Liposarcoma of the pleural cavity.

Liposarcoma rarely occurs in the pleura or thoracic cavity, and few reports appear in the literature. We hypothesized that combining clinicopathologic, immunohistochemical, and fluorescence in situ hybridization methods would allow definite diagnoses. Using formalin-fixed, paraffin-embedded blocks, we examined 6 atypical lipomatous tumor/well-differentiated liposarcomas (ALT/WDLPS), 5 dedifferentiated liposarcomas (DDLPSs), 2 pleomorphic liposarcomas, and 1 myxoid liposarcoma (MLPS). We used the Kaplan-Meier method and the Wilcoxon test for survival analysis for prognostic factor evaluation. Histologically, ALT/WDLPS was composed of a relatively mature adipocytic proliferation, accompanied by some lipoblasts. DDLPS exhibited round-to-oval tumor cells with a high nucleus-to-cytoplasm ratio that had proliferated in nests, accompanied in case 10 by some giant cells but no fatty cells. The pleomorphic type contained a varying proportion of pleomorphic lipoblasts. MLPS displayed uniform round- to oval-shaped cells and small signet-ring lipoblasts in a myxoid stroma. Immunohistochemically, 11 (79%), 11 (79%), and 10 (71%) of 14 cases were positive for S-100, p16, and CDK4, respectively. Six of the 14 cases (43%) were positive for MDM2 and adipophilin. One case of ALT/WDLPS and 3 cases of DDLPS exhibited MDM2 amplification by fluorescence in situ hybridization (Vysis LSI MDM2 SpectrumGreen Probe plus Vysis CEP 12 SpectrumOrange probe). ALT/WDLPS was the most favorable type for survival, while adipophilin tended to be a negative prognostic factor for pleural liposarcoma. For a firm diagnosis of liposarcoma in the pleura, immunohistochemistry for CDK4, MDM2, and adipophilin together with MDM2 gene amplification by fluorescence in situ hybridization may be an important diagnostic tool.

Diagnosing pleural effusions using mass spectrometry-based multiplexed targeted proteomics quantitating mid- to high-abundance markers of cancer, infection/inflammation and tuberculosis.

Pleural effusion (PE) is excess fluid in the pleural cavity that stems from lung cancer, other diseases like extra-pulmonary tuberculosis (TB) and pneumonia, or from a variety of benign conditions. Diagnosing its cause is often a clinical challenge and we have applied targeted proteomic methods with the aim of aiding the determination of PE etiology. We developed a mass spectrometry (MS)-based multiple reaction monitoring (MRM)-protein-panel assay to precisely quantitate 53 established cancer-markers, TB-markers, and infection/inflammation-markers currently assessed individually in the clinic, as well as potential biomarkers suggested in the literature for PE classification. Since MS-based proteomic assays are on the cusp of entering clinical use, we assessed the merits of such an approach and this marker panel based on a single-center 209 patient cohort with established etiology. We observed groups of infection/inflammation markers (ADA2, WARS, CXCL10, S100A9, VIM, APCS, LGALS1, CRP, MMP9, and LDHA) that specifically discriminate TB-PEs and other-infectious-PEs, and a number of cancer markers (CDH1, MUC1/CA-15-3, THBS4, MSLN, HPX, SVEP1, SPINT1, CK-18, and CK-8) that discriminate cancerous-PEs. Some previously suggested potential biomarkers did not show any significant difference. Using a Decision Tree/Multiclass classification method, we show a very good discrimination ability for classifying PEs into one of four types: cancerous-PEs (AUC: 0.863), tuberculous-PEs (AUC of 0.859), other-infectious-PEs (AUC of 0.863), and benign-PEs (AUC: 0.842). This type of approach and the indicated markers have the potential to assist in clinical diagnosis in the future, and help with the difficult decision on therapy guidance.

Survivin is a negative prognostic factor in malignant pleural effusion.

