Article

Calretinin staining patter aids in the differentiation of mesothelioma from adenocarcinoma in serous effusions

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Abstract

Background: The differentiation between malignant mesothelioma and adenocarcinoma based on morphology alone can be a diagnostic challenge. The majority of the available antibodies recognize molecules expressed by adenocarcinoma whereas to the authors' knowledge specific markers for mesothelial cells are lacking. Calretinin, a calcium-binding protein, has been reported to be a selective marker for mesothelioma and largely is absent from adenocarcinoma on histologic material. The results with cytologic preparations have been inconsistent. Methods: To evaluate the specificity of calretinin in differentiating mesothelioma from adenocarcinoma in cytologic preparations, 21 paraffin embedded cells blocks of serous effusions from 15 patients with metastatic adenocarcinoma and 16 cell blocks from 9 patients with malignant mesothelioma were stained with a monoclonal antibody against calretinin. The immunoreactivity was evaluated blindly by two observers. Positive staining was defined as nuclear and cytoplasmic staining with or without intense membranous decoration. The former resulted in a characteristic "fried egg" appearance. Results: Calretinin staining was positive in all but 2 cases of mesothelioma (14 of 16 cases; 87.5%). The latter contained predominantly spindle-shaped neoplastic mesothelial cells in the cell block preparations. All adenocarcinoma specimens were classified as negative for calretinin staining; 9 (42.9%) lacked any immunoreactivity and 12 (57.1%) showed weak, sparse, coarse, granular cytoplasmic staining without nuclear or membranous staining. Benign reactive mesothelial cells, when observed in association with adenocarcinoma, also showed the characteristic "fried egg" appearance. The difference in the staining pattern of calretinin between cells of mesothelial origin and adenocarcinoma cells was statistically significant. Conclusions: Calretinin is a useful marker in differentiating mesothelioma of the epithelial type from adenocarcinoma in serous effusions. The "fried-egg" appearance or cytoplasmic and nuclear staining pattern is characteristic of cells of mesothelial origin.

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... 5,18,29,36, 39 -41,43-51 "Positive" mesothelial markers also have been introduced as part of the panel, including OV632, thrombomodulin, CK 5/6, calretinin, HBME-1, N-cadherin, CD44S, and WT1, also with varying results. 5,18,20,24,25,29,30,[35][36][37]39,40,43,[45][46][47][48][49][51][52][53][54][55][56][57][58][59][60][61][62][63] Finally, there is speculation that markers such as p53, E-cadherin, and MOC-31 may be useful for the identification of reactive mesothelial cells. 43,46,47,64 In our laboratory, we routinely use an antibody panel comprised of B72.3, BerEP4, CA 19-9, and calretinin for the differentiation of ACA from MM in effusions. ...
... 18,30,39,47,54,59,79,80 Antibodies to this protein are strongly immunoreactive with malignant and benign mesothelial cells, as evidenced by both a cytoplasmic and nuclear type of staining pattern, described by one group as a "fried-egg" appearance ( Fig. 10). 62 In ACAs, this antibody generally is nonimmunoreactive or, rarely weakly, immunoreactive, exhibiting only a cytoplasmic pattern. 30,54,79,80 Differences in sensitivity and specificity exist, as many of the published studies have been performed using polyclonal antibodies from various manufacturers. ...
... 30,54 Numerous effusion studies using various types of cyto- logic preparations including cell blocks, cytospins, and Papanicolaou-stained smears of effusions also have shown calretinin to be a sensitive and specific marker of reactive and malignant mesothelial cells, with positivity rates of 88 -100% for MM and 80 -100% for reactive effusions. 18,[60][61][62]80 We have found anticalretinin to be invaluable in a panel of markers used in diagnostic effusion cytology, particularly when there is a need to distinguish between single ACA cells and benign reactive mesothelial cells. ...
Article
Cytology plays a pivotal role in the diagnosis of pleural effusions. In many cases, immunocytochemistry (ICC) is required to elucidate the etiology of the atypical cells. Effusions are samples that present unique problems for ICC. To date there is no standardization of ICC methods for effusions and cytology in general. The authors review the most commonly used cytologic preparations, fixatives, and antibodies used in effusion ICC. Through the utilization of cell block preparations and a panel of antibodies appropriate for the differential diagnosis in question, ICC conditions utilized in surgical pathology can be most closely replicated. ICC may provide reliable insights into various diagnostic dilemmas in effusion cytology, provided that laboratory standardization practices are followed.
... The recent Calretinin antibodies ( Figure 2B) require both nuclear and cytoplasmic staining to support a diagnosis of mesothelioma [26]. There are earlier reports of only nuclear staining with "fried egg appearance" [27,28]. Cytoplasmic staining alone should be interpreted negatively [27]. ...
... There are earlier reports of only nuclear staining with "fried egg appearance" [27,28]. Cytoplasmic staining alone should be interpreted negatively [27]. In effusions, the sensitivity of calretinin in detecting mesothelioma ranges from 81 to 100% [26,29,30]. ...
... Calretinin can be expressed in breast carcinomas [31], and a weak cytoplasmic staining is reported in variety if other adenocarcinomas [27,28]. Some studies have shown calretinin positivity in squamous cell carcinoma (SCC) of the lung ranging from 40 to 100% [27,32]. ...
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For the clinicians with non-pathology background, first encountering the patients with pleural or peritoneal effusions, mesothelioma is only one statistically rare but clinically significant option of many differential diagnoses. This review aims to help the clinicians and broad life science audiences to understand step by step the possibilities and shortcomings of pathological diagnosing of mesothelioma, including the basic technical requirements. The first cytomorphology evaluation of pleural and peritoneal effusions in routinely stained smears enables in most cases only to identify cells suspicious for malignancy. The recent guidelines of epithelioid mesothelioma cytologic diagnosis and reporting emphasize immunochemistry (IC) in the cell blocks is mandatory whenever a diagnosis of malignancy is clinically entertained and/or cytologically suspected. The IC workup is challenging, since there is no fixed antibody panel, but multiple questions must be solved, such as 1) confirm the mesothelial or epithelial origin of isolated atypical cells and cell clusters; 2) delineate their benign or malignant nature; and 3) discriminate mesothelioma from other malignancies and metastatic disease. The rationale of the most widely clinically used IC markers is given and illustrated by the examples. The final confirmation of mesothelioma diagnosis and establishing its subtype and grade is possible only in the histological samples.
... Penelitian yang dilakukan oleh Barberis dengan pewarnaan anticalretinin menunjukan 100% positif pada mesothelioma maligna dan 23% yang positif pada metastasis adenokarsinoma pada cairan pleura. Sebaliknya penelitian yang dilakukan Simsir 27% positif pada reaktif mesothelial, 35% pada mesothelioma maligna dan 48% positif pada adenokarsinoma (Chhieng et al., 2000). Menurut Nelson calretinin dapat positif 6-10% pada adenokarsinoma paru, 31-38% pada serous karsinoma dan 0-4% pada renal sel karsinoma, sedangkan squamous sel karsinoma dari paru calretinin dapat positif pada 23-39% kasus dengan tingkat positivitas yang berbedabeda (Ordonez, 2005). ...
... Blok dipotong dan dilakukan pengecatan immunositokimia calretinin. Ekspresi calretinin merupakan skor ekspresi calretinin positif pada sel ganas, dengan pemeriksaan imunositokimia, berdasarkan persentase sel ganas yang tercat pada inti dan sitoplasma (Ensani et al., 2011;Roberts et al., 2001;Foster et al., 2001), yaitu negatif = tidak tampak sel ganas yang tercat, positif I = 1-25% tercat pada sel ganas, positif II = 26-50% tercat pada sel ganas, positif III = 51-100% tercat pada sel ganas (Chhieng et al., 2000;Roberts et al., 2001;Foster et al., 2001;Ordonez et al., 2006). Dengan melihat pada 10 lapang pandang menggunakan pembesaran 400x secara acak, kemudian dibaca oleh dua dokter spesialis Patologi Anatomi secara blindly selanjutnya dibandingkan dengan menggunakan test Kappa. ...
... Pada 14 sampel yang bereaksi positif menunjukan sel-sel tersebut mengandung calretinin, dengan tingkat positivitas yang berbeda-beda sedangkan pada 6 sampel yang negatif menunjukan sel-sel tersebut tidak mengandung kalretinin. Secara teoritis calretinin terdapat terutama pada sel-sel saraf pusat dan perifer juga pada sel mesothel normal maupun keganasan (Kuznicki et al., 1995), Namun pada beberapa penelitian menunjukan adenocarcinoma, squamous cell carcinoma atau tumor lain juga dapat bereaksi positif pada pewarnaan dengan calretinin (Chhieng et al., 2000;Foster et al., 2001). ...
Article
Metastasis adenokarsinoma adalah kasus yang paling sering dijumpai pada efusi pleura yang disebabkan oleh keganasan. calretinin sampai saat ini digunakan sebagai penanda untuk sel mesothel baik jinak maupun ganas. Calretinin digunakan terutama untuk membedakan mesothelioma dari suatu karsinoma atau metastasis keganasan lainnya. Namun beberapa penelitian membuktikan bahwa kalretinin tidak hanya positif untuk sel-sel mesothel tetapi dapat pula pada keganasan lain seperti metastasis adenokarsinoma atau squamous sel karsinoma. Penelitian ini bertujuan membuktikan terdapat ekspresi kalretinin pada efusi pleura dengan gambaran sitomorfologi suatu adenokarsinoma. Jenis penelitian ini adalah deskriptif observasional dengan desain cross sectional. Terdapat 20 blok sitologi dari efusi pleura yang memenuhi kriteria kemudian dilakukan pengecatan calretinin. Ekspresi positif ditandai dengan tercatnya inti dan sitoplasma sel yang dicurigai ganas. Ekspresinya dibaca oleh 2 orang spesialis Patologi Anatomi secara blindly. Ekspresi calretinin negatif pada 6 sampel, positif I pada 8 sampel, positif II pada 2 sampel dan positif III pada 4 sampel. Terdapat ekspresi calretinin pada pada efusi pleura dengan gambaran sitomorfologi suatu adenokarsinoma dengan tingkat positivitas yang berbeda-beda, hal ini menunjukan bahwa kalretinin bukan merupakan penanda yang spesifik dan sensitif untuk sel-sel mesothel baik jinak maupun ganas.Kata kunci: Calretinin, Sitologi efusi, Adenokarsinoma
... Immunocytochemistry (ICC) analysis is one such easily performed technique and uses a panel of markers. [4][5][6][7][8][9][10][11][12][13][14][15][16] However, there is a huge lacuna in achieving an accurate panel of immunomarkers as a diagnostic aid in solving the problems. Several antibodies have been tried in the differentiation of reactive mesothelial (RM) cells from metastatic ACs; unfortunately, no single marker is so far 100% specific and sensitive for neither AC cells nor RM cells. ...
... Several antibodies have been tried in the differentiation of reactive mesothelial (RM) cells from metastatic ACs; unfortunately, no single marker is so far 100% specific and sensitive for neither AC cells nor RM cells. [4][5][6][7][8][9][10][11][12][13][14][15][16][17][18] Hence, the present study evaluated the efficacy of four markers that include MOC-31, epithelial membrane antigen (EMA), calretinin (CAL), and mesothelin (MES) to differentiate AC cells from RM cells in serous effusions. ...
... The reported incidence of CAL positive in ACs ranged from 5-10% including colonic and ovarian ACs. [4,8,[10][11][12] In the current study, one ovarian AC (2.4%) showed positivity for CAL; the staining pattern was focal, weak, and less intense when compared with that of RM cells; and similar to previous studies. [8,10] In the present study, MES was expressed in all except one RM cases with strong membranous staining pattern with Sn of 97.62%. ...
Article
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Introduction Although cytological examination helps in diagnosis of malignancy in serous effusion, at times it is difficult to differentiate atypical reactive mesothelial cells from adenocarcinoma (AC) cells. To resolve this problem, various ancillary methods have been used. Immunocytochemistry (ICC) is one such commonly used technique in which various panel of antibodies has been tried. Unfortunately, so far no unique marker is available to solve this issue. Hence, the present study evaluates the efficacy of four antibody panel comprising of MOC-31, epithelial membrane antigen (EMA), calretinin (CAL), and mesothelin (MES) to solve this problem. Materials and Methods Forty-two cases suspected of malignant effusion in pleural/peritoneal fluid and 42 cases of reactive effusion were included. Cytospin smears were prepared and stained with Giemsa stain for cytomorphological diagnosis. Cytospin smears and cell blocks were made forICC. ICC for MOC-31, EMA, CAL, and MES was performed. Results Among the suspected malignant effusion cases, 30 cases were AC and 12 cases were suspicious for malignancy by cytomorphology. MOC31 demonstrated 100% sensitivity (Sn) and 95.24% specificity (Sp), and EMA had 88.1% Sn and 92.86% Sp for AC cases. CAL demonstrated 100% and 97.62%, and MES 97.62% and 88.1% Sn and Sp in reactive mesothelial cells, respectively. Conclusion In conclusion, combination of MOC-31 and CAL as a limited panel will be helpful in giving an appropriate diagnosis in difficult cases and thereby, help in patient management. In addition, ICC on cytospin smears gave results similar to cell blocks, and if standardised cytospin is simple technique to perform, unlike cell blocks.
... Similar to carcinoma-defining markers, the mesothelial markers calretinin and WT-1 exhibit variable sensitivity and specificity rates; these markers also do not facilitate distinction between benign and malignant mesothelial cells. Although calretinin appears to have very high sensitivity for mesothelial cells, [33][34][35] a small percentage of adenocarcinomas can show calretinin reactivity. 33,35 WT-1 appears to have high sensitivity for exfoliated mesothelial cells, 6 but is positive in serous carcinomas of ovarian and peritoneal sites. ...
