Pleural Lavage Fluid Research Articles

Pressurized IntraThoracic Aerosol Chemotherapy (PITAC) directed therapy of patients with malignant pleural effusion and pleural metastasis

Abstract Objectives Pressurized IntraThoracic Aerosol Chemotherapy (PITAC) has been suggested as a new therapy for patients with malignant pleural effusion (MPE) and/or pleural metastasis (PLM). The patients have a poor prognosis with a median survival of 3 to 12 months. We present feasibility, patient safety, and cytological/histological response assessment in PITAC-treated patients with MPE and/or PLM. Methods Patients eligible for PITAC and treated at Odense PIPAC Center were included. PITAC was performed in lateral decubitus or prone position under double-lumen endotracheal tube ventilation to allow exclusion of the lung if necessary. After positioning of the ultrasound-guided trocar, the second trocar is inserted by video-assisted thoracoscopy. MPE was evacuated and measured. Pleural lavage was performed if no or small amounts of MPE were present. MPE or pleural lavage fluid was analyzed cytologically. Visible PLM was biopsied and sent for histology assessment using a four-tiered Thoracic Regression Grading Score (TRGS). After a walkthrough of the safety checklist, the chemotherapy was nebulized followed by 30 min of passive diffusion. The chemotherapy and chemotherapy-saturated air was evacuated through a closed bag and ventilation system. Results We report data on 11 intended PITACs in five patients. Nine PITACs were completed and two PITACs were discontinued due to intraoperative complications or technical reasons. Response evaluation was available in three patients: one showed complete response (TRGS 1) and another stable disease (TRGS 2). Cytology was available from two patients: one showed conversion from malignant to benign. The 30-day mortality was zero. Conclusions PITAC appears to be safe and feasible.

Ventilator-induced Lung Injury Promotes Inflammation within the Pleural Cavity.

Mechanical ventilation contributes to the morbidity and mortality of patients in intensive care, likely through the exacerbation and dissemination of inflammation. Despite the proximity of the pleural cavity to the lungs and exposure to physical forces, little attention has been paid to its potential as an inflammatory source during ventilation. Here, we investigate the pleural cavity as a novel site of inflammation during ventilator-induced lung injury. Mice were subjected to low or high tidal volume ventilation strategies for up to 3 hours. Ventilation with a high tidal volume significantly increased cytokine and total protein levels in BAL and pleural lavage fluid. In contrast, acid aspiration, explored as an alternative model of injury, only promoted intraalveolar inflammation, with no effect on the pleural space. Resident pleural macrophages demonstrated enhanced activation after injurious ventilation, including upregulated ICAM-1 and IL-1β expression, and the release of extracellular vesicles. In vivo ventilation and invitro stretch of pleural mesothelial cells promoted ATP secretion, whereas purinergic receptor inhibition substantially attenuated extracellular vesicles and cytokine levels in the pleural space. Finally, labeled protein rapidly translocated from the pleural cavity into the circulation during high tidal volume ventilation, to a significantly greater extent than that of protein translocation from the alveolar space. Overall, we conclude that injurious ventilation induces pleural cavity inflammation mediated through purinergic pathway signaling and likely enhances the dissemination of mediators into the vasculature. This previously unidentified consequence of mechanical ventilation potentially implicates the pleural space as a focus of research and novel avenue for intervention in critical care.

Can cell-free DNA (cfDNA) in pleural lavage serve as a predictive and prognostic biomarker among surgically treated Stage I-III a nonsmall cell lung cancer (NSCLC)? A pilot study.

