Journal of Cytology
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   Table of Contents - Current issue
Coverpage
April-June 2019
Volume 36 | Issue 2
Page Nos. 75-135

Online since Friday, March 8, 2019

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ORIGINAL ARTICLES  

A comparative study for selectivity of micronuclei in cervical exfoliated cells on chronic boron effects p. 75
Hatice Yıldırım, Aslı Göker, Hülya Demirci, Tevfik Güvenal, Mehmet Korkmaz
DOI:10.4103/JOC.JOC_185_17  
Background: Cervical cancer is one of the most frequent malignancies in women. Micronucleus (MN) testing has gained popularity as a biomarker in early diagnosis of many types of cancer. Aims: This study aims to investigate the role of MN testing on early detection of cervical cancer and the effect of boron exposure on cervical cells. Settings and Design: The study population comprised women who were diagnosed to be human papillomavirus (HPV)-positive and had atypical squamous cells of undetermined significance (ASCUS) as cervical cytology in a cervical screening project. A total of 15 HPV-positive and 36 ASCUS patients were identified. Randomly selected 20 women were selected from boron-rich region (n = 10) and nonboron region (n = 10). Materials and Methods: Cervical swab specimens were dyed using Papanicolaou (PAP) and May–Grünwalds–Giemsa (MGG) techniques, and MN count in 1000 cells was performed. The results were statistically evaluated. Statistical Analysis Used: Data were analyzed using Statistical Package for the Social Sciences (SPSS) version 13. Quantitative data were presented as mean ± standard deviation. MN test scoring was compared using Mann–Whitney U-test. Results: Boron content of urine was measured to be 3.02 ± 1.45 and 0.98 ± 0.42 mg/day in boron-rich and nonboron regions, respectively. When MN counts were compared according to PAP and MGG staining in HPV- and ASCUS-positive women, there was statistically no significant difference (P > 0.05). Disregarding regions, HPV/control and HPV/ASCUS cases stained with PAP and MGG had statistically significant difference in MN count (P < 0.05). Conclusion: These findings suggest that MGG and PAP staining gives similar results with regard to MN count. On the other hand, it has been shown again that HPV induces MN and causes genomic instability.
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Cytology microarray on cell block preparation: A novel diagnostic approach in fluid cytology p. 79
Arghya Bandyopadhyay, Soumi Bhattacharyya, Shreosee Roy, Kaushik Majumdar, Kaushik Bose, Anup K Boler
DOI:10.4103/JOC.JOC_15_17  
Background: The cytological examination of serous body effusions to diagnose and stage malignancy is well accepted in clinical medicine. Conventional smear (CS) and cell block (CB) study has to be complemented with immunohistochemistry (IHC) for a definitive diagnosis of malignancy and also to differentiate it from reactive mesothelial cells. Cytology microarray (CMA) is a modification of tissue microarray which involves core needle biopsy of multiple cell blocks and embedding it in a single block. Aim: The aim of this study was to assess the effectiveness of IHC technique in CMA for rapid diagnosis of malignancy and to reduce the cost of testing. Materials and Methods: In this study, 82 pleural fluids were collected and subjected to CS and CB study followed by IHC in CMA blocks. Six commonly used antibodies were applied to confirm malignancy and diagnose the primary. Results: Nineteen cases were diagnosed as malignancy by CB method. MOC-31 confirmed adenocarcinoma deposit in 67% cases of which 44% were proved to be of lung primary by TTF1. Conclusions: IHC on CMA blocks of effusion fluids is a very effective technique that can significantly reduce the cost of testing by >70%.
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Clinical and cyto-morphological characterization of triple negative breast cancer p. 84
Chayanika Kala, Mohd Athar, Sanjay Kala, Lubna Khan, Ramendra K Jauhari, Amitabh Satsangi
DOI:10.4103/JOC.JOC_47_18  
Objective: Triple negative breast cancer (TNBC), despite being the uncommon subtype, contributes a major portion to mortality and associated with poor prognosis. The purpose of this study was to evaluate the cytological criteria for the diagnosis of TNBC through fine-needle aspiration cytology (FNAC). Material and Method: Clinical, cytological, histological, and immunohistochemical (IHC) data of 256 patients were evaluated, and patient were classified as TNBC and non-TNBC phenotype by IHC. All cytological specimens were reviewed for 12 criteria: cellularity, tubule/gland formation, syncytial clusters, large bare nuclei, nuclear atypia, chromatin pattern, cell borders, nucleolus, cytoplasm, lymphocytic infiltrate, calcification, and necrosis. The Fischer's exact test was used to show test association. Result: Out of 256 patients, 82 patients were TNBC, and 174 patients were non-TNBC. TNBC phenotype showed statistically significant association to cellularity, tubule/gland formation, syncytial cluster formation, bare nuclei, nuclear atypia, cell borders, lymphocyte infiltration, and necrosis. Conclusion: FNAC can be helpful in making diagnosis of TNBC and along with ER, PR, HER2 characterization, helpful in planning treatment strategy, saving time, manpower, and resources in the patient management.
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Comparative analysis of liquid based and conventional cytology smears in fine needle aspirates from breast lesions p. 89
Vasundhara Sharma, Veena Gupta, Padam Parmar, Promil Jain, Deepak Thakran, Rajeev Sen
DOI:10.4103/JOC.JOC_61_18  
Context: Liquid-based cytology (LBC) is being extensively used for the evaluation of both gynecological and non-gynecological specimens. Suspension of cells in monolayer makes better morphological assessment possible. Along with this, inherent morphological changes such as altered, reduced, or lost background material, fragmented cell clusters, smaller cell size, nucleolar prominence, etc., need to be considered. Aim: Present study was aimed at comparative evaluation of utility of LBC versus conventional smear (CS) in assessing breast lesions and whether it can be used as an alternative to conventional preparation. Settings and Design: Present study was a prospective study in which 75 cases of breast fine-needle aspiration cytology from patients with palpable breast lumps constituted the study group. Material and Methods: The first pass was used for CS and LBC; a second pass was given. The representative CS and LBC smears were compared using several criteria. Statistics: Each feature was scored individually and evaluated statistically using Wilcoxon's signed rank test on the SPSS program. Results: A statistically significant difference was found in informative background and background blood-debris, whereas the difference was not statistically significant in other features such as cellularity, cytoarchitectural pattern, presence of monolayer, and nuclear and cytoplasmic details. Conclusion: LBC is a promising technique in the field of cytology. It has the potential to decrease the number of slides screened per case and decrease the turn-around-time.
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Hormone receptors and human epidermal growth factor (HER2) expression in fine-needle aspirates from metastatic breast carcinoma – Role in patient management p. 94
Issam M Francis, Rabeah Abbas Altemaimi, Bushra Al-Ayadhy, Preeta Alath, Mohammed Jaragh, Fatma Jasem Mothafar, Kusum Kapila
DOI:10.4103/JOC.JOC_117_18  
