A case report of primary central nervous system lymphoma with immune deficiency/disorder setting diagnosed by cerebrospinal fluid cytology.

Cerebrospinal fluid (CSF) cytology of primary central nervous system lymphoma arising in the immune deficiency/dysregulation setting (IDD-PCNSL) has not been described. This study presented a case of IDD-PCNSL-DLBCL, a GCB phenotype who was successfully diagnosed by CSF cytology in conjunction with ICC, ISH, FCM and clinical information.

Cerebrospinal fluid cytology in a case of epithelioid glioblastoma.

Epithelioid glioblastoma (eGB) is a rare GB subtype exhibiting characteristic morphology and genetic alterations. The efficacy of BRAF and MEK-1/2 inhibitors is demonstrated in eGB treatment, and therefore, considering eGB is important to enhance patient care and prognosis.

Pulmonary Langerhans Cell Histiocytosis: Diagnosis in Bronchoalveolar Lavage Liquid-Based Cytology Samples.

INTRODUCTION: Pulmonary Langerhans cell histiocytosis (PLCH) is a rare interstitial lung disease characterized by the accumulation of Langerhans cells within the lung tissue. The diagnosis of PLCH traditionally involves clinical, radiological, and lung biopsy histopathological evaluations. CASE PRESENTATION: We present 2 cases where the diagnosis of PLCH was confirmed through the analysis of bronchoalveolar lavage (BAL) fluid cytology using immunoperoxidase technique, highlighting the significance of this minimally invasive technique in the diagnostic process. Clinical and radiological examination suggested advanced interstitial lung disease characterized by a fibrocystic pattern in both cases. The cytologic analysis of the BAL fluid revealed typical histiocytes with longitudinal grooves and eosinophils, which was better seen on liquid-based cytology (LBC) smears. ICC with CD1a, Langerin, and S-100 confirmed the diagnosis of PLCH. CONCLUSION: Detecting PLCH through the examination of BAL cytology poses challenges, yet it is achievable, particularly with the assistance of LBC and ICC.

Diagnostic utility of transfer learning by using convolutional neural network for cytological diagnosis of malignant effusions.

INTRODUCTION: Cytological analysis of effusion specimens provides critical information regarding the diagnosis and staging of malignancies, thus guiding their treatment and subsequent monitoring. Keeping in view the challenges encountered in the morphological interpretation, we explored convolutional neural networks (CNNs) as an important tool for the cytological diagnosis of malignant effusions. MATERIALS AND METHODS: A retrospective review of patients at our institute, over 3.5 years yielded a dataset of 342 effusion samples and 518 images with known diagnoses. Cytological examination and cell block preparation were performed to establish correlation with the gold standard, histopathology. We developed a deep learning model using PyTorch, fine-tuned it on a labelled dataset, and evaluated its diagnostic performance using test samples. RESULTS: The model exhibited encouraging results in the distinction of benign and malignant effusions with area under curve (AUC) of 0.8674, F-measure or F1 score which denotes the harmonic mean of precision and recall, to be 0.8678 thus, demonstrating optimal accuracy of our CNN model. CONCLUSION: The study highlights the promising potential of transfer learning in enhancing the clinical pathology laboratory efficiency when dealing with malignant effusions.

Intraperitoneal Dedifferentiated Liposarcoma: Unveiling a Rare Diagnosis in Peritoneal Fluid Cytology - A Case Report with Literature Review.

INTRODUCTION: Sarcomas presenting as malignant effusions are rare, and diagnosing them on fluid cytology requires expertise and clinicoradiological correlation as cells undergo morphological changes, mimicking carcinoma or mesothelioma. CASE PRESENTATION: We present a case of a 70-year-old man with abdominal distention and pain, initially suggestive of carcinoma on peritoneal fluid cytology. However, subsequent analysis with immunohistochemistry on the cell block revealed diffuse nuclear positivity for MDM2, leading to the diagnosis of dedifferentiated liposarcoma. CONCLUSION: The cytological diagnosis of dedifferentiated liposarcoma is challenging and requires a high index of suspicion, with clinicoradiological correlation. Utilizing immunohistochemistry on cell block samples enhances diagnostic accuracy and guides appropriate patient management.