BACKGROUND: Survivin is a well-known member of the inhibitor of apoptosis family, and has been related to increased tumour aggressivity, both in tissue and in pleural fluid. OBJECTIVES: In patients with malignant pleural effusion, we sought to investigate the changes in pleural fluid survivin concentrations induced by talc instillation into the pleural space. Those changes were also examined in relation to pleurodesis outcome and patient survival. METHODS: We investigated 84 patients with malignant pleural effusion who underwent talc pleurodesis. Of them, 32 had breast cancer, 25 lung cancer and 27 had mesothelioma. Serial samples of pleural fluid were obtained before thoracoscopy (baseline) and 24 hours thereafter. RESULTS: Survivin levels were successfully quantified in all pleural fluid samples, and they were significantly higher in samples obtained after thoracoscopic talc poudrage compared with baseline (P < .001). Patients with higher pleural fluid survivin levels at baseline had a significantly poorer pleurodesis outcome (P = .004). A 30 pg/mL cut-off for baseline survivin in pleural fluid predicted failure of pleurodesis with a 54% sensitivity and 79% specificity (P = .009). Moreover, median postpleurodesis survival of patients with baseline survivin levels ≥30 pg/mL was 4 months (range: 0.1-38), compared with 13 months (range: 0.1-259) in patients below that cut-off (P < .001). CONCLUSION: Elevated pleural fluid survivin concentrations are useful to predict failure of pleurodesis and are associated with shorter survival in patients with malignant pleural effusion.

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

BACKGROUND: 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. METHODS: 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). RESULTS: 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. CONCLUSIONS: 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.

[Difficulties in classifying body cavity fluids into transudate/ exudate depending on the various diagnostic criteria]. / Trudnosci w klasyfikacji plynów z jam ciala na przesiek/wysiek na podstawie róznych kryteriów diagnostycznych.

Disease processes may impair the production and reabsorption of fluid from in the body cavities, which results in its excessive accumulation. AIM: The aim of the study was the evaluation of difficulties regarding the classification of fluids from the body cavities into transudate/exudate observing the following: Light's criteria, total fluid protein concentration, and total protein ratio (TP ratio) and lactate dehydrogenase ratio (LDH ratio). MATERIALS AND METHODS: Retrospective analysis was conducted on pleural (N=314), peritoneal (N=114) and pericardial (N=10) fluids, which were tested for the total protein concentration and LDH activity both in fluid and serum and calculated on TP ratio and LDH ratio. RESULTS: Based on the total protein concentration, 278 fluids from pleural cavity were classified as an exudate; 36 as a transudate. Applying the Light's criteria 240 fluids were classified as an exudate; the remaining 74 fluids were classified as a transudate. Based on TP and LDH ratios, 229 fluids from pleural cavity were classified as an exudate; 85 as a transudate. Depending on the total protein concentration, 35 fluids from the peritoneal cavity were classified as an exudate; 79 as a transudate. Applying the Light's criteria 54 fluids were classified as an exudate; the remaining 60 fluids were classified as a transudate. Based on TP and LDH ratios, 22 fluids from peritoneal cavity were classified as an exudate; 92 as a transudate. Analysis of pericardial fluids, depending on the total protein concentration classified 9 of them as an exudate and 1 as a transudate. The same results were obtained by applying Light's criteria. Based on TP and LDH ratios, 7 fluids from pericardial cavity were classified as an exudate; 3 - as a transudate. CONCLUSIONS: Applying the Light's criteria or the total protein concentration in differential diagnostics of fluids from the body cavities resulted in qualification more of them as an exudates as compared to the analysis of the same fluids depending on the TP and LDH ratios. It can be assumed that some of the transudative/exudative fluids were incorrectly classified. Performed analysis suggest that more adequate criteria of the classification of fluids from the body cavities into transudate/exudate are of great importance.

Pleural gas analysis for the identification of alveolopleural fistulae.

