Long-term tissue cultures of human pleural effusions: A cytological follow-up

Summary A cytological follow-up was performed on long-term cultures of 53 metastatic pleural effusions and 34 cases of mesothelial hyperplasia. Cells were grown in Leighton tubes with up to 11 changes of cover slip in the same tube. Cell morphology and arrangement was followed for up to 1 year. Morphological cariteria are of no use in recognizing malignant cells with any degree of certainty. As to cell distribution, several types of heaping up are observed both in benign and malignant pleural effusions. In malignant cases, patches of cells in an epithelial-like arrangement are seen heaping up from their substrate. They can be recovered from the used medium by cytocentrifugation. Such behavior may explain the numerous unsuccessful attempts to establish epithelial cell lines, as the clusters of epithelial cells are progressively lost in the supernates. Supported in part by Contract INSERM ATP 76-1-493 and by a grant from the Ligue Nationale Fran?aise contre le Cancer.     

Mass spectrometric characterization and physiological actions of VPNDWAHFRGSWamide, a novel B type allatostatin in the crab, Cancer borealis

The neural networks in the crustacean stomatogastric ganglion are modulated by neuroactive substances released locally into the neuropil of the stomatogastric ganglion and by circulating hormones released by neuroendocrine structures including the pericardial organs. Using nanoscale liquid chromatography coupled to electrospray ionization quadrupole-time-of-flight mass spectrometry, we have identified and sequenced a novel B type allatostatin (CbAST-B1), VPNDWAHFRGSWamide, present in the pericardial organs of the crabs, Cancer borealis, and Cancer productus. We describe the physiological actions of CbAST-B1 on the pyloric rhythm of the stomatogastric ganglion of the crab, Cancer borealis. CbAST-B1 reduces the pyloric network frequency in a dose-dependent manner. The effect of bath-applied CbAST-B1 depends on the preceding physiological state of the preparation. Surprisingly, despite marked amino-acid sequence dissimilarity between the novel CbAST-B1 and the A type allatostatin family of peptides (AST-A), the physiological effects of CbAST-B1 are similar to those of AST-A.     

自发性气胸患者术后并发胸腔积液与肺部感染、胸水葡糖糖水平的相关性分析

目的:探讨自发性气胸患者术后并发胸腔积液与肺部感染、胸水葡糖糖(Glu)水平的相关性,为自发性气胸患者术后并发胸腔积液的预防诊治提供参考。方法:选择我院2016年6月至2018年3月收治的96例行胸腔镜手术的自发性气胸患者作为研究对象,按照术后是否并发胸腔积液将其分为对照组和观察组两组,其中对照组患者56例,术后无并发症,观察组患者40例,术后并发胸腔积液。采用单因素分析法对两组患者的一般资料进行分析,并通过Pearson分析法对上述资料进行相关性分析。对术后肺部感染患者的病原菌分布及构成比进行分析,观察两组患者手术前后外周血炎症因子变化。结果:观察组患者感染率、住院时间、术后恢复时间、胸水Glu水平明显高于对照组,差异显著具有统计学意义(P0.05);对上述具有显著性差异的一般资料进行Pearson分析显示,肺部感染与SP患者术后并发胸腔积液呈现正相关,胸水Glu与SP患者术后并发胸腔积液呈现负相关(P0.05)。17例感染病例中,革兰氏阴性菌为11例,构成比64.71%,革兰氏阳性菌为6例,构成比35.29%,无真菌感染病例发生。两组患者治疗前各外周血炎症因子水平差异不具有统计学意义,具有可比性(P0.05);两组患者治疗后降钙素原(PCT)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)以及白细胞介素-10(IL-10)水平均明显升高,且观察组患者上述指标升高更为显著,差异具有统计学意义(P0.05)。结论:自发性气胸患者术后并发胸腔积液与胸水Glu水平呈现负相关,与肺部感染呈现正相关,且肺部感染患者中革兰氏阴性菌相对较多,对自发性气胸患者术后并发胸腔积液临床诊治具有一定的借鉴意义。     

The usefulness of a panel of immunostains in the diagnosis and differentiation of metastatic malignancies in pericardial effusions

