Pleural Effusion Protein Research Articles

The derivation of predictive equations to speculate the post-mortem interval using cases with over 20-mL pleural effusion: A preliminary study

Often, pleural effusion is noted at autopsy when the cause of death is drowning or diseases such as heart, renal and liver failure. Several studies have established a correlation between the concentrations of electrolytes in pleural effusion and the post-mortem interval (PMI) or those concentrations and drowning site. The present study aims to investigate the relationship between the amount of pleural effusion, concentrations of electrolytes and total protein in pleural effusion, by integrated interpretation using various factors such as the deceased's gender, age, cause of death, drowning site, PMI and body temperature. We included 40 cadavers (26 male, 14 female) with >20-mL pleural effusion, which were categorised into four groups as follows: freshwater drowning; brackish water drowning; seawater drowning (drowning group); and not drowning. An equation derived to assess the lung weight revealed that the drowning site affected the lung weight. An equation for the amount of pleural effusion in the drowning group for the first time revealed that the amount of pleural effusion was directly proportional to the PMI. Using an equation to assess the PMI, we could estimate the PMI within 13.0–13.2 h in cases with >20-mL pleural effusion. Despite a small number of cases in the present study, we attained exciting results from the integrated statistical analysis.

The diagnostic role of pentraxin-3 in the differential diagnosis of pleural effusions

Discrimination of pleural effusion etiology is not always easy in clinical practice. Pentraxin-3 (PTX-3) is a new acute- phase protein. The aim of this study was to investigate the role of PTX-3 in the differential diagnosis of pleural effusions. This prospective study enrolled all consecutive patients from two tertiary hospitals who underwent diagnostic or therapeutic thoracentesis. In a cohort of 149 subjects with pleural effusion, including transudates and malignant (MPE), tuberculous (TPE), and parapneumonic effusion (PPE), serum and pleural effusion PTX-3 concentration measurements were performed using ELISA. Serum and pleural effusion protein, lactate dehydrogenase, C-reactive protein (CRP), and adenosine deaminase levels were also assessed. Of these patients, 34 had transudates, 29 had PPE, 63 had MPE, and 23 had TPE. There was a weak correlation between pleural effusion PTX-3 level and serum CRP (P < 0.01). There was a significant difference in pleural PTX-3 levels between the exudative effusion groups (P < 0.01). The median pleural effusion PTX-3 was significantly higher in patients with PPE (11.2 ng/mL, 2–17.8) than MPE (4.7 ng/mL, 1.8–13.9) and TPE (3.1 ng/mL, 2.0–4.1). At a cut-off point of 5.89 ng/mL, PTX-3 had the best discriminatory power for PPE versus other exudative effusions (sensitivity: 86.2%, specificity: 87.7%). The exudative effusion group had a significantly different pleural effusion/serum PTX-3 ratio (P = 0.03). PTX-3 concentration in pleural effusion was elevated without a significant correlation with serum PTX-3 in PPE. These results may suggest that PTX-3 is a local acute-phase reactant and may allow discrimination of PPE from other exudative effusions.

1809P - Development and validation of a deep learning model using biomarkers in pleural effusion for prediction of malignant pleural mesothelioma

Background: Malignant pleural mesothelioma (MPM) is an aggressive asbestos-related disease that is challenging to diagnose. The value of tumour biomarkers in pleural effusion fluid is limited in patients with the disease. We have developed and validated a deep learning model that can predict a diagnosis of MPM using pleural tumour biomarkers and patient characteristics. Methods: This retrospective cohort study included patients who underwent thoracentesis for undiagnosed pleural effusion at a single tertiary medical centre between September 2014 and August 2016. The diagnosis was established by two independent physicians who were blinded to the pleural effusion data. A deep learning model was constructed to differentiate MPM from other diseases and evaluated using biomarkers in pleural effusion (carcinoembryonic antigen, cytokeratin 19 fragment, soluble mesothelin-related peptides, lactate dehydrogenase), total protein in pleural effusion, and patient age and sex as input parameters. Missing data were handled by single imputation. The model consisted of three hidden layers and was trained for 4000 steps. The data were divided into a training set and a test data set and processed using TensorFlow 1.7.0 and Python 3.6 software. The performance of the model was evaluated by accuracy and the area under the receiver-operating characteristic curve (AUROC). Results: Twenty-eight of the 188 patients who underwent thoracentesis were diagnosed to have MPM and divided into a training data set (containing first 150 records with 20 MPM patients) and a test data set (38 records with 8 MPM patients). The accuracy values for the training and test data sets were 0.99 and 0.97, respectively, and the respective AUROC were 1.00 and 0.92. Conclusions: Our deep neural network model had good diagnostic accuracy for MPM and may help in making a definitive diagnosis when there is an indication for invasive pleural biopsy. Legal entity responsible for the study: Hyogo Prefectural Amagasaki General Medical Center. Funding: Has not received any funding. Disclosure: All authors have declared no conflicts of interest.

