Effusion Samples Research Articles

Complement activation in pediatric patients with recurrent acute otitis media

ObjectiveOtitis media (OM) is one of the most common childhood diseases. The relative contribution of complement activation in protection and pathogenesis during OM remains largely unknown. The purpose of this study was to investigate the beneficial and pathogenic contributions of complement activation in the middle ear of pediatric patients with recurrent acute otitis media (rAOM), and therefore to provide a rational approach to prevent sequelae of OM such as hearing loss. MethodsTwenty children undergoing pressure equalization tube placement with or without adenoidectomy for rAOM were enrolled in the study. Bacterial cultures, enzyme-linked immunosorbent assay (ELISA) for complement components and cytokines and western blot for complement activation were performed on middle ear effusion (MEE) and serum samples. The levels of complement C3a, C5a and sC5-b9 in MEEs and serum samples were compared. The levels of these factors were also examined in regards to length of episode. Pearson's correlation coefficients were calculated on variables between C5a and IL-6 or IL-8. Complement gene expression in human middle ear epithelial (HMEE) cells induced by otopathogens was evaluated. Data were analyzed with Student's t test or the Mann–Whitney rank sum test. In all cases, a P value of <0.05 was set as the measure of significance. ResultsOur data demonstrated that the complement classical/lectin, alternative and terminal pathways were activated in the middle ear of children with rAOM. Increased complement components of C3a, C5a and sC5-b9 in MEEs were detected in patients with the episode lasting more than six weeks. There was a strong correlation between C5a and IL-6 or IL-8 in the MEEs. Additionally, otopathogens induced enhanced gene expression of factor B and C3 in HMEE cells, which is beneficial for host defense against invading pathogens. ConclusionOur studies provided important new insights on how complement activation contributes to inflammatory process during rAOM. Knowledge of the activity of the complement pathway in patients with rAOM may stimulate the development of new strategies to prevent middle ear inflammatory tissue destruction by directing treatment to specific pathways within the complement cascade.

Diagnostic Value of Superoxide Dismutase in Tuberculous and Malignant Pleural Effusions

The aim of this study was to investigate the diagnostic value of superoxide dismutase (SOD) in tuberculous pleural effusions (TPEs) and malignant pleural effusions (MPEs). Pleural effusion (PE) samples from 100 patients were classified on the basis of diagnosis as TPE (n=57) and MPE (n=43). The activity of SOD was determined by pyrolgallol assay. A significant difference was observed in SOD activity (P<0.01) between TPE and MPE, levels of being significantly higher in TPE compared to MPE. With a threshold value of 41 U/L, the area under the ROC curve was 0.653, SOD had a sensitivity of 61.4% and a specificity of 61.0% for differential diagnosis. Thus, SOD activity in PE was not a good biomarker in differentiating TPE and MPE. To the best of our knowledge, five SOD isoforms may be present in PE. Identification of which SOD contributes to the difference of SOD level between TPE and MPE is very important for illustrating mechanisms and improving the differential diagnostic value.

Efficacy of two fluorescence in situ hybridization (FISH) probes for diagnosing malignant pleural effusions

It is difficult to differentiate tumor cells in pleural fluid from reactive benign mesothelium. Fluorescence in situ hybridization (FISH) can increase diagnostic accuracy. Two hundred pleural fluid samples were analyzed by using FISH probes for chromosomes 11 and 17. Histological analysis was used to diagnose cancer. Clinical, radiological, and histological data were used to exclude malignancy. Eighty-two pleural effusion samples had positive cytology, 51 were benign, and 67 were atypical, but inconclusive. The 82 positive cases were confirmed to be malignant. Among the 51 negative cytology cases, videothoracoscopy-guided pleural biopsy revealed malignancy in three; aneuploid cells were detected by FISH in all cases. In 43 of the 67 cases with inconclusive cytology, malignancy was confirmed based on histology and fluorescence in situ hybridization. One case of parapneumonic effusion with no evidence of cancer during clinical follow-up had a suspicious cytology and positive fluorescence in situ hybridization result. The remaining 23 cases had no histological, radiological, clinical, or genetic evidence of malignancy. This study demonstrated that cytogenetic analysis of fresh pleural fluid samples using only two FISH probes is a valuable ancillary method for the identification of malignant pleural effusion, particularly in cases in which oncotic cytology is inconclusive.