Pericardial effusions are not uncommon in patients with an advanced malignancy Rarely malignancies may present initially with a pericardial effusion. Cytological examination of pericardial fluid may be valuable in differentiation of these cases. However, a metastatic tumour in serous effusion may not always show the functional differentiation of the primary tumour. In such a situation, although a wide range of special studies have been suggested for the diagnosis of malignancy we have found the use of a panel of a few common immunostains to be useful in confirming or suggesting the site of a primary tumour. The material for this study consisted of 76 pericardial fluids obtained between January 1991 and October 1998 from 46 males (mean age 59 years) and 30 females (mean age 52 years). Metastatic malignancy was diagnosed in 22 of the 76 patients and in 7/22 cases pericardial effusions were the initial presentation. The subsequent follow-up in the seven cases revealed adenocarcinoma of lung (n = 2), small cell anaplastic carcinoma of lung (n = 1), squamous cell carcinoma lung (n = 1), melanoma leg (n = 1), non-Hodgkin's lymphoma retroperitoneal lymph nodes (n = 1) and carcinoma of the breast (n = 1). Of the remaining 15 cases with a known history of malignancy, eight had cancers (three adeno; two small cell; one poorly differentiated, and two squamous cell types) of the lung; breast (n = 3); colon (n = 1); melanoma (n = 2) and non Hodgkin's lymphoma (n = 1). Immunostains which were useful in the diagnosis were EMA, CEA, cytokeratin, B72.3, HMB45, vimentin, S100, LCA, L26 and kappa and lambda light chains.     

The value of calretinin and cytokeratin 5/6 as markers for mesothelioma in cell block preparations of serous effusions

Objective: To determine the value of calretinin and cytokeratin (CK) 5/6 in discriminating mesothelioma from adenocarcinoma in serous effusion specimens. Methods: A total of 101 recent, histologically or clinically confirmed malignant effusions with immunostained cell block preparations were reviewed. The cases consisted of 34 mesotheliomas and 67 adenocarcinomas. This included 17 ascitic fluid and 84 pleural fluid samples. The adenocarcinomas included metastatic carcinomas from the breast (12), lung (19), stomach (3), colon (1), pancreas (2), ovary (6) endometrium (1) and 23 histologically confirmed metastases from unknown primary sites. The cases were assessed as negative or positive (>5% of cells stained). The staining pattern was recorded as cytoplasmic, cell membrane, nuclear or cytoplasmic and nuclear staining. Results: Calretinin staining was present in 97% (33/34) of the mesothelioma cases with a majority of them showing both cytoplasmic and nuclear staining (29/33). Only 3% (2/67) of adenocarcinomas were positive for calretinin, one being a lung adenocarcinoma and the other an adenocarcinoma of unknown primary site in an ascitic fluid. Cytokeratin 5/6 staining was also present in 33/34 (97%) of mesothelioma cases. Six (9%) adenocarcinomas were positive, including metastases from the lung (1), breast (1), ovary (2) and unknown primary site (2). Four of the six adenocarcinoma cases positive for CK5/6 were in ascitic fluids. No cases of mesothelioma were negative for both calretinin and CK5/6. Only one adenocarcinoma case, (which was from unknown primary site in an ascitic fluid sample), was positive for both markers. Conclusions: The results confirm that calretinin and CK 5/6 are useful markers for mesothelioma in effusion specimens. CK5/6 staining may be less useful for peritoneal fluid specimens where metastatic adenocarcinomas may be more likely to express the antigen. Further study of ascitic/peritoneal specimens is warranted. However, positive staining, particularly for both antigens, is highly indicative of a mesothelial origin for cells. The two markers make a useful addition to EMA and the panel of adenocarcinoma markers routinely applied to effusion specimens.     