... Although calretinin appears to have very high sensitivity for mesothelial cells, [33][34][35] a small percentage of adenocarcinomas can show calretinin reactivity. 33,35 WT-1 appears to have high sensitivity for exfoliated mesothelial cells, 6 but is positive in serous carcinomas of ovarian and peritoneal sites. Thus, in the case of malignant peritoneal effusions, particularly in female patients, Pax-8 is helpful in distinguishing m€ ullerian adenocarcinomas from malignant mesothelioma. ...
Article
Adenocarcinoma can be challenging to distinguish from malignant mesothelioma in effusions, and this distinction often requires ancillary studies and clinical correlation. Immunohistochemistry for claudin-4, a tight-junction-associated protein, has recently been shown to distinguish adenocarcinoma from malignant mesothelioma, mostly in surgical specimens. Our aim was to validate and assess the immunoreactivity profile of claudin-4 in a large series of malignant effusions. We evaluated 159 malignant effusions (84 adenocarcinomas and 75 malignant mesotheliomas). Claudin-4 immunohistochemistry was performed on cell-block paraffin sections and scored for staining intensity, staining pattern (cytoplasmic versus membranous), and percentage of positive tumor cells. Appropriate positive and negative controls were used throughout. All cases of mesothelioma were negative for claudin-4 (0 of 64). Eighty-three of 84 cases of adenocarcinoma were positive (99%); 1 case of serous carcinoma was negative. Most adenocarcinomas showed strong and diffuse membranous staining (71 of 84; 84%); 12 cases (14%) showed membranous staining of moderate intensity. The overall sensitivity for adenocarcinoma was 99% (83 of 84). Claudin-4 immunohistochemistry effectively distinguishes adenocarcinoma from malignant mesothelioma with high sensitivity and specificity in the evaluation of malignant effusions. Cancer (Cancer Cytopathol) 2013. © 2013 American Cancer Society.
... Calretinin, a 29-kD calcium-binding protein, is expressed normally in neurons of the central and peripheral nervous system 8 . An increasing number of studies have shown the ability of this antibody as a biomarker for the diagnosis of MM in effusion specimens [9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26] . Systematic analysis of these data may be valuable to finally confirm the application potential of calretinin as a marker for MM. ...
... As consistent staining pattern of antibody is important for diagnostic test, sensitivity analyses were performed based on different cut-off staining patterns. 14 studies [10][11][13][14][15][16][17][18][19][20][21][22][23]25 reported test results with the cut-off of presenting nuclear staining. The pooled sensitivity and specificity were 0.94 (95% CI: 0.89-0.97) ...
Article
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Numerous studies have investigated the utility of calretinin in differentiating malignant mesothelioma (MM) from metastatic carcinoma (MC) in serous effusions. However, the results remain controversial. The aim of this study is to determine the overall accuracy of calretinin in serous effusions for MM through a meta-analysis of published studies. Publications addressing the accuracy of calretinin in the diagnosis of MM were selected from the Medline (Ovid), PubMed, the Cochrane Library Database and the Web of Science. Data from selected studies were pooled to yield summary sensitivity, specificity, positive and negative likelihood ratio (LR), diagnostic odds ratio (DOR), and receiver operating characteristic (SROC) curve. Statistical analysis was performed by Meta-Disc 1.4 and STATA 12.0 softwares. 18 studies met the inclusion criteria and the summary estimating for calretinin in the diagnosis of MM were: sensitivity 0.91 (95%CI: 0.87-0.94), specificity 0.96 (95%CI: 0.95-0.96), positive likelihood ratio (PLR) 14.42 (95%CI: 7.92-26.26), negative likelihood ratio (NLR) 0.1 (95%CI: 0.05-0.2) and diagnostic odds ratio 163.03 (95%CI: 54.62-486.63). The SROC curve indicated that the maximum joint sensitivity and specificity (Q-value) was 0.92; the area under the curve was 0.97. Our findings suggest that calretinin may be a useful diagnostic tool for confirming MM in serous effusions.
... Calretinin (CAL) is a 29-KD calcium binding protein that is normally expressed normally in neurons of central and peripheral nervous 13 system. CAL is both a sensitive and specic marker of reactive and neoplastic mesothelial cells in cytologic cell block preparations. ...
Article
Background: Distinction between reactive mesothelial cells, malignant mesothelioma and carcinoma is challenging in both biopsy and cytologic material. This study was conducted to differentiate benign/ reactive mesothelial proliferation from malignant mesothelial proliferations and metastatic adenocarcinoma by using immunohistochemical (IHC) markers Desmin (DES), Epithelial membrane antigen (EMA) and Calretinin (CAL) in pleural uid cell block (CB) preparations. A two year descriptive study (Oct.2016- Sept.2018). 46 pleural uids samples sentMethods : to the Dept. of Pathology, RIMS for routine examination and histopathological examination by CB preparation were studied using IHC markers EMA,DES and CAL following H & E stain. Out of 46 cases, 9(19.6%) cases were diagnosed as benign, 23(50.0%) as reactive andResults: 14(30.4%) as adenocarcinoma on H & E section by CB preparations within an age range 34 to 80 years (Mean±SE, 60.32±12.13). Following IHC staining with EMA, DES & CAL, 11(23.9%) cases were conrmed as benign, 17(37.0%) as reactive, 16(34.8%) as adenocarcinoma and 2(4.3%) cases as malignant mesothelioma. This study showed that EMA, DES and CAL helpful in conrming benign or reactive mesothelialConclusions: and malignant mesothelial with epithelial cells which will be helpful in providing appropriate diagnosis in difcult cases and provide better patient management.
... In addition, cytologic smears also suffer with the limitation of inability to evaluate multiple immunomarkers simultaneously.[1718] Some authors have recommended single antibodies, mostly calretinin.[21223031] We do not agree with the use of a single monoclonal antibody for this purpose, which has therapeutic as well as prognostic implications. ...
Article
Differentiation between reactive, but morphologically atypical, mesothelial cells and adenocarcinoma in effusions can be problematic. Elaborate immunohistochemical panels have been devised. Techniques like DNA analysis, flow/image cytometry, and K-ras mutation analysis are research oriented and difficult to perform in routine, especially in resource-poor centers. We evaluated the efficacy of a limited two-antibody panel comprising calretinin and Ber-EP4 on cytospin and cell block preparations, in 100 effusion samples. Fifty cases of reactive mesothelial hyperplasia and 50 cases of adenocarcinoma diagnosed by cytomorphology in ascitic/pleural fluid specimens over a 2-year period were assessed. The diagnoses were confirmed by clinical/histopathologic correlation. Cytospin smears were made in all. Cell blocks were prepared, wherever adequate fluid was available. Immunocytochemistry (ICC) for calretinin and Ber-EP4 was performed. Forty-five of the reactive effusion cases (90%) were calretinin reactive and Ber-EP4 negative. Among the adenocarcinoma cases, 49 (98%) were calretinin negative but Ber-EP4 positive. Thus, both calretinin and Ber-EP4 had a high sensitivity (90% and 98%, respectively), as well as a high specificity (100% and 86%, respectively). In the 21 reactive mesothelial cases, whose cell blocks were made, results were comparable to those on cytospin. However, of the 19 adenocarcinoma cases in which cell blocks were prepared, all were Ber-EP4 immunopositive except for three, which were positive on cytospin, implying false-negative results on cell blocks. A limited panel of two monoclonal antibodies, calretinin and Ber-EP4, may be useful in cytology, as a "primary antibody panel", for accurate diagnosis and patient management. Additionally, ICC can be performed easily on cytospin preparations, which gave results comparable to cell blocks in our study.
... Our results showed that E-cadherin of the exfoliated cells were valuable for the diagnosis of malignant effusions, with a high sensitivity, specificity, positive predictive value, negative predictive value rate similar to other studies. [22,23] In some previous studies, 85% of adenocarcinoma cells have reacted to E-cadherin marker, which almost complies with our findings (88%). [23,24] 3 adenocarcinoma samples, which reacted negatively to the E-cadherin, included 2 metastatic poorlydifferentiated colonic adenocarcinoma toward peritoneum and 1 metastatic poorly-differentiated gastric adenocarcinoma to pleura. ...
Article
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One of the problems in studying serous effusion cytological samples is differentiation of reactive mesothelial cells from metastatic adenocarcinoma cells. In this study, the immunohistochemical diagnostic value of E-cadherin and fibronectin markers for differentiation of these 2 groups of cells was studied. 50 cell block samples prepared from serous effusions were examined. Based on clinical and histological studies, 25 cases had primary carcinoma, and the other 25 were proved to be benign effusion cases. All the cases were studied for E-cadherin and fibronectin immunostaining using an envision technique. Statistical analyzes were performed employing Chi-square and exact Fisher tests, using SPSS software (version 16). 24 of the 25 benign cases were stained with fibronectin and 2 with E-cadherin, whereas from among the 25 metastatic cases, 2 reacted to fibronectin and 22 to E-cadherin. Considering the staining of the 2 markers under conditions that the cells were stained with fibronectin but not with E-cadherin, positive predictive value (PPV) and negative predictive value (NPV) to identify reactive mesothelial cells were 100% and 92.5% while under conditions that had not been stained with fibronectin but with E-cadherin, PPV and NPV to detect adenocarcinoma cells were 95.2% and 82.1%, respectively. Employing this short panel can be helpful for better differentiation of adenocarcinoma and reactive mesothelial cells in serous fluids.
... In this case, positivity for anti-calretinin and anti-cytokeratin 5/6 were helpful for the diagnosis of mesothelioma. There were convincing evidences that cytoplasmic and nuclear stainings for calretinin were largely observed in malignant mesotheliomas, but not in other adenocarcinomas 10,11 . Positive reactions for cytokeratin 5/6 also suggested mesothelioma 12 . ...
Article
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A 61-year-old woman came to the hospital with dyspnea and pleural effusion on chest radiography. She underwent repeated thoracentesis, transbronchial lung biopsy, bronchoalveolar lavage, and thoracoscopic pleural biopsy with talc pleurodesis, but diagnosis of her was uncertain. Positron emission tomography showed multiple lymphadenopathies, so she underwent endobronchial ultrasound-guided transbronchial needle aspiration of mediastinal lymph nodes. Here, we report a case of malignant pleural mesothelioma that was eventually diagnosed by endobronchial ultrasound-guided transbronchial needle aspiration. This is an unusual and first case in Korea.
... O autor conclui que os melhores marcadores para serem utilizados na diferenciação entre mesotelioma epitelial e adenocarcinoma são a calretinina e a citoqueratina 5/6, que são expressas nos mesoteliomas e não nos adenocarcinomas; e o antígeno carcinoembrionário, que é expresso nos adenocarcinomas e não nos mesoteliomas epiteliais. Apesar de existirem algumas controvérsias na literatura (5,6,22) , a hiperplasia mesotelial reacional é geralmente negativa para o antígeno da membrana epitelial ou apresenta fraca positividade, é negativa para o antígeno carcinoembrionário e reage positivamente com a calretinina; o mesotelioma é difuso e intensamente positivo para o antígeno da membrana epitelial e a calretinina, sendo negativo para o antígeno carcinoembrionário. No presente estudo, a imuno-histoquímica foi indicada para a tentativa de diferenciar hiperplasia mesotelial reacional, mesotelioma e adenocarcinoma em 11% dos casos usando calretinina, antígeno carcinoembrionário e antígeno de membrana epitelial. ...
Article
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OBJETIVO: avaliar o motivo para indicação e analisar o grau de auxílio da imuno-histoquímica (IHQ) para o diagnóstico de neoplasias e lesões pseudoneoplásicas. MATERIAL E MÉTODO: avaliação retrospectiva observacional e descritiva de 4.459 casos submetidos a análise no laboratório de IHQ do Departamento de Patologia do Jackson Memorial Hospital, Universidade de Miami, Estados Unidos, em 1999. RESULTADOS: 3.706 casos possuíam dados suficientes para o desenvolvimento dos objetivos propostos. Em 19% dos casos a IHQ foi utilizada para determinar fatores prognósticos ou índices proliferativos; em 17% dos casos teve como objetivo identificar microrganismos, células, estruturas ou materiais; e em 64% dos casos teve aplicação diagnóstica propriamente dita. Em 835 casos desta última categoria a IHQ contribuiu para um diagnóstico específico em 83% das vezes e diminuiu o número de diagnósticos diferenciais em 12%. Em 5% das vezes a IHQ não auxiliou o patologista devido a exigüidade de algumas amostras, presença de necrose extensa ou indiferenciação extrema de algumas neoplasias. Os principais problemas de diagnóstico diferencial para os quais a IHQ foi utilizada foram: determinar o local de origem de carcinomas e adenocarcinomas e diferenciar entre hiperplasia mesotelial reacional, mesotelioma e adenocarcinoma, entre outros. Foram utilizados 4,1 anticorpos por caso, em média. CONCLUSÕES: Quando bem indicada e aplicada, a imuno-histoquímica é um método diagnóstico complementar útil em 95% dos casos e muitas vezes contribui fundamentalmente para as condutas cirúrgica e terapêutica. Amostras muito exíguas ou necróticas e neoplasias extremamente indiferenciadas são situações que comprometem o exame imuno-histoquímico e seus resultados. Quando utilizada de maneira direcionada aos principais diagnósticos diferenciais, a técnica apresenta uma relação custo/benefício alta.