Downregulation of cell‐free miR‐198 as a diagnostic biomarker for lung adenocarcinoma‐associated malignant pleural effusion

Circulating cell-free microRNAs (miRNAs) are potential cancer biomarkers. The aim of this study was to identify miRNAs that are differentially expressed between benign pleural effusion (BPE) and lung adenocarcinoma-associated malignant pleural effusion (LA-MPE). The expression level of cell-free miRNA was investigated in 107 patients with pleural effusion. Microarrays were used to screen 160 miRNAs in a discovery set comprising 20 effusion samples (ten BPEs and ten LA-MPEs). Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to validate the profiling results obtained for the discovery set and those obtained for a validation set comprising 42 BPEs and 45 LA-MPEs. The area under the receiver operating characteristic curve (AUC) was used to evaluate the diagnostic performance of the identified miRNAs and other common tumor markers, such as carcinoembryonic antigen (CEA) and cytokeratin fragment (CYFRA) 21-1. Microarray profiling showed that miR-198 was significantly downregulated in LA-MPE compared with BPE (p = 0.002). The miRNA microarray analysis results were confirmed by qRT-PCR (p < 0.001) using the validation set. The AUCs for miR-198, CEA and CYFRA 21-1 in the validation set were 0.887, 0.898 and 0.836, respectively. The diagnostic performance of miR-198 was comparable with that of CEA, but better than that of CYFRA 21-1. The AUC for all three markers combined was 0.926 (95% confidence interval, 0.843-0.973) with a sensitivity of 89.2% and a specificity of 85.0%. The present study suggests that cell-free miR-198 from patients with pleural effusion might have diagnostic potential for differentiating LA-MPE from BPE.

Abstract PR5: Intravital multiphoton imaging reveals multicellular streaming as a crucial component of in vivo cell migration in human breast tumors

Abstract Metastasis is the main cause of death in breast cancer patients. Cell migration is an essential component of almost every step of the metastatic cascade, especially the early step of invasion inside the primary tumor. We have used intravital multiphoton microscopy to visualize the in vivo migration and invasion of human breast tumor cells in live primary tumors. We used two different tumors for this study: a. orthotopic xenografts of the highly metastatic breast cancer cell line MDA-MB-231, and b. low passage orthotopic xenografts of breast tumor cells isolated from a patient pleural effusion sample (TN1). The TN1 tumors were only passaged in vivo and never cultured. Both tumor cells were engineered to stably express GFP (green fluorescence protein) and were visualized in vivo based on their fluorescence emission. Collagen I matrix was visualized by second harmonic generation, while injected fluorescent dyes were used for visualization of blood vessels and stromal cells in the primary tumors. High-resolution 4D imaging of these two human tumors in vivo showed that they both shared common patterns of high-speed migration: a. cells moving as single entities irrespective of their neighboring cells, and b. multiple cells following each other in the same direction and in a single file but without cohesive cell junctions (i.e. multicellular streaming). We found that although the two tumors were derived from diverse genetic backgrounds, they exhibited almost identical proportions of either single or streaming cells, suggesting our observations may be generally applied to human breast cancer in vivo cell migration. Cells moving as multicellular streams were overall more frequent in both tumors, with approximately 60% of total motile cells moving as streams. Critically, we found that only multicellular streaming and not random migration of single cells was significantly associated with proximity to blood vessels. Macrophages were co-migrating with the tumor cells in the multicellular streams in both human tumors analyzed, and functional inhibition of macrophages in vivo by clodronate liposomes significantly decreased the frequency of multicellular streams, while increasing the frequency of single moving cells. Macrophage inhibition also significantly abrogated intravasation, as measured by the number of circulating tumor cells in the bloodstream of the treated mice. Since we found that macrophage inhibition reduced streaming migration while enhancing single cells migration, we deduce that multicellular streaming is the type of in vivo migration that most likely directly contributes to intravasation in human breast tumors. Finally, we analyzed the gene expression profile of the migratory tumor cells from both the MDA-MB-231 and the TN1 tumors and found that they both exhibited multiple gene expression changes in genes regulating cell motility. These gene expression changes were largely common in the two tumors and coordinated to regulate the basic actin motility machinery, leading to the same end phenotype of activated actin polymerization and enhanced migration. Our data is the first direct visualization of in vivo migration of live patient-derived breast tumor cells and the first assessment of the type of cell motility involved in tumor cell dissemination in a human tumor cell derived breast tumor. This abstract is also presented as Poster A51. Citation Format: Antonia Patsialou, Jose Javier Bravo-Cordero, Yarong Wang, Huiping Liu, Michael F. Clarke, John S. Condeelis. Intravital multiphoton imaging reveals multicellular streaming as a crucial component of in vivo cell migration in human breast tumors. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Invasion and Metastasis; Jan 20-23, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;73(3 Suppl):Abstract nr PR5.

