Use of ThinPrep monolayer technique and cytospin preparation in urine cytology: a comparative analysis

We compared the ThinPrep (TP) technique to the cytospin (CS) preparation in the cytological diagnosis of urine by processing 79 specimens by these two techniques. Ten cases were positive for malignancy (six high grade (HG)/carcinoma in situ; four low grade (LG) transitional cell carcinomas (TCC)). Forty-eight cases were within normal limits (59%) and 21 cases had atypical cytological features (19%). The TP technique was better in terms of a cleaner background with fewer obscuring inflammatory cells and blood and with a more even distribution of cells. In general, the cytomorphology was comparable in both techniques. However, in cases with malignancy, CS was relatively superior in the cytomorphologic details; in TP, the diagnostic cells were mostly dispersed as single cells with loss of architectural features and were difficult to find. Artifactual empty spaces and air-drying were more frequently present in TP. In cases contaminated with squamous cells, the urothelial cells were difficult to find in TP. Screening time was comparable for both techniques. In conclusion, to avoid false-negative diagnosis, CS would be complementary to the TP technique in malignant cases and, in particular, those with low cellularity.     

The high post-test probability of a cytological examination renders further investigations to establish a diagnosis of epithelial malignant pleural mesothelioma redundant

The aim of the study was to establish in a prospective and blinded manner the diagnostic yield of morphology, immunocytochemistry (ICH) and electron microscopy (EM) in the cytological analysis of malignant pleural mesothelioma (MPM). Pleural fluid from consecutive patients, 14 with a histologically proven MPM, 12 with a malignant pleuritis due to adenocarcinoma (AC), and 13 with a reactive pleural effusion (RM), was separately analyzed. Smears were incubated with monoclonal antibodies (Tag72, Ber-Ep4, anti-CEA, EMA). These were considered suggestive for MPM when only EMA stained positive, for AC when three out of four markers stained positive, and for RM when no marker stained positive. The post-test probability of the morphological, ICH, and EM analysis were 92, 100, 92% or MPM, 91, 100, 86% for AC, and 88, 88, 90% for RM, respectively. We concluded that the high post-test probability of a combined morphological and ICH diagnosis of MPM warrants to cease further diagnostic procedures in these patients. Electron microscopy did not add to accuracy of diagnosis.     

Diagnostic efficacy of electron microscopy and pleural effusion cytology for the distinction of pleural mesothelioma and lung adenocarcinoma

The diagnosis of malignant pleural mesothelioma (MPM) is challenging and requires immunohistochemistry or electron microscopy assays to specifically differentiate MPM from lung adenocarcinoma. An ultrastructural study of fresh tissue is considered to be the “gold standard.” In most cases, the first diagnostic approach is performed on pleural effusion, and in some patients, this is the only available sample for diagnosis. The aim of the present study is to evaluate if an examination of pleural effusion samples based on electron microscopy (EMpe) is a useful tool for the differential diagnosis of MPM and lung adenocarcinoma. An EMpe study was performed in 25 pleural effusion samples. Histological and immunohistochemical markers confirmed the diagnosis of either mesothelioma (5) or adenocarcinoma (20). Of the five cases that were diagnosed with mesothelioma, two samples (40%) showed cells with “bushy” microvilli, which are characteristic of mesothelioma, by EMpe, and three were acellular (60%). Of the 20 cases of adenocarcinoma, EMpe showed cells with short microvilli in 9 (45%), and 11 were acellular (55%). EMpe identifies unequivocal morphological changes that are useful for the differential diagnosis of MPM or adenocarcinoma when the pleural effusion sample contains evaluable tumor cells.     

Lymphohistiocytoid mesothelioma of the pleura: A case report with cytological findings

Lymphohistiocytoid malignant mesothelioma is an infrequent variant of sarcomatoid mesothelioma representing approximately 0.5–3.3% of malignant mesotheliomas. It has been related to asbestos exposure. The tumor is characterized by a diffuse large histiocyte‐like cells proliferation mixed with an inflammatory infiltrate of lymphocytes and plasma cells. Its cytological diagnosis is difficult. We present a case of a 67‐year‐old female with lymphohistiocytoid mesothelioma involving the left pleura. The cytological, histological, and immunohistochemical features are discussed. Diagn. Cytopathol. 2013. © 2011 Wiley Periodicals, Inc.     