Introduction: Estrogen receptors (ER), progesterone receptors (PR), and epidermal growth factor (HER2) are prognostic and predictive factors for breast carcinoma. We determined them by immunohistochemistry (IHC) on cell blocks from fine-needle aspirates (FNA) of metastatic breast carcinoma to axillary lymphnodes and compared them with that reported in the primary breast carcinoma (PBC) to document any change in their expression for future management. Materials and Methods: ER, PR, and HER2 by IHC and HER2 oncogene by fluorescent in-situ hybridization (FISH) were studied on cell blocks of FNA of axillary lymphnodes in 53 of 94 PBC cases from 2012 to 2016. Results: In 25 of 38 (65.8%) ER, PR negative PBC the metastasis on FNA was ER, PR+, whereas the 15 (28.3%) ER, PRPBC remained negative. In 10 of 11 (91%) of HER2-IHC+, PBC the metastatic tumor was HER2-IHC+. 7 of 32 (21.9%) HER2-IHC negative PBC were HER2-IHC+ in metastatic tumor. HER2-FISH was performed in 37 cases on FNA. Six of 37 were HER2 amplified/positive, whereas 9 and 19 remained equivocal and negative for HER2 copy number, and 3 were not interpretable. All the 6 HER2-FISH+ cases were positive by IHC. In our study, 34.2% of ER, PR+ cases of PBC became ER, PR– in the metastatic tumor and 21.9% of HER2-IHC negative PBC became HER2-IHC+ in the metastatic aspirate. Conclusion: ER, PR, and HER2 by IHC in cell blocks of metastatic lymphnodes are reliable. Change in receptor (34.2%) and HER2 status (21.9%) was documented, which is of clinical significance as these patients warrant a change of management.
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Diagnostic efficacy of ultrasound-guided fine needle aspiration combined with the bethesda system of reporting p. 101
K Ajitha Kumari, Poonam D Jadhav, Chaya Prasad, NV Smitha, Annie Jojo, VD Manjula
DOI:10.4103/JOC.JOC_168_16  
Background: Image-guided fine needle aspiration cytology (FNAC) is emerging as an important diagnostic tool in the evaluation of thyroid swellings. Aim: This study aims to assess the efficacy of ultrasound (US)-guided FNAC combined with “The Bethesda system' of reporting as a primary screening test for all thyroid lesions. Settings and Design: A prospective cohort study was made and all the US-guided FNACs done were followed up to find out the histopathological diagnoses wherever surgery was done. Materials and Methods: In all, 1050 patients who underwent US-guided FNAC were studied during a period of 1 year. Age, sex, cytological features, and histological diagnoses were analyzed. Statistical analyses of all the findings were done to derive conclusions. Results: Of the 1050 patients, only 10.5% underwent surgery. Higher than expected rate (as per the Bethesda system) of malignancy was noted with the so-called grey zone lesions. The test results revealed a high level of sensitivity, specificity, and diagnostic accuracy. Conclusion: The study showed that The Bethesda System of Reporting Thyroid Cytology provides effective communication between clinician and pathologists thereby enabling clear management strategies. We also concluded that US-guided FNACs offer better results compared with palpation-guided FNACs.
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Image-guided fine needle aspiration cytology of intrathoracic lesions p. 106
Annie E Abraham, Pooja K Suresh, HB Sridevi, Kausalya K Sahu, Deepa Adiga, Jessica Minal, Santosh Rai, Vishak Acharya
DOI:10.4103/JOC.JOC_187_17  
Background: Percutaneous, image-guided transthoracic fine needle aspiration cytology (TTFNAC) is a rapid, yet accurate, and well-established diagnostic method used in the cytological evaluation of intrathoracic lesions. The study was done to determine the utility of image-guided TTFNAC in diagnosis of intrathoracic lesions. Subjects and Methods: A retrospective analysis of all cases who underwent image-guided TTFNAC of a suspected intrathoracic lesion, in a tertiary care hospital was done over a period of 3 years. Results: During the study period, 124 cases of image-guided FNAC of intrathoracic lesions were obtained. The mean age at presentation was 60.5 years with M:F: 3.6:1. Neoplastic lesions (71.5%) outnumbered the nonneoplastic lesions (28.5%). The most common tumor was adenocarcinoma (25%) followed by squamous cell carcinoma (SCC, 11%), and small cell carcinoma (5%). There was one case each of anaplastic carcinoma, plasmacytoma, bronchoalveolar carcinoma, and non-Hodgkin lymphoma (NHL). Most of the lesions were found on the right side and upper lobe. Among the mediastinal lesions, we found two cases of thymoma and one case each of NHL)/primitive neuroectodermal tumor (PNET), NHL, and small cell carcinoma metastasis to lymph node followed by ten cases of inflammatory lesions and seven cases of tuberculosis (TB). Conclusion: Image-guided TTFNAC of intrathoracic lesions is a safe method when done by well-trained medical personnel with lesser rate of complications. An early accurate diagnosis of malignancy can be made based on the cytological features; however, further subtyping of the malignancy may sometimes be difficult due to overlapping cytological features. TTFNAC can be a diagnostic tool for identifying nonneoplastic lesion such as TB. Hence, image-guided FNAC aids in early diagnosis and management of patients with intrathoracic lesions.
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Utility of DNA-specific stains in micronuclei assay as a marker of genotoxicity in oral potentially malignant disorders and oral squamous cell carcinoma p. 111
Saurabh Juneja, Sonal Katyal, Ajit Singh Rathore, Devi Charan Shetty, Ankita Tandon, Pankaj Jain
DOI:10.4103/JOC.JOC_230_16  
Background: The term oral potentially malignant disorder (OPMD) was recommended to refer to precancer as it conveys that not all disorders described under this term may transform into cancer. Oral squamous cell carcinoma (OSCC) arises through an accumulation of genetic alterations, deoxy ribonucleic acid (DNA) changes, and epigenetic alterations. Thus, a simple yet a sensitive and specific test for early diagnosis is the need of an hour. The micronuclei (MN) assay in exfoliated epithelial cells is potentially an excellent biomarker to detect chromosome loss or malfunction of mitotic spindle. Aim of the Study: To compare the frequency of MN in exfoliated cells from oral mucosa exposed to genotoxic agents using different staining procedures and to observe the incidence of micronucleus in potentially malignant and malignant lesions. Materials and Methods: The study was undertaken to observe the cytogenetic damage in the exfoliated buccal cells of 75 cases of tobacco-related PMDs, OSCC and control subjects (25 cases from each group) and were evaluated with nonspecific May-Grünwald Giemsa stain and DNA-specific Feulgen stain. The results were statistically determined using SPSS version 17.0. Results: Correlation analyses in the present study depicted that MN frequency was significantly more in oral squamous cell carcinoma than OPMDs and normal group (P < 0.05). Giemsa-stained slides correlated significantly with karyorrhexis, karyolysis, condensed chromatin, and binucleates, whereas no such correlations were found with DNA-specific stains. Conclusion: Malignant transformation is accompanied by loss of cell capacity to evolve to death in situations of DNA damage. These findings indicate that nuclear anomalies may be misinterpreted as MN with nonspecific DNA stains and lead to false-positive results in studies with cells of epithelial origin.
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SYMPOSIUMS Top

