Serous effusion fluid is one of the most commonly encountered specimens in routine cytopathology practice. It provides invaluable information about the patient and the clinical status; but to get the most of it, specimen handling and processing must be carried out properly. Cytomorphology is the basis of a successful analysis which should complemented by ancillary tests when needed. A wide spectrum of ancillary techniquesranging from immunocytochemistry and flow cytometry to different assays of molecular pathologycan be applied to serous effusions. This article describes the acquisition and management of serous effusion fluids, methods for preservation and transportation, different techniques of cytopreparation, application of immunocytochemistry, flow cytometry, and fluorescence in-situ hybridization (FISH), as well as DNA extraction for polymerase chain reaction (PCR) and next generation sequencing (NGS). Principles of bio-banking of effusion samples are also discussed which is getting more important in correlation with the developments in personalized medicine. K E Y W O R D S bio-banking, effusion cytology, fluorescence in situ hybridisation, immunocytochemistry, molecular tests, preparation techniques, serous effusion 1 | INTRODUCTION Serous effusion fluid-pleural effusion, ascites and pericardial effusion-is one of the most commonly examined specimen types in cytopathology. Serous effusions occur in the context of a wide spectrum of benign and malignant diseases. In many clinical situations, effusion fluid is the first sample obtained and analysed, and, more often than not, this sample is the key to precise and timely diagnosis. Classical cytomorphology is still the basis of the application of all ancillary techniques. Immunocytochemistry can be accepted as part of the routine procedure since it has been used as a complementary method for years to increase the diagnostic accuracy of effusion cytology. All ancillary tests, including the modern techniques of molecular pathology, can be applied to serous effusions in routine workflow. It is a rapidly evolving field; however, some basics do not change, such as the selection of the right area/sample for DNA extraction or for fluorescence in situ hybridisation (FISH) analysis. This paper intends to be a practical handout for readers who deal with effusion samples. The acquisition and management of effusion samples are considered; preparation techniques for morphological examination and application of ancillary tests are discussed. The last part of the text deals with bio-banking of effusion specimens, which deserves sincere attention for the future of effusion cytology, regarding the new therapy options and research facilities. | ACQUISITIONEffusion fluid can be obtained in three ways:• by thoracentesis, paracentesis, pericardiocentesis with aspiration of fresh fluid into a syringe or vial,
Malignant mesothelioma (MM) is a rare form of cancer. Its histopathological diagnosis is very difficult, as it exhibits a number of different appearances that can be misinterpreted as metastatic invasion or atypical hyperplasia. Thus, there is an urgent need to identify adequate markers to distinguish between benign and malignant cells, allowing the implementation of appropriate therapies and, possibly, specific directed therapies. MM, like other tumors, show an increase in glucose uptake, due to high rates of glycolysis, inducing an intracellular overload of acids. In this context, monocarboxylate transporters (MCTs) emerge as important players, by mediating the transmembranar co-transport of lactate with a proton, thereby, regulating pH and allowing continuous glycolysis. Importantly, proper MCT expression and activity depend on its co-expression with a chaperone, CD147, which is associated with poor prognosis in cancer. Twenty-two samples including reactive mesothelial cells, MM, and atypical mesothelial hyperplasias were evaluated for immunoexpression of MCT1, MCT4, and CD147. Expression of these proteins was compared with GLUT1 as a new promising marker for MM. Although MCT isoforms were not differentially expressed in the two types of cytological specimens, CD147, as GLUT1, was almost exclusively expressed in MM. Both MCT1 and MCT4 are not able to discriminate between mesothelial reactive cells and mesothelial malignant cells, while CD147 was able to distinguish these two proliferations. If confirmed, besides being a good marker for identification of MM, CD147 may also be a target for therapeutical strategies in this rare type of tumor.
Eccrine porocarcinoma (EP), although rare, is widely recognized as the most common malignant sweat gland tumor. EP typically grows slowly and usually is cured by surgical excision with clear margins. An elevated mortality rate, however, is observed when regional lymph nodes are involved. We herein describe cytohistologic findings in a case of metastatic EP. An 86-year-old man with a history of EP of the left lateral ankle and squamous cell carcinoma in situ (Bowen's disease) of the penis presented with enlarged left inguinal lymph nodes. A superficial fine-needle aspiration (FNA) was performed and demonstrated a hypercellular sample with discohesive clusters and/or individual tumor cells. The tumor cells were round or oval with most of the cells showing dense, refractile cytoplasm. Intracytoplasmic vacuoles were readily appreciated in some of the cells. Nuclear enlargement, high N/C ratio, nuclear hyperchromasia, bi- and multinucleation, and prominent nucleoli were seen. A diagnosis of metastatic eccrine porocarcinoma was rendered. Enlarged retroperitoneal lymph nodes were detected and CT-guided left retroperitoneal core biopsy was performed 1 week later. The biopsy revealed features consistent with metastatic eccrine porocarcinoma.
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