Journal of Cytology
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   2021| October-December  | Volume 38 | Issue 4  
    Online since November 22, 2021

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P16INK4a/ki67 immunocytochemistry in improving the predictive value for high grade cervical intraepithelial (≥CIN2) neoplasia in pap smear
G Vinoth Kumar, Anne Jennifer Prabhu, Ajit Sebastian, Raghavendran , Priya Abraham, Abraham Peedicayil
October-December 2021, 38(4):180-185
Introduction: Cervical cytology has limited sensitivity to detect cervical pre-cancerous lesions. High-risk human papillomavirus (hr-HPV) DNA testing has high sensitivity but its specificity is limited. This study was done to assess the utility of p16INK4a/ki-67 dual stained cytology in improving the predictive value for high-grade cervical (CIN2+) lesions. Aim/Objective: To assess the significance of P16/Ki-67 immunocytochemistry in improving the predictive value for high-grade cervical intraepithelial (≥CIN 2+) lesions on Pap smear. Material and Methods: This was a prospective diagnostic study that included 93 patients with ASC-US/LSIL/ASC-H and HSIL on thin prep cervical smears and who also underwent hr-HPV DNA test and colposcopy-guided biopsy. Biopsy was the gold standard against which the performance of P16INK4a/Ki-67 and hr-HPV results were compared. Results: In women of all ages, sensitivity of (96.8%) hr-HPV test and p16/Ki-67 dual immunocytochemistry (≥1 positive cell) were similar and negative predictive value (NPV) was (97.1% vs. 97.9%) but the latter test showed better specificity (69.4% vs. 53.2%) and positive predictive value (PPV, 61.2% vs. 50.8%) for ≥CIN 2 lesions. A higher cut off of at least 10 positive cells gives a higher specificity and PPV, with slightly decreased sensitivity and NPV. Conclusion: Because high-risk HPV test has a high sensitivity and NPV, whereas P16/Ki-67 dual immunocytochemistry (≥10 positive cells) has a high specificity and PPV, the latter can be recommended as an ancillary test in hr-HPV-positive women to reduce the number of women going for colposcopy and biopsies.
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Diagnostic utility and accuracy of scrape cytology in evaluation of neoplastic lesions
Tanya Sharma, Sheetal Singhal, Neelkamal Kapoor
October-December 2021, 38(4):186-190
Background: Frozen section, intraoperative fine needle aspiration cytology (FNAC) and imprint cytology have been used traditionally by the surgical pathology laboratories for rapid intraoperative diagnosis of tumors. Scrape cytology is a modification of imprint cytology that involves lightly scraping or brushing cells from freshly cut surface of the surgically removed tumor specimens. The present study was carried out to evaluate the utility of scrape cytology in diagnostic evaluation of tumors. Material and Methods: A prospective study was carried out at a tertiary care teaching hospital in Central India. A total of 50 consecutively received specimens in the Department of Pathology and Laboratory Medicine with clinical or radiological suspicion of neoplasia were included in the study. Scrape smears were prepared from freshly cut surface of tumor specimens received unfixed or in 10% formalin. Results: Overall, the scrape cytology diagnosis was concordant with final histopathological diagnosis in 48 out of 50 cases with a diagnostic accuracy of 96%. The sensitivity was 90.9% (CI-58.72% to 99.77%) and specificity was 97.4%(CI-86.52% to 99.94%). Positive predictive value and negative predictive value were 90.91% and 97.44%, respectively. Conclusion: Scrape cytology is a cost-effective and reasonably accurate cytodiagnostic technique for rapid tumor diagnosis. Scrape cytology can be a useful supplementary tool to frozen section, and may be applied for rapid diagnosis where frozen section technique is not available. The material and knowledge obtained from scrape cytology of routinely received histopathological specimens can be utilized as a teaching material and may help unveil diagnostic cytopathological features of infrequent cytologically encountered lesions.