Validation of a novel alcohol-free preservative solution in comparison with conventional prefixation on quality of serous body fluid smears.

BACKGROUND: We describe a novel alcohol-free preservative composed of glucose, mannitol, disodium hydrogen orthophosphate, thymol, and distilled water (glucose-mannitol-disodium dihydrogen orhtophosphate-thymol [GMDT] preservative) in appropriate proportion as an alternative to alcohol prefixation (APF) of body fluids. OBJECTIVES: To assess the cytomorphologic preservation and staining quality of serous body fluid smears generated by GMDT preservative and compare it with smears processed by standard 50% APF. METHODOLOGY: The study comprised 151 effusion samples. Each sample was equally divided into four tubes. Equal volumes of APF and GMDT preservatives were added to the first two tubes and left at room temperature for 24 h. Similarly, the corresponding preservatives were added to the third and fourth tubes and stored for 48 h. Two smears were prepared from the centrifuged sediments of each tube (all four tubes) and stained with May-Grünwald Giemsa and Papanicolaou (Pap) stains. Using a three-tiered scoring system, the smear examination was blinded to assess the extent of cellular preservation and the staining quality by two cytotechnologists and two cytopathologists. Statistical analysis was performed by STATA 16.0. RESULTS: Samples processed with the GMDT preservative at 24 h showed better cytoplasmic preservation and smear background, while nuclear features and staining quality showed no difference between the two preservatives. Mild cytoplasmic and nuclear degenerative changes were noted with the GMDT at 48 h, while all four parameters remained similar with APF at 24 and 48 h. CONCLUSIONS: The newly developed alcohol-free, GMDT preservative, could be a feasible and cost-effective alternative to 50% APF, preferably when samples are processed within 24 h.

Reevaluation of ovarian cyst fine-needle aspiration cytology: A 10-year institutional experience.

OBJECTIVES: Fine-needle aspiration (FNA) of ovarian cyst fluid remains useful for certain clinical circumstances despite low sensitivity and potential safety concerns. The current study aimed to reevaluate the performance of ovarian cystic fluid cytology following American College of Obstetricians and Gynecologists guidelines using a single-institution cohort. METHODS: A total of 507 ovarian cyst FNA cases from 2013 to 2023 were reviewed. Patients' demographics and clinical and radiologic information were collected through the electronic database. The performance was calculated using corresponding surgical pathology diagnosis as the gold standard. RESULTS: Overall, cytologic diagnoses were nondiagnostic (ND), negative for malignancy (NFM), atypical (ATY), suspicious for malignancy (SFM), and malignant (M) in 5 (1.0%), 478 (94.3%), 14 (2.7%), 2 (0.4%), and 8 (1.6%) cases, respectively. Among 349 specimens (68.8%) that had a corresponding surgical pathology, the rate of malignancy (including borderline tumors) was 1.2% (4 of 325) in NFM, 72.7% in ATY (8 of 11), and 100% in both SFM (2 of 2) and M (8 of 8) specimens. Considering NFM and ATY as negative results and SFM and M as positive results, overall, the sensitivity of ovarian cystic fluid cytology was 45.4% and the specificity was 100%. CONCLUSIONS: As an uncommon test, ovarian cystic fluid cytology has moderate sensitivity and high specificity. Despite limitations, ovarian cystic FNA cytology remains a valuable diagnostic tool in certain aspects.

Role of Ancillary Techniques in Reporting Serous Fluid Cytology - "Redefining Categories, Refining Diagnosis".