DNA extraction and polymerase chain reaction p. 116
Nalini Gupta
DOI:10.4103/JOC.JOC_110_18  
DNA extraction and polymerase chain reaction (PCR) are the basic techniques employed in the molecular laboratory. This short overview covers various physical and chemical methods used for DNA extraction so as to obtain a good-quality DNA in sufficient quantity. DNA can be amplified with the help of PCR. The basic principle and different variants of PCR are discussed.
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Core needle biopsy versus fine needle aspiration cytology in bone and soft tissue tumors p. 118
Bharat Rekhi
DOI:10.4103/JOC.JOC_125_18  
Fine needle aspiration cytology (FNAC) for diagnosis of musculoskeletal tumors is well discussed and debated. The current evidence restricts its application in recurrent and metastatic lesions, with core needle biopsy as the acceptable technique/gold standard for the primary diagnosis of these challenging tumors. However, there are several studies showing reasonable sensitivity and specificity in primary diagnosis of bone and soft tissue tumors, especially in differentiating benign from malignant tumors. There is an acceptable limitation in the exact subtyping of soft tissue tumors, based on cytology smear examination, in the absence of ancillary techniques. Nonetheless, cytology constitutes an optimal material for triaging cases for ancillary techniques, such as immunocytochemistry, immunohistochemistry, molecular cytogenetics, and molecular diagnosis, in the form of smears and cell block preparations. This review focuses on the strengths and limitations of cytology versus core needle biopsy in the diagnosis of bone and soft tissue tumors, with the current evidence in the form of published studies, including the authors' experience.
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Lung molecular cytopathology: EGFR and beyond p. 124
Deepali Jain
DOI:10.4103/JOC.JOC_135_18  
Lung cancer (LC) is the leading cause of cancer-related mortality. Unfortunately, most patients of LC present at the advanced stage of the disease with a poor prognosis and 1-year survival of less than 20%. At the advanced stage of the disease, surgical resection cannot be possible, hence small biopsy or cytology specimens remain a choice for their correct diagnosis. The recognition of molecular drivers has revolutionized the treatment paradigm of non-small cell lung cancer (NSCLC) with introduction of tyrosine kinase inhibitors. Epidermal growth factor receptor (EGFR) gene mutations were identified, first, to be targeted in NSCLC followed by activating fusions in anaplastic lymphoma kinase (ALK) and rearrangements in c-ros oncogene 1 (ROS1) genes. In addition, the encouraging progress of immunotherapy in patients with NSCLC has been associated with predictive biomarker testing in the form of programmed death ligand-1 (PD-L1) immunohistochemistry assay. To test for these alterations, accurate biomarker testing is needed from biopsy or cytology specimens. In this brief review, testing of biomarkers is discussed using cytology specimens.
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CME Top