PURPOSE OF REVIEW: The method for identification of alveolopleural fistulae (APF) by visual inspection of air bubbles in the chest drainage system has several limitations and suffers from poor accuracy. Here we discuss the use of a novel technique of pleural gas analysis in the identification and management of APF. RECENT FINDINGS: We found that pleural gas analysis has higher sensitivity and specificity than visual inspection in identifying APF. Additionally, we demonstrated that intrapleural gas milieu impacts lung healing and reduction of intrapleural carbon dioxide can promote resolution of APF. SUMMARY: Pleural gas analysis is a novel technique to identify and manage APF. Integration of gas analysis in chest drainage systems would provide a more objective method for managing chest tubes and providing a favorable pleural gas environment for lung healing.

[Value of Cell Block in the Diagnosis of Malignant Pleural Effusion].

Malignant pleural effusion (MPE ) is due tumor which arises from the mesothelium or metastases from tumor origniating other sites. In large, for undiagnosed unilateral pleural effusions, the most frequent and important diagnosis to be established or excluded is malignancy. Cell block is prepared from residual fluid which is centrifuged or is naturally sedimenting to obtain clots at the bottom of the container. The cell block technique is simple, relatively non-invasive, reproducible and has a high yield for malignant plerual effusion. It plays an important role in the diagnosis, guiding the treatment of maligant pleural effusion. Herein, we summarize the technologys which make the cell block, the differential diagnostic value when multiple sections of the cell block are processed for immunhistochemistry, advantages in the diagnosis of malignant pleural effusion, the clinical value of gene screening in cell block. The aim of this article is to discuss the value of cell block in diagnosis of maligant pleural effusion.

Pleural Gas Analysis for Detection of Alveolopleural Fistulae.

PURPOSE: Visual inspection (VI) of bubbles in the chest drainage unit does not differentiate a true leak of alveolopleural fistula (APF) from a false leak. We hypothesized that detection of elevated levels of carbon dioxide, increase in oxygen content, or both, in pleural gas upon the administration of supplemental oxygen would accurately identify APF. DESCRIPTION: Prospective study comparing pleural gas analysis (GA) with VI to detect APF after surgical lobectomy (n = 50). EVALUATION: APF was found in 22 (44%) patients at the time of analysis. VI revealed air bubbles in 31 (62%) patients, indicating the presence of APF, of whom 12 (38.7%) were false leaks. VI failed to identify APF in 3 (6%) patients that resulted in post-tube removal pneumothorax. By contrast, GA accurately demonstrated APF in 21 patients, with only one false negative and no false positives. GA demonstrated better sensitivity (95.5% vs 86.4%), specificity (100% vs 57.1%), positive predictive value (100% vs 61.3%), and negative predictive value (96.6% vs 84.2%) compared to VI. CONCLUSIONS: Pleural gas analysis is an effective technique to detect APF and can facilitate timely and safe chest tube removal.

WGA-Alexa transsynaptic labeling in the phrenic motor system of adult rats: Intrapleural injection versus intradiaphragmatic injection.

BACKGROUND: Intrapleural injection of CTB-Alexa 488, a retrograde tracer, provides an alternative labeling technique to the surgically invasive laparotomy required for intradiaphragmatic injection. However, CTB-Alexa 488 is incapable of crossing synapses restricting the tracer to the phrenic nuclei and the intercostal motor nuclei in the spinal cord. NEW METHOD: Intrapleural injection of WGA-Alexa 488, a transsynaptic tracer, provides a method to label the respiratory motor pathway in both the spinal cord and medulla. Intradiaphragmatic injection of WGA-Alexa 594 and vagal nerve injections of True blue were used to confirm the phrenic nuclei and to differentiate between the rVRG and the NA in the medulla. RESULTS: Following intrapleural injection, WGA-Alexa 488 was retrogradely transported to the phrenic nuclei and to the intercostal motor nuclei. Subsequently WGA-Alexa 488 was transsynaptically transported from the phrenic motoneurons to the pre-motor neurons in the rVRG that provide the descending drive to the phrenic neurons during inspiration. In addition WGA-Alexa 488 was identified in select cells of the NA confirmed by a dual label of both WGA-Alexa 488 and True blue. COMPARISON WITH EXISTING METHOD: WGA-Alexa 488 demonstrates retrograde transsynaptic labeling following intrapleural injection whereas the previous method of injecting CTB-Alexa 488 only demonstrates retrograde labeling. CONCLUSIONS: Intrapleural injection of WGA-Alexa fluor conjugates is an effective method to transsynaptically label the phrenic motor system providing an alternative for the invasive laparotomy required for intradiaphragmatic injections. Furthermore, the study provides the first anatomical evidence of a direct synaptic relationship between rVRG and select NA cells.

