mild complications. Whereas relatively minimal invasive techniques, such as thoracentesis, image-guided pleural biopsy or blind pleural biopsy, can yield sufficient cell or tissue material to establish the diagnosis of the underlying condition, more definite invasive diagnostic and therapeutic procedure, such as thoracoscopy, may be required for accurate sampling and diagnosis, and further provide real-time treatment options in same procedure. If thoracoscopy is considered the gold standard for the diagnosis is a fact in case. The current review aims to provide informations on thoracoscopy indications in benign pleural diseases according to up to date publications.
Topics: Chylothorax; Cost-Benefit Analysis; Humans; Image-Guided Biopsy; Pleura; Pleural Effusion; Sensitivity and Specificity; Thoracentesis; Thoracoscopy; Tuberculosis, Pleural
PubMed: 30578625
DOI: 10.1111/crj.12983 -
Pneumologie (Stuttgart, Germany) Feb 2022The diagnosis of pleural tuberculosis remains a clinical challenge due to the paucibacillary nature of disease. Medical thoracoscopy remains the gold standard in...
INTRODUCTION
The diagnosis of pleural tuberculosis remains a clinical challenge due to the paucibacillary nature of disease. Medical thoracoscopy remains the gold standard in diagnosing tuberculous pleuritis.
OBJECTIVE
To establish the diagnostic yield of sago-seed thoracoscopic appearance of pleura in tuberculosis and its correlation with histopathology, tissue AFB culture and tissue Xpert MTB/Rif assay.
METHODS
All consecutive patients with lymphocytic exudative pleural effusion, who fulfilled inclusion criteria of the study underwent medical thoracoscopy under local anesthesia and pleural tissue was sent for histopathology, AFB culture and Xpert MTB/Rif assay. Chronic granulomatous inflammation on histopathology and response to anti-tuberculous treatment was taken as reference standard for diagnosis of tuberculous pleurisy.
RESULTS
A total of 249 patients were included in the study, out of which 168 had effusion secondary to tuberculosis. Sago-like nodules visualized on thoracoscopy had a sensitivity of 58.9 %, specificity of 92.6 % and diagnostic accuracy of 69.88 % for pleural tuberculosis. There is a strong association between the presence of sago-like nodules and detection of mycobacterium tuberculosis on Xpert MTB/Rif assay and AFB culture of pleura (-value 0.007).
CONCLUSION
Sago seed nodules on pleura have a high positive predictive value for tuberculous pleurisy. In high endemic countries patients with this finding on thoracoscopy can be commenced on anti-tuberculous treatment before histopathology or culture results are available.
Topics: Biopsy; Humans; Mycobacterium tuberculosis; Pleural Effusion; Sensitivity and Specificity; Tuberculosis, Lymph Node; Tuberculosis, Pleural
PubMed: 34847611
DOI: 10.1055/a-1666-5851 -
Journal of Medical Microbiology May 2021Pleural infections cause major morbidity and mortality, particularly amongst paediatric and elderly populations. The aetiology is broad, but pleural culture fails to...
Pleural infections cause major morbidity and mortality, particularly amongst paediatric and elderly populations. The aetiology is broad, but pleural culture fails to yield a causative pathogen in approximately 40 % of cases. Alternative pathogen identification methods are therefore required. The aim of the study was to investigate the yield from and impact on patient care when performing 16S rRNA PCR on culture-negative pleural fluid specimens and to determine whether any individual laboratory parameters were associated with a positive 16S rRNA PCR result. We conducted a study on 90 patients with suspected pleural infection, who had a culture-negative pleural fluid specimen, which underwent 16S rRNA PCR analysis between August 2017 and June 2019. This study was undertaken at a large NHS Trust in London, UK. Thirty-one per cent of culture-negative pleural fluid specimens tested by 16S rRNA PCR yielded a positive PCR result. Our data demonstrated that 16S rRNA PCR detected a significantly higher proportion of (<0.0001) and fastidious, slow-growing and anaerobic pathogens (=0.0025) compared with culture-based methods. Of the 25 16S rRNA PCR results that were positive for a causative pathogen, 76 % had a direct impact on clinical management. No single laboratory variable was found to be associated with a positive 16S rRNA PCR result. The findings from our real-world evaluation highlight the importance of 16S rRNA PCR in confirming pleural infection when the aetiology is unknown, and its direct, positive impact on clinical management.