... O autor conclui que os melhores marcadores para serem utilizados na diferenciação entre mesotelioma epitelial e adenocarcinoma são a calretinina e a citoqueratina 5/6, que são expressas nos mesoteliomas e não nos adenocarcinomas; e o antígeno carcinoembrionário, que é expresso nos adenocarcinomas e não nos mesoteliomas epiteliais. Apesar de existirem algumas controvérsias na literatura (5,6,22) , a hiperplasia mesotelial reacional é geralmente negativa para o antígeno da membrana epitelial ou apresenta fraca positividade, é negativa para o antígeno carcinoembrionário e reage positivamente com a calretinina; o mesotelioma é difuso e intensamente positivo para o antígeno da membrana epitelial e a calretinina, sendo negativo para o antígeno carcinoembrionário. No presente estudo, a imuno-histoquímica foi indicada para a tentativa de diferenciar hiperplasia mesotelial reacional, mesotelioma e adenocarcinoma em 11% dos casos usando calretinina, antígeno carcinoembrionário e antígeno de membrana epitelial. ...
Article
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BACKGROUND: Immunohistochemistry (IHC) is a valuable tool in diagnostic surgical pathology. We evaluated the frequency of IHC use and its contribution to the final diagnosis of tumors and pseudotumors. METHODS: A retrospective study of the 4,459 cases received in 1999 for immunoperoxidase study at the Immunohistochemistry Laboratory, Department of Pathology, University of Miami/Jackson Memorial Hospital, USA, was performed. RESULTS: 3,706 cases yielded all data necessary for the study. In 19% of cases IHC was used for localization of predictive tumor markers or evaluation of proliferation indicators; 17% of cases performed IHC to identify organisms or acellular structures and in 64% of cases IHC examination was to aid the pathologists in differential diagnosis of tumors. In 835 cases of the latter category, IHC helped the pathologists to render a specific diagnosis in 83% of instances. In 12% IHC narrowed down the diagnostic possibilities to two or three entities. In the remaining 5% of cases, IHC had no contribution to the final diagnosis due to limited diagnostic material, extensive necrosis or lack of tumor differentiation. The main differential diagnosis dilemmas included determination of the site of origin of carcinomas, differentiation between reactive mesothelial hyperplasia, mesothelioma and adenocarcinoma, and demonstration of cell phenotype in undifferentiated neoplasms. The average of antibodies per case was 4.1. CONCLUSIONS: Immunohistochemistry is a valuable tool in diagnostic surgical pathology capable of delineating the nature of disease in 95% of cases. In some instances, IHC is essential for the treatment decision-making. On the other hand, limited diagnostic material, extensive necrosis or lack of tumor differentiation can interfere in IHC performance. A well-designed differential diagnosis-driven utilization of the technique is extremely cost-effective.
... The main diagnostic difficulty is differentiating MPM from adenocarcinoma (2). Among the aforementioned markers, calretinin, a vitamin D-dependent calcium-binding protein, is the most definitive in differentiating mesothelioma from adenocarcinoma (26); calretinin is largely present in mesothelioma, but absent in adenocarcinoma. Renal cell carcinoma may also complicate the diagnosis of MPM; calretinin and cytokeratin are positively expressed in epithelioid mesothelioma, but not in renal cell carcinoma (8). ...
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Malignant mesothelioma is a rare type of cancer, most commonly associated with exposure to asbestos. Mesothelioma of the peritoneum, the membrane lining the abdominal cavity, is extremely rare. The current study reports the case of a 60-year-old female who presented with intestinal fistula, recurrent incisional hernia and abdominal infection, with no history of asbestos exposure, and was diagnosed with clear cell MPM. Computed tomography scans of the abdomen revealed extensive small bowel adhesions and massive peritoneal effusion. Histological examination of biopsy specimens indicated a diagnosis of malignant peritoneal mesothelioma with clear cell morphology. A laparotomy was performed, with subsequent resection of the bowel with fistula. Follow-up examination performed at 1-year post-surgery revealed that the patient was alive and in generally good health.
... 4 Calretinin is a vitamin D-dependent calcium-binding protein that is expressed in mesothelial cell lines; even benign mesothelial cells may show the characteristic "fried-egg" staining pattern. 10 Cytokeratins specifically permit identification of the epithelial origin based on the expression profile. Immunohistochemistry confirmed the mesothelial origin of the splenic cyst in our case as evidenced by the positive calretinin and low molecular weight cytokeratin and weakly positive CA 19-9 staining in the cyst wall. ...
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A young adult in her third decade presented with a 2-week history of catching left upper abdominal pain and was detected to have a cystic lesion occupying almost the entire spleen. Laparoscopic total splenectomy was carried out, and the cyst wall revealed a true mesothelial cyst with no squamous metaplasia. The various aspects of mesothelial cysts, including immunophenotyping and treatment modalities, are briefly discussed.
... Expression of calretinin, a marker for mesothelial cells [24] was observed in cytoplasm of OSE cells (Fig 2c, 3) and positively correlates with cyto-keratins 7 and 8 expression (p <0.01). ...
... This is similar to what was obtained by Seda et al., [26] who found that no positive results were observed in other malignant tumors used in the study rather than MPM. David et al., [27] found the same result, as all adenocarcinoma specimens were classified as negative for calretinin staining. ...
... Several markers have been described in the literature as important for the discrimination between benign, primary and secondary malignancies in serous effusions. Relevant markers may be epithelial markers (cytokeratin 7, 8 and 20, EMA), lymphoid (LCA, CD5, CD15, CD20, CD68, kappa and lambda light chains), mesenchymal (vimentin, desmin) and mesothelial markers (calretinin), as well as other specific biomarkers (CEA, B72.3, Oc125, S100, E-Cadherin, Ber-Ep4, HMB-45, HMFG2, Her-2Neu, estrogen and progestron receptor, chromogranin, thyreoglobuline) [1,3,4,8,10,19,20,22,24,26]. Gupta et al. recently described a panel of common immunostains (EMA, CEA, Cytokeratin, B72.3, HMB45, Vimentin, S100, LCA, L26, and kappa and lambda light chains) to be useful in confirming or suggesting the site of the primary tumor [10]. ...
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Cytological examination is a valuable diagnostic tool in case of a serous effusion. The first manifestation of malignancy may be an effusion of the pleural, pericardial, or peritoneal cavity, especially in carcinoma of the ovary, or lung, and malignant mesothelioma. In other malignancies effusions may occur in the course of the disease. The contribution by Mother by et al. in this issue of ACP focuses on the contribution of image and flow cytometry to establish the presence or absence of malignancy in serous effusions. They point out that the sensitivity of DNA image cytometry in equivocal effusions may be as high as 87.5%, and that for the detection of malignancy, DNA image cytometry is superior to flow cytometry.
... and CAM2) directly from the malignant ascites of ovarian cancer patients (Supplemental Table 1; supplemental material available online with this article; https://doi.org/10.1172/JCI146186DS1). Immunostaining confirmed that the primary mesothelial cells were positive for the mesothelial markers calretinin (8,23,24), cytokeratin, and vimentin (25) and were negative for markers of other cell types, including immune, endothelial, and fibroblast cells (Supplemental Figure 1A). Primary mesothelial cells displayed a char-OPN in the ovarian cancer microenvironment and that therapeutic targeting of OPN may be an effective strategy for enhancing platinum sensitivity in ovarian cancer. ...
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Ovarian cancer is the leading cause of gynecological malignancy-related deaths, due to its widespread intraperitoneal metastases and acquired chemoresistance. Mesothelial cells are an important cellular component of the ovarian cancer microenvironment that promote metastasis. However, their role in chemoresistance is unclear. Here, we investigated whether cancer-associated mesothelial cells promote ovarian cancer chemoresistance and stemness in vitro and in vivo. We found that osteopontin is a key secreted factor that drives mesothelial-mediated ovarian cancer chemoresistance and stemness. Osteopontin is a secreted glycoprotein that is clinically associated with poor prognosis and chemoresistance in ovarian cancer. Mechanistically, ovarian cancer cells induced osteopontin expression and secretion by mesothelial cells through TGF-β signaling. Osteopontin facilitated ovarian cancer cell chemoresistance via the activation of the CD44 receptor, PI3K/AKT signaling, and ABC drug efflux transporter activity. Importantly, therapeutic inhibition of osteopontin markedly improved the efficacy of cisplatin in both human and mouse ovarian tumor xenografts. Collectively, our results highlight mesothelial cells as a key driver of ovarian cancer chemoresistance and suggest that therapeutic targeting of osteopontin may be an effective strategy for enhancing platinum sensitivity in ovarian cancer.
... A negative result from a panel of adenocarcinoma-associated antibodies (e.g., Ber-EP4, CEA, MOC 31) is the standard approach in many laboratories, often combined with recently proposed mesothelioma-associated antibodies (e.g., calretinin, thrombomodulin, CD44H). [1][2][3][4] The value of cytokeratin (CK) immunophenotyping of the tumor cells in pleural effusions 5 or pleural biopsies 6 also has been studied. The most often proposed cytokeratin antibody is CK5 (CK5-6), considered as one of the most specific mesothelioma-associated antibodies, and seldom expressed by adenocarcinomas. ...
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... There was scattered PAX-8 positivity, a nonspecific finding. CK5/6 (epithelial markers), WT-1 (a marker for mesothelioma) [7 ,8] , D2-40 [9] , calretinin [10] , p40 [11] , estrogen receptor (see Discussion section), progesterone receptor (PR), STAT-6 (see Discussion section), Ckit [12] , ERG [13] , and CD1a [14] stains were negative in the lesional cells. Markers for melanoma (HMB45, Melan-A, and S100 [15 ,16] ) were negative. ...
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Pulmonary adenomyomas are rare adenomyomatous hamartomas. In the few cases described in the literature, these benign tumors are encapsulated by lung parenchyma. We describe a case of a 59 year-old woman with acetylcholine receptor antibody-negative myasthenia gravis and a right cardiophrenic mass initially thought to be a thymoma. Histopathology surprisingly revealed a pulmonary adenomyoma which involved the mediastinal fat at the cardiophrenic angle. Keywords: Pulmonary adenomyoma, Harmatoma, Adenofibroma, Solitary fibrous tumor, Myasthenia gravis, Thymoma
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The presence of malignant cells in peritoneal washings leads to classification as International Federation of Gynecology and Obstetrics stage IC or higher in ovarian carcinomas and at least International Federation of Gynecology and Obstetrics stage IIIA in endometrial carcinomas. Unfortunately, the morphologic examination of cytologic specimens has not proven to be a sensitive or specific diagnostic tool. Malignant cells might be few in number and might be unrecognized among a large population of mesothelial cells and/or macrophages, or reactive mesothelial cells might be misinterpreted as neoplastic cells leading to unnecessary chemotherapy. To evaluate the main pitfalls in the evaluation of peritoneal washings in patients with gynecologic malignancies and analyze the ancillary studies that might be helpful to achieve the correct diagnosis with an emphasis on immunocytochemistry. A comprehensive review of the literature was performed. Peritoneal effusions may represent major challenges to the pathologist and can have important clinical implications. Immunostains for epithelial markers such as B72.3, MOC-31, and Ber-EP4 represent the best available markers to identify epithelial cells. Caution is advised to not overdiagnose endometriosis or endosalpingiosis as adenocarcinoma.
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(CHEST 2009; 135:578–582) A n 81-year-old man presented with persistent dyspnea and left pleuritic chest pain for 2 months following a left lower lobe pneumonia treated at another institution. He denied undergoing thoracentesis or pleural catheter drainage during treatment of the pneumonia. He reported resolution of cough, and denied hemoptysis, fever, night sweats, or weight loss. His exposure history included working on a farm as a teenager and in iron mines for 4 years in the 1950s, and retail sales work thereafter. He was a life-long nonsmoker, had no significant travel history outside of Minnesota, and his only pets were two cats and a dog. He specifically denied exposure to tuberculosis, asbestos, or silica. His medical history was significant for hypertension, gastroesophageal reflux disease, and prostatic adenocarcinoma (Gleason grade 3 4, score 7), treated with radiation therapy 5 years previous, with a currently normal serum prostate specific antigen. His medications included atenolol, felodipine, hydrochlorothiazide, lisinopril, omeprazole, and entericcoated acetylsalicylic acid. Physical examination revealed small mobile cervical adenopathy, decreased breath sounds at the left lung base, and no edema or
Article
To evaluate the use of a panel of markers to differentiate adenocarcinoma and the reactive/inflammatory process in fluid cytology, we stained 29 formalin-fixed, paraffin-embedded cell blocks of effusion fluid from patients with metastatic adenocarcinoma and 24 cell blocks from patients with benign effusion with mucicarmine and antibodies to carcinoembryonic antigen (CEA), B72.3, and calretinin. Positive staining with CEA, B72.3, and mucicarmine was seen in 22 (76%), 20 (69%), and 18 (62%) adenocarcinoma cases, respectively. All except 1 adenocarcinoma was negative for calretinin. No benign cases were positive for B72.3 and mucicarmine. In 1 benign case, scattered epithelial cells demonstrated weak positivity for CEA. The majority of combinations were 100% specific for adenocarcinoma. The highest sensitivity (86%) for adenocarcinomas was achieved with the staining combination of negative for calretinin and positive for any adenocarcinoma marker (CEA, B72.3, or mucicarmine). The use of a panel of markers that recognize adenocarcinoma and mesothelial cells is useful in the differential diagnosis between metastatic adenocarcinoma and the reactive/inflammatory process. The profile of positive staining with at least one of the adenocarcinoma markers and negative calretinin staining is highly specific and sensitive for identifying adenocarcinoma in fluid cytology.