High pneumococcal serotype specific IgG, IgG1 and IgG2 levels in serum and the middle ear of children with recurrent acute otitis media receiving ventilation tubes

Recurrent acute otitis media (AOM), frequently caused by Streptococcus pneumoniae, is a major paediatric health problem. A reduced antibody response against pneumococcal polysaccharides may contribute to an increased susceptibility to AOM. Using a multiplex bead-based assay we measured IgG, IgG1 and IgG2 levels against 11 pneumococcal polysaccharides in serum samples from 166 children below 3 years of age with a history of at least 3 episodes of acute otitis media receiving ventilation tubes, and 61 healthy controls. Pneumococcal serotype specific IgG was also determined in 144 middle ear effusion samples. Pneumococcal serotype specific IgG, IgG1 and IgG2 levels were similar in children with or without AOM, except for IgG and IgG1 levels against serotype 5, which were significantly higher in children with a history of frequent AOM (IgG: 137.5μg/ml vs. 84.0μg/ml; p=0.02; IgG1: 24.5μg/ml vs. 18.2μg/ml; p=0.05). The age-related development of pneumococcal serotype-specific IgG, IgG1 and IgG2 levels was similar in children with or without a history of AOM. Pneumococcal serotype specific IgG was present in middle ear effusion and these levels correlated significantly with serum titres. Children with a history of frequent AOM receiving ventilation tubes do not have a deficient IgG, IgG1 or IgG2 response against pneumococcal polysaccharides, either induced by vaccination or due to natural exposure. The strong correlation between IgG levels in serum and the middle ear suggests parenteral pneumococcal conjugate vaccination induces antibodies in the middle ear which may therefore contribute to reducing the burden of AOM.

Determination of Three Aromatic Amino Acids in Pleural Effusion by HPLC and Its Application to Differential Diagnosis

A HPLC method was developed for the simultaneous determination of tyrosine,phenylanine and tryptophan in pleural effusion and applied to the differential diagnosis of pleural effusion causes.Pleural effusion was employed for the experimental conditions optimizing and the results were as follows: using VertexTM AQ C18(250 mm×4.6 mm,5 μm) as separation column,acetonitrile-10 mmol/L ammonium acetate(adjust pH to 5.6 with acetic acid) as the mobile phase with a gradient elution at the flow rate of 1.0 mL/min and detection wavelength at 210 nm and 278 nm.The result showed that good linear relationship was achieved between the chromatographic peak area and the concentration of each component.The linear correlation coefficient of these three substances was above 0.999.The relative standard deviation of seven parallel injections was less than 3.2%.The limits of detection(LOD) of these three substances were 0.50,0.17 and 0.20 μmol/L,respectively.Pleural effusion samples were precipitated with methanol,then centrifuged to remove protein residue and finally analyzed by HPLC.The recoveries were ranged from 80.7% to 101.1%,the relative standard deviation of five parallel injection was less than 8.7%.The method was applied to the analysis of 72 patients with lung cancer(20 patients) and benign lung diseases(52 patients).The results indicated that the difference of these three amino acids in different patients was statistically significant and had certain clinical value in differential diagnosis of pleural effusion cause.