Cytological features of lung adenocarcinoma with micropapillary pattern in the pleural or pericardial effusion: analysis of 5 cases

Micropapillary pattern is a distinct histopathological pattern, and usually shows a high frequency of lymphatic invasion and lymph node metastases. This pattern is also reported in lung adenocarcinoma, however, only one cytological report of lung adenocarcinoma with micropapillary pattern has been reported. In this study, we analyzed the cytological features of this type of carcinoma in the pleural or pericardial effusion. This study was comprised of 5 consecutive cases of lung adenocarcinoma with micropapillary pattern, in which the tumor cells were present in the pleural or pericardial effusion and whose diagnoses were histopathologically confirmed. The characteristic cytological findings in the pleural or pericardial effusion were as follows: i) tightly cohesive small nests of tumor cells showing papillary structure without fibrovascular core, ii) these nests were comprised of approximately 5-20 tumor cells, iii) cauliflower-like and acinar-like structures were also observed, iv) intracytoplasmic vacuoles were observed in 40% of the cases, and v) the neoplastic cells had large round to oval nuclei containing coarse chromatin and occasional conspicuous nucleoli. It has been reported that the presence of micropapillary structure and intracytoplasmic vacuolation are also characteristic cytological features of micropapillary carcinoma of the urinary bladder, therefore, they are thought to be common cytological features of carcinomas with micropapillary pattern. Consequently, detection of these features can lead to a cytodiagnosis of lung adenocarcinoma with micropapillary pattern in the pleural or pericardial effusion. Recognition of these features is important because this type of tumor shows an aggressive clinical course.     

内科胸腔镜在提高胸腔积液诊断率中的价值

目的评价内科胸腔镜在提高胸腔积液病因诊断中的价值及内科胸腔镜的检查时机。方法连续收集渗出性胸腔积液患者311例,其中男性194例,女性117例;年龄17～90岁,平均年龄58.3岁。评估胸腔积液细胞学对恶性胸水的诊断率。对27例经胸水细胞学未能诊断的患者和2例胸水细胞学恶性胸腔积液但未准确分型的患者进行了内科胸腔镜检查,评估胸腔镜检查联合胸水细胞学检查对胸腔积液诊断率及恶性胸水的准确诊断率的影响。结合不同次数的胸腔积液细胞学的阳性率及相关支出,评估内科胸腔镜的检查时机。结果311例渗出性胸腔积液患者经胸水细胞学诊断恶性胸腔积液的患者为80例;231例胸水细胞学未确诊的患者结合各自情况,经过内科胸腔镜检查和/或支气管镜、或肺动脉CT造影等检查,并结合临床资料系统分析后,诊断如下:恶性胸腔积液106例,良性胸腔积液91例,未确诊病例34例。胸水细胞学检查的确诊率为28.9%(80/277)。29例接受了内科胸腔镜检查患者中,25例患者直接确诊,诊断率为86.2%。内科胸腔镜检查使该组病例的确诊率提高到37.9%(105/277),与单纯胸腔积液细胞学检查结果比较差异有统计学意义(P=0.024)。3次及以上的胸腔积液细胞学检查不能较2次胸腔积液细胞学检查提高更多的诊断率(P=0.156),且其支出超过内科胸腔镜的支出。结论内科胸腔镜不仅较胸腔积液细胞学检查能提高恶性胸腔积液的诊断率,更能提高胸腔积液总体诊断率。对2次胸腔积液细胞学检查无结果者,如无禁忌证,应积极进行内科胸腔镜检查。     

The sensitivity of cytologic evaluation of pleural fluid in the diagnosis of malignant mesothelioma

Pleural malignant mesothelioma (MM), which is an aggressive neoplasm with a high mortality, frequently manifests initially as pleural effusions. The sensitivity of cytologic examination for its diagnosis varies widely in literature and most of the figures are from earlier studies with conventional cytologic preparations. The objective of this study was to provide the current evidence on the role and sensitivity of cytologic examination of pleural fluid in the diagnosis of MM. We reviewed the cytologic findings in pleural effusions of a large series of histologically proven MM (234 cases) diagnosed in our institution between 2001 and 2008. Of all cases, 154 (66%) had cytologic material examined. A specific diagnosis of MM was rendered or suspected in 53% (79 patients). The lowest sensitivity (20%) was noticed in sarcomatoid MM cases. MM was favored over adenocarcinoma in 97% of patients with positive cytologic findings that have been confirmed with immunohistochemistry. In this series, five cases were inadequate and five cases were initially reported as atypical, whereas 65 cases (44%) were reported as negative for malignancy. On review of the cytology slides, only four cases were upgraded from benign to suspicious compared to four cases downgraded from suspicious to atypical but no significant improvement to the diagnosis could be made on revision. These data suggested that a cytologic diagnosis contributed useful information in patients with epithelioid and biphasic pleural MM. Limitations of the cytologic examination of MM should also be acknowledged. Diagn. Cytopathol. 2010;38:874–879. © 2010 Wiley‐Liss, Inc.     