Article
The objective of the study is to estimate the expression of some antibodies in the metastatic adenocarcinomas, malignant epithelial mesotheliomas, and reactive mesothelial cells in serous effusions and to choose effective panel to the differential diagnosis. Totally 113 effusion cytology samples (80 pleural fluid, 30 ascitic, and 3 pericardial fluid) from 60 cases of metastatic adenocarcinoma (ACA), 18 cases of malignant epithelial mesothelioma (MM), and 35 cases of reactive mesothelium (RM) were included in this study. The cytological diagnoses of these cases were confirmed by histopathology or clinical datas. Smears and cell blocks were prepared for each case. Immunocytochemical study was performed on the cell block sections. The sensitivity of E-cadherin, CEA, MOC-31, and Ber-EP4 for adenocarcinoma was 86.7%, 80%, 70%, and 76.4%, respectively. The specificity was 98.1%, 96.2%, 92.5%, and 86.8%, respectively. The sensitivity of calretinin, HBME-1, and thrombomodulin for RM/MM was 83%, 79.2%, and 47.2% respectively. The specificity was 88.3%, 21.7%, and 70%, respectively. The expression of E-cadherin, CEA, MOC-31, Ber-EP4, calretinin, and thrombomodulin showed significant difference between ACA and RM/MM (P < 0.01). The reactivity of EMA and Des showed significant difference between RM and MM (P < 0.01). In our opinion, the antibody panel that consists of E-cadherin, CEA, calretinin, and thrombomodulin should be the best for differential diagnosis between metastatic adenocarcinomas and RM/MM in serous effusions. EMA and Des should be used to differentiate malignant epithelial mesothelioma and reactive mesothelial cells. EMA positive and Des negative favor MM, while Des positive and EMA negative favor RM.
Article
The distinction of mesothelial cells in cytologic samples is often a diagnostic challenge. This is particularly true in potentially malignant effusions in which reactive mesothelial cells may simulate adenocarcinoma (ACA) cells, and in the differentiation of ACA vs. mesothelioma. We sought to determine the superior antibody for the positive identification of mesothelial cells in these circumstances. Cell block sections of 25 reactive and 8 malignant mesothelioma effusions were immunostained with an avidin-biotin procedure, using antibodies to HBME-1 and calretinin. No pretreatment of samples was necessary for the HBME-1-stained slides; microwave antigen retrieval was performed on all slides stained for calretinin. A negative control was performed on each sample. The staining intensity of tumor cells was scored on a scale of 0–3+, with the proportion of immunoreactive cells categorized as <25%, 25–50%, 50–75%, and >75%. The predominant staining pattern for HBME-1 was surface, with rare samples also exhibiting cytoplasmic staining as well. The calretinin-staining pattern was cytoplasmic, with peripheral condensation/prominence and accompanying nuclear staining. All samples were immunoreactive with both antibodies. Fifty-five percent (18/33) of samples showed significantly stronger immunoreactivity with calretinin than with HBME-1; 45% (15/33) of samples showed equivalent staining with the two markers. None of the samples in this study showed stronger immunoreactivity with HBME-1 than with calretinin. Sixty-one percent (20/33) of samples stained with HBME-1 at a moderate (2+) intensity. Fifty-five percent (18/33) of samples stained with calretinin at a strong (3+) intensity. While only 12% of samples showed >75% immunoreactivity for HBME-1, 58% of samples showed >75% of cells immunoreactive for calretinin. Calretinin is the preferred marker in identifying mesothelial cells in cytologic samples, showing the highest sensitivity for mesothelial cells, as evidenced by a more intense staining reaction in a higher percentage of cells than with HBME-1. Diagn. Cytopathol. 2001;25:158–161. Published 2001 Wiley-Liss, Inc.
Article
Renal cell carcinoma (RCC) is known for its unpredictable behavior. RCC rarely involves serosal surfaces and, when present, can be difficult to distinguish from mesothelial cells in cytologic preparations. Immunohistochemical stains are frequently used with effusion cytology; however, RCCs express traditional glandular antigens less frequently than other adenocarcinomas. We investigated the utility of typical immunohistochemical stains for identifying effusion involvement by RCC, along with more specific RCC markers. The cytology databases from two academic institutions were searched for all effusions involved by RCC with retrievable cell-block material. A four-marker immunohistochemical panel we generally use for distinguishing adenocarcinoma from mesothelial proliferations was then applied (calretinin, WT1, MOC31, and B72.3). In addition, each case was stained for RCC antigen, CD10, and PAX2. Eleven cases of RCC involving serous effusions were identified: six conventional clear-cell RCCs, three papillary RCCs, and two RCCs, not otherwise specified. Neoplastic cells were positive for MOC-31 in 3 of 11 cases, RCC antigen in 5 of 11 cases, and CD10 in 10 of 11 cases. RCC cells were negative for B-72.3, WT1, and calretinin in all cases. Background mesothelial cells showed high-background cytoplasmic staining for PAX-2; all RCC tumor cells were negative or equivocal. A conventional panel used for the diagnosis of adenocarcinoma in fluids will fail to detect most cases of metastatic RCC, particularly clear-cell RCC. Additional antibodies, such as those to CD10 and RCC, may be helpful to identify these tumors. PAX2 shows high background in mesothelial cells, which makes interpretation of nuclear staining difficult.
Article
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. Diagn. Cytopathol. 2002;26:61–66. Published 2002 Wiley-Liss, Inc.
Article
To determine the true incidence of Müllerian and mesothelial lymph node involvement in serous and mucinous borderline ovarian tumors (BLOT) with serial sectioning and immunohistochemistry. Formalinfixed, paraffin-embedded lymph node blocks from patients with serous (N = 21) and mucinous (N = 5) BLOT who underwent lymphadenectomy between 1995 and 2002 were serially sectioned at 5 microm levels with 3 consecutive sections taken at surface, 125 microm and 475 microm. One slide from each level was stained with hematoxylin-eosin (H-E), cytokeratin (AE1-AE3, DAKO) and calretinin (DAKO). Lymph node involvement was defined as epithelioid cells recognized by H-E and confirmed by immunoreaction with keratin (Müllerian) and calretinin (mesothelial) or identified by immunohistochemistry alone. The results obtained by serial sectioning and immunohistochemistry were compared with those obtained by routine histologic examination at the time of the original surgery. A total of 240 lymph nodes (215 from patients with serous and 25 from patients with mucinous BLOT) were examined. Original pathologic examination identified lymph node involvement in 29/215 lymph nodes from 21 patients with serous BLOT. Twelve of the 21 patients with serous BLOT (57%) and none of the 5 patients with mucinous BLOT (0%) demonstrated Müllerian lymph node involvement. Serial sectioning and keratin immunostaining identified Müllerian involvement in 4 (1.6%) and 10 (4.2%) additional nodes not diagnosed in original sections, respectively. However, no additional node-positive patients were identified. Mesothelial involvement was identified in 2 patients (2/26, 7.6%). Patients with serous BLOT have a high incidence of Müllerian lymph node involvement. Distinction between Müllerian and mesothelial differentiation may require immunohistochemical study. Compared with routine histologic examination, serial sectioning and immunohistochemical examination yield a higher number of involved lymph nodes.
Article
It has been shown that detection of the Wilms tumor susceptibility gene 1 protein (WT1) has diagnostic utility in the distinction of mesothelioma from adenocarcinoma in tissue sections of pleural tumors. This immunohistochemical study evaluates the effectiveness of WT1 as a marker for malignant mesothelioma in paraffin sections of cell block preparations derived from effusion specimens. The authors evaluated 111 cell blocks for WT1 immunoreactivity, including 14 mesotheliomas and 97 metastatic adenocarcinomas from various sites. Nuclear reactivity for WT1 was observed in all samples of mesothelioma. However, only 22 of 97 samples (23%) of metastatic adenocarcinoma, nearly all of which were of ovarian origin (91%), exhibited nuclear reactivity for WT1. In 14 other samples (most of pulmonary derivation), WT1 staining restricted to the cytoplasm was observed for some tumor cells and was regarded as nonspecific. Based on this staining profile, WT1 represents an effective marker for mesotheliomas in cell block preparations and can aid in its distinction from pulmonary adenocarcinoma. In assessment of effusion specimens with metastatic carcinoma, nuclear reactivity for WT1 is highly suggestive of an ovary primary tumor
Chapter
Peritoneal washing cytology has been used in the evaluation of gynecologic malignancies for many decades. FIGO introduced peritoneal washing as part of the staging of ovarian carcinomas in 1975. The presence of malignant cells in serous effusions indicates spread of disease beyond the organ of origin and is associated with significant therapeutic and prognostic implication. This chapter discusses the cytomorphologic criteria for the diagnosis of ovarian tumors in liquid cytology, potential pitfalls, and ancillary methods that may be helpful in characterizing cell types. KeywordsCytology-Effusion-Pelvic washing-Peritoneal washing - Ovarian tumors-Peritoneum
Article
Differentiating reactive mesothelial cells (RMs) from metastatic adenocarcinoma cells (MAC) in serous fluids based on cytomorphologic features alone can be very challenging. Various immunocytochemical (ICC) markers have been used to maximize the diagnostic accuracy, however, cytopathologists still encounter difficulties in effusion cytologic diagnosis. The aim of this study was to evaluate previous and recent ICC stains to identify the most sensitive and specific markers and the best panel for differentiating RM from MAC. Cell block sections from 41 MAC and 43 RM effusions cases were subjected to ICC staining for MOC-31, BerEp4, carcinoembryonic antigen (CEA), calretinin, HBME-1, CK5/6, and D2-40. For the MAC cases, the sensitivity of BerEp4, MOC-31, and CEA was 82.9, 92.6, and 17%, respectively, and the specificity was 95.3, 93, and 100%, respectively. For the RM cases, the sensitivity of calretinin, CK5/6, D2-40, and HBME-1 was 95.3, 27.9, 58.1, and 93%, respectively, and the specificity was 70.7, 73.1, 75.6, and 82.9%, respectively. The results show that BerEp4 and MOC-31 are highly sensitive and specific for detecting MAC, whereas calretinin and HBME1 are highly sensitive but only modestly specific for detecting RM cases (P < 0.05). Forced entry logistic regression revealed that using MOC-31, BerEp4, HBME-1, and calretinin, is an excellent panel for making correct diagnosis with 97.6% sensitivity in detecting MAC and 90.7% specificity in detecting RM. We conclude that adding a panel of MOC-31, BerEp4, calretinin, and HBME-1 immunostains to routine cytomorphologic features can greatly enhance the diagnostic accuracy of serous effusions.
Article
The aim of this study was to investigate the occurrence of (pre)neoplastic lesions in overtly normal Fallopian tubes from women predisposed to developing ovarian carcinoma. The presence of (pre)neoplastic lesions was scored in histological specimens from 12 women with a genetically determined predisposition for ovarian cancer, of whom seven tested positive for a germline BRCA1 mutation. A control group included 13 women. Immunohistochemistry was used to determine the expression of p21, p27, p53, cyclin A, cyclin D1, bcl-2, Ki67, HER-2/neu, and the oestrogen and progesterone receptors. Loss of heterozygosity (LOH) analysis on the BRCA1 locus was also assessed on dysplastic tissue by PCR studies. Of the 12 women with a predisposition for ovarian cancer, six showed dysplasia, including one case of severe dysplasia. Five harboured hyperplastic lesions and in one woman no histological aberrations were found in the Fallopian tube. No hyperplastic, dysplastic or neoplastic lesions were detected in the Fallopian tubes of control subjects. In the cases studied, morphologically normal tubal epithelium contained a higher proportion of Ki67-expressing cells (p=0.005) and lower fractions of cells expressing p21 (p<0.0001) and p27 (p=0.006) than in the control group. Even higher fractions of proliferating cells were found in dysplastic areas (p=0.07) and accumulation of p53 was observed in the severely dysplastic lesion. Expression patterns of other proteins studied, including the hormone receptors, were similar in cases and controls. One subject, a germline BRCA1 mutation carrier, showed loss of the wild-type BRCA1 allele in the severely dysplastic lesion. In conclusion, the Fallopian tubes of women predisposed to developing ovarian cancer frequently harbour dysplastic changes, accompanied by changes in cell-cycle and apoptosis-related proteins, indicating an increased risk of developing tubal cancer. Copyright © 2001 John Wiley & Sons, Ltd.
Article
Full-text available
O mesotelioma maligno é uma neoplasia rara associada, em 80 % dos casos, a exposição prolongada a asbestos, existindo um período de latência de 20 a 50 anos. O seu tratamento é quase sempre paliativo, dada a extensão da doença aquando do diagnóstico, sendo o tumor pouco sensível à quimioterapia e à radioterapia. A sobrevivência média varia entre 4 e 18 meses, raramente ultrapassando os 5 anos.
Article
Definitive cytopathological interpretation of some of the effusion fluids may not be possible based on cytomorphological evaluation alone. As discussed in other reviews, this is due to various reasons specifically applicable to effusion fluids including remarkably wide morphologic spectrum of reactive mesothelial cells overlapping with some well to moderately differentiated metastatic carcinoma. The challenge is subject to various factors including level of interpreter training or experience, institutional demographics (such as type of prevalent diseases, predominant sex and age group), technical advances in ancillary support, and expertise in cytopreparatory processing. In such cases immunohistochemistry performed on cell-block sections is simple objective adjunct with or without other ancillary techniques. Ongoing increase in number of immunomarkers along with rabbit monoclonal antibodies with relatively higher affinity is further refining this field. SCIP (subtractive coordinate immunoreactivity pattern) approach, discussed as separate dedicated review article, facilitates refined interpretation of immunoreactivity pattern in coordinate manner on various serial sections of cell-blocks. However, many variables such as delay after specimen collection, specimen processing related factors including fixation and storage; ambient conditions under which paraffin blocks are archived (for retrospective testing); antigen retrieval method; duration of antigen retrieval step; antibody clone and dilution; and antibody application time are common with application of immunohistochemistry in other areas. This review is dedicated to highlight technical aspects including processing of effusion specimens for optimum immunocytochemical evaluation along with commonly used immunomarkers in effusion cytopathology. This review focuses on the technical and general information about various immunomarkers.