The role of cell-free DNA (cfDNA) in operable nonsmall cell lung cancer (NSCLC) is unclear. This study was aimed to evaluate the feasibility for identification of cfDNA in pleural lavage fluid and its correlation with plasma in resectable NSCLCs. Consecutively resected NSCLCs were evaluated for cfDNA levels in preoperative plasma (PLS1), intraoperative pleural-lavage (PLV) and postoperative (at 1 month) plasma sample (PLS2). CfDNA was isolated and measured quantitatively by qPCR in a TaqMan probe-detection approach using the human β-actin gene as the amplifying target. All (n = 34) except one were negative for malignant cells in PLV cytology. CfDNA could be isolated from all the three samples (PLS1, PLV, and PLS2) successfully in each patient. The median cfDNA levels in PLS1, PLV and PLS2 were 118 ng/mL (IQR 61-158), 167 ng/mL (IQR 59.9-179.9) and 103 ng/mL (IQR 66.5-125.4) respectively. The median follow-up was 34.1 months (IQR 25.2-41.6). A significant overall-survival (OS) and disease-free survival (DFS) were recorded for patients with cfDNA level cut-offs at 125, 170, and 100 ng/mL, respectively for PLS1, PLV, and PLS2. Patients with raised cfDNA in PLS1(>125 ng/mL) and PLV(>170 ng/mL) had significantly poorer 2-year OS, p = 0.005 and p = 0.012, respectively. The hazards (OS) were also higher for those with raised cfDNA in PLV (HR = 5.779, 95%CI = 1.162-28.745, p = 0.032). PLV(>170 ng/mL) had increased pleural recurrences (p = 0.021) and correlated significantly with poorer DFS at 2-years (p = 0.001) with increased hazards (HR = 9.767, 95% CI = 2.098-45.451, p = 0.004). Multivariable analysis suggested higher cfDNA in PLV as a poor prognostic factor for both OS and DFS. Among patients with operable NSCLC, it is feasible to identify cfDNA in pleural lavage and correlate PLV cfDNA with pleural recurrences and outcomes.

Pleural inflammatory response, mesothelin content and DNA damage in mice at one-year after intra-pleural carbon nanotube administration

Many in vitro and in vivo studies have shown that exposure to carbon nanotubes (CNTs) is associated with inflammation, oxidative stress and genotoxicity, although there is a paucity of studies on these effects in the pleural cavity. In the present study, we investigated adverse outcomes of pleural exposure to multi-walled CNTs (MWCNT-7, NM-401 and NM-403) and single-walled CNTs (NM-411). Female C57BL/6 mice were exposed to 0.2 or 5 µg of CNTs by intra-pleural injection and sacrificed one-year post-exposure. Exposure to long and straight types of MWCNTs (i.e. MWCNT-7 and NM-401) was associated with decreased number of macrophages and increased number of neutrophils and eosinophils in pleural lavage fluid. Increased protein content in the pleural lavage fluid was also observed in mice exposed to MWCNT-7 and NM-401. The concentration of mesothelin was increased in mice exposed to MWCNT-7 and NM-411. Levels of DNA strand breaks and DNA oxidation damage, measured by the comet assay, were unaltered in cells from pleural scrape. Extra-pleural effects were seen in CNT exposed mice, including enlarged and pigmented mediastinal lymph nodes (all four types of CNTs), pericardial plaques (MWCNT-7 and NM-401), macroscopic abnormalities on the liver (MWCNT-7) and ovaries/uterus (NM-411). In conclusion, the results demonstrate that intra-pleural exposure to long and straight MWCNTs is associated with adverse outcomes. Certain observations such as increased content of mesothelin in pleural lavage fluid and ovarian/uterine abnormalities in mice exposed to NM-411 suggests that exposure to SWCNTs may also be associated with some adverse outcomes.

Difference in carcinogenicities of two different vapor grown carbon fibers with different physicochemical characteristics induced by intratracheal instillation in rats