Moxifloxacin pharmacokinetics and pleural fluid penetration in patients with pleural effusion.

The aim of this study was to evaluate the pharmacokinetics and penetration of moxifloxacin (MXF) in patients with various types of pleural effusion. Twelve patients with empyema/parapneumonic effusion (PPE) and 12 patients with malignant pleural effusion were enrolled in the study. A single-dose pharmacokinetic study was performed after intravenous administration of 400 mg MXF. Serial plasma (PL) and pleural fluid (PF) samples were collected during a 24-h time interval after drug administration. The MXF concentration in PL and PF was determined by high-performance liquid chromatography, and main pharmacokinetic parameters were estimated. Penetration of MXF in PF was determined by the ratio of the area under the concentration-time curve from time zero to 24 h (AUC24) in PF (AUC24PF) to the AUC24 in PL. No statistically significant differences in the pharmacokinetics in PL were observed between the two groups, despite the large interindividual variability in the volume of distribution, clearance, and elimination half-life. The maximum concentration in PF (CmaxPF) in patients with empyema/PPE was 2.23±1.31 mg/liter, and it was detected 7.50±2.39 h after the initiation of the infusion. In patients with malignant effusion, CmaxPF was 2.96±1.45 mg/liter, but it was observed significantly earlier, at 3.58±1.38 h (P<0.001). Both groups revealed similar values of AUC24PF (31.83±23.52 versus 32.81±12.66 mg·h/liter). Penetration of MXF into PF was similarly good in both patient groups (1.11±0.74 versus 1.17±0.39). Despite similar plasma pharmacokinetics, patients with empyema/parapneumonic effusion showed a significant delay in achievement of PF maximum MXF levels compared to those with malignant effusion. However, in both groups, the degree of MXF PF penetration and the on-site drug exposure, expressed by AUC24PF, did not differ according to the type of pleural effusion.

Serum and pleural fluid N-Terminal-Pro-B-Type natriuretic peptide concentrations in the differential diagnosis of pleural effusions.

Currently, new biomarkers like N-Terminal-Pro-B-Type natriuretic peptide (NT-proBNP) have been used in the differential diagnosis of pleural effusions. In our study, we aimed to investigate the diagnostic value of NT-proBNP, especially in cardiac originated pleural effusions. Forty-five patients with pleural effusions were included in the study. NT-proBNP levels and biochemical markers involved in the Light's criteria were analyzed in pleural fluid and serums of the patients. Pleural fluid culture, AFB smear, cytology were performed where they were indicated according to the clinical evaluation. In patients, to whom cardiac pathology was considered to be; cardiological evaluation and echocardiography were also done. Thirty-eight pleural effusions were exudative and, 7 were transudative according to the Light's criteria. Final diagnosis were malignant effusion in 13, infection (tuberculosis/pneumonia) in 10, congestive heart failure in 21, and other conditions related with pleural effusion in 1 of the patients. Median (25th to 75th percentiles) NT-proBNP levels of serum and pleural fluid due to congestive heart failure (CHF) were 4747 pg/mL (931-15754) and 4827 pg/mL (1290-12.430) while median NT-proBNP levels of serum and pleural fluid related with non-cardiac reasons were 183 pg/mL (138-444) and 245 pg/mL (187-556) respectively. NT-proBNP levels of serum and pleural fluid were significantly high in CHF (p< 0.001 for both). When four groups were compared serum and pleural fluid NT-proBNP levels were highest in the CHF group which was followed by malignancy, infection and others (p< 0.001 for both). Fourteen of 21 patients who were accepted to have congestive heart failure as the final diagnosis by a cardiological evaluation had an exudative pleural fluid according to the Light's criteria. Serum and pleural fluid NT-proBNP levels were higher in transudates and this reached statistically significance for pleural fluid (p= 0.009). We suggest that measurement of pleural fluid NT-proBNP is a smart approach and pleural fluid NT-proBNP can reflect cardiac origin of effusions better than serum NT-proBNP and Light's criteria.

