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• Cell block cytology material is reliable for EGFR testing with high concordance. • Cytology material showed lower adequacy rates than biopsy or excision specimens. • Effusion fluid had the highest mutation detection rate among all cytology specimens and was superior to pleural biopsy. • EGFR mutation detection rate for lung and lymph node aspirates were non-inferior to tissue biopsy or excision. Cytology is integral to lung cancer diagnosis. Aspiration and exfoliative fluid specimens represent valuable tumor material for molecular testing. In this study, a large retrospective cohort of EGFR tests was reviewed to address the adequacy, detection and discrepancy rate in tests performed with cytology material. EGFR tests performed from 2013 to 2022 were reviewed and classified by the modality of obtaining tissue and by tissue type. EGFR tests for tissue specimens were performed on unstained sections of paraffin-embedded material on glass slides. Adequacy and types of mutation(s) detected were analysed. Cases where multiple EGFR testing was performed on the same patient were reviewed for discordance. There were 5,504 tests retrieved, with 1,855, 3,607 and 42 performed on cytology, surgical and blood specimens. Lung and excision specimens were more often adequate (p < 0.001). Cytology material showed lower adequacy rates (p < 0.01). EGFR detection (positive) rate was higher in pleural fluid compared to biopsy (59.8 % vs 50.7 %, p = 0.022), but similar between lung and lymph node cytology and non-cytology specimens. Effusion fluid specimens had the highest adequacy (81.5 %) and detection rate (59.3 %) among cytology specimens (p < 0.001). Four (4.4 %) cases showed discordant results in cytology specimens. Two were false negatives in the non-cytology material. Only in one case was cytology material genuinely discrepant. The remaining discordance was attributed to the interval treatment effect. The findings support that EGFR testing in cell block is reliable and complements tissue material. In addition, pleural fluid appears to be superior to pleural biopsies for molecular testing. [ABSTRACT FROM AUTHOR]

Accumulation of fluid in serous cavities as effusions may have to be drained for therapeutic and diagnostic indications. As compared to many other procedures, the technicality of paracentesis procedures is relatively easy. As a result, effusion fluids comprise a significant proportion of specimens in most of the laboratories, including those in community settings. Because of relative complexities in the cytopathologic evaluation of effusion fluids, application of appropriately standardized protocol is critical for achieving optimum results by applying standardized steps from handling of specimens during the initial stages of collection to the final interpretation phase. Understanding various limitations and challenges during collection and processing phases by all the personnel involved, including clinicians, pathologists, and technologists is critical for optimum diagnostic yield. This review highlights various cytopreparatory techniques applicable to effusion cytology in one place. It is organized by projecting the details predominantly in the form of different tables and figures including summary of the recommended protocols with reagents and stains used. Also included is a sample of cytopathology report based on the approach discussed in this series. [ABSTRACT FROM AUTHOR]

Background: Feline Infectious Peritonitis (FIP) is a fatal disease of cats that can be very difficult to definitively diagnose antemortem. Multiplex fluorescent immunocytochemical (MF‐ICC) assays are emerging as useful diagnostic tests in veterinary medicine, particularly for fluid samples. Objective: We aimed to develop and optimize an MF‐ICC assay to detect feline coronavirus within macrophages, with the primary goal of determining the allowable/recommended sample storage conditions for clinical use of this assay. Methods: A feline macrophage cell line was infected with the FIP virus. Following harvest into EDTA tubes (simulating typical clinical collection of effusion), cells were stored at 4℃, 22℃, and 37℃. For each temperature condition, slides for MF‐ICC were made at 0, 1, 2, 3, and 5 days post‐collection. To assess the stability of immunoreactivity following fixation, freshly harvested infected cells were fixed onto slides and maintained at 4℃ for 1, 2, 4, and 12 weeks. All slides were analyzed by MF‐ICC for the presence of mononuclear cells with co‐expression of vimentin and coronaviral antigen. Results: MF‐ICC confirmed that cells tested positive for coronavirus at 4℃ through 3 days post‐harvest, 22℃ through 48 hours post‐harvest, and 37℃ through 24 hours post‐harvest. The MF‐ICC assay was successfully performed on fixed slides through the 12‐week time point. This assay also demonstrated positive results on a clinical sample of abdominal fluid from a cat later confirmed to have FIP. Conclusions: The MF‐ICC assay described here offers a potentially specific and relatively stable antemortem diagnostic test for feline infectious peritonitis. Evaluation of this assay in clinical samples is ongoing. [ABSTRACT FROM AUTHOR]