BackgroundCarbon fibers are high aspect ratio structures with diameters on the submicron scale. Vapor grown carbon fibers are contained within multi-walled carbon tubes, with VGCF™-H commonly applied as a conductive additive in lithium-ion batteries. However, several multi-walled carbon fibers, including MWNT-7, have been reported to induce lung carcinogenicity in rats. This study investigated the carcinogenic potential of VGCF™-H fibers in F344 rats of both sexes with the vapor grown carbon fibers VGCF™-H and MWNT-7 over 2 years. The carbon fibers were administered to rats by intratracheal instillation at doses of 0, 0.016, 0.08, and 0.4 mg/kg (total doses of 0, 0.128, 0.64, and 3.2 mg/kg) once per week for eight weeks and the rats were observed for up to 2 years after the first instillation.ResultsHistopathological examination showed the induction of malignant mesothelioma on the pleural cavity with dose-dependent increases observed at 0, 0.128, 0.64, and 3.2 mg/kg in rats of both sexes that were exposed to MWNT-7. On the other hand, only two cases of pleural malignant mesothelioma were observed in the VGCF™-H groups; both rats that received 3.2 mg/kg in male. The animals in the MWNT-7 groups either died or became moribund earlier than those in the VGCF™-H groups, which is thought related to the development of malignant mesothelioma. The survival rates were higher in the VGCF™-H group, and more carbon fibers were observed in the pleural lavage fluid (PLF) of the MWNT-7 groups. These results suggest that malignant mesothelioma is related to the transfer of carbon fibers into the pleural cavity.ConclusionsThe intratracheal instillation of MWNT-7 clearly led to carcinogenicity in both male and female rats at all doses. The equivocal evidence for carcinogenic potential that was observed in male rats exposed to VGCF™-H was not seen in the females. The differences in the carcinogenicities of the two types of carbon fibers are thought due to differences in the number of carbon fibers reaching the pleural cavity. The results indicate that the carcinogenic activity of VGCF™-H is lower than that of MWNT-7.

Utility of chest imaging in the diagnosis and management of patients with visceral leishmaniasis: A systematic review.

Visceral leishmaniasis remains a deadly parasitic disease with diagnostic complexities. Currently, point-of-care chest imaging is gaining momentum in the diagnosis of infectious diseases. Respiratory symptoms are common in visceral leishmaniasis. Here we aimed to systematically synthesize the evidence on the utility of chest imaging on the diagnosis and management of patients with visceral leishmaniasis. We searched PubMed, Scopus, Web of Science, ScienceDirect, and Google Scholar databases for studies reporting chest imaging findings in patients with visceral leishmaniasis, published in English from database inception to November 2022. We used the Joanna Briggs Institute checklists to evaluate the risk of bias. The protocol of this systematic review was registered with the Open Science Framework: https://doi.org/10.17605/OSF.IO/XP24W. Of 1792 studies initially retrieved, 17 studies with 59 participants were included. Of the 59 patients, 51% (30) had respiratory symptoms and 20% (12) were human immunodeficiency virus co-infected. Chest X-ray, high-resolution computed tomography, and chest ultrasound findings were available for 95% (56), 93% (55), and 2% (1) of the patients, respectively. The most common findings were pleural effusion (20%; 12), reticular opacities (14%; 8), ground-glass opacities (12%; 7), and mediastinal lymphadenopathies (10%; 6). High-resolution computed tomography was more sensitive than chest X-ray and detected lesions that were lost on chest X-ray, 62% (37) versus 29% (17). In almost all cases, regression of the lesions was observed with treatment. Microscopy of pleural or lung biopsy detected amastigotes. Polymerase chain reaction yield was better in pleural and bronchoalveolar lavage fluids. A parasitological diagnosis from pleural and pericardial fluid was possible in AIDS patients. Overall, the risk of bias was low. Visceral leishmaniasis patients frequently had abnormal findings on high-resolution computed tomography. Chest ultrasound is a useful alternative in resource-limited settings to aid in diagnosis and subsequent treatment follow-up, especially when routine tests yield negative results despite clinical suspicion.

Assessment of the toxicity and carcinogenicity of double-walled carbon nanotubes in the rat lung after intratracheal instillation: a two-year study