Comparison of three commonly used cytologic preparations in effusion immunocytochemistry.

Discrepant results in effusion immunocytochemistry are often the result of specimen processing. Smears, cytospins, cell blocks, and monolayer preparations have all been used in various published studies; thus, there is no consistency in the immunostaining process for cytology to compare with the surgical pathology "gold standard" results. We sought to evaluate optimal specimen preparation for the immunostaining of effusion samples. Fourteen reactive and 15 malignant effusion samples (various epithelial/mesothelial neoplasms) were each prepared in three forms: air-dried cytospins (postfixed in ethanol), formalin-fixed, paraffin-embedded cell blocks, and liquid-based thin-layer (ThinPrep, CYTYC, Boxborough, MA) processing. All slides were immunostained with antibodies commonly used in effusion cytology: HBME-1, calretinin, E-cadherin, BerEP4, B72.3, LeuM1, and CA19-9. Cytospin and ThinPrep samples performed in a similar manner: high background staining was encountered in 66% of cases, most evident in three-dimensional clusters of cells. In addition, membrane staining patterns were difficult to interpret. Cell blocks provided the best milieu for morphologic interpretation, with less background staining (only 17% of cases) and results that most closely approximated those reported in the surgical pathology literature. The cost per test for cell block immunocytochemistry was also the most economical for our laboratory.     

CD44H expression in reactive mesothelium, pleural mesothelioma and pulmonary adenocarcinoma

Malignant mesotheliomas are known to produce hyaluronic acid, in contrast to most pulmonary adenocarcinomas which produce neutral mucin. CD44H is the major cell surface receptor for hyaluronic acid. The aim of this study was to investigate immunohistochemically the expression of this antigen in reactive mesothelium, pleural mesothelioma and pulmonary adenocarcinoma and to assess its diagnostic utility in distinguishing the two tumours. Diffuse and intense membranous CD44H immunoreactivity was seen in 15 of 20 (75%) mesotheliomas and in all 20 biopsies of reactive mesothelium. In contrast, focal (<10% tumour) expression of CD44H was seen in only three of 20 (15%) pulmonary adenocarcinomas. We advocate the use of CD44H as a positive mesothelial marker for incorporation alongside other established immunohistochemical markers used to distinguish mesothelioma from adenocarcinoma.     

The comparative accuracy of different pleural biopsy techniques in the diagnosis of malignant mesothelioma

Aims:  To evaluate the diagnostic accuracy of closed and open pleural biopsies in diagnosing malignant pleural mesothelioma.
Methods and results:  The autopsy study group comprised 45 malignant mesotheliomas. All prior pleural biopsy investigations were reviewed. Forty-one of 45 (91%) had had an antemortem diagnosis of malignant mesothelioma. In these 41 cases, 57 prior diagnostic pleural biopsies had been performed [36 closed needle biopsies: 31 blind; five computed tomography (CT)-guided and 21 open pleural biopsies]. For definitive diagnosis open pleural biopsy yielded a sensitivity of 95% and specificity of 100%. For definitive diagnosis closed blind pleural biopsies yielded a sensitivity of 16% and specificity of 94%. Thirty-two per cent of 'blind' biopsies were inadequate. CT-guided pleural biopsies yielded a definitive diagnostic accuracy of 100% (5/5). Biopsy specimen size was important in obtaining a positive definitive diagnosis. Diagnosis was attained in 75% of specimens >10 mm in size compared with 8% <10 mm in size.
Conclusions:  Overall, all procedures had utility but definitive diagnostic accuracy for 'blind' closed pleural biopsy was low (16%), dependent on biopsy specimen size and tumour subtype. Sarcomatoid subtype malignant mesothelioma yielded the lowest diagnostic accuracy. For all subtypes of malignant mesothelioma, open pleural biopsy produced the highest diagnostic accuracy (100% sensitivity, 95% specificity).     