Lymphoid disorders associated with HHV-8/KSHV infection: facts and contentions

Following the demonstration in 1994, that Kaposi's sarcoma (KS) was associated with a novel virus (KSHV or HHV-8) belonging to the lymphotropic herpes family, this virus was also found in certain lymphoid neoplasias of immunodeficient (HIV⁺) and immune competent hosts. The association of HHV-8/KSHV infection is now well established with primary effusion lymphoma (PEL) or body cavity based lymphoma (BCBL) and multicentric Castleman's disease (MCD) of the plasma cell type. A possible pathogenic role of HHV-8/KSHV in other lymphoid tumours including primary central nervous system lymphoma (PCNSL) and multiple myeloma (MM) as well as some atypical lymphoproliferations and sarcoidosis has also been suggested, but this is at present a controversial matter, or not confirmed. SeveralHHV-8/KSHV genes, including potential oncogenes, genes homologous to various cellular genes and growth factors have been incriminated in the pathogenesis of KS and PEL/BCBL, but a common pathogenic mechanism for the clearly diverse proliferations represented by PEL, MCD and KS is at present not evident.     

An investigation of adequate volume for the diagnosis of malignancy in pleural fluids

S. C. Thomas, L. R. R. Davidson and M. E. McKean An investigation of adequate volume for the diagnosis of malignancy in pleural fluids Objective: The cytological examination of pleural effusions is an important investigation in the diagnosis of malignancy. Maximizing the chances of identifying malignant effusions is therefore desirable. Recent Royal College of Pathologists guidelines, based on the British Society for Clinical Cytology codes of practice, have suggested that a minimum of 20 ml of pleural fluid is required for diagnostic purposes. Methods & Results: We examined 2155 pleural fluids received over a 6‐year period in order to define a minimum required volume for adequacy. By examining the plateau phase of a graph of threshold volumes for initial samples received for each patient (n = 1584) we determine that a minimum fluid volume of 25 ml is required and that more than 50 ml does not improve sensitivity. Conclusion: Between 25 and 50 ml of fluid are required for the adequate assessment of pleural effusions for malignancy.     

The diagnostic and molecular characteristics of malignant mesothelioma and ovarian/peritoneal serous carcinoma

B. Davidson The diagnostic and molecular characteristics of malignant mesothelioma and ovarian/peritoneal serous carcinoma Malignant mesothelioma and ovarian/peritoneal serous carcinoma are two of the most common tumours affecting the serosal cavities. Unlike other malignant tumours diagnosed at this anatomical site, such as lung and breast carcinoma, malignant mesothelioma and serous carcinoma share a common histogenesis, may be difficult to differentiate morphologically, and co‐express many of the diagnostic markers used by cytopathologists in effusion diagnosis. Selected markers have nevertheless shown sufficient sensitivity and specificity to differentiate serous carcinoma from malignant mesothelioma effectively. Recently, our group applied high throughput technology to the identification of new markers that may aid in differentiating these two cancer types and validated several of these markers in follow‐up studies. This review will present current data regarding the diagnostic and biological aspects of malignant mesothelioma and ovarian/peritoneal serous carcinoma.     

Use of narrow‐range peptide IEF to improve detection of lung adenocarcinoma markers in plasma and pleural effusion

In this study we applied narrow‐range peptide IEF to plasma or pleural effusion prior to LC/MS/MS. Two methods for narrow‐range IEF were run; IPG strips and free‐flow electrophoresis. Data from this study was compared with cell line data to evaluate the method performance in body fluids. To test the methods potential in quantitative biomarker discovery studies, plasma and pleural effusion from patients with lung adenocarcinoma (n=3) were compared with inflammatory pleuritis (n=3) using iTRAQ quantification. Using narrow‐range IEF on the peptide level we were able to identify and quantify 282 proteins in plasma and 300 proteins in pleural effusion. These body fluid proteomes demonstrated high degree of overlap; however, more proteins significantly differently altered levels related to adenenocarcinoma were found in pleural effusion compared with plasma, suggesting enrichment of lung tissue‐related proteins in pleural effusion. Nine proteins were chosen for initial validation with Western blot, and one protein (NPC2) was chosen for further validation using imunohistochemistry. Overall, the quantitative results from IEF/LC/MS/MS showed good correlation with the results from Western blot and imunohistochemistry, showing the potential of this methodology in quantitative biomarker discovery studies.