Analgesic and anti-inflammatory activities and mechanisms of 70% ethanol extract of Zygophyllum macropodum in animals

ObjectiveTo screen the preliminary phytochemical components, to investigate the acute oral toxicity, the analgesic and anti-inflammatory effects, and to analyze inflammatory factors on mice or rats of 70% ethanol extract of Zygopgyllum macropodum aerial parts (ZME). MethodsPreliminary phytochemical screening was carried out by color reaction. Acute oral toxicity was investigated by body weight, relative organ weight and other toxic signs. Acetic acid induced writhing and hot plate test were used to determine analgesic effect. Acetic acid induced vascular permeability and carrageenan induced paw edema were used to confirm anti-inflammation. Protein in pleural effusion, prostaglandin E2 (PGE2) and tumor necrosis factor alpha (TNF-α) in serum of pleuritic rats induced by carrageenan were analyzed to explore the action mechanisms. The test groups received ZME with 100, 300, 600 mg/kg, the positive control with aspirin (ASP) 200 mg/kg for mice, and ZME with 70, 210, 420 mg/kg and ASP 150 mg/kg for rats orally. The control (C) or negative control (NC) groups received 2% Tween 80 of 10 mL/kg orally. ResultsZME contain flavonoids, saponins, phenols and tannins, steroids, terpenoids, fats and oils, glycosides, carbohydrates, and reducing sugar, but no alkaloids. The lethal dose 50% (LD50) of ZME was greater than 2000 mg/kg and no toxic or deleterious effects and death during 14 d. Oral administration 300 and 600 mg/kg of ZME produced analgesic and anti-inflammatory effects significantly (P < 0.05 or P < 0.001) vs NC. It can reduce the writhing number, prolong the heat resisting time, suppress the permeability of the capillary wall increasing, mitigate the paw edema, reduce the content of protein in pleural effusion, and reduce PGE2 and TNF-α in blood. ConclusionsZME possessed analgesic and anti-inflammatory activities which related to inhibit the production of protein, PGE2 and TNF-α. The LD50 of ZME treated orally to mice was greater than 2000 mg/kg.

Proteome profiling reveals novel biomarkers to identify complicated parapneumonic effusions

Patients with pneumonia and parapneumonic effusion (PPE) have elevated mortality and a poor prognosis. The aim of this study was to discover novel biomarkers to help distinguish between uncomplicated PPE (UPPE) and complicated PPE (CPPE). Using an iTRAQ-based quantitative proteomics, we identified 766 proteins in pleural effusions from PPE patients. In total, 45 of these proteins were quantified as upregulated proteins in CPPE. Four novel upregulated candidates (BPI, NGAL, AZU1, and calprotectin) were selected and further verified using enzyme-linked immunosorbent assays (ELISAs) on 220 patients with pleural effusions due to different causes. The pleural fluid levels of BPI, NGAL, AZU1, and calprotectin were significantly elevated in patients with CPPE. Among these four biomarkers, BPI had the best diagnostic value for CPPE, with an AUC value of 0.966, a sensitivity of 97%, and a specificity of 91.4%. A logistic regression analysis demonstrated a strong association between BPI levels > 10 ng/ml and CPPE (odds ratio = 341.3). Furthermore, the combination of pleural fluid BPI levels with LDH levels improved the sensitivity and specificity to 100% and 91.4%, respectively. Thus, our findings provided a comprehensive effusion proteome data set for PPE biomarker discovery and revealed novel biomarkers for the diagnosis of CPPE.