BackgroundConsidering the expanding industrial applications of carbon nanotubes (CNTs), safety assessment of these materials is far less than needed. Very few long-term in vivo studies have been carried out. This is the first 2-year in vivo study to assess the effects of double walled carbon nanotubes (DWCNTs) in the lung and pleura of rats after pulmonary exposure.MethodsRats were divided into six groups: untreated, Vehicle, 3 DWCNT groups (0.12 mg/rat, 0.25 mg/rat and 0.5 mg/rat), and MWCNT-7 (0.5 mg/rat). The test materials were administrated by intratracheal-intrapulmonary spraying (TIPS) every other day for 15 days. Rats were observed without further treatment until sacrifice.ResultsDWCNT were biopersistent in the rat lung and induced marked pulmonary inflammation with a significant increase in macrophage count and levels of the chemotactic cytokines CCL2 and CCL3. In addition, the 0.5 mg DWCNT treated rats had significantly higher pulmonary collagen deposition compared to the vehicle controls. The development of carcinomas in the lungs of rats treated with 0.5 mg DWCNT (4/24) was not quite statistically higher (p = 0.0502) than the vehicle control group (0/25), however, the overall incidence of lung tumor development, bronchiolo-alveolar adenoma and bronchiolo-alveolar carcinoma combined, in the lungs of rats treated with 0.5 mg DWCNT (7/24) was statistically higher (p < 0.05) than the vehicle control group (1/25). Notably, two of the rats treated with DWCNT, one in the 0.25 mg group and one in the 0.5 mg group, developed pleural mesotheliomas. However, both of these lesions developed in the visceral pleura, and unlike the rats administered MWCNT-7, rats administered DWCNT did not have elevated levels of HMGB1 in their pleural lavage fluids. This indicates that the mechanism by which the mesotheliomas that developed in the DWCNT treated rats is not relevant to humans.ConclusionsOur results demonstrate that the DWCNT fibers we tested are biopersistent in the rat lung and induce chronic inflammation. Rats treated with 0.5 mg DWCNT developed pleural fibrosis and lung tumors. These findings demonstrate that the possibility that at least some types of DWCNTs are fibrogenic and tumorigenic cannot be ignored.

7P Predictive and prognostic value of cell-free DNA in plasma and pleural lavage among surgically treated adenocarcinomas of the lung (ADCL)

Unlike advanced ADCL, the role of cfDNA in operable ADCL is unclear and this study was aimed to evaluate the feasibility for identification of cfDNA in pleural lavage fluid and its correlation with plasma in resectable ADCL.

Pyopneumothorax from coinfection by Trichomonas tenax and Geotrichum capitatum in a child from China: a case report

BackgroundTrichomonas tenax may appear in the oral cavity of humans due to poor dentition or oral hygiene. Pyopneumothorax is a serious complication of lower respiratory tract infections that very rarely can be caused by a trichomonad species in predisposed individuals. We report a rare case of pleurisy due to T. tenax with coinfection by a fungus.Case presentationWe describe a 16-year-old patient with cerebral palsy who presented with severe pyopneumothorax. T. tenax was identified by microscopic examination of the pleural effusion and next-generation sequencing. We also identified Geotrichum capitatum in the pleural effusion and bronchoalveolar lavage fluid cultures. Treatment with voriconazole and metronidazole successfully eliminated these pathogens and relieved the clinical symptoms. A literature review indicated this is the first reported case of pleurisy due to T. tenax with coinfection by a fungus.ConclusionThe rarity of pyopneumothorax caused by T. tenax coinfection with a fungus should not be overlooked in the clinic. These patients should be and treated in a timely manner.

Folliculin haploinsufficiency causes cellular dysfunction of pleural mesothelial cells

Birt–Hogg–Dubé syndrome (BHDS), an autosomal dominant inheritance disease caused by folliculin (FLCN) mutations, is associated with lung cysts and spontaneous pneumothorax. The possibility of FLCN haploinsufficiency in pleural mesothelial cells (PMCs) contributing to development of pneumothorax has not yet been clarified. Electron microscopy revealed exposed intercellular boundaries between PMCs on visceral pleura and decreased electron density around the adherens junctions in BHDS. To characterize cellular function of PMCs in BHDS patients (BHDS-PMCs), during surgery for pneumothorax, we established the flow cytometry-based methods of isolating high-purity PMCs from pleural lavage fluid. BHDS-PMCs showed impaired cell attachment and a significant decrease in proliferation and migration, but a significant increase in apoptosis compared with PMCs from primary spontaneous pneumothorax (PSP) patients (PSP-PMCs). Microarray analysis using isolated PMCs revealed a significant alteration in the expression of genes belonging to Gene Ontology terms “cell–cell adhesion junction” and “cell adhesion molecule binding”. Gene set enrichment analysis demonstrated that CDH1, encoding E-cadherin, was identified in the down-regulated leading edge of a plot in BHDS-PMCs. AMPK and LKB1 activation were significantly impaired in BHDS-PMCs compared with PSP-PMCs. Our findings indicate that FLCN haploinsufficiency may affect the E-cadherin-LKB1-AMPK axis and lead to abnormal cellular function in BHDS-PMCs.