High-sensitivity detection of carbohydrate antigen 15-3 using a gold/zinc oxide thin film surface plasmon resonance-based biosensor.

We report that gold/zinc oxide (Au/ZnO) nanocomposite films were effectively employed to enhance the performance of surface plasmon resonance (SPR) for the detection of tumor markers. Carbohydrate antigen 15.3 (CA15-3), a tumor marker for breast cancer, was chosen as a model analyte. We analyzed intensity response to the samples at various concentrations (0.0125 U/mL to 160 U/mL) in pleural fluid to evaluate the detection capability of the SPR biosensor based on Au/ZnO thin films. The linear range extended from 1 to 40 U/mL with a correlation coefficient of R(2) = 0.991 and a limit of detection reaching 0.025 U/mL at a signal-to-noise ratio of 3:1. Compared with the degree of the shift in SPR intensity induced by the specific binding event between antibody and antigen, the change of intensity on the Au/ZnO layers was increased by at least 2 fold over that on the gold/chromium (Au/Cr) layers. In addition, we determined that the Au/ZnO layers allowed for a detection limit 4 times lower than the Au/Cr layers, which are in widespread use as the sensing interfaces in current SPR-based detectors. In conclusion, the use of Au/ZnO films greatly enhanced the SPR signal yield for this bimolecular interaction and showed high sensitivity.

Protein S-100 is present in extracerebral fluids before and after cardiac surgery in children.

BACKGROUND: Protein S-100 is found in extracerebral sources. The aim of our study was to examine the content of protein S-100 in native pericardial fluid as well as in postoperative extracerebral fluids after cardiac surgery in children. METHODS: We conducted a prospective study in 90 children measuring protein S-100 concentration in pericardial fluid directly after opening the pericardial sac before starting with cardiac surgery. Postoperatively, we examined pleural, peritoneal and pericardial fluid. RESULTS: Pericardial fluid sampled directly after opening the pericardial sac has a protein S-100 content of 3.2 (1.3-5.4) microg/L. Postoperatively, protein S-100 content was 0.89 (0.56-2.6) microg/L in pleural effusion, 0.14 (0.1-1.1) microg/L in peritoneal fluid and 2.75 (2.2-24.4) microg/L in pericardial fluid. The protein S-100 concentration in pericardial fluid before and after cardiac operation did not differ significantly. Pericardial protein S-100 concentrations were significantly higher than pleural and peritoneal protein S-100 concentrations. CONCLUSIONS: Protein S-100 is present in extracerebral fluids before and after cardiac surgery in children. The time point of fluid withdrawal after the operation did not influence the protein S-100 concentration.

Cell-free DNA concentration in pleural fluid and serum: quantitative approach and potential prognostic factor in patients with cancer and pleural effusions.