Proteome screening of pleural effusions identifies IL1A as a diagnostic biomarker for non-small cell lung cancer

Non-small cell lung cancer (NSCLC) is a common malignant disease, and in ∼10–20% of patients, pleural effusion is the first symptom. The pleural effusion proteome contains information on pulmonary disease that directly or indirectly reflects pathophysiological status. However, the proteome of pleural effusion in NSCLC patients is not well understood, nor is the variability in protein composition between malignant and benign pleural effusions. Here, we investigated the different proteins in pleural effusions from NSCLC and tuberculosis (TB) patients by using nano-scale liquid chromatography-tandem mass spectrometry (nLC-MS/MS) analysis. In total, 363 proteins were identified in the NSCLC pleural effusion proteome with a low false discovery rate (<1%), and 199 proteins were unique to NSCLC. The proteins in the NSCLC patients’ pleural effusion were involved in cell adhesion, proteolysis, and cell migration. Furthermore, interleukin 1 alpha (IL1A), a protein that regulates tumor growth, angiogenesis, and metastasis, was significantly more abundant in the NSCLC group compared to the TB group, a finding that was validated with an ELISA assay.

Differential proteome profiling of pleural effusions from lung cancer and benign inflammatory disease patients

The pleural effusion proteome has been found containing information that directly reflects pathophysiological status and represents a potential diagnostic value for pulmonary diseases. However, the variability in protein composition between malignant and benign effusions is not well understood. Herein, we investigated the changes of proteins in pleural effusions from lung adenocarcinoma and benign inflammatory disease (pneumonia and tuberculosis) patients by two-dimensional difference gel electrophoresis (2D-DIGE). Twenty-eight protein spots displayed significantly different expression levels were positively identified by MALDI-TOF-MS representing 16 unique proteins. Five identified protein candidates were further validated and analyzed in effusions, sera or tissues. Among them, hemopexin, fibrinogen gamma and transthyretin (TTR) were up-regulated in cancer samples. The effusion concentration of serum amyloid P component (SAP) was significantly lower in lung cancer patients than in benign inflammatory patients, but no differences were found in sera samples. Moreover, a Jumonji C (JmjC)-domain-containing protein, JMJD5, was observed to be down-regulated in malignant effusions, lung cancer tissues and cancer cells. These results shed light on the altered pleural effusion proteins as a useful and important complement to plasma or other routine clinical tests for pulmonary disease diagnosis.

Soluble mesothelin-related protein in pleural effusion from patients with malignant pleural mesothelioma

Malignant pleural mesothelioma (MPM) is a highly aggressive neoplasm primarily arising from surface serosal cells of the pleura and is strongly associated with asbestos exposure. Patients with MPM often develop pleural fluid as initial presentation. However, cytological diagnosis using pleural fluid is usually difficult and has limited utility. A useful molecular marker for differential diagnosis particularly with lung cancer (LC) is urgently needed. The aim of the present study was to investigate the diagnostic value of soluble mesothelin-related protein (SMRP) in pleural fluid. Pleural fluids were collected from 23 patients with MPM, 38 with LC, 26 with benign asbestos pleurisy (BAP), 5 with tuberculosis pleurisy (TP) and 4 with chronic heart failure (CHF), and the SMRP concentration was determined. All data were analyzed by using non-parametric two-sided statistical tests. The median concentration of SMRP in MPM, LC, BAP, TP and CHF were 11.5 (range 0.90-82.80), 5.20 (0.05-36.40), 6.65 (1.45-11.25), 3.20 (1.65-6.50) and 2.03 (1.35-2.80) nmol/l, respectively. The SMRP concentration was significantly higher in MPM than in the other diseases (P=0.001). The area under the ROC curve (AUC) values of the MPM diagnosis was 0.75 for the differential diagnosis from the other groups. Based on the cut-off value of 8 nmol/l, the sensitivity and specificity for diagnosis of MPM were 70.0 and 68.4%, respectively. These results indicate that the SMRP concentration in pleural fluid is a useful marker for the diagnosis of MPM.