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.

Comparison of the clinical benefits for non-small cell lung cancer patients between different volume of pleural lavage fluid following video-assisted thoracoscopic lobectomy and systematic mediastinal lymph node dissection: study protocol for a randomized controlled trial

BackgroundPleural lavage is regularly performed before closing the chest wall in pulmonary surgeries to prevent pleural implantation of tumor cells and postoperative infection. However, scant data could be found in the literature regarding the optimal regimen for performing pleural lavage. To establish a proper volume of pleural lavage, we herein designed a protocol for a randomized controlled trial.MethodsA total of 400 participants with non-small cell lung cancer undergoing video-assisted thoracoscopic surgery (VATS) lobectomy and systematic mediastinal lymph node dissection (MLND) will be randomly assigned to one of two groups: group A (500 mL pleural lavage fluid) and group B (3000 mL pleural lavage fluid). The primary outcomes include the levels of leukocytes, neutrophils, and inflammatory factors on the first postoperative day. The secondary outcomes include (i) the levels of leukocytes, neutrophils, and inflammatory factors on the second and third postoperative days; (ii) the incidence of postoperative fever on the first, second, and third postoperative days; (iii) the volumes of chest drainage within the first 3 operative days, the duration of drainage, and postoperative hospitalization; and (iv) the incidence of postoperative complications (incision infection, pain, atelectasis, hemorrhage, etc.) and the incidence of pleural effusion requiring thoracic puncture or drainage within 30 days after surgery. The main content of the analysis includes effectiveness and safety analysis. We will perform subgroup analyses to identify potential influence factors.DiscussionAs far as we know, this will be the first randomized controlled trial to compare the clinical outcomes between different volumes of pleural lavage fluid following VATS and MLND. Findings from this trial will determine the appropriate amount of pleural lavage before chest wall closure.Trial registrationThis study was registered with the Chinese Clinical Trial Registry ( on 17 March 2019. ChiCTR 1900021950).

Clinicopathological significance of microRNA-21 in extracellular vesicles of pleural lavage fluid of lung adenocarcinoma and its functions inducing the mesothelial to mesenchymal transition.

BackgroundPre‐resection pleural lavage cytology is useful to predict tumor recurrence and the prognosis of lung cancer patients. Recently, extracellular vesicles (EVs) isolated from effusion specimens have come under the spotlight, and several studies showed that microRNA in EVs is associated with prognosis. MicroRNA‐21 (miR‐21) is a representative onco‐microRNA, and miR‐21 in EVs (EV‐miR‐21) promotes cancer dissemination by inducing mesothelial to mesenchymal transition (MMT) in the peritoneal cavity. In this study, we isolated EVs from pleural lavage fluid and focused on EV‐miR‐21 as a diagnostic factor with a relationship to pleural dissemination.MethodsThe Cancer Genome Atlas dataset comprising of 448 cases of lung adenocarcinoma, tissue microarray of 144 cases of lung adenocarcinoma, and pleural lavage fluid of 41 cases was used to examine miR‐21 expression levels. The function of EV‐miR‐21 was investigated in vitro.ResultsThe miR‐21 expression level in primary sites was associated with a poor prognosis and correlated with pleural invasion of adenocarcinoma. EV‐miR‐21 levels in pleural lavage fluid were associated with positive cytology and pleural invasion in the primary sites, even in cytology‐negative cases. In vitro studies demonstrated that EV‐miR‐21 induces the MMT. Mesothelial cells in the MMT showed functions similar to cancer‐associated fibroblasts, which are an important stromal component in primary sites and disseminated pleural lesions.ConclusionsEV‐miR‐21 in pleural lavage fluid is important as a diagnostic and prognostic factor. Moreover, EV‐miR‐21 induces the MMT, which can form premetastatic niches of dissemination in the pleural cavity.