Background: The "International System of Reporting Serous Fluid Cytology (TIS)" together with cytomorphology promotes the use of ancillary techniques to resolve difficulties in reporting serous fluid cytology. Objective: To classify serous effusion fluid samples received at our department in line with "TIS", indicating the risk of malignancy (ROM), and directing appropriate usage of ancillary testing. Materials and Methods: Prospective study carried out from October 2021 to September 2022. The study included all pleural, ascitic, and pericardial fluid samples, reported according to 'TIS'. Flow cytometry and immunocytochemistry were ancillary methods utilized to assist in reporting. Cases with available history and convincing correlations didn't require further assessment. Results: A total of 1200 serous effusion samples were evaluated including 604 pleural, 591 ascitic, and 5 pericardial fluid samples. After categorization, there were 23 samples in non-diagnostic (ND, 1.9%), 575 in negative for malignancy (NFM, 47.91%), 44 in atypia of undetermined significance (AUS, 3.66%), 64 in suspicious for malignancy (SFM, 5.33%), and 494 in malignant category (MAL, 41.16%). Ancillary studies were beneficial in the recategorization of 26% (11/44) AUS cases, 29.6% (19/64) SFM cases, and it helped refine tumor characteristics in 35.42% (175/494) cases categorized as malignant. Final ROM calculated for each category: ND 25%, NFM 18.6%, AUS 66.6%, SFM 88%, and MAL 100%. Conclusion: Serous fluid is an easily obtainable sample that can provide opportunities for ancillary testing with clinical implications. In AUS and suspicious category although, diagnostic yield is increased however, a larger number of cases are required to obtain definite results.

Utility and performance of cell blocks in cerebrospinal fluid cytology: Experience at two teaching hospitals.

BACKGROUND: Cytology cell blocks (CBs) are not routinely made for cerebrospinal fluid (CSF) specimens. The goal of this study was to identify when CSF CB preparation improves diagnostic performance. MATERIALS AND METHODS: Under institutional review board approval, a retrospective review of CSF cytology cases was conducted at a tertiary university-based hospital and an affiliated county hospital. Patient history, CSF volume, final diagnosis, use of stains, and whether the CB was contributory was determined from the cytopathology report. CSF nucleated cell count data was obtained from the medical record. RESULTS: A total of 69 CSF specimens with CBs from January 2006 to March 2023 were identified from 61 patients. The median CSF volume was 8 mL (interquartile range, 4-13 mL; range, 1-800 mL), with immunohistochemical stains performed on 29 (42%) cases. Per cytology report, CB was contributory in 23 cases (33%), not contributory in 34 cases (49%), and not discussed in 12 cases (17%). The median volume was 8 mL for cases in which CB was contributory, not contributory, or not discussed. There was no difference in average nucleated cell counts between cases in which CB was contributory versus not contributory (73.9 vs. 40.0, p = .175). CONCLUSIONS: CBs for CSF samples were contributory in a subset (33%) of cases. The authors were unable to identify any specific pre-analytic factors, including specimen volume and average nucleated cell counts, for cases in which CB was contributory. Further evaluation is needed to identify if there are scenarios in which CSF CBs should be routinely prepared.

Utility of anticoagulation, pre-smearing and post-smearing hemolytic techniques on morphological assessment and reproducibility in fluid cytology.

Objective: Knowledge of proper collection, storage, preservation, and processing techniques is critical to ensuring proper handling and analysis of fluid cytology specimens. This study was conducted to determine the effect of anticoagulation, pre-smearing acetic acid treatment technique, and saline rehydration technique on morphological assessment, reproducibility, and reporting in fluid cytology. Material and Methods: The study was carried out in the cytopathology laboratory over 2 months (April-May 2022), where 100 effusion samples were analyzed. At least 20-40 mL of fluid was collected in heparinized and non-heparinized containers for each patient. Samples were processed in cytospin and stained with Giemsa and Papanicolaou stains. For 70 hemorrhagic specimens, an extra smear was prepared from the sediment and subjected to the saline rehydration technique as per the Indian Academy of Cytologists (IAC) guidelines. Seventy-three hemorrhagic specimens whose quantity received was more than 35 mL were subjected to the pre-smearing technique. These smears were evaluated for (a) the presence or absence of blue background/any other background staining, (b) cellularity, (c) cell morphology and (d) the presence/absence of microclots. Results: Heparinized samples showed no compromise in cellular morphology or cellularity although a blue background was observed in an occasional case. The pre-smearing technique had less background hemorrhage and preserved cell characteristics. The post-smearing saline rehydration technique did not compromise the cellularity but distorted morphology and showed background staining. Conclusion: The pre-smearing acetic acid treatment showed better-preserved cellularity and cytomorphology with the absence of background staining when compared to the post-smearing saline rehydration technique.