Role of EBUS-TBNA in the diagnosis of tuberculosis and sarcoidosis p. 128
Valliappan Muthu, Nalini Gupta, Ritesh Agarwal, Sahajal Dhooria
DOI:10.4103/JOC.JOC_150_18  
Endobronchial ultrasound (EBUS)-guided transbronchial needle aspiration (TBNA) has diagnostic accuracy and negative predictive value for the diagnosis of mediastinal tubercular lymphadenitis, especially when culture and cytological diagnosis is combined with high clinical suspicion. Both sarcoidosis and tuberculosis (TB) demonstrate granulomatous inflammation on cytology. Diagnosis of sarcoidosis in regions with a high burden of TB is challenging. We conducted a prospective study in 179 cases of suspected granulomatous pathology in mediastinal lymphadenopathy cases to evaluate the role of EBUS-TBNA in diagnosis of sarcoidosis and TB. It was found that extensive caseous necrosis, acid-fast bacilli positivity on Ziehl–Neelsen staining, and/or microbiological culture positivity have high positive predictive value for the diagnosis of TB.
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IMAGES IN CYTOPATHOLOGY Top

Cytologic features of pulmonary langerhans cell histiocytosis in bronchial washing: A report of two cases p. 131
Myunghee Kang, Seung Yeon Ha, Mee Sook Roh
DOI:10.4103/JOC.JOC_80_18  
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LETTERS TO EDITOR Top

Briskly motile Balantidium-like ciliate morphologically resembling Chilodonella spp. in urine sediment p. 133
Ujjawal Khurana, Kaushik Majumdar, Tadepalli Karuna, Rajnish Joshi, Neelkamal Kapoor
DOI:10.4103/JOC.JOC_21_18  
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Intraoperative flow cytometry for diagnosis of central nervous system lesions p. 134
George A Alexiou, George Vartholomatos, Kalliopi Stefanaki, Georgios S Markopoulos, Athanasios P Kyritsis
DOI:10.4103/JOC.JOC_45_18  
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