Diagnostic Value and Prognostic Significance of Pleural C-Reactive Protein in Lung Cancer Patients with Malignant Pleural Effusions

PurposeC-reactive protein (CRP) has been implicated in various inflammatory and advanced malignant states. Increased serum CRP (s-CRP) levels have been shown to be associated with independent prognostic factors for survival in patients with advanced lung cancer. However, only few studies have focused on the role of CRP in pleural effusions. This study aimed to evaluate the diagnostic and prognostic value of pleural CRP (p-CRP) in lung cancer patients with malignant pleural effusion (MPE).Materials and MethodsPleural effusion (PE) samples were collected from patients with MPE (68 lung cancers; 12 extrathoracic tumors), and from 68 patients with various benign conditions (31 with pneumonia; 37 with tuberculosis). Concentrations of p- and s-CRP were measured by enzyme-linked immunosorbent assay. CRP level in pleural fluid and its association with survival were examined.Resultsp-CRP levels correlated with s-CRP levels (r=0.82, p<0.0001). For the differential diagnosis of MPE and benign PE, the area under the receiver operating characteristic curve was greater for p-CRP (0.86) than for s-CRP (0.77). High p-CRP expression significantly correlated with shorter overall survival (p=0.006). P-CRP was independent prognostic factor significantly associated with overall survival on multivariated analysis (p=0.0001). The relative risk of death for lung cancer patients with high p-CRP levels was 3.909 (95% confidence interval, 2.000-7.639).ConclusionP-CRP is superior to s-CRP in determining pleural fluid etiology. Quantitative measurement of p-CRP might be a useful complementary diagnostic and prognostic test for lung cancer patients with MPE.

Real-Time Polymerase Chain Reaction Versus Conventional PCR: A Comparison between two Methods for the Detection of Fusobacterium Nucleatum in Saliva, Nasopharyngeal Secretion and Middle Ear Effusion Samples

ABSTRACTOral and middle ear infections are biofilm related infections. Fusobacterium nucleatum is considered to be a key species in dental biofilm formation. The aim of this study was to compare conventional and real-time PCR, for the detection of F. nucleatum in saliva, nasopharyngeal secretion and middle ear effusion samples. Sixty samples were collected from 20 patients with otitis media with effusion. Species-specific 16S rRNA primers were used for conventional PCR and species-specific primers and TaqMan probes were used for real-time PCR analysis. Real-time PCR had a better performance than conventional PCR in detecting bacteria in the analyzed samples. The tested real-time PCR assay can identify and quantify F. nucleatum in biofilm samples with a high degree of sensitivity when compared with conventional PCR techniques.

A decrease in galectin-1 (Gal-1) levels correlates with an increase in anti-Gal-1 antibodies at the synovial level in patients with rheumatoid arthritis