Background: Identifying malignant cells in effusion fluid is vital in staging and management of cancers. Differentiating reactive mesothelial cells from malignant cells in effusion fluid is a challenging task and there is an ongoing need for simpler and cost effective tool to aid the diagnosis. Micronucleus is an additional smaller nucleus in the cytoplasm, formed by chromosomes or chromosomal fragments formed during cell division. Aims: The aim of this study was to assess the significance of micronucleated cell in effusion fluids to distinguish adenocarcinomatous from reactive mesothelial effusions. Materials and Methods: Thirty cases of unequivocal malignant effusion fluids and 30 benign cases with reactive mesothelial cells as control were studied. Number of microucleated cells present per1000 well-preserved cells in Leishman-stained smears were counted. Results: Mean (±SD) micronucleated score in malignant and benign effusions were 15.77 ± 9.78 and 1.87 ± 1.78, respectively. The median scores were 13 and 2, respectively. Mann–Whitney test showed that this difference was statistically significant (P < 0.001). This study revealed that there was a significant difference in micronucleus scoring between benign and malignant effusions. Conclusions: Micronucleus score can be used as an additional biomarker in the interpretation of routinely stained cytosmears.

Filariasis is endemic in developing countries with highest prevalence in India. Microfilarias are infrequently reported during cytological examination of body cavity fluids. Its association with neoplasm is still rare, with only a few reported cases of co-existent microfilaria with neoplasm. The malignancies reported in association with malignant pleural effusions are mostly carcinomas. Finding microfilaria in a pleural effusion is a very rare event even in the regions endemic for this organism. Its association with a lymphoma is even rarer. On extensive search of literature we could not find any case report of filarial pleural effusion associated with lymphoma. We here report a case of microfilaria in malignant pleural effusion secondary to lymphoma.

OBJECTIVES: The role of autophagy in the pathophysiology of otitis media with effusion (OME) remains unclear, particularly regarding the difference between pediatric and adult patients. The present study analyzed the expression levels of autophagy-associated mRNAs in effusion fluids obtained from pediatric and adult patients with OME. MATERIALS and METHODS: Middle ear fluid samples were collected from 76 pediatric patients and 41 adult patients with OME, and the levels of mRNAs encoding autophagy-related genes were measured using real-time reverse transcription-polymerase chain reaction. The relationships between the levels of autophagy-associated mRNAs and the frequency of ventilation tube insertion, the characteristics of middle ear fluid, and the results of bacterial culture were analyzed. RESULTS: Autophagy-associated mRNAs were present in the effusion fluid of all patients. The level of Beclin-1 mRNA was significantly lower in pediatric than in adult patients, regardless of the frequency of surgery or fluid characteristics (p<0.05). CONCLUSION: Autophagy-associated mRNAs were expressed in effusion fluids of both pediatric and adult patients with OME. However, the level of Beclin-1 mRNA was significantly lower in the effusion fluid of pediatric than adult patients. [ABSTRACT FROM AUTHOR]

Background: Identifying malignant cells in effusion fluid is vital in staging and management of cancers. Differentiating reactive mesothelial cells from malignant cells in effusion fluid is a challenging task and there is an ongoing need for simpler and cost effective tool to aid the diagnosis. Micronucleus is an additional smaller nucleus in the cytoplasm, formed by chromosomes or chromosomal fragments formed during cell division. Aims: The aim of this study was to assess the significance of micronucleated cell in effusion fluids to distinguish adenocarcinomatous from reactive mesothelial effusions. Materials and Methods: Thirty cases of unequivocal malignant effusion fluids and 30 benign cases with reactive mesothelial cells as control were studied. Number of microucleated cells present per 1000 well-preserved cells in Leishman-stained smears were counted. Results: Mean (±SD) micronucleated score in malignant and benign effusions were 15.77 ± 9.78 and 1.87 ± 1.78, respectively. The median scores were 13 and 2, respectively. Mann-Whitney test showed that this difference was statistically significant (P < 0.001). This study revealed that there was a significant difference in micronucleus scoring between benign and malignant effusions. Conclusions: Micronucleus score can be used as an additional biomarker in the interpretation of routinely stained cytosmears. [ABSTRACT FROM AUTHOR]