Prewarming effect of transcutaneous acupoint electrical stimulation preconditioning in patients undergoing elective video-assisted thoracoscopic lobectomy

To observe the prewarming effect of transcutaneous acupoint electrical stimulation (TAES) preconditioning of Dazhui (GV14) and Mingmen (GV4) in patients undergoing elective video-assisted thoracoscopic lobectomy, so as to determine whether TAES can improve intraoperative hypothermia. A total of 80 patients undergoing elective video-assisted thoracoscopic lobectomy were randomly divided into TAES group (40 cases) and control group (40 cases). Before surgery, all the patients were transferred to the fixed area of an anesthetic preparation room by using a surgery cart carrying the same temperature sheets and quilts before surgery. TAES (2 Hz/100 Hz, 20-30 mA) was applied to Dazhui (GV14) and Mingmen (GV4) for 30 min for patients of the TAES group and the same sheet electrodes of EA stimulator were only attached to GV14 and GV 4 without electrical current transmission for patients in the control group. Then, these patients in the two groups were transferred to the operation room and treated by total intravenous anesthesia, and their anesthetic depth was monitored with bispectral index (BIS, between 45-60) and end-tidal carbon dioxide tension (PETCO2, between 30-45 mmHg). The auricular tympanic temperature was monitored, and when the temperature was below 35.5 ℃, forced-air blanket was used to warm the patient as the remedial measure. The same temperature of operation room, surgical drape, infusion solution and pleural lavage fluid were controlled. The patients' body temperature in the preparation room and operation room during surgery, incidence of hypothemia, blood pressure (BP), heart rate (HR), duration of anesthesia, duration of operation, blood loss volume, urine output, total infusion volume, recovery (awaking) time, and chills during recovery were recorded. The body temperature of patients in the TAES group was significantly higher than that in the control group at the time of entering the operation room (P<0.05). The incidence of chills during recovery was obviously lower in the TAES group (3/40,7.5%) than in the control group (7/40, 17.5%, P<0.05), and the recovery time was significantly shorter in the TAES group than in the control group (P<0.05). There were no significant differences between the two groups in the incidence of intraoperative hypothermia, the duration of anesthesia and operation, blood loss volume, urine output, total infusion volume, BP and HR (P>0.05). TAES preconditioning of GV14 and GV4 can produce prewarming effect before anesthesia, shorten the awaking time and reduce the incidence of chills in the recovery period in patients undergoing elective video-assisted thoracoscopic lobectomy.

The Ecto-5'-Nucleotidase/CD73 Inhibitor, α,β-Methylene Adenosine 5'-Diphosphate, Exacerbates Carrageenan-Induced Pleurisy in Rat.

The ecto-5’-nucleotidase (ecto-5’NT/CD73) represents a crucial enzyme for endogenous adenosine generation. Several findings have shown that CD73 plays an important role in regulating vascular permeability and immune cell function. Adenosine 5’-(α,β-methylene)diphosphate (APCP) is a CD73 inhibitor, widely used as pharmacological tool to investigate the role of CD73/adenosine pathway in several in vitro and in vivo models, although it has been also shown to inhibit other ectoenzymes involved in adenosinergic pathway. Here, we evaluated the effect of APCP in the development of inflammation in carrageenan-induced pleurisy model. We found that treatment with APCP (400 µg/rat) significantly increased cell accumulation, exudate formation, and pro-inflammatory cytokine content into the pleural cavity in the acute phase (4 h) of inflammation, with no differences in the sub-acute phase (72 h) except for the regulation of monocyte chemotactic protein-1 levels. In addition, cells collected by pleural lavage fluids of APCP-treated rats, 4 h following carrageenan injection, showed increased ability to migrate in vitro, both in presence and in absence of N-formyl-L-methionyl-L-leucyl-L-phenylalanine as chemotactic stimulus, compared to cells obtained by control rats. Our results demonstrate that APCP exacerbates the early phase of carrageenan-induced pleurisy by controlling pleural effusion and polymorphonuclear migration in vivo and ex vivo. This effect is likely dependent upon CD73 inhibition, although an inhibitory effect of other ectoenzymes cannot be ruled out.