Reliably making the primary diagnosis of mesothelioma utilizing serous fluid cytology specimens: an institutional experience.

INTRODUCTION: The diagnosis of mesothelioma has historically been challenging, especially on serous fluid cytology (SFC). Distinguishing between reactive and neoplastic mesothelial cells can be difficult on cytomorphology alone. However, additional ancillary tests, such as BRCA1 associated protein-1 immunohistochemistry and fluorescence in situ hybridization for cyclin-dependent kinase inhibitor 2A deletion, can provide a sensitive and highly specific method of proving malignancy. MATERIALS AND METHODS: SFC specimens diagnosed as mesothelioma, suspicious for mesothelioma (SM), and atypical mesothelial cells (AMCs) since 2012 were identified by querying the laboratory information system. Clinical data and pathologic parameters were gathered. RESULTS: One hundred ten cases of mesothelioma, SM, and AMC were identified. Of these, 61 cases had a definitive diagnosis of mesothelioma on SFC. Average age at SFC diagnosis was 67 years (26-87 years), with most patients being male (67%). Out of the 61 cases, 11 cases (18%) had an initial diagnosis of mesothelioma made on SFC specimens, with 5 of these 11 cases being in patients that never received a histologic diagnosis of mesothelioma. Ancillary studies were utilized in all 11 cases. An initial diagnosis of metastatic mesothelioma was made on SFC in 9 cases (15%). For 6 of these 9 cases, the SFC diagnosis was the sole diagnosis of metastatic mesothelioma without a companion histologic diagnosis. In addition, 15 cases were diagnosed as SM, with 11 of these cases following a definitive mesothelioma diagnosis. Thirty-four cases were diagnosed as AMC, with 27 cases following a definitive mesothelioma diagnosis. CONCLUSIONS: The diagnosis of mesothelioma can be reliably made on SFC with the appropriate cytomorphology criteria and/or confirmatory ancillary testing.

Myelomatous meningitis diagnosed by cerebrospinal fluid cytology examination.

Myelomatous meningitis diagnosed by CSF cytology. The combined use of cytology with immunocytochemistry can identify the presence of multiple myeloma cells in cerebrospinal fluid specimens.

The diagnostic accuracy and prognostic implication of pelvic and peritoneal fluid cytology specimens in ovarian clear cell carcinoma.

INTRODUCTION: Ovarian clear cell carcinoma (OCCC) is a rare subtype of ovarian epithelial carcinoma. Patients with low-stage disease have an excellent prognosis, while the prognosis for those with high-stage disease is poor. Neoplastic cells in abdominopelvic washings upstages the patient to at least FIGO 1C3. Positive cytology confers a worse prognosis when compared to similar stage patients with negative cytology. This study aims to investigate the diagnostic performance of abdominopelvic fluid cytology specimens in cases with pure OCCC and reaffirm the importance of accurate cytologic detection and its impact on patient prognosis. MATERIALS AND METHODS: The laboratory information system was queried to identify all patients treated for ovarian clear cell carcinoma at our institution over a period of 20 years with a companion abdominopelvic fluid cytology specimen at the time of surgical resection. Cases were sorted by the FIGO stage of the corresponding oophorectomy specimen. Cytology results, patient demographics, fluid volume, immunohistochemical results, and follow-up data were recorded. RESULTS: A total of 143 cases were identified. The overall detection rate was 38%, with 54 of 143 cases positive for malignancy. Cytologic detection rates increased as FIGO stages increased. Fifty percent of stage 1C cases were upstaged on cytology alone. Ascites fluids performed better among stage 1 cases compared to pelvic wash specimens (77% detection rate versus 23%). Stage 1 patients with positive cytology trended towards a worse prognosis compared to those with negative cytology. CONCLUSIONS: Positive cytology in low stage cases of OCCC has significant prognostic and therapeutic implications. Our large cohort further underscores the importance of accurate cytologic detection and subsequent staging in this setting.