BACKGROUND The cytological diagnosis of malignant mesothelioma (MM) on serous effusion is challenging due to significant morphologic overlap with reactive mesothelial cells and adenocarcinoma. One of the morphologic features of MM in effusion cytology, small orangiophilic squamous-like cells (SOSLC), has received little attention. To the best of the authors' knowledge, the current study is the first to assess the sensitivity and specificity of SOSLC in serous effusion specimens from histology-proven MM cases along with those of reactive mesothelial and adenocarcinoma cases. METHODS A total of 130 cases of pleural (86 cases) and peritoneal (44 cases) effusion cytology cases (30 with histology-proven MM, 41 with adenocarcinoma, and 59 with reactive mesothelial cells) were studied. The presence or absence of SOSLC was recorded in each case. RESULTS The cytological diagnoses of the 30 histology-proven MM cases included 1) atypical mesothelial cells, favor reactive (4 cases); 2) atypical mesothelial cells, suspicious for or cannot exclude mesothelioma (18 cases); and 3) positive for MM (8 cases). SOSLC were found in 10 of the 30 MM effusion cases (33.3%), 1 of the 41 adenocarcinoma cases (2.4%), and 5 of the 59 reactive mesothelial cell cases (8.5%). SOSLC were more likely to be present in MM effusions compared with either adenocarcinoma ( P < .0001) or reactive mesothelial cell ( P < .02) effusions. All 10 cases of MM with SOSLC were from pleural fluids. One case of peritoneal serous adenocarcinoma had SOSLC and 5 cases of reactive mesothelial cells in peritoneal fluid were found to have SOSLC. CONCLUSIONS Although not sensitive, the presence of SOSLC is quite specific for MM in pleural fluid cytology specimens. Finding this morphological feature in pleural fluid should alert the pathologist to a possible diagnosis of MM. Cancer (Cancer Cytopathol) 2014;122:70-75. © 2013 American Cancer Society. [ABSTRACT FROM AUTHOR]

Filariasis is endemic in developing countries with highest prevalence in India. Microfilarias are infrequently reported during cytological examination of body cavity fluids. Its association with neoplasm is still rare, with only a few reported cases of co-existent microfilaria with neoplasm. The malignancies reported in association with malignant pleural effusions are mostly carcinomas. Finding microfilaria in a pleural effusion is a very rare event even in the regions endemic for this organism. Its association with a lymphoma is even rarer. On extensive search of literature we could not find any case report of filarial pleural effusion associated with lymphoma. We here report a case of microfilaria in malignant pleural effusion secondary to lymphoma. [ABSTRACT FROM AUTHOR]

Background: The expression of programmed cell death ligand-1 (PD-L1) is a biomarker in patients with non-small-cell lung carcinoma (NSCLC). Patients with advanced-stage NSCLC receive a variety of molecular genetic tests, possibly resulting in insufficient tissue for immunoassay of PD-L1. Thus, to determine whether effusion fluid specimens are a reliable alternative to tissue specimens for PD-L1 testing, we compared the results of PD-L1 immunostaining using body-fluid cell blocks and tumor tissues., Methods: PD-L1 immunostaining was performed in 62 paired samples of cytology cell blocks (ie, immunocytochemistry) and tumor tissues (ie, immunohistochemistry) from 36 patients using the E1L3N, SP142, and SP263 anti PD-L1 antibody clones. Of the 62 cytology specimens, 50 were from malignant effusion fluid. PD-L1 expression was scored as the percentage of tumor cells with clear membranous staining., Results: A strong positive correlation was observed between the immunostains on cytology cell blocks and tumor tissue (Pearson's correlation coefficient, R = .804, P < .001). When the score was categorized as <1%, ≥1% and <10%, ≥10% and <50%, and ≥50%, the overall concordance rate was 74.2% (46/62, Cohen's k = 0.568). After dichotomizing the cases using cutoff values of 1%, 10%, and 50%, the concordance rates were 84% to 100% for both adenocarcinoma and squamous cell carcinoma. The concordance rate was higher in patients with NSCLC with an EGFR mutation and using the SP263 rather than the E1L3N clone., Conclusion: The results of PD-L1 immunostaining of cell blocks, particularly from effusion fluid, reflect the PD-L1 expression status of NSCLC tissue., (© 2020 Wiley Periodicals, Inc.)