Comparative pulmonary toxicity of a DWCNT and MWCNT-7 in rats

Very little is known about the in vivo toxicity of inhaled double-walled carbon nanotubes (DWCNTs). In the present study, we compared the pulmonary toxicity of DWCNT to MWCNT-7, a well-known multi-walled carbon nanotube. Rats were divided into six groups: untreated, vehicle, low-dose DWCNT, high-dose DWCNT, low-dose MWCNT-7, and high-dose MWCNT-7. The test materials were administered by intra-tracheal intra-pulmonary spraying (TIPS) every other day for 15 days: the low-dose and high-dose groups were administered final total doses of 0.25 and 0.50 mg/rat of the test material. The animals were sacrificed 1 and 6 weeks after the final TIPS administration. Six weeks after the final TIPS administration, rats administered MWCNT-7 had high levels of macrophage infiltration into the lung with dense alveolar wall fibrous thickening throughout the lung; significant elevation of lactate dehydrogenase activity, alkaline phosphatase activity, and total protein concentration in the bronchioalveolar lavage fluid; an increase in the pulmonary cell PCNA index; slightly elevated levels of 8-OHdG DNA adducts in lung tissue DNA; a small but significant increase in protein concentration in the pleural cavity lavage fluid and an increase in the visceral mesothelial cell PCNA index. None of these parameters was increased in rats administered DWCNT. The primary lesion in rats administered DWCNT was scattered formation of granulation tissue containing internalized DWCNT fibers. Our data indicate that DWCNT has lower pulmonary and pleural toxicity than MWCNT-7.

Experimental Model of Human Malignant Mesothelioma in Athymic Mice.

Malignant pleural mesothelioma (MPM) is a thoracic aggressive cancer caused by asbestos exposure, which is difficult to diagnose and treat. Here, we characterized an in vivo orthotopic xenograft model consisting of human mesothelioma cells (designed as H2052/484) derived from a pleural NCI-H2052 tumor injected in partially immunodeficient athymic mice. We assessed tumor formation and tumor-dependent patterns of inflammation. H2052/484 cells conserved their mesothelioma phenotype and most characteristics from the parental NCI-H2052 cells. After intra-thoracic injection of H2052/484 cells, thoracic tumors developed in nearly all mice (86%) within 14 days, faster than from parental NCI-H2052 cells. When the mice were euthanized, the pleural lavage fluid was examined for immune cell profiles. The pleural immune cell population increased with tumor development. Interestingly, the proportion of myeloid-derived suppressor cell and macrophage (especially CD206+ M2 macrophages) populations increased in the pleural fluid of mice with large mesothelioma development, as previously observed in immunocompetent mice. This reliable orthotopic model recapitulates human mesothelioma and may be used for the study of new treatment strategies.

The Significance of SCC and CEA mRNA in the Pleural Cavity After Lymphadenectomy in Esophageal Cancer Patients who Underwent Preoperative Treatment.

This report focuses on the surgical manipulation and spread of cancer cells. Our previous study suggested an association between a poor prognosis and positive pleural lavage cytology after resection of esophageal cancer without preoperative treatment. However, little is known regarding the clinical significance of the lavage procedure in esophageal cancer patients who undergo preoperative treatment. A cohort of 94 patients with squamous cell carcinoma of the esophagus who underwent esophagectomy with radical lymph node dissection was prospectively analyzed for free cancer cells in the pleural cavity after mediastinal lymphadenectomy. Reverse transcription-polymerase chain reaction was performed to detect free cancer cells in the pleural lavage fluid by measuring squamous cell carcinoma-related antigen (SCC) and carcinoembryonic antigen (CEA). Forty-two patients (44.7%) were positive for SCC after thoracic lymphadenectomy, and 15 patients (15.9%) were positive for CEA. SCC positivity was significantly associated with venous invasion (p=0.037) and with the clinical response to preoperative treatment (p=0.001). Furthermore, SCC positivity was associated with poor prognosis compared with negative SCC (p=0.026). Multivariate analysis revealed that SCC positivity was an independent prognostic factor. Regarding recurrence patterns, SCC positivity tended to be associated with hematogenous recurrence (p=0.063). Conversely, positive CEA was not associated with any clinicopathological finding, treatment response, prognosis, or recurrence pattern. Tumor spillage during the evaluated surgical manipulation was assessed in esophageal cancer patients who underwent preoperative treatment. Tumor spillage as evaluated by SCC mRNA was associated with a poor prognosis.