Diagnosis of peritoneal metastasis in an unruptured hepatocellular carcinoma on ascitic fluid cytology: A rare scenario with brief review of literature.

Hepatocellular carcinoma (HCC) is the most common primary liver malignancy in adults occurring in a background of cirrhosis. Peritoneal dissemination of HCC is an unusual presentation with an incidence of 2%-16%. Peritoneal metastasis of an unruptured HCC is extremely uncommon. Despite low yield, ascitic fluid cytology serves as a valuable tool for diagnostic evaluation in a patient of cirrhosis with suspicion of malignant transformation. We present a rare case scenario in an elderly female with cirrhosis where the diagnosis of peritoneal metastasis was established on ascitic fluid cytology and confirmed by immunocytochemistry. This report illustrates the unique clinical presentation of an unruptured HCC with its cytological features and a brief review of literature.

Liposarcoma Involving Serous Fluid Cavities-A Case Series Illustrating Clinical Implications and the Diagnostic Role of Exfoliative Cytology.

Introduction: Cytological diagnosis of sarcomas requires detailed cytomorphological assessment and integration of immunocytochemistry and/or molecular testing. The role of exfoliative cytology, as compared to aspiration cytology, is less understood. This case series describes well-differentiated/dedifferentiated liposarcomas in effusions, with cytomorphological features, ancillary test results and clinical outcomes detailed. Methods: A computerized search of the department pathology archives was performed for sarcomatous effusions with histological diagnosis or clinical history of well-differentiated/dedifferentiated liposarcoma. Clinical progress, cytology slides, immunocytochemistry and molecular test results were reviewed. Results: Six patients were identified. In 5 patients with clinical follow up, 4 (80%) were deceased within 5 months of malignant effusion. One patient was alive with 12 years disease-free survival after radical resection with adjuvant radiotherapy. Three patients showed dedifferentiation on histology, and high-grade (dedifferentiated) tumor cells were present in effusion cytology of 2 patients. Two showed well-differentiated components only on biopsy, but high-grade (dedifferentiated) tumor cells were identified in cytology. The high-grade tumor cells displayed marked nuclear irregularity, enlargement, size variation, with macronucleoli and multinucleation. Well-differentiated lipomatous components were demonstrated in 4 patients (66.7%), comprising of multivacuolated lipoblasts and atypical lipocytes. CDK4 and MDM2 immunoreactivity in all 3 cases with cell blocks, and CDK4 and MDM2 amplification in one were successfully demonstrated. Conclusion: Lipomatous and dedifferentiated components can be sampled and cytomorphologically identified on effusion fluids of liposarcomas, with sufficient cellularity for immunocytochemistry and molecular testing. Although generally associated with poor prognosis, long disease-free survival with sarcomatous effusion is possible with radical surgery and adjuvant treatment.

Bacillary Positive Tuberculous Body Fluid Smears: A Perspective on How Fast to Use Acid Fast.

Background Tuberculous effusions are common. Classically, they are described as bacteria poor and lymphocyte rich. Our experience, however, has been more varied. We compiled this rare group of bacteria-positive tuberculous fluids to document their cytologic spectrum and to look for possible predictors of bacillary positivity. Methods Fifty-one cases of bacillary positive fluids were identified and their clinicopathological details were noted. Per case, the smear background was assigned as either clear, caseous, suppurative, granular proteinaceous or frankly hemorrhagic. Fine, punched-out vacuoles in the smear background were also noted. The bacillary load in each case was classified from scanty to 3+. Eventually, the clinicopathologic variables were tabulated for frequency and studied for any association with bacillary presence. Results Only 19 of the 51 patients had a history of tuberculosis. Retropositive patients comprised a small proportion (9.8%) and did not always indicate strong (3+) bacillary positivity. The granular proteinaceous background was the most frequent (35%) pattern. Only a suppurative background was associated with strong bacillary positivity. Fine vacuoles were seen almost always with caseous and granular proteinaceous backgrounds but without statistical significance. Conclusion Tuberculous effusions can have diverse smear backgrounds, not necessarily one rich in caseous material. When tuberculosis is known or clinically suspected, non-classical findings such as abundant neutrophils or suppurative background should not dissuade one from requisitioning mycobacterial stains. In fact, acid-fast stains should probably routinely accompany Giemsa slides of clinically idiopathic effusions in endemic areas since it is still the cheapest and fastest method for a conclusive diagnosis.