Article
Due to the remarkably wide morphologic spectrum of reactive mesothelial cells, some of the effusion fluids may be difficult to interpret with objective certainty by cytomorphology alone. Cytomorphology of well to moderately differentiated adenocarcinomas (responsible for the bulk of malignant effusions) may overlap with floridly reactive mesothelial cells. Even mesotheliomas including diffuse malignant epithelioid mesothelioma, are usually cytomorphologically bland without unequivocal features of malignancy. The intensity of challenge depends on the interpreter’s training or experience level, institutional demographics of patients (such as type of prevalent diseases, predominant sex and age group), technical support, and quality of cytopreparatory processing. In general immunocytochemistry is valuable adjunct to facilitate objective interpretation with or without other ancillary techniques as indicated. An increasing number of immunomarkers is further refining the contribution of immunohistochemistry to this field. However, application of immunohistochemistry to effusion fluids is relatively challenging because of many variables. Multiple factors such as delay after specimen collection, specimen processing related factors including fixation and storage; ambient conditions under which paraffin blocks are archived (for retrospective testing); antigen retrieval method; duration of antigen retrieval step; antibody clone and dilution; and antibody application time are identical to application of immunohistochemistry in other areas. The significant challenge related to the potential compromization of the immunoreactivity pattern due to exposure to non-formalin fixatives / reagents is also applicable to effusion fluid specimens. The immunoreactivity results would be compared and corelated with cumulative metadata based on the reported studies performed and validated on formalin-fixed paraffin-embedded tissue sections. Deviating from such protocols may lead to suboptimal results, which is not uncommon in clinical practice with potential compromization of patient care and related liability. Because of this, it is critical to perform immunocytochemistry on formalin-fixed cell-block sections only. In addition, unless the interpretation criteria for immunohistochemical evaluation of effusion fluids are not modified specifically, it may not be productive in resolving some challenging cases. However, this aspect is not well elaborated in the literature. A basic and critical challenge is finding and locating the cells of interest in cell-block sections of effusion fluids. A unique approach is to choose a fundamental immunopanel which highlight the mesothelial and inflammatory cells in reactive effusion fluids to create the basic map. This allows detection of a ‘second-foreign’ population which can be immunocharacterized further with the help of subtractive coordinate immunoreactivity pattern (SCIP) approach elaborated here.
Article
Background: The cytologic evaluation of serous effusions to distinguish malignant cells from reactive mesothelial cells (RMCs)was an enormous challenge. The purpose of this study was to investigate the diagnostic value of glucose transporter 1 (GLUT1) and calretinin (CR) in serous effusions of patients with malignant and in order to significantly ameliorate the diagnostic accuracy. Methods: The expressions of GLUT1 and CR were measured by streptavidin-peroxidase (S-P) immunocytochemical technique in serous effusions of 183 patients with malignant and in 95 patients with benign diseases. Results: The positive ratio of GLUT1 was 91.8% (168/183) in serous effusions from patients with malignant and 5.3% (5/95) in benign diseases, they had a significant difference (P < .01). CR was expressed 89.5% (85/95) in benign diseases and 6.6% (12/183) in malignant, it also showed an important difference (P < 0.01). The combination of GLUT1 + CR revealed the best efficiency: the sensitivity and specificity were 100% and 98.9%, respectively. Conclusion: Immunocytochemical staining for GLUT1 and CR may be used as a complementary tool for the detection of malignant effusions and help to make a distinction between cancer cells and RMCs. The combination of GLUT1 and CR with immunocytochemistry stained can be achieved a higher diagnostic performance.
Article
The differential diagnosis in cellular effusions with cytological atypia often includes malignant mesothelioma (MM), reactive mesothelial proliferation, and malignancies of metastatic origin, particularly carcinomas. The International Reporting System for Serous Fluid recently established guidelines for reporting MM. In conjunction with the cytomorphologic evaluation, the role of immunochemistry (IC) was emphasized as a very useful tool in the workup of serous fluids, especially with the availability of novel markers. Utilizing a panel of markers, IC allows the characterization of the cells, whether mesothelial or not, and when mesothelial origin is established, IC can frequently assist in delineating its benign or malignant nature. IC can also confirm metastatic disease, allowing the identification of the primary origin in most cases. This review summarizes the current status of IC and its role in the diagnosis of MM and its differential diagnosis in serous fluids.
Article
Context.—The presence of malignant cells in peritoneal washings leads to classification as International Federation of Gynecology and Obstetrics stage IC or higher in ovarian carcinomas and at least International Federation of Gynecology and Obstetrics stage IIIA in endometrial carcinomas. Unfortunately, the morphologic examination of cytologic specimens has not proven to be a sensitive or specific diagnostic tool. Malignant cells might be few in number and might be unrecognized among a large population of mesothelial cells and/or macrophages, or reactive mesothelial cells might be misinterpreted as neoplastic cells leading to unnecessary chemotherapy. Objective.—To evaluate the main pitfalls in the evaluation of peritoneal washings in patients with gynecologic malignancies and analyze the ancillary studies that might be helpful to achieve the correct diagnosis with an emphasis on immunocytochemistry. Data Sources.—A comprehensive review of the literature was performed. Conclusions.—Peritoneal effusions may represent major challenges to the pathologist and can have important clinical implications. Immunostains for epithelial markers such as B72.3, MOC-31, and Ber-EP4 represent the best available markers to identify epithelial cells. Caution is advised to not overdiagnose endometriosis or endosalpingiosis as adenocarcinoma.
Book
Rock iguanas of the West Indies are considered to be the most endangered group of lizards in the world. They are a flagship species in the Caribbean and on most islands are the largest native land animals. Unfortunately, human encroachment and introduced animals have brought this species to the brink of extinction. Cyclura: Natural History, Husbandry, and Conservation of the West Indian Iguanas is the first book to combine the natural history and captive husbandry of these remarkable reptiles, while at the same time outlining the problems researchers and conservationists are battling to save these beautiful, iconic animals of the Caribbean islands. Authors Jeffrey Lemm and Allison Alberts have been studying West Indian iguanas for nearly 20 years in the wild and in captivity; their experiences with wild iguanas and their exquisite photos of these charismatic lizards in the wild make this book a must-have for reptile researchers, academics and enthusiasts, as well as anyone interested in nature and conservation. Includes chapters with contributions by leading experts on rock iguana taxonomy, nutrition, and diseases Features color photos of all taxa, including habitat and captive shots Provides easily understandable and usable information gleaned from experience and hands-on reptile research.
Article
Metastatic urothelial carcinoma (UC) to serous effusion (SE) is extremely rare and its cytomorphological features have only been described in case reports. In this study, we searched the pathology database at University of Michigan for SEs due to metastatic UC in the last 20 years. A total of 25 cases from 20 patients with clinically and pathologically confirmed metastatic UC in SEs were retrieved. The specimens consisted of 15 pleural, 8 peritoneal, and 2 pericardial effusions. Smears were reviewed and evaluated for the following features: cellularity, single cells, cell clusters or short cords, cell wrapping, "windows" between the cells, two-tone cytoplasm, cytoplasmic vacuoles, signet ring cells, nuclear to cytoplasmic (N/C) ratio, nuclear hyperchromasia, irregular nuclear membrane, nuclear centricity, double or multiple nuclei, nucleoli, anaplastic cells and mitosis. Our results showed that UC manifested in SEs predominantly as a single cell population with or without clusters or short cords, and frequently exhibited the "cell wrapping" of two or more cells. Individual UC cell in SEs exhibited nuclear enlargement with increased N/C ratio, irregular nuclear membranes, hyperchromatic coarse chromatin and frequently prominent nucleoli. Double or multinucleated cells, cells with vacuolated cytoplasm or signet ring appearance were also frequently present. Our results demonstrated that while certain features could suggest the diagnosis of UC, the cytomorphological features are not specific and often overlap with those of reactive mesothelium, mesothelioma, metastatic adenocarcinoma, or squamous cell carcinoma in SEs. Accurate diagnosis of UC rests on the combination of clinical history, cytomorphologic features and appropriate immunohistochemical panel. Diagn. Cytopathol. 2012 © 2012 Wiley Periodicals, Inc.
Article
BACKGROUND Cytology plays a pivotal role in the diagnosis of pleural effusions. In many cases, immunocytochemistry (ICC) is required to elucidate the etiology of the atypical cells. Effusions are samples that present unique problems for ICC. To date there is no standardization of ICC methods for effusions and cytology in general.METHODS The authors review the most commonly used cytologic preparations, fixatives, and antibodies used in effusion ICC.RESULTSThrough the utilization of cell block preparations and a panel of antibodies appropriate for the differential diagnosis in question, ICC conditions utilized in surgical pathology can be most closely replicated.CONCLUSIONSICC may provide reliable insights into various diagnostic dilemmas in effusion cytology, provided that laboratory standardization practices are followed. Cancer (Cancer Cytopathol) 2001;93:293–308. © 2001 American Cancer Society.
Article
Background The cytologic assessment of pleural effusions to distinguish carcinoma cells from reactive mesothelial cells is particularly challenging. The aim of this study was to investigate the diagnostic value of monoclonal antibody (MOC-31) and calretinin in pleural fluid of patients with lung cancer to significantly improve the diagnostic accuracy.Methods The expressions of MOC-31 and calretinin were detected by means of S-P immunocytochemical technique in pleural effusions of patients with lung cancer (n = 92) and in patients with benign lung disease (n = 70).ResultsThe positive rate of MOC-31 in pleural fluid was 90.2% (83/92) from patients with lung cancer and 2.9% (2/70) from patients with benign lung diseases, showing a significant difference (P < 0.01). Calretinin was expressed 87.1% (61/70) in benign lung diseases and 6.5% (6/92) in lung cancer, also showing a significant difference (P < 0.01). The optimal combination for assay was MOC-31 + calretinin: Sensitivity and specificity were 100 and 98.6%, respectively.ConclusionMOC-31 and calretinin are of important clinical value for diagnosing and differentially diagnosing the cancer cells in pleural fluid of patients with lung cancer. Diagn. Cytopathol. 2014. © 2014 Wiley Periodicals, Inc.
Article
L’immunocytochimie est une technique d’appoint utile au typage de la plupart des épanchements néoplasiques des séreuses. Les auteurs décrivent les différentes techniques utilisables: étalements directs après centrifugation, cytocentrifugation, inclusion des culots, cytologie en milieu liquide. La fiabilité des immunomarquages dépend de l’expérience et des mises au point faites par chaque utilisateur. Les panels d’anticorps les plus couramment utilisés associent marqueurs des carcinomes et des cellules mésothéliales.
Article
This paper addresses well-known problem areas in the cytodiagnosis of serous effusions, relating to the distinction between adenocarcinoma, malignant mesothelioma and highly reactive mesothelial cells. The cytomorphology of these three entities has been scrutinized extensively by light microscopy; the majority of serous effusions are readily placed in one of the three categories based on astute assessment of cytological features and architecture of cell groupings. Nevertheless, a small, apparently irreducible number of cases remains in which overlapping cytological details make definitive diagnosis impossible. This occurs both histologically and cytologically. Adjunctive techniques such as histochemistry and electron microscopy were used in the past; however, it was not until the introduction of immunochemical analysis that diagnosis of exuberant serous lesions became more objective. The early generation of markers was directed primarily at the identification of adenocarcinoma, negative staining of both malignant mesothelioma and reactive mesothelial cells being accepted by exclusion. It is only in the last 4–5 years that positive markers of mesothelium have become commercially available and widely used. Chief amongst these agents are Calretinin and the Cadherins, which have brought a clarity, not previously anticipated, to bear on diagnosis in serous lesions, both histologically and cytologically. Future techniques such as measurement of ploidy and proliferation markers are addressed. It can now be stated that, with judicious use, definitive diagnosis of serous effusions can be accomplished cytopathologically in the majority of cases.
Article
The diagnosis of metastatic cancer in peritoneal fluid is of great importance for the patient and the attending physician. A cytopathologist's responsibility is twofold: (1) to accurately identify malignant cells; (2) to interpret tumor type and if possible the site of its origin even in the absence of complete clinical history of other clues. The difficulty in the diagnosis of metastatic neoplasms in peritoneal fluid is due to 2 factors: (1) abnormal mesothelial cells or macrophages may simulate cancer cells, or may conceal tumor cells; and (2) peritoneal fluid constitutes a natural and hitherto inadequately explored medium of cell culture, in which neoplastic cells may proliferate free of the boundaries imposed upon them by the framework of organs and tissues. Immunocytochemistry (ICC) and molecular techniques are essential to establish an accurate diagnosis. From a great many points of view malignant peritoneal fluid is suitable for continuous study of cancer cells, thus providing knowledge about biologic aspects of human solid tumors.
Article
Numerous new immunohistochemical markers that can be used in the diagnosis of mesothelioma have recently become available. As a result, new panels of antibodies that could be useful for distinguishing between epithelioid mesotheliomas and adenocarcinomas have been proposed. However, great differences of opinion exist regarding the individual value of some of these markers, especially when compared with those whose value has already been established. This article provides a critical review of the currently available information on those markers that could be useful in the diagnosis of epithelioid mesotheliomas or whose utility remains controversial. A practical approach to the diagnosis of these tumors is also provided.