The prognostic value of carcinoembryonic antigen levels in blood and intraoperative pleural lavage fluid in non-small-cell lung cancer.

IntroductionThere is no specific marker for lung cancer, but, in some lung cancer types, carcinoembryonic antigen (CEA) can reach high levels in the blood and pleural fluid.AimThis study investigated the relationship of CEA levels in blood (CEAB) and intraoperative pleural lavage fluid (CEAP) in non-small-cell lung cancer (NSCLC) with the type, stage, and extent of lung cancer.Material and methodsA total of 50 patients, who underwent surgery at our clinic due to NSCLC (group I) or benign lung pathology (group II), were assessed. For this prospectively designed study, 25 consecutive patients were included in each group, and their CEAB and CEAP levels were investigated.ResultsWhen the levels of CEAP were compared, the average value of group I (1.35 ng/ml) was significantly higher than the average value of group II (0.04 ng/ml) (p = 0.027). When CEA levels were examined separately, and average values were taken according to surgical pathology results, both CEAB and CEAP levels of adenocarcinoma patients were found to be higher than those of the other groups. This difference was only significant for the level of CEAP (p = 0.026).ConclusionsAlthough the average CEAB levels of patients with adenocarcinoma were higher than those of patients with other histopathological types, this difference was not statistically significant. However, we found that CEAP levels were significantly higher in patients with adenocarcinoma. These results have led us to consider that CEAP elevation is a more sensitive marker than the elevation of CEAB.

Pleural inhibition of the caspase-1/IL-1β pathway diminishes profibrotic lung toxicity of bleomycin.

BackgroundIdiopathic and toxic pulmonary fibrosis are severe diseases starting classically in the subpleural area of the lung. It has recently been suggested that pleural mesothelial cells acquire a myofibroblast phenotype under fibrotic conditions induced by TGF-β1 or bleomycin. The importance and role of inflammation in fibrogenesis are still controversial. In this work, we explored the role of IL-1β/caspase-1 signaling in bleomycin lung toxicity and in pleural mesothelial cell transformation.MethodsC57BL/6 mice were intravenously injected with either bleomycin or nigericin or NaCl as control. In vitro, the Met5A cell line was used as a model of human pleural mesothelial cells.ResultsIntravenous injections of bleomycin induced lung fibrosis with histologically-proven peripheral distribution, collagen accumulation in the pleural and subpleural area, and overexpression of markers of myofibroblast transformation of pleural cells which migrated into the lung. These events were associated with an inflammatory process with an increase in neutrophil recruitment in pleural lavage fluid and increased caspase-1 activity. TGF-β1 was also overexpressed in pleural lavage fluid and was produced by pleural cells following intravenous bleomycin. In this model, local pleural inhibition of IL-1β with the IL-1β inhibitor anakinra diminished TGF-β1 and collagen accumulation. In vitro, caspase-1 inhibition interfered with Met5A cell transformation into the myofibroblast-like phenotype induced by bleomycin or TGF-β1. Moreover, nigericin, a caspase-1 activator, triggered transformation of Met5A cells and its intra-pleural delivery induced fibrogenesis in mice.ConclusionsWe demonstrated, after intravenous bleomycin injection in mice, the role of the pleura and highlighted the key role of IL-1β/caspase-1 axis in this fibrogenesis process.Electronic supplementary materialThe online version of this article (doi:10.1186/s12931-016-0475-8) contains supplementary material, which is available to authorized users.