Background: Several studies have confirmed that galectin-1 (Gal-1) plays a role in controlling the immune response because of its pro-apoptotic effect. Although studies based on a rheumatoid arthritis (RA) mouse model have suggested a crucial role for Gal-1 in inflammation, clinical data are lacking. We have detected the presence of autoantibodies against galectins in blood, but their physiological meaning remains unknown.Objectives: To compare plasma and synovial levels of Gal-1 in RA patients and in healthy controls, and correlate them with clinical parameters.Methods: Plasma and synovial (non-arthritic knee effusion) samples were collected from RA patients and healthy donors. All patients were receiving treatment with steroids and/or disease-modifying anti-rheumatic drugs (DMARDs). A blood sample was taken at a baseline visit to determine plasma anti-cyclic citrullinated peptide (anti-CCP) antibodies, tumour necrosis factor alpha (TNF-α), Gal-1, and anti-Gal-1 autoantibodies.Results: Although plasma levels of Gal-1 were similar in patients and controls, the concentration of Gal-1 was significantly reduced in the synovial fluid of patients with RA. This reduction was not correlated with TNF-α or C-reactive protein (CRP) levels. However, the decrease in synovial Gal-1 correlated with a significant increase in anti-Gal-1 autoantibodies and anti-CCP antibody titres, suggesting a physiological effect of autoantibodies limiting the amount Gal-1 and potentially blocking its biological effect in RA patients.Conclusion: Gal-1 levels were significant reduced at the synovial level in RA patients, possibly as a consequence of the increase in anti-Gal-1 autoantibodies.

Subclassification of lymphoproliferative disorders in serous effusions

Rare studies have reported the application of multiple ancillary tests to the diagnosis of lymphoproliferative disorder in serous effusions. In the current study, the authors evaluated the effectiveness of using an algorithm for the triage of serous effusions and the contribution of ancillary studies to achieve a specific subtype of lymphoproliferative disorder. Serous effusion samples that had a final diagnosis of lymphoproliferative disorder or suspicious for lymphoma were selected from cases that were diagnosed between 2001 and 2010. Data were collected on patient and sample characteristics as well as results from immunophenotype and molecular studies. In total, 168 serous effusions were identified from 110 patients. The most common site of involvement was the pleural cavity (n = 133) followed by the peritoneal cavity (n = 30) and pericardial cavity (n = 5). The volume of serous effusions ranged from 2 mL to 1000 mL (mean, 238 mL). In 42 patients (38.2%), serous effusions were the primary source of diagnosis. In 129 patients who had a diagnosis of LPD, either generic (n = 82) or specific (n = 47) ancillary tests were performed as a single test in 58 samples (67.4%) or as a combination of multiple studies in 19 samples (23.2%). Immunophenotyping was successful in almost all samples that had a specific subtype with 16 B-cell and 4 T-cell lymphomas being diagnosed. More samples with a specific subtype of lymphoma underwent molecular tests compared with those who had a generic diagnosis (19.1% vs 13.4%). Successful, specific subtyping of lymphoproliferative disorders was achieved in approximately 33% of cases that were tested for ancillary studies following an approach for the triage and aliquoting of serous effusion samples.

Testing for HER2 in Breast Cancer: Current Pathology Challenges Faced in Canada

This review is designed to highlight several key challenges in the diagnosis of human epidermal growth factor receptor 2 (her2)-positive breast cancer currently faced by pathologists in Canada: Pre-analysis issues affecting the accuracy of her2 testing in non-excision sample types: core-needle biopsies, effusion samples, fine-needle aspirates, and bone metastasesher2 testing of core-needle biopsies compared with surgical specimensCriteria for retesting her2 status upon disease recurrenceLiterature searches for each topic were carried out using the medline, Embase, International Pharmaceutical Abstracts, and biosis databases. In addition, the congress databases of the American Society of Clinical Oncology (2005-2011) and the San Antonio Breast Cancer Symposium (2007-2011) were searched for relevant abstracts.All authors are expert breast pathologists with extensive experience of her2 testing, and several participated in the development of Canadian her2 testing guidelines. For each topic, the authors present an evaluation of the current data available for the guidance of pathology practice, with recommendations for the optimization or improvement of her2 testing practice.