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The transmission (TEM) and scanning electron microscopic (SEM) features of cancer cells and benign cells from human effusions were described. Both in the TEM and the SEM the following cell types were indentified; mesothelial cells, histiocytes, lumphocytes, eosinophilic leukocytes, neutrophilic leukocytes, plasma cells, erthrocytes and cancer cells. The ultrastructure of malignant cells originating from cancers most commonly metastisizing to the serous body cavities was described in detail. Both the TEM and the SEM clearly demonstrated fundamental differences between the surfaces of the cancer cells and benign cells in body cavity fluids: cancer cells were covered by numerous microvilli, while non-neoplastic cells were not. The approach described in this paper can be used for purposes of cytologic diagnosis in difficult preselected cases and may also have some important theoretical implications.
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One of the great challenges in the cytodiagnosis of effusions is the distinction between reactive mesothelium/histiocytes and cancer cells. This is notably true in patients having undergone radiation and/or chemotherapy. To establish whether monoclonal antibodies (MoAbs) could be used as reliable diagnostic adjuvants, the authors retrospectively and blindly studied 60 cases diagnosed by standard cytologic criteria (malignant, benign, and equivocal), with a panel of seven readily available MoAbs (cytokeratins, vimentin, EMA, B72.3, alpha-CEA, HMFG-2, and Leu-M1) and the lectin Ulex europaeus I. All 18 (100%) malignant cases showed reactivity with EMA and HMFG, whereas 17 (95%) and 11 (61%) reacted with B72.3 and alpha-CEA, respectively. Combinations of (1) EMA + B72.3, (2) EMA + alpha-CEA, and (3) EMA + alpha-CEA + B72.3 displayed positivity in 17 (95%), 11 (61%), and 10 (56%) malignant cases, respectively. Of the 18 benign cases, 7 reacted with HMFG and 2 each with EMA and B72.3. Only one case (5.5%) reacted with both EMA and B72.3. Based on these results, the 24 equivocal cases were regrouped into 14 malignant and 10 benign cases. Follow-up effusions obtained within the ensuing three months in all these patients allowed the authors to unequivocally confirm the diagnosis in all but five. The combination of EMA and B72.3 MoAbs detected malignant cells in 95% of the cases, with a 3.5% incidence of false positive cases in this study. A panel of EMA, B72.3, and alpha-CEA MoAbs should prove the most useful and simple approach to the correct diagnosis in most questionable effusions. Some of the potential pitfalls are discussed.
Article
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Keratin profiles of exfoliated mesothelial and adenocarcinoma cells were determined using antisera to different molecular weight keratins (45, 46, 55, 63 kdaltons) and the immunoperoxidase technic. Most metastatic adenocarcinomas in effusions stained for low (45, 46 kdaltons) and intermediate (55 kdaltons) molecular weight keratins but were negative for 63 kdalton keratin. In contrast, most reactive and malignant mesothelial cells in effusions stained strongly for 63 kdalton keratin and keratins of lower molecular weight. This is the first report of high molecular weight (greater than 60 kdaltons) keratin in exfoliated cells of nonepidermal origin. Differences in staining for 63 kdalton keratin between mesothelial and adenocarcinoma cells may help to distinguish these cells in effusions.
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This is a report of SEM studies of several cell types observed in pleural and ascitic fluids of patients with a variety of disorders. Prior to scanning, the identity of the cells was determined by light microscopic studies performed by a simple and inexpensive method developed by the authors. Differences in surface configuration of mesothelial cells, macrophages, leukocytes, and cancer cells were described and discussed. Irregular surface microvilli of high density were the predominant surface feature of cells of metastatic cancers of various origins and types, except for oat-cell carcinoma. Mesothelial cells had variable surface configuration with the majority of cells covered by blebs. Macrophages were primarily characterized by veil-like ridges. Detailed description of the various cell types under various circumstances and a discussion of the possible significance of the surface features in reference to cell biology conclude this review.
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To document that a polyclonal antiserum to calretinin, a 29-kd calcium-binding protein, consistently decorates normal and tumor mesothelial cells in cytologic preparations. Thirty-three archival cytologic specimens from eight patients with histologically confirmed malignant mesothelioma and 13 from patients with metastatic serous effusions were destained and then immunostained with anticalretinin antiserum. For investigation of cell suspensions, four pleural fluids were incubated with anticalretinin antiserum. After cytocentrifugation the specimens were stained in accordance with the alkaline phosphatase anti-alkaline phosphatase (APAAP) method. For electron microscopic examination the cell suspensions were then incubated with gold-labeled antirabbit antibody. The diagnostic sensitivity of this new immunocytochemical approach reached 100% for the eight malignant mesotheliomas investigated. Only 3 of the 13 adenocarcinomas metastatic to the serous membranes included in this study were weakly reactive, accounting for 81% specificity. Binding of anticalretinin antiserum to living mesothelial cells was consistently documented in all four cases investigated. Calretinin is a very useful marker for positive identification of normal and tumor mesothelial cells in serous effusions.
Article
HBME-1 monoclonal antibody has recently become available as a suggested immunohistochemical tool for the positive identification of malignant mesothelioma. In this study, the HBME-1 antibody was immunohistochemically evaluated on selected normal tissues and in 540 epithelial and nonepithelial tumors of different organs. Most mesotheliomas (25 of 29) showed a strong reactivity, with either a luminal or a cytoplasmic pattern. However, less differentiated mesotheliomas with sarcomatous features as well as sarcomatous areas of otherwise more differentiated mesotheliomas were negative. With regard to carcinomas, about half of the adenocarcinomas of the lung showed reactivity that was usually less conspicuous than the reactivity observed in mesothelioma. Consistently positive adenocarcinomas included endometrial and ovarian serous carcinomas. Thyroid lesions reacted differently: papillary and follicular carcinomas were strongly positive, whereas follicular adenomas and normal tissues were usually negative, suggesting that HBME-1 may be helpful in the evaluation of thyroid lesions. While gastric and pancreatic adenocarcinomas showed positivity in a minority of cases, tumors from the breast only rarely showed significant reactivity. Adenocarcinomas of colon, kidney, and prostate were almost invariably negative. Among soft-tissue and bone tumors, chordoma and cartilaginous tumors were strongly positive, and synovial sarcoma showed reactivity in the epithelial component. Other sarcomas and melanomas were negative. On the basis of our findings, HBME-1 monoclonal antibody is helpful in the differential diagnosis of mesothelioma, but because adenocarcinomas of many sites, including lung, are sometimes strongly positive, this antibody has to be used in a panel with other antibodies. We believe that the HBME-1 antibody could replace one of the antibodies that is typically negative in mesothelioma and positive in carcinoma. Interestingly, HBME-1 appears to be useful in subtyping lesions of certain organs, primarily thyroid tumors, soft-tissue and bone tumors of cartilaginous differentiation, and those with epithelial traits.
Article
Mesotheliomas and metastatic adenocarcinomas involving the pleura are frequently difficult to distinguish by light-microscopic and histochemical methods. In a double-blind study, we have compared ultrastructural features of 10 mesotheliomas of epithelial type and 10 adenocarcinomas from the lung, breast, and upper GI tract, i.e., sites known to give rise to metastases which mimic mesothelioma. Mesotheliomas were observed to have a significantly greater microvillus length/diameter ratio (LDR) than adenocarcinomas (p < 0.01) and more abundant intermediate filaments (p < 0.001). Mesotheliomas had more complex microvilli than adenocarcinomas, whereas adenocarcinomas had rootlets (2/10 cases) and lamellar inclusion bodies (2/10 cases), both of which were absent in the mesotheliomas. This study provides quantitative and qualitative ultrastructural features of potential utility in the differential diagnosis of pleural mesotheliomas and adenocarcinomas.
Article
The immunohistochemical diagnosis between epithelial mesothelioma and adenocarcinoma is currently based on the use of a panel of antibodies to adenocarcinoma-associated antigens and a few antibodies to mesothelial-associated antigens. Since the introduction of epitope retrieval methods, the sensitivity of many antibodies has been enhanced. Thus, a reevaluation of the mesothelioma/adenocarcinoma diagnostic panel becomes necessary. We studied 268 paraffin-embedded formalin-fixed tumor samples that included 57 epithelial mesotheliomas and 211 adenocarcinomas of various origins, comparing an extensive antibody panel with and without heat-induced epitope retrieval (HIER). Marked increase in the sensitivity of several antibodies, with no loss of specificity, was found when HIER was used. After statistical analysis, the antibodies to the epithelial glycoproteins carcinoembryonic antigen, BerEp4, and Bg8 emerged as the best discriminators between adenocarcinoma and epithelial mesothelioma within the entire panel. The mesothelium-associated antibodies, HBME-1, calretinin, and thrombomodulin were less sensitive and less specific than the former, although they were found to be useful on certain cases. Antibodies to cytokeratins and vimentin, although of minor diagnostic value in this context, may be helpful to evaluate the quality of antigen preservation. This study confirms the value of immunohistochemistry to accurately distinguish mesothelioma from adenocarcinoma when an antibody panel approach is used. The addition of heat-induced epitope retrieval methods increases the effectiveness of the procedure and is recommended for most of the antibody panel members.
Article
Diagnostic Surgical Pathology, Editor: Stephen S. Sternberg; Lippincott, Williams and Wilkins, Philadelphia, 1999.
Article
A novel gene of the calmodulin superfamily, encoding a 29-kD neuronal protein here named "calretinin," has been isolated as a cDNA clone from chick retina. The encoded sequence includes four putative calcium-binding sites and a fusion protein binds calcium. The most similar protein known is the 28-kD intestinal calcium-binding protein, calbindin (58% homology). Both genes date from before the divergence of chicks from mammals. The distribution of calretinin and calbindin mRNAs in chick tissues has been mapped using RNA gel blots and in situ hybridization. RNAs from both genes are abundant in the retina and in many areas of the brain, but calretinin RNA is absent from intestine and other nonneural tissues. Calretinin and calbindin are expressed in different sets of neurons throughout the brain. Calretinin RNA is particularly abundant in auditory neurons with precisely timed discharges.
Article
The histologic types of lung cancer in 855 patients (747 men and 107 women) from three hospitals and one international study of insulation workers were evaluated. Of these, 196 cases had asbestos exposure. About one half of the cases were diagnosed from surgical slides and one half from autopsy slides. Squamous cell carcinoma constituted the largest percentage of tumor types and was found with the same frequency in exposed and nonexposed groups. Small cell carcinoma was found in 25% of the exposed and in 15% of the nonexposed patients. Upper lung sites were involved in about two thirds of the cases with asbestos exposure and lower lobes in the other one third. There was little difference in histologic type in cases regardless of whether upper or lower lobes were involved. Cigarette smokers who smoked until their cancer diagnosis showed no difference in histologic type by amount smoked, and slight but not statistically significant differences from ex-cigarette smokers.
Article
The distinction between reactive mesothelial cells (RMC), malignant mesothelioma (MM), and metastatic adenocarcinoma (ACA) in pleural effusions may be impossible based on morphology alone. E-cadherin, N-cadherin, and calretinin are newly described immunocytochemical markers which can potentially be utilized for facilitating this distinction. E-cadherin and N-cadherin are calcium-dependent intercellular adhesion molecules expressed in epithelial cells and mesenchymal/mesothelial cells, respectively. The differential expression of E-cadherins in epithelial cells and N-cadherins in mesothelial cells has been utilized to differentiate reactive mesothelial cells, MMs and ACAs. Calretinin is a calcium-binding protein within the family of EF-hand proteins. It is abundantly expressed in peripheral and central nervous tissues, and has been shown to consistently immunoreact with mesothelial cells. We studied cell block sections from 77 pleural effusions (22 RMC, 26 MM, and 29 ACA) to investigate the potential immunocytochemical use of anti-E-cadherin, anti-N-cadherin, and anti-calretinin antibodies for differentiating between RMC, MM, and ACA in pleural effusions. A modified avidin-biotin peroxidase complex (ABC) method was used. E-cadherin immunostaining was observed in 14% of RMC, 46% of MMs, and 97% of ACAs. A distinct membrane staining pattern was seen in ACAs. The pattern of staining was cytoplasmic in all reactive RMC and varied from membrane to cytoplasmic in MMs. Anti-N-cadherin immunoreacted with 77% of RMC, 35% of MMs, and 48% of ACAs. Twenty-seven percent of RMC, 58% of MMs, and 31% of ACAs immunoreacted with anti-calretinin. Based on these results, we conclude that anti-E-cadherin is a potentially useful marker in the distinction of ACA cells from RMC. However, it is not as useful for the distinction of ACA and MM. Anti-N-cadherin and anti-calretinin did not reliably distinguish between reactive mesothelial, MM, and ACA cells in pleural effusions. Diagn. Cytopathol. 1999:20:125–130. Published 1999 Wiley-Liss, Inc.