Electrolyte analysis of pleural effusion as an indicator of drowning in seawater and freshwater

It is important for forensic pathologists to determine the diagnosis of drowning as well as the site of drowning. In a previous study, we propose that analysis of electrolytes in pleural effusion from rats may be useful for determining whether drowning has occurred in seawater or freshwater. To test this proposal, we measured the concentration of sodium, potassium and chloride ions and total protein in pleural effusion from 40 autopsy cases: 24 involving seawater drowning, 9 freshwater drowning and 7 no drowning. The concentrations of sodium and chloride ions in pleural effusion showed a significant difference between seawater drowning and freshwater drowning. The concentration of potassium ions and total protein showed no difference between each group, although they increased in proportion to the postmortem interval in cases of both seawater and freshwater drowning. These results are almost same as our previous study and, thus, the quantitative analysis of electrolytes in pleural effusion may be useful for determining whether drowning has occurred in seawater or freshwater.

Electrolyte analysis in pleural effusion as an indicator of the drowning medium

In medico-legal autopsies for drowned bodies, the location of drowning needs to be determined. To investigate the usefulness of electrolyte analysis in pleural effusion as an indicator of the location where the deceased has drowned, we determined the concentrations of electrolytes in the pleural effusion of rats drowned in four kinds of water. The concentrations of sodium and chloride ions in the pleural effusion of rats that drowned in seawater were significantly greater than those of rats that drowned in freshwater at both 1 day and 3 days after drowning. The concentration of potassium ions in pleural effusion 1 day after drowning showed no difference between each group, although it then increased from 1 to 3 days after seawater drowning, whereas it decreased from 1 to 3 days after freshwater drowning. The concentration of total protein in pleural effusion increased from 1 to 3 days after drowning, however, there was no significant difference in the concentration of total protein in pleural effusion between each group at either 1 day or 3 days after drowning. These results suggest that analysis of electrolytes in pleural effusion may be useful for determining whether drowning has occurred in seawater or in freshwater.

Clinical significance of immunohistochemical detection of p53 protein in pleural effusion and bronchial specimen.

To explore the usefulness of p53 immunohistochemical staining as a marker of malignancy in the diagnosis of lung cancer. The expression of p53 in 57 samples of pleural effusion and 69 bronchial biopsy specimens was investigated by immunohistochemical method. p53 was well preserved in the samples of pleural effusion. p53 positive rate was 0%(0/27) ,33.3 %(2/6) and 54.2%(13/24) respectively in cytologically benign , suspicious but not conclusively malignant and malignant pleural effusion. Benign pleural effusion referred as tuberculous and other inflammatory effusion. p53 poisitive rate was 0%(0/23) ,14.3 %(1/7) and 35.9 % (14/39) in histologically benign , atypical proliferating and malignant bronchial biopsy sample. The cases with p53 immunoreactivity were proved to have lung cancer after follow-up in cytologically suspicious but not conclusively malignant pleural effusion and histologically atypical proliferating bronchial biopsy sample. p53 immunostaining could be useful as a marker of malignancy in the cytologic examination of pleural effusion and bronchial biopsy sample. The presence of p53 immunoreactivity in these samples is strongly suggestive of malignancy , though its absence does not exclude neoplasms.

The search for tumor-associated proteins in pleural effusions by means of monoclonal antibodies and a dot blot assay.

Liquid moiety of 61 pleural effusions was tested for tumor-associated proteins (TAP) by means of a dot immunobinding (dot blot) assay (DIA) and a panel of monoclonal antibodies (Moabs). The sensitivity of the assay was checked using a purified, serially diluted carcinoembryonic antigen (CEA) preparation and an anti-CEA monoclonal IgG system. The latter was examined using both DIA and enzyme-linked immunosorbent assay ELISA solid phase assays in simulated conditions that mimicked the protein content of effusions. Finally, the results of DIA were compared to the immunohistochemistry carried out on cell sediments from the same effusions with similar Moabs. It was found that the prevalence of several TAPs, including CEA, epithelial membrane antigen (EMA), vimentin, tenascin, and Thomsen Friedenreich antigen, was significantly higher in the malignant effusions than in the nonmalignant ones. A total, larger than 2, of detected TAPs in a given fluid, was found almost exclusively in malignant effusions (p less than 0.0001). The detection limit of the DIA for a CEA was determined at 5 ng/ml, while for the ELISA it was 1 ng/ml. Several TAPs, especially the CEA, could be detected in parallel tests, carried out on the liquid moiety and the cell sediments of malignant effusions. The evaluation of selected TAPs in pleural effusions by dot blot assay may be of clinical value.