Adenocarcinoma cells in effusion cytology as a diagnostic pitfall with potential impact on clinical management: A case report with brief review of immunomarkers

Distinguishing metastatic carcinoma cells from reactive mesothelial cells in effusion samples is often challenging based on morphology alone. Metastatic carcinoma cells in fluid samples may mimic reactive mesothelial cells due to overlapping cytological features. We report a case of a pleural effusion in a 51-year-old female patient with a medical history significant for bilateral ovarian tumors and peritoneal implants diagnosed as serous tumor of borderline malignant potential. The effusion was composed almost entirely of adenocarcinoma cells that morphologically mimicked reactive mesothelial cells. The diagnosis of metastatic adenocarcinoma was made after a wide immunostaining panel of antibodies. Recognizing metastatic adenocarcinoma cells in effusion samples can be challenging and an accurate diagnosis may have significant impact on clinical management as demonstrated by this case.

Viral and bacterial pathogens identification in children hospitalised for severe pneumonia and parapneumonic empyema.

Pneumonia is caused by respiratory bacteria and/or viruses. Little is known if co-infections are an aggravating factor in hospitalised children with severe pneumonia. We studied the impact of respiratory pathogens on the severity of pneumonia. Between 2007 and 2009, 52 children hospitalised with a well-documented diagnosis of community-acquired pneumonia (CAP), with or without parapneumonic empyema (PPE), were enrolled in the study. The patients were classified into 2 groups: CAP + PPE (n = 28) and CAP (n = 24). The identification of respiratory viruses and bacteria in nasopharyngeal aspirates and pleural effusion samples were performed using conventional bacterial techniques and molecular assays. Using real-time multiplex PCR and antigen detection, Streptococcus pneumoniae was the main agent identified in 76% of the cases by molecular tests and BinaxNOW® in pleural fluid. A total of 8% of pleural fluid samples remained undiagnosed. In nasopharyngeal aspirates, rhinovirus, parainfluenza viruses, human metapneumovirus, and respiratory syncytial virus were detected in both CAP and CAP + PPE populations; however, the percentage of viral co-detection was significantly higher in nasopharyngeal aspirates from CAP + PPE patients (35%) compared with CAP patients (5%). In conclusion, viral co-detection was observed mainly in patients with more severe pneumonia. Molecular biology assays improved the pathogens detection in pneumonia and confirmed the S. pneumoniae detection by BinaxNOW® in pleural effusion samples. Interestingly, the main S. pneumoniae serotypes found in PPE are not the ones targeted by the heptavalent pneumococcal conjugate vaccine.

Potential implication of new torque teno mini viruses in parapneumonic empyema in children

An unexplained increase in the incidence of parapneumonic empyema (PPE) in pneumonia cases has been reported in recent years. The present study investigated the genetic and biological specifications of new isolates of torque teno mini virus (TTMV) detected in pleural effusion samples from children hospitalised for severe pneumonia with PPE.A pathogen discovery protocol was applied in undiagnosed pleural effusion samples and led to the identification of three new isolates of TTMV (TTMV-LY). Isolated TTMV-LY genomes were transfected into A549 and human embryonic kidney 293T cells and viral replication was assessed by quantitative real-time PCR and full-length genome amplification. A549 cells were further infected with released TTMV-LY virions and the induced-innate immune response was measured by multiplex immunoassays.Genetic analyses of the three TTMV-LY genomes revealed a classic genomic organisation but a weak identity (<64%) with known sequences. We demonstrated the in vitro replication of TTMV-LY in alveolar epithelial cells and the effective release of infectious viral particles. We also showed a selective production of inflammatory mediators in response to TTMV infection.This study reports the description of replicative TTMV-LY isolated from parapneumonic effusions of children hospitalised with PPE, suggesting a potential role of the virus in the pathogenesis of pneumonia.