Article
The diagnostic value of an immunoperoxidase panel composed of antisera to carcinoembryonic antigen (CEA), epithelial membrane antigens (EMA), and high- and low-molecular-weight cytokeratins was evaluated on 39 consecutive pleural and peritoneal fluid specimens and correlated with routine cytologic and histochemical studies. The cases were classified into two groups: malignant (epithelial and small-cell undiffer-entiated carcinomas) and benign effusions. We found that the CEA and EMA antisera identified 91% and 100% of the epithelial malignancies, respectively. Mucin stains were positive in only 41% (mucicarmine) and 59% (Alcian blue) of these cases. The battery of cytokeratin markers identified malignant cells from 45%-100% of the case but showed considerable positive staining of mesothelial cells. A selective review of the literature is presented along with recommendations for technical improvement in immunoperoxidase studies. We believe that an immunoperoxidase panel can be of considerable value in the cytologic diagnosis of epithelial malignancies in effusions. The panel composed of CEA and EMA can prove helpful in a routine cytology practice. Although the cytokeratin marker identified malignant cells, the concomitant immunostaining of mesothelial cells limits its utility. The commercially available panel can be a potential aid in improving the accuracy of serous fluid cytologic examination by decreasing both false-positive and false-negative diagnoses and thereby helping to prevent delays in diagnosis and treatment. Diagn Cytopathol 1987;3:134–140.
Article
Malignant mesothelioma is an uncommon, but increasingly important, neoplasm. The existing English-language medical literature concerning non-asbestos-related malignant mesotheliomas was reviewed for evidence of other agents associated with the induction of malignant mesothelioma. Both animal and human data were reviewed. In most reviews of malignant mesothelioma, there are a significant proportion of cases without documented asbestos exposure (range, 0% to 87%). Furthermore, there are several fairly well-documented agents other than asbestos that induce malignant mesothelioma in animals, and strong evidence exists that such is the case in man. In reviews of malignant mesothelioma, the percentage of cases with asbestos exposure varies, but a significant number are apparently not asbestos related. It is believed that sufficient evidence exists to suggest that nonasbestos agents can induce malignant mesotheliomas in man, and additional epidemiologic studies in this area are needed.
Article
BACKGROUND The distinction between malignant mesothelioma (MM) and adenocarcinoma (ACA) in cytologic specimens frequently is difficult, often requiring immunocytochemistry to support the diagnosis. Recent reports have proposed the utilization of antibodies to mesothelial cell clone HBME-1 and thrombomodulin (TM), because they are immunoreactive in MM and less commonly reactive in ACA. Immunoreactivity for the monoclonal antibody CA 19-9 has been observed in many ACAs and reportedly is absent in MM.METHODS In this study, immunostaining was performed on formalin fixed, paraffin embedded cell blocks from effusions or fine-needle aspirations using the avidin-biotin-peroxidase method. Thirty-eight MMs and 49 ACAs were tested using antibodies to CA 19-9, HBME-1, and TM.RESULTSAnti-CA 19-9 stained only 1 of the 37 cases of MM tested (3%), but stained 24 of the 49 cases of ACA (49%). Anti-HBME-1 stained 34 of 38 cases of MM (89%), and 28 of 43 cases of ACA tested (65%). Anti-TM stained 24 of 36 cases of MM (67%), and 21 of 40 cases of ACA tested (53%).CONCLUSIONSCA 19-9 has utility as part of an immunocytochemical panel for distinguishing ACA from MM, because a positive staining reaction would make the diagnosis of MM unlikely. Although HBME-1 and TM can identify MM positively, each frequently is detected in ACA, thus limiting the utility of these antibodies in cytologic specimens.Cancer (Cancer Cytopathol) 1998;84:101-8. © 1998 American Cancer Society.
Article
The cytomorphologic and cytochemical investigation of 59 samples of pleural and peritoneal effusions with malignant tumor cells was performed and the results were compared. The diagnosis was confirmed histologically in most cases. The cytomorphologic method gave very good results not only in the determination of malignant tumor cells but also in the differentiation of various kinds of tumors. A diagnosis was made of differentiated carcinoma (adenocarcinoma and squamous carcinoma) in 40 cases, undifferentiated carcinoma in five, sarcoma or hemoblastosis in nine and undifferentiated malignant tumor in five cases. The results of cytochemical reactions investigated demonstrated a significant difference between the cells of carcinoma and sarcoma, and in various kinds of carcinoma the most important difference was found between the positive reaction of the alkaline phosphatase and negative reaction of the acid phosphatase in the cells of ovarina and uterine carcinoma. Although we consider the morphologic examination of effusions the most important for the diagnosis of malignancy, the cytochemical methods can sometimes be helpful in a more precise differentiation.
Article
Ca2+ ions intervene during different phases of the progression of the cell cycle, but only one calcium-binding protein, calmodulin, has been shown to be associated with dividing cells. We therefore screened cancer cells for the presence of other related calcium-binding proteins. Using molecular biological and immunohistochemical techniques we show that human tumor cells of epithelial origin, express calretinin. Calretinin immunoreactivity can be demonstrated at precise moments of the cell cycle and, in particular, in phase G1 and during mitosis. During mitosis calretinin is localized both in the cytoplasm and in the mitotic spindle. In the cytoplasm we find calretinin after prophase and until telophase. In the spindle apparatus, calretinin is already present in cells in prometaphase and persists in all the succeeding mitotic phases. It is associated with the kinetochore microtubules but, in contrast to calmodulin, also with the polar microtubules. The role that calretinin plays in well-defined moments of the cell cycle of these cells is as yet unknown, but our results strongly suggest that, in collaboration with other molecules, calretinin intervenes in the dynamic phenomena regulating the separation of the chromosomes.
Article
Immunoperoxidase histochemical staining of cryostat sections from human tumor tissues revealed that a murine monoclonal antibody (MAb), K1, can distinguish epithelial mesotheliomas from lung adenocarcinomas. All of 15 epithelial-type mesotheliomas and all four mixed type mesothelioma samples, but none of 23 lung adenocarcinomas with different degrees of histologic differentiation demonstrated reactivity with antibody K1. Of the cell populations in each mesothelioma tested, 80% to 100% showed strong and homogeneous staining with MAb K1. Immunofluorescence analysis of live cultured cells from an epithelioid mesothelioma (H-meso) and several lung carcinoma cell lines as well as a pleural effusion of a patient with mesothelioma also showed selective reactivity of K1 with the mesothelioma cells. These data indicate that K1 can be useful as a mesothelial cell marker for the differential pathological diagnosis of the epithelial form of mesothelioma; K1 may also be useful in the study of the pathogenesis, immunodiagnosis, and immunotherapy of epithelial-type and mixed-type human malignant mesothelioma.
Article
Most compensations for asbestos-related deaths secondary to cancer center around mesothelioma and bronchogenic carcinoma. The differential diagnosis between mesothelioma and adenocarcinoma is a common and troublesome one, necessitating the correlation between clinical history, radiographic findings, and pathologic examination of tissues and cells. We describe a multimodal approach based on the use of routine and special stains, immunocytochemistry, and electron microscopy for distinguishing between mesothelioma and adenocarcinoma. Once a malignant diagnosis is arrived at by careful pathological examination, the tumor is classified as mesothelioma if mesothelial cells are identified as the constituent cells of the neoplasm. Mesothelial cells are recognized by (1) their main ultrastructural features: slender and elongated microvilli, abundant intermediate filaments, and lacking secretory granules; and (2) their characteristic immunocytochemical reactivity: positivity for cytokeratin, EMA, and vimentin, and negativity for carcinoembryonic antigen (CEA), B72-3, Leu-M1, and other gland-cell markers. A variety of methods have been attempted in an effort to distinguish between reactive and malignant mesothelial cells. In practice, however, such distinction depends more on experience and expertise than in any fool-proof ancillary tests. A number of these tests are discussed along with the illustration of classical and unusual examples of mesothelioma and other pleural tumors.
Article
Differentiating mesothelioma, reactive mesothelium, and adenocarcinoma in serous effusions is often difficult, despite the application of ancillary techniques in support of the traditional cytomorphologic criteria. A polyclonal antimesothelial-cell antibody recently developed by our group was evaluated as a histogenetic marker on a series of primary (n = 12) and metastatic (n = 12) malignant effusions. Immunostaining was performed on paraffin sections from cell blocks. All mesothelioma effusions stained positive for the antibody, whereas, in contrast, all metastatic carcinoma specimens failed to react. These results (100 percent specificity and 100% sensitivity for mesothelioma) provide a basis for a reliable use of the antibody in the cytologic examination of suspicious or malignant serous effusions.
Article
The cytologic diagnosis of malignancy in serous effusions can be challenging. An immunocytochemical (ICC) panel using commercially available antibodies (to carcinoembryonic antigen [CEA], epithelial membrane antigen [EMA], B72.3, Leu-M1, cytokeratin [CK], leukocyte common antigen [LCA], S-100 protein, and vimentin) was applied to cell blocks fixed in methyl Carnoy's solution that were from 55 consecutive pleural, peritoneal, and pericardial fluid specimens. The results were correlated with data from clinical records and routine cytologic studies. Final cytologic diagnoses included 26 of adenocarcinoma and 1 of mesothelioma. The remaining 28 cases were considered to be benign (reactive) proliferations. EMA, CEA, B72.3, and Leu-M1 were present in 96%, 77%, 58%, and 42% of adenocarcinomas, respectively. These determinants were absent in the mesothelioma and the reactive effusions, although anti-CEA yielded strong background staining of inflammatory cells. The CK markers identified malignant cells in 93% of cases, but consistently stained mesothelial cells as well. Antivimentin strongly labeled mesothelial cells in all cases, with weak to absent staining of malignant cells. In 3 of 26 carcinoma cases (12%), the ICC panel identified malignant cells that were not recognized initially on routine cytologic examination. In 1 of 26 cases (4%), the panel was falsely negative. Use of this approach can improve the diagnostic accuracy of cytologic examination of serous fluids. The ICC panel is especially helpful when atypical mesothelial proliferation is present, or in cases that are clinically suspect for malignancy, but cytologically negative because there are only a few malignant cells, or those that are cytologically bland.
Article
We have isolated a new monoclonal antibody (MAb), K1, that reacts with an epitope on the surface of human ovarian carcinoma cells. This antibody was generated by immunization of mice with periodate-treated human ovarian carcinoma (OVCAR-3) cells. These mice had been previously made tolerant with normal human kidney membranes. Spleen lymphocytes from these mice were selected prior to fusion using a panning purification method on living OVCAR-3 cells. Initial screening of surface-reactive clones was performed in a single day using immunofluorescence on living OVCAR-3 cells, and secondary screening was performed using immunoperoxidase histochemistry on cryostat sections of normal human tissues and human tumors. The K1 clone was subcloned and identified as an IgM isotype, but was subsequently isotype-switched to IgG1K using a panning selection method. When evaluated by immunohistochemistry, the antigen reactive with K1 was found in many ovarian non-mucinous tumors, as well as in squamous tumors of the esophagus, and cervical cancer. The only normal adult human tissues showing uniform reactivity with K1 were the mesothelia of the peritoneal, pleural and pericardial cavities. There was also limited reactivity with epithelia of the trachea, tonsil and Fallopian tube. A similar tissue reactivity for K1 was found in tissues from cynomolgus monkeys. K1 reacted with many of the same tissues and tumors as the previously identified antibody OC125, but several lines of evidence indicate that K1 reacts with a different epitope and probably a different molecule, when compared to OC125. This evidence included assays employing immunofluorescence competition, double-label immunofluorescence, and solid-phase and live-cell radioimmunoassays. Since our data indicate that the antigen reactive with the K1 antibody is a new molecular species, we have named the antigen CAK1. Unlike the shed antigen CA125, CAK1 was only cell-associated and was not found in the supernatant of cultured OVCAR-3 cells or in the blood of ovarian cancer patients. The K1 antibody may be useful as a targeting agent for therapy and in the diagnosis of ovarian carcinoma, as well as some other human cancers.
Article
In pleural of ascitic effusions the cytomorphological distinction of adenocarcinoma cells, reactive mesothelial cells, and malignant mesothelioma cells often causes a diagnostic dilemma. The value of immunocytochemistry was investigated on cytological smears of 24 well-established cases of malignant mesothelioma, a selected series of 31 metastatic adenocarcinomas, and 20 smears of patients without known malignancy. In these smears we scored the immunoreactivity with a panel of four monoclonal antibodies. In addition to antibodies for epithelial membrane antigen (EMA) and carcinoembryonic antigen (CEA), the monoclonal antibody MOC31 and the ovarian carcinoma specific antibody OV632 were incorporated in the panel. With none of these four antibodies was immunostaining of reactive mesothelial cells found. CEA- and MOC31-positive tumour cells were frequent in metastatic adenocarcinomas, but occurred rarelay in malignant mesothliomas. EMA-positive tumour cells were in all metastatic adenocarcinomas (100 per cent) and in most malignant mesotheliomas (83 per cent). In addition to the expected reactivity of OV632 with ovarian carcinomas, 22 of 24 malignant mesotheliomas contained immunopositive tumour cells, while only a small proportion of non-ovarian adenocarcinomas reacted with this antibody. This selective staining of malignant mesothelioma cells, but not reactive mesothelial cells, with OV632 now permits the positive identification of malignant mesothelioma cells in male patients.
Article
The immunohistochemical profile (i.e., carcinoembryonic antigen, keratin proteins, epithelial membrane antigen, human milk fat globule-derived antigen, and mucin) of paraffin-embedded cell blocks of 20 malignant effusions from patients with malignant mesothelioma was compared with that of 39 malignant effusions from patients with metastatic adenocarcinoma to determine whether these markers distinguished between these tumor types. Twenty-three adenocarcinomas (59 per cent) stained for mucin. Immunoreactivity for carcinoembryonic antigen (CEA) was observed in 28 adenocarcinomas (72 per cent). All were immunoreactive for keratin proteins, and 29 adenocarcinomas (74 per cent), including seven that were mucin and CEA negative and exhibited a "peripheral predominant" staining pattern for keratin proteins. By contrast, none of the mesotheliomas stained for mucin or for CEA, and, although all were immunoreactive for keratin proteins, none demonstrated a peripheral predominant pattern of staining. Epithelial membrane antigen and milk fat globule-derived antigen were identified in the majority of both mesotheliomas and adenocarcinomas. Neither staining intensity nor pattern of reactivity of these markers clearly distinguished the tumors. This study of cell blocks of serous effusions suggests that staining for mucin, immunoreactivity for carcinoembryonic antigen, and a peripheral predominant pattern of reactivity for keratin proteins represent highly characteristic markers of adenocarcinomas, which identify the majority of these tumors (38 of 39) and allow their distinction from malignant mesotheliomas.