PURPOSE: The presence of pleural effusions in patients with tumors is often indicative of locally advanced or metastatic disease, and detection of malignancy in effusion samples frequently leads to a disease upstaging. Our purpose was to quantify the DNA in pleural effusion and serum in patients presenting pleural effusion in order to assess the potential prognostic impact. PATIENTS AND METHODS: The DNA level was determined by amplifying hRNase P in paired samples of serum and pleural fluid in 70 consecutive patients with cancer showing pleural effusion. A group of 30 patients without cancer was included. The correlation between serum and pleural DNA was calculated. Survival curves according to serum and pleural DNA were analyzed. RESULTS: Median DNA concentrations were greater in patients with neoplasia than in patients without malignancy: 105 ng/mL versus 40 ng/mL (P = 0.001) in serum samples, respectively; 93 ng/mL versus 21 ng/mL (P = 0.001) in pleural fluids, respectively. A positive correlation between serum and pleural levels was confirmed (r = 0.3; P < 0.05). Median survival time for patients with serum DNA < or = 105 ng/mL was 11.03 months in contrast to only 3.63 months for patients with higher values (P = 0.036). Accordingly, median survival time for patients with pleural DNA < or = 93 ng/mL was 12.3 months versus only 4.63 months in case of higher levels (P = 0.027). CONCLUSION: This study shows that there is a strong correlation between higher levels of free DNA in pleural fluid or serum and malignancy. Survival is worse for patients with higher DNA levels in serum and pleural fluid.

Vascular leakage in severe dengue virus infections: a potential role for the nonstructural viral protein NS1 and complement.

BACKGROUND: Vascular leakage and shock are the major causes of death in patients with dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Thirty years ago, complement activation was proposed to be a key underlying event, but the cause of complement activation has remained unknown. METHODS: The major nonstructural dengue virus (DV) protein NS1 was tested for its capacity to activate human complement in its membrane-associated and soluble forms. Plasma samples from 163 patients with DV infection and from 19 patients with other febrile illnesses were prospectively analyzed for viral load and for levels of NS1 and complement-activation products. Blood and pleural fluids from 9 patients with DSS were also analyzed. RESULTS: Soluble NS1 activated complement to completion, and activation was enhanced by polyclonal and monoclonal antibodies against NS1. Complement was also activated by cell-associated NS1 in the presence of specific antibodies. Plasma levels of NS1 and terminal SC5b-9 complexes correlated with disease severity. Large amounts of NS1, complement anaphylatoxin C5a, and the terminal complement complex SC5b-9 were present in pleural fluids from patients with DSS. CONCLUSIONS: Complement activation mediated by NS1 leads to local and systemic generation of anaphylatoxins and SC5b-9, which may contribute to the pathogenesis of the vascular leakage that occurs in patients with DHF/DSS.

Comparison of surfactant lipids between pleural and pulmonary lining fluids.

Saturated phospholipids (PCs), particularly dipalmitoylphosphatidylcholine (DPPC), predominate in surfactant lining the alveoli, although little is known about the relationship between saturated and unsaturated PCs on the outer surface of the lung, the pleura. Seven healthy cats were anesthetized and a bronchoalveolar lavage (BAL) was performed, immediately followed by a pleural lavage (PL). Lipid was extracted from lavage fluid and then analyzed for saturated, primarily dipalmitoylphosphatidylcholine (DPPC), and unsaturated PC species using high-performance liquid chromatography (HPLC) with combined fluorescence and ultraviolet detection. Dilution of epithelial lining fluid (ELF) in lavage fluids was corrected for using the urea method. The concentration of DPPC in BAL fluid (85.3+/-15.7 microg/mL) was significantly higher (P=0.021) than unsaturated PCs ( approximately 40 microg/mL). However, unsaturated PCs ( approximately 34 microg/mL), particularly stearoyl-linoleoyl-phosphatidylcholine (SLPC; 17.4+/-6.8), were significantly higher (P=0.021) than DPPC (4.3+/-1.8 microg/mL) in PL fluid. These results show that unsaturated PCs appear functionally more important in the pleural cavity, which may have implications for surfactant replenishment following pleural disease or thoracic surgery.