The negative enrichment by immunomagnetic beads for tumor cells from malignant pleural effusions

To establish a method (negative enrichment by immunomagnetic beads) for detection of tumor cells in pleural effusions and to evaluate the sensitivity and specificity of the method for clinical application. Five, 10, 20, 50 and 100 A549 (lung adenocarcinoma) cells were labeled with DAPI and added into 20 ml pleural effusions [containing (1 - 10)×10(6)cells] from heart failure patients, followed by immunomagnetic negative enrichment method. Recovered cancer cells were enumerated using a fluorescent microscope. Tumor cells were enriched from pleural effusion samples by means of density gradient centrifugation and negative enrichment by immunomagnetic beads method, followed by identification with cytology analysis (Wright's Giemsa's staining), immunofluorescence staining (IF) and fluorescence in situ hybridization (FISH) using centromere DNA probes of chromosome 7 and 8. Cytology, IF and FISH evaluations were performed in 53 pleural effusion samples, including 36 cases of malignant disease (25 male and 11 female patients aging 40 to 78 years, mean age (63 ± 9) and 17 cases of benign disease (8 male and 9 female patients aging 25 to 81 years, mean age (53 ± 18). After DAPI staining and mixing with pleural effusions from heart failure patients, the cell recovery rates of A549 cells evaluated under fluorescence microscope were 75%, 78%, 82%, 85%, 88%, and the average recovery rate was 81.6%. Using negative enrichment method and density gradient centrifugation combined with cytology analysis, the positive rates of tumor cells in 36 malignant pleural effusion samples were 81% (29/36) and 61% (22/36), respectively (χ(2) = 4.00, P = 0.039). Using negative enrichment method combined with IF, the positive rate of CK18(+), DAPI(+), CD(45)(-) cells was 100%. Moreover, using negative enrichment method combined with FISH analysis, the positive rate of tumor cells was 86% (31/36), much higher than that using density gradient centrifugation combined with cytology analysis (χ(2) = 5.818, P = 0.012). In 17 cases of benign pleural effusions, using negative enrichment method combined with IF, the positive rate was 100%. But other methods didn't find cancer cells from benign pleural effusions. It was applicable to enrich tumor cells from pleural effusions using negative enrichment method by immunomagnetic beads. This method combined with cytology analysis or FISH significantly enhanced the sensitivity and specificity of tumor cell detection in pleural effusions. But it was difficult to distinguish cancer cells from mesothelial cells using immunofluorescence staining with CK18, DAPI and CD(45) label. More specific markers were needed to recognize tumor cells from pleural effusions.

Diagnostic Value of Interleukin 21 and Carcinoembryonic Antigen Levels in Malignant Pleural Effusions

The aim of this study was to evaluate the diagnostic value of interleukin 21 (IL-21) and carcinoembryonic antigen (CEA) in tuberculous pleural effusions (TPEs) and malignant pleural effusions (MPEs). Pleural effusion samples from 103 patients were classified on the basis of diagnosis as TPE (n=51) and MPE (n=52). The concentration of IL-21 was determined by ELISA. Lactate dehydrogenase (LDH), adenosine dehydrogenase (ADA) and CEA levels were also determined in all patients. A significant difference was observed in the levels of ADA and CEA (P<0.01), but not in the levels of LDH (P>0.05) between TPE and MPE. The concentration of IL-21 in MPE was significantly higher compared to TPE (P<0.01). With a threshold value of 4.32 pg/ml, IL-21 had a sensitivity of 76.9% (40/52) and a specificity of 80.4% (41/51). Combined detection of IL-21 and CEA had a sensitivity of 69.2% (36/52) and a specificity of 92.2% (47/51). These two markers can contribute to the differential diagnosis of MPEs.