Article
Recognition of malignant effusion relies heavily on cytologic examination despite the difficulty of distinguishing atypical mesothelial hyperplasia from metastatic carcinoma. The combination of CEA, EMA, vimentin, keratin, high-molecular-weight cytokeratin (HMWK), low-molecular-weight cytokeratin (LMWK), and Alcian blue was tested in 51 cytologic specimens of pleural, peritoneal, and pericardial effusions. These showed metastatic carcinoma in 38 cases (ovary, 14; lung, 8; breast, 7; GI, 4; endometrium, 4; bladder, 1) and mesothelial processes in 13 (hyperplasia, 9; mesothelioma, 4). Strong positivity for EMA (92%), CEA (90%), and Alcian blue (71%) was noted in metastatic carcinoma but not in the mesothelial processes. Keratin was positive in all cases of mesothelioma but occurred also in mesothelial hyperplasias (44%) and metastatic carcinomas (47%). In mesothelial cells, HMWK was consistently stronger than LMWK, whereas in adenocarcinoma the reverse was true. There was no difference in the degree or distribution of positivity of any of the markers among the various primary sites of the neoplasms. Our findings are consistent with the view that immunocytochemistry with a battery of antibodies is useful in the recognition of malignant effusions but cannot, as yet, determine the site of origin of metastatic neoplasms.
Article
Traditionally, diffuse epithelial mesotheliomas are mainly identified at the ultrastructural level by the numerous, long, wavy-appearing surface microvilli. By electron microscopy of a series of diffuse mesotheliomas of varying subtype (epithelial, biphasic, sarcomatous, and poorly differentiated), it can be demonstrated that the differentiation of this specialized surface organelle is quite variable even in well-differentiated lesions. The presence of only a few, scattered, short microvilli does not exclude a diagnosis of epithelial mesothelioma, particularly if historical, surgical, and radiologic findings support this diagnostic conclusion. Indeed, even the complete absence of surface microvilli is compatible with a diagnosis of diffuse epithelial mesothelioma. It is important to become aware of the spectrum of tumor cell differentiation in serosal tumors, as all of the fine structural diagnostic criteria in mesotheliomas are expressed to varying degrees in individual cases.
Article
A novel gene of the calmodulin superfamily, encoding a 29-kD neuronal protein here named "calretinin," has been isolated as a cDNA clone from chick retina. The encoded sequence includes four putative calcium-binding sites and a fusion protein binds calcium. The most similar protein known is the 28-kD intestinal calcium-binding protein, calbindin (58% homology). Both genes date from before the divergence of chicks from mammals. The distribution of calretinin and calbindin mRNAs in chick tissues has been mapped using RNA gel blots and in situ hybridization. RNAs from both genes are abundant in the retina and in many areas of the brain, but calretinin RNA is absent from intestine and other nonneural tissues. Calretinin and calbindin are expressed in different sets of neurons throughout the brain. Calretinin RNA is particularly abundant in auditory neurons with precisely timed discharges.
Article
Serous effusions from 50 patients suspected of having cancer were subjected to cytogenetic analysis by the use of a simple direct technique. Within 24 hours the specimens were reported as unsatisfactory or as positive or negative for malignancy, according to specific chromosomal criteria, which included hypodiploidy. Reports were sent directly to referring clinicians, who also received independently derived findings from routine cytologic study. The value of cytogenetic analysis was clearly shown in this situation, in which the chromosomal findings took the form of an independent second opinion in clinical assessment. There were no false-positive results in any of the 16 cases diagnosed by cytogenetic analysis. Furthermore, in six of these, the initial cytologic report was equivocal, and the repeat sample from three of these failed to yield material suitable for a definite cytologic diagnosis of malignancy. Negative cytogenetic findings also supported the use of cytogenetic analysis as an adjunct to cytologic examination: 23 of the 24 negative cytogenetic reports proved to be accurate, whereas in 8 of these cases the first cytologic report was equivocal. There was one cytogenetic false negative, in which, after retrospective analysis, chromosomal criteria of malignancy were found.
Article
To improve the accuracy of diagnosis, 82 cases of malignant mesothelioma were studied for mucosubstance production by the periodic acid Schiff reaction after diastase digestion (DPAS) and the colloidal iron (CI) reaction before and after hyaluronidase. 26 carcinomas of various origins, similarly treated, were compared. The tumors had been fixed and processed in routine fashion. The reactions were regarded as positive only when there was a significant number of stained cytoplasmic vacuoles and/or stained secretion in tubular or glandular lumina or in spaces between epithelial cells. Scattered intracytoplasmic granules and positive material in the stroma or in mesenchymal tumors were not considered diagnostic. Approximately half of the mesotheliomas were CI positive. All but two showed complete or partial removal of the CI positive secretion by hyaluronidase. Thus, almost half of the mesotheliomas were demonstrated to contain hyaluronic acid and almost 33% of these were shown to contain another type of acid mucosubstance as well. A parallelism appeared to exist between the degree of histologic differentiation and the production of hyaluronic acid. The production of neutral mucosubstance, as indicated by the DPAS, was not conclusively demonstrated in any mesothelioma. Each staining method showed a small number of equivocal reactions. Twelve of 26 carcinomas were DPAS and CI positive but hyaluronidase had no effect on the CI reaction.
Article
Two localized and two diffuse pulmonary mesotheliomas were studied with both light and electron microscopes. The diffuse mesotheliomas showed features and characteristics of mesothelial cells in the electron microscope, whether lining spaces or within solid parts of the tumor. The localized mesothelioma showed predominance of poorly differentiated cells and fibroblasts. The relationship between the localized and diffuse mesotheliomas is not clear, but the former could be a group of variegated tumors. It appears that a definite electron microscopic diagnosis of diffuse pulmonary mesothelioma can be made at least in some of the surgically removed tissues suspected to be diffuse mesothelioma by clinical and light microscopic findings.
Article
The histologic types of lung cancer in 855 patients (747 men and 107 women) from three hospitals and one international study of insulation workers were evaluated. Of these, 196 cases had asbestos exposure. About one half of the cases were diagnosed from surgical slides and one half from autopsy slides. Squamous cell carcinoma constituted the largest percentage of tumor types and was found with the same frequency in exposed and nonexposed groups. Small cell carcinoma was found in 25% of the exposed and in 15% of the nonexposed patients. Upper lung sites were involved in about two thirds of the cases with asbestos exposure and lower lobes in the other one third. There was little difference in histologic type in cases regardless of whether upper or lower lobes were involved. Cigarette smokers who smoked until their cancer diagnosis showed no difference in histologic type by amount smoked, and slight but not statistically significant differences from ex-cigarette smokers.
Article
The histological diagnosis of mesothelioma may be difficult, especially when only a small biopsy is available. The epithelial form of mesothelioma can closely resemble an adenocarcinoma, so much so that it is often difficult or impossible to distinguish between them by light microscopy. This paper examines the results of staining 30 cases of the epithelial form of mesothelioma by various techniques for mucus. The slides were evaluated according to the intensity and extent of staining, the color of the reaction, the location of the staining (intra- or extra-cellular), and the reproducibility and reliability of the technique.
Article
A panel of seven monoclonal antibodies was applied to smears of cell deposit from 70 pleural and peritoneal fluids, using an immunoalkaline phosphatase (IAP) procedure. The cases were chosen to show typical cytological patterns, both benign and malignant, and in this way the diagnostic value of the method could be assessed. The antibodies used were 2D1 (anti-leucocyte), Ca 1, HMFG-2 (anti-milk fat globule membrane), LE61 and M73 (both anti-intermediate filament antibodies), anti-CEA, and K92 (anti-keratin). The anti-leucocyte antibody was found useful for distinguishing lymphoma from carcinoma. Anti-CEA gave positive reactions in 80% of carcinoma cases and was not seen to react with any other cell types. Ca 1 was positive with some cells in 95% of carcinoma cases, but mesothelial cells reacted with it in two cases. A strong reaction with the anti-milk fat globule membrane antibody was very constant in carcinoma but was also seen in mesothelial cells in 30% of benign effusions. The anti-keratin reacted with malignant cells in only a small proportion of cases. The antibodies against epithelial intermediate filaments reacted equally strongly with benign mesothelial cells and carcinoma cells, but gave negative reactions with lymphoma cells. It is concluded that a suitably chosen panel of monoclonal antibodies can be of great value in identifying neoplastic cells in serous effusions.
Article
Mesotheliomas and metastatic adenocarcinomas involving the pleura are frequently difficult to distinguish by light-microscopic and histochemical methods. In a double-blind study, we have compared ultrastructural features of 10 mesotheliomas of epithelial type and 10 adenocarcinomas from the lung, breast, and upper GI tract, i.e., sites known to give rise to metastases which mimic mesothelioma. Mesotheliomas were observed to have a significantly greater microvillus length/diameter ratio (LDR) than adenocarcinomas (p less than 0.01) and more abundant intermediate filaments (p less than 0.001). Mesotheliomas had more complex microvilli than adenocarcinomas, whereas adenocarcinomas had rootlets (2/10 cases) and lamellar inclusion bodies (2/10 cases), both of which were absent in the mesotheliomas. This study provides quantitative and qualitative ultrastructural features of potential utility in the differential diagnosis of pleural mesotheliomas and adenocarcinomas.
Article
Malignant mesothelioma is an uncommon, but increasingly important, neoplasm. The existing English-language medical literature concerning non-asbestos-related malignant mesotheliomas was reviewed for evidence of other agents associated with the induction of malignant mesothelioma. Both animal and human data were reviewed. In most reviews of malignant mesothelioma, there are a significant proportion of cases without documented asbestos exposure (range, 0% to 87%). Furthermore, there are several fairly well-documented agents other than asbestos that induce malignant mesothelioma in animals, and strong evidence exists that such is the case in man. In reviews of malignant mesothelioma, the percentage of cases with asbestos exposure varies, but a significant number are apparently not asbestos related. It is believed that sufficient evidence exists to suggest that non-asbestos agents can induce malignant mesotheliomas in man, and additional epidemiologic studies in this area are needed.
Cells from body cavity effusions from 35 patients were examined by scanning electron microscopy using both secondary electron imaging and backscatter electron imaging (BEI). This technique allows simultaneous viewing of the same cell for histological identification and for surface configuration. Mesothelial cells, macrophages, lymphocytes, neutrophils and malignant cells could be identified by their nuclear details and their surface features. Mesothelial cells had either smooth surfaces or blebs with few microvilli. Lymphocytes and neutrophils had similar surface features but could be easily identified by BEI. Macrophages had lamellar cytoplasm. Malignant cells were usually large and occurred singly or more frequently in groups. Their irregular chromatin pattern was easily recognized by BEI and was clearly different from mesothelial cells. This technique uses micropore filters to collect the cells. It is a rapid and simple procedure which offers advantages over previously described techniques using glass coverslips.
Article
Flow cytometry allows rapid and accurate analysis of the deoxyribonucleic acid (DNA) content of a large number of cells. In solid tumors, the presence of aneuploidy has been shown to correlate wall with the presence of neoplastic cells. Both cytologic examination and DNA analysis by flow cytometry were performed on pleural effusions from 33 patients. Results of the two examinations were in agreement in 10 of 12 malignant pleural effusion (two false-negatives) and in 20 of 21 benign effusions. One patient with cirrhosis, ascites and Nocardia pneumonia had hypodiploid cells (false-positive) in the pleural fluid. All patients who had a malignancy, but whose pleural effusion proved to be due to a benign cause, had cells with normal DNA content in their pleural effusion. DNA analysis using flow cytometry can be rapidly performed and is highly specific and sensitive. The finding of hyperdiploid cells is highly suggestive of malignancy.
Article
ME1 is a monoclonal antibody which is generated by the use of a mesothelioma cell line (SPCIII). The antibody has a preferential reaction to antigens on mesothelial and mesothelioma cells. In a prospective study we determined the reactivity in frozen sections from malignant mesotheliomas (two cases, positive controls), lung tumours (115 cases) and other malignant tumours (23 cases). The two malignant mesotheliomas were immunoreactive in most of the tumour cells. The reaction was strong, often with a diffuse staining of the cytoplasm and in some tumour cells there was heavy staining of the cell membrane. Five adenocarcinomas of the lung (9%), one large cell carcinoma (10%) and 18 squamous cell carcinomas of the lung (41%) were positive (defined as tumours containing more than 10% positive tumour cells with a strong reaction). The same was true for seven out of 23 (30%) extrapulmonary malignancies. The overall nosologic specificity of ME1 was 76%. Twenty out of the 26 ME1-positive lung tumours and six out of seven ME1-positive extrapulmonary malignancies were also positive for one or more markers, which is considered characteristic of carcinomas. The six negative lung tumours were squamous cells carcinomas and the negative extrapulmonary tumour was a meningeoma; all of them with a morphology different to malignant mesothelioma. In conclusion, when frozen sections are available, ME1 might be useful in the differential diagnosis of malignant tumours. However, a positive reaction is not specific for malignant mesothelioma.