A fine decision tree consisted of CK5/6, IMP3 and TTF1 for cytological diagnosis among reactive mesothelial cells,metastatic adenocarcinoma of lung and non-lung origin in pleural effusion

The utility of combination with CK5/6, IMP3 and TTF1 to differentiate among reactive mesothelial cells (RMs), metastatic adenocarcinoma of lung (LAC) and non-lung (NLAC) origin was investigated by using immunocytochemistry (ICC) and conventional PCR (C-PCR) in pleural effusion. A total of 108 cell blocks (32 RMs, 51 LAC and 25 NLAC were evaluated by ICC for CK5/6, IMP3 and TTF1 protein expression. In addition, we further performed C-PCR for amplification of CK5/6, IMP3 and TTF1 DNA from 28 specimens (9 MAC and 7 RMs, 6 LAC and 6 NLAC) for molecular diagnosis. CK5/6 staining was observed in the majority of reactive specimens (78.1%) and was rare in adenocarcinoma cells (14.5%), whereas the opposite was true for IMP3 and TTF1. We found a high frequency of TTF1 positivity (76.5%) in LAC, but not in NLAC (4.0%); while there was no significant difference of IMP3 expression in LAC (88.2%) and NLAC (88.0%). The 487 bp DNA fragments of IMP3 was expected to be amplified in 6/9 of adenocarcinoma cases showed negative in ICC; and the 394 bp DNA fragments of CK5/6 was also expected to be amplified in 4/7 of RMs cases showed negative in ICC. Consistent with ICC results, there was significant difference of TTF1 expression in the LAC and NLAC compared with IMP3 expression. The combination with CK5/6, IMP3 and TTF1 immunostaining appears to be useful to improve the accuracy of cytological diagnoses between RMs, metastatic adenocarcinoma of lung and non-lung origin in pleural effusion. In addition, C-PCR may act as a useful supplemental approach for ICC, especially negative cases in ICC for differential cytological diagnosis.     

Concurrent pleural infiltration by chronic lymphocytic leukemia and adenocarcinoma of unknown primary site diagnosed by effusion cytology

Synchronous malignancies in a pleural effusion are rare. A case of concurrent pleural infiltration by adenocarcinoma of unknown primary site and chronic lymphocytic leukemia (CLL) is presented in this case study, which was diagnosed by effusion cytology. Pleural effusion is not an uncommon complication in patients with B‐CLL. Even in a pleural effusion rich in monoclonal lymphocytes, the presence of a second cancer must be excluded because this can be the main cause of mortality. The role of cytology in such cases is of paramount importance. Diagn. Cytopathol. 2014;42:151–155. © 2012 Wiley Periodicals, Inc.     

联合抗体在胸水细胞块肺腺癌细胞和增生性间皮细胞中的表达及意义

目的：探讨联合抗体的运用在鉴别胸水肺腺癌细胞和反应性增生间皮细胞中的意义。方法：胸水细胞块切片后行免疫组织化学sP法检N25例胸水转移性肺腺癌细胞和20例反应性增生间皮细胞中甲状腺转录因子．1、细胞角蛋白-7、间皮细胞及钙结合蛋白表达。结果：甲状腺转录因子-1、细胞角蛋白．7、间皮细胞和钙结合蛋白在胸水肺腺癌细胞和反应性增生间皮细胞中的表达比较差异有统计学意义fP〈0．05）。甲状腺转录因子．1和细胞角蛋白-7联合标记胸水肺腺癌细胞的敏感性为88％,特异性为100％;间皮细胞和钙结合蛋白联合标记胸水增生性间皮细胞的敏感性为60％,特异性为95％。结论：联合应用甲状腺转录因子．1、细胞角蛋白-7、间皮细胞和钙结合蛋白抗体检测胸水细胞块对鉴别肺腺癌细胞和反应性增生间皮细胞具有较高的应用价值。     

Measurement of telomere length in cells from pleural effusion: Asbestos exposure causes telomere shortening in pleural mesothelial cells

We estimated the telomere lengths of neoplastic and non‐neoplastic mesothelial cells and examined their correlation with asbestos exposure and the expression of markers of mesothelial malignancy. Cell blocks of pleural effusion obtained from 35 cases of non‐neoplastic disease (NN), 12 cases of malignant mesothelioma (MM) and 12 cases of carcinomatous effusion due to lung adenocarcinoma (LA) were examined. Fifteen of the 35 NN cases had pleural plaques (NNpp+) suggestive of asbestos exposure, and the other 20 cases had no pleural plaques (NNpp‐). Telomere length was measured using the tissue quantitative fluorescence in situ hybridization method, and expressed as normalized telomere‐to‐centromere ratio. NN cases had significantly longer telomeres than MM (P < 0.001) and LA (P < 0.001) cases. Telomeres in NNpp+ cases were slightly shorter than those of NNpp‐ cases (P = 0.047). MM and LA showed almost the same telomere length. NN cases with shorter telomeres tended to show aberrant expression of epithelial membrane antigen (EMA), CD146, glucose transporter 1 (GLUT1) and IGF‐II messenger RNA‐binding protein 3 (IMP3). These results suggest that telomere shortening and subsequent genetic instability play an important role in the development of MM. Measurement of telomere length of cells in pleural effusion might be helpful for earlier detection of MM.