Topics: Adolescent; Adult; Aged; Aged, 80 and over; Child; Child, Preschool; Female; Humans; Infant; Male; Middle Aged; Pleura; Pleural Diseases; Pneumococcal Infections; Polymerase Chain Reaction; RNA, Ribosomal, 16S; Retrospective Studies; Streptococcus pneumoniae; United Kingdom; Young Adult
PubMed: 34038341
DOI: 10.1099/jmm.0.001366 -
The Indian Journal of Tuberculosis Jul 2021Residual pleural opacity (RPO) is a common radiographic sequela in patients with tubercular pleural effusion at the end of the treatment. This study was designed to find...
INTRODUCTION
Residual pleural opacity (RPO) is a common radiographic sequela in patients with tubercular pleural effusion at the end of the treatment. This study was designed to find out the risk factors associated with residual pleural opacity (RPO).
MATERIALS & METHODS
This was a prospective longitudinal study performed to analyse data of 56 patients (46 males & 10 females) who were diagnosed as tubercular pleural effusion and treated for the same between 1st Jan 2019 to 30th March 2020. Chest X-ray posteroanterior & Lateral view was done (performed) at 0 and 6 months of treatment to quantify the amount of pleural effusion and measured the residual pleural opacity at the end of the treatment. RPO included both non resolving pleural effusion as well as residual pleural thickening (RPT). All statistical analysis was done using SPSS version 20.0 (SPSS Inc., Chicago, IL, USA). Multivariate logistic regression was performed to explore the association of risk factors and Residual pleural opacity. The statistical significance level was set at 0.05 (two-tailed).
RESULTS
The incidence of Residual pleural opacity (RPO) at the end of 6 months of antituberculosis treatment was 53.57% (30/56)). The study patients were divided into RPO and non- RPO group. Male gender had significantly higher incidence of RPO (93.3% vs 69.2% P = 0.01)). Patients with RPO group had significantly more cough and weight loss as compared to non RPO group (96.6% vs 65.3% P = 0.002 and 60% vs 23% P = 0.005). The proportion of patients who underwent therapeutic aspiration and gained weight of more than 5kg during treatment (19.5% vs 7.6% P = 0.02 & 46.6% vs 7.6% P = 0.001) was significantly higher in RPO group. A significantly lower protein, glucose and higher LDH level in pleural fluid was observed in the RPO group compared to non-RPO group (P = 0.006, P = 0.01, P = 0.001)). No significant difference was found in the pleural fluid ADA, lymphocyte, neutrophil levels between the two groups (p > 0.05). Logistic regression analysis showed that the male gender, low pleural fluid glucose, presence of cough and weight loss were associated with significantly increased risk of residual pleural opacity and thickening (p < 0.05).
CONCLUSION
Tubercular pleural effusion is associated with residual pleural opacity in more than half of the patients. Male gender and low glucose levels in pleural fluid was associated with increased risk of residual pleural opacity.
Topics: Adult; Antitubercular Agents; Duration of Therapy; Female; Glucose; Humans; Incidence; India; Longitudinal Studies; Male; Organ Size; Pleura; Pleural Cavity; Pleural Effusion; Radiography, Thoracic; Risk Factors; Sex Factors; Treatment Outcome; Tuberculosis, Pleural
PubMed: 34099203
DOI: 10.1016/j.ijtb.2020.12.012 -
Thoracic Surgery Clinics Nov 2021Empyema may occur in the pleural space after pulmonary resection. Subsequent bacterial contamination results in infection and development of frank empyema.... (Review)
Review
Empyema may occur in the pleural space after pulmonary resection. Subsequent bacterial contamination results in infection and development of frank empyema. Pneumonectomy-surgical removal of the entire lung-is the treatment of choice for centrally located bronchogenic carcinoma, diffuse malignant mesothelioma, and chronic inflammatory lung diseases with destroyed lung from pulmonary tuberculosis, fungal infections, and bronchiectasis. In the uncomplicated case, on the pneumonectomy side, the diaphragm becomes elevated as the air-fluid level decreases with chest wall deformation and gradual disappearance of hydrothorax. The pneumonectomy space is at potential risk for getting infected from bacterial contamination and developing empyema.