The Value of a Negative Pleural Fluid Cytology and Biopsy in Diagnosing Malignant Pulmonary Lesions.

Over 2 million patients developed lung cancer in 2018, and lung malignancy is responsible for an estimated 1.8 million deaths worldwide. Lung cancer diagnosis usually occurs after suspicious symptoms or incidental radiologic findings on chest imaging when the cancer is probably in an advanced stage. Therefore, initial evaluation, diagnosis, staging, and prompt treatment of lung cancer are required to improve pulmonary malignancies' morbidity and mortality rate. Unfortunately, the size of the tumor, the time of imaging, the quality and quantity of pleural fluid, and pleural biopsy all contribute to diagnostic difficulties in evaluating a lung lesion, leaving even the most astute clinician occasionally perplexed. We discuss a case of a female with lung cancer whose diagnosis was challenging because of a negative pleural biopsy, despite initial radiographic imaging suggesting a lung lesion.

The diagnostic significance of cerebrospinal fluid cytology and circulating tumor DNA in meningeal carcinomatosis.

Objective: The objective of this research is to investigate the clinical application value of cerebrospinal fluid (CSF) cytology and circulating tumor DNA (ctDNA) in lung adenocarcinoma (LUAD) meningeal metastasis-meningeal carcinomatosis (MC), and to further explore the possible molecular mechanisms and drug treatment targets of LUAD meningeal metastasis by next-generation sequencing (NGS). Methods: We retrospectively analyzed LUAD with MC in 52 patients. CSF cytology was carried out using the slide centrifugation precipitation method and May-Grüwald-Giemsa (MGG) staining. Tumor tissue, plasma and CSF ctDNA of some MC patients were detected by NGS. Results: Of the 52 MC patients, 46 (88.46%) were positive for CSF cytology and 34 (65.38%) were positive for imaging, with statistically significant differences in diagnostic positivity (P < 0.05). In 32 of these patients, CSF cytology, cerebrospinal fluid ctDNA, plasma ctDNA and MRI examination were performed simultaneously, and the positive rates were 84.38, 100, 56.25, and 62.50% respectively, the difference was statistically significant (P < 0.001). Analysis of the NGS profiles of tumor tissues, plasma and CSF of 12 MC patients: the mutated gene with the highest detection rate was epidermal growth factor receptor (EGFR) and the detection rate were 100, 58.33, and 100% respectively in tumor tissues, plasma and CSF, and there were 6 cases of concordance between plasma and tissue EGFR mutation sites, with a concordance rate of 50.00%, and 12 cases of concordance between CSF and tissue EGFR mutation sites, with a concordance rate of 100%. In addition, mutations not found in tissue or plasma were detected in CSF: FH mutation, SETD2 mutation, WT1 mutation, CDKN2A mutation, CDKN2B mutation, and multiple copy number variants (CNV), with the most detected being CDKN2A mutation and MET amplification. Conclusion: CSF cytology is more sensitive than traditional imaging in the diagnosis of meningeal carcinomatosis and has significant advantages in the early screening and diagnosis of MC patients. CSF ctDNA can be used as a complementary diagnostic method to negative results of CSF cytology and MRI, and CSF ctDNA can be used as an important method for liquid biopsy of patients with MC, which has important clinical significance in revealing the possible molecular mechanisms and drug treatment targets of meningeal metastasis of LUAD.