Comparing the results of VATS pleurectomy and talc pleurodesis with small sized catheter in randomised patients with malign pleural effusion

Objectives: Pleural effusion occurs as a result of detoriation in equilibrium between absorption and secretion. We aimed to investigate the clinical responses of talk pleurodesis and pleurectomy with video assisted thoracoscopic surgery (VATS) in patients with malignant pleural effusion. Materials and methods: Forty-five patients with malignant pleural effusion between June 2007 and June 2008 were included in this study. Thoracentesis was performed in order to study the biochemical, microbiologic, cytological analysis. Glucose, total protein, albumin, lactic dehydrogenases and cytological examination were studied in effusion and blood sample simultaneously. Cases were classified into two groups; in group I (n=25), small calibrated catheter and talk pleurodesis were performed. In group II (n=20), VATS pleurectomy was performed. Results: There were 32 females and 13 males with mean age 51.58 (27-75) years. Diagnosis was made with cytologic examination of pleural fluid that was aspirated with thoracentesis. Transudate- exudate discrimination was done according to Light’s criteria. Success rates of the groups were as complete response; 84% (n=21) in group I, 85% (n=17) in group II. Unsuccessful response was 16% (n=4) in group I and 15% (n=3) in group II. There was no statistically significant difference between hospital stay and tube duration of groups when compared to each other. Conclusion: The treatment of malignant pleural effusion is palliative due to poor prognosis. The purpose is to eliminate dyspnea in patients with short survival time. There was no significant difference between the VATS pleurectomy and talc pleurodesis that has been known as effective sclerosant agent.

Low dose macrolide administration for long term is effective for otitis media with effusion in children

To explore the clinical effect and possible mechanism of macrolides on otitis media with effusion in children. Children with otitis media with effusion were recruited and prescribed for macrolides according to a designed scheme, and followed up for 8-12 weeks. Middle ear effusion samples were collected from the participants to assess the presence of biofilm. Macrolides were found to have significant effect on the therapy of early stage otitis media with effusion for 88.7-92.5% of participants compared with 50.9-60.3% in control group after 8-12 weeks. Almost 72.1% of chronic otitis media with effusion patients recovered after an 8 weeks' course low dose macrolides prescription. Biofilm occurred in 30.8% (4/13) of middle ear effusion. The results indicate that macrolides are effective for otitis media with effusion in children. Bacterial biofilm may play an important role in the pathogenesis of otitis media with effusion.

Abstract 5346: Tumor cell heterogeneity and chromosomal abnormality in malignant pleural effusion (MPE) samples of lung cancer

Abstract Tumor heterogeneity is a hallmark of cancer and different types and stages of cancer express various cell surface markers and gene expression profiles. To understand tumor cell heterogeneity in lung cancer, we have developed a model using primary cells from malignant plural effusion (MPE) from lung cancer patients. Using this system we have identified CD44high expressing primary tumor cells within the MPE that form colonies in soft agar in vitro, and tumors in SCID/NOD mice. RT-PCR analysis revealed higher expression of cancer stem cell markers, Bmi-1, hTERT, EZH2, and OCT-4 in CD44high cells than the CD44low cells. Thus, our results indicate that primary tumor cells of MPE derived from lung cancer patients show heterogeneity with respect to their tumorigenic potential. To determine whether we could detect cytogenetic abnormalities in these samples, we performed karyotypic analysis of the MPE samples. Fibroblast cell line GM 05399 served as a control for these studies. Hyperdiploidy was observed in all three MPE samples and in the control cell line H 2122. Abnormalities of chromosome 1p were observed seen in all the samples that led us to perform FISH (fluorescence in situ hybridization) studies using a dual color 1p36 and 1q25 probe (control). We found LOH (loss of heterozygosity) at 1p36 in two MPE samples and rearrangements of both 1p/1q regions in the third MPE sample. Cell line H 2122 contained one normal chromosome 1 and an unbalanced translocation of unknown origin at 1p36 consistent with deletion of 1p. Our studies therefore suggest that this 1p36 deletion could result in the inactivation of a tumor suppressor gene and other sequences that include miRNA 34a also mapped to this locus. Chromosomal rearrangements further indicate that cells with multiple phenotypes including those related to metastasis, and chemo-radiation resistance might be present in the MPE samples. These studies provide a good basis suggesting that MPE would serve as a useful model system for the identification of genes related to the progression of lung cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5346. doi:1538-7445.AM2012-5346