Topics: Bronchial Fistula; Empyema, Pleural; Humans; Lung Diseases; Pleural Diseases; Pneumonectomy; Sepsis
PubMed: 34696853
DOI: 10.1016/j.thorsurg.2021.08.001 -
The Lancet. Respiratory Medicine Jun 2024Extended pleurectomy decortication for complete macroscopic resection for pleural mesothelioma has never been evaluated in a randomised trial. The aim of this study was... (Randomized Controlled Trial)
Randomized Controlled Trial Comparative Study
BACKGROUND
Extended pleurectomy decortication for complete macroscopic resection for pleural mesothelioma has never been evaluated in a randomised trial. The aim of this study was to compare outcomes after extended pleurectomy decortication plus chemotherapy versus chemotherapy alone.
METHODS
MARS 2 was a phase 3, national, multicentre, open-label, parallel two-group, pragmatic, superiority randomised controlled trial conducted in the UK. The trial took place across 26 hospitals (21 recruiting only, one surgical only, and four recruiting and surgical). Following two cycles of chemotherapy, eligible participants with pleural mesothelioma were randomly assigned (1:1) to surgery and chemotherapy or chemotherapy alone using a secure web-based system. Individuals aged 16 years or older with resectable pleural mesothelioma and adequate organ and lung function were eligible for inclusion. Participants in the chemotherapy only group received two to four further cycles of chemotherapy, and participants in the surgery and chemotherapy group received pleurectomy decortication or extended pleurectomy decortication, followed by two to four further cycles of chemotherapy. It was not possible to mask allocation because the intervention was a major surgical procedure. The primary outcome was overall survival, defined as time from randomisation to death from any cause. Analyses were done on the intention-to-treat population for all outcomes, unless specified. This study is registered with ClinicalTrials.gov, NCT02040272, and is closed to new participants.
FINDINGS
Between June 19, 2015, and Jan 21, 2021, of 1030 assessed for eligibility, 335 participants were randomly assigned (169 to surgery and chemotherapy, and 166 to chemotherapy alone). 291 (87%) participants were men and 44 (13%) women, and 288 (86%) were diagnosed with epithelioid mesothelioma. At a median follow-up of 22·4 months (IQR 11·3-30·8), median survival was shorter in the surgery and chemotherapy group (19·3 months [IQR 10·0-33·7]) than in the chemotherapy alone group (24·8 months [IQR 12·6-37·4]), and the difference in restricted mean survival time at 2 years was -1·9 months (95% CI -3·4 to -0·3, p=0·019). There were 318 serious adverse events (grade ≥3) in the surgery group and 169 in the chemotherapy group (incidence rate ratio 3·6 [95% CI 2·3 to 5·5], p<0·0001), with increased incidence of cardiac (30 vs 12; 3·01 [1·13 to 8·02]) and respiratory (84 vs 34; 2·62 [1·58 to 4·33]) disorders, infection (124 vs 53; 2·13 [1·36 to 3·33]), and additional surgical or medical procedures (15 vs eight; 2·41 [1·04 to 5·57]) in the surgery group.
INTERPRETATION
Extended pleurectomy decortication was associated with worse survival to 2 years, and more serious adverse events for individuals with resectable pleural mesothelioma, compared with chemotherapy alone.
FUNDING
National Institute for Health and Care Research (NIHR) Health Technology Assessment programme (15/188/31), Cancer Research UK Feasibility Studies Project Grant (A15895).
Topics: Humans; Female; Male; Pleural Neoplasms; Middle Aged; Aged; Mesothelioma; Treatment Outcome; United Kingdom; Pleura; Mesothelioma, Malignant; Combined Modality Therapy; Adult; Antineoplastic Combined Chemotherapy Protocols; Lung Neoplasms
PubMed: 38740044
DOI: 10.1016/S2213-2600(24)00119-X