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How to mimic a histological sample slide for RNAscopeTM applications from BAL cytological specimens
Denise Morotti, Emma Caroli, Valeria Forlani, Massimiliano Cadamuro, Andrea Gianatti
October-December 2021, 38(4):231-232
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Extranodal NK/T cell lymphoma presenting as premaxillary swelling: A cytological diagnosis
Pallavi Sinha, Pritika Kushwaha, Kanika Gupta, Meeta Singh, Shyama Jain
October-December 2021, 38(4):233-234
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Traveling with ROSE in EBUS-TBNA – Experience from a tertiary care hospital in South India
Gramani Arumugam Vasugi, Koushik Muthuraja Mathivanan, Swaminathan Rajendiran, Sandhya Sundaram, Irfan I Ayub
October-December 2021, 38(4):175-179
Aims and Objectives: Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) has emerged as a precise modality for tissue sampling of mediastinal and hilar lesions adjacent to the proximal airway. This study aims to determine the diagnostic efficacy, sensitivity, and specificity of rapid-on site evaluation (ROSE) in EBUS-TBNA. Materials and Methods: This is a retrospective study that included 100 patients who underwent EBUS-TBNA of paratracheal and mediastinal lymph nodes in a tertiary care hospital in South India between March 2018 and March 2020. After the procedure, the diagnostic yield from the nodes sampled was transferred to slides that were stained with rapid hematoxylin and eosin (H and E), and then onsite evaluation was done. The tissue derived was also processed for histopathologic examination in all cases. ROSE was performed to assess sample adequacy and to arrive at a preliminary diagnosis. In patients suspected of tuberculosis, the sample was collected for GeneXpert evaluation as well. Results: Of the 100 cases studied, 51 were males and 49 were females. The age distribution was between 3 and 78 years. Forty-seven cases were diagnosed as granulomatous lymphadenitis, 13 as metastatic malignancies, 33 as reactive lymphadenitis, 3 as atypical cells, and 1 case was diagnosed as a cystic lesion. The diagnostic yield was not adequate for evaluation in three cases. Diagnostic yield was obtained in the first two passes where the lymph nodes were more than 2 cm in size. More diagnostic passes were required in lymph nodes less than 2 cm and those located between and adjacent to major vessels. The onsite diagnosis was correlated with the final histopathologic diagnosis. Conclusion: ROSE serves as a useful adjunct to reduce procedure time and enhance sample collection and triaging, and reduces the need for further invasive testing.
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Comparative evaluation of clinical, cytological and microbiological profile in abdominal vs. cervical lymph nodal tuberculosis with special emphasis on utility of Auramine-O staining
Harveen K Gulati, Michael Mawlong, Arushi Agarwal, Karen R Ranee
October-December 2021, 38(4):191-197
Context: Extrapulmonary tuberculosis (EPTB) especially abdominal lymph nodal tuberculosis (LNTB) poses a unique diagnostic challenge. The clinical, cytological, and microbiological profiles, especially with respect to the use and role of Auramine –O (AO) stain, are not as well characterized in abdominal LNTB as cervical LNTB and were evaluated in the present comparative study. Subjects and Methods: This study was conducted in the Department of Pathology of a tertiary care hospital in Shillong, Meghalaya in 540 clinical suspected cases of tuberculosis who underwent FNAC. The smears were submitted for Leishman's stain for cytological analysis, along with ZN and Auramine O stain for demonstration of the organism, analyzed, and scored and the results were compared with culture wherever available. The results from abdominal and cervical lymph nodal tuberculosis were compared using Microsoft Excel and SPSS software. Results: Out of 540 cases, most were tuberculosis (266) followed by reactive lymphadenitis (162), malignancy, and acute necrotizing lesion. On comparing, abdominal lymph nodes (n = 163) were more likely to reveal cheesy/purulent material macroscopically, necrotizing lymphadenitis along with ZN stain and Auramine positivity (P < 0.05) while cervical lymph nodes (n = 66) revealed a higher proportion of granulomatous lymphadenitis and culture positivity (P < 0.05). The sensitivity, NPV, and diagnostic accuracy of AO stain (85.9%, 48.0%, and 62.3%) were higher as compared to ZN stain (47.4%, 39.3%, and 51.9%) with culture as the gold standard. The combined sensitivity of Ziehl Neelsen stain and Auramine stain was 92.05%. Conclusion: Cytological and microbiologic features of abdominal LNTB differ from cervical LNTB. Moreover, AO stain increases the smear positivity, is almost twice as sensitive as ZN stain and should be used as an adjunct in cytological material wherever available.
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Should liquid based cytology (LBC) be applied to thyroid fine needle aspiration cytology samples?: Comparative analysis of conventional and LBC smears
Swati Mahajan, Arvind Rajwanshi, Radhika Srinivasan, Bishan Dass Radotra, Naresh Panda
October-December 2021, 38(4):198-202
Context: Liquid-based cytology (LBC) is a cyto-preparatory technique that may be applied to fine-needle aspiration (FNA) samples. However, its efficacy over conventional smears (CS) in thyroid is controversial. Aims: The aim of this study was to compare CS versus LBC (SurePath, BD) preparations in thyroid FNA samples for diagnostic efficacy. Settings and Design: Prospective case-control study using split sample analysis in 200 non-consecutive cases of thyroid FNA reported as per the Bethesda system. Methods and Material: Detailed cytomorphological features were evaluated in CS and LBC preparations. Cellularity was scored as 0–3. Based on diagnostic efficacy, they were categorized into three groups:(i) CS and LBC equivalent for diagnosis, (ii) CS better than LBC, and (iii) LBC better than CS for providing the diagnosis. Statistical Analysis Used: Paired t test for cellularity scores and descriptive for diagnostic efficacy. Results: There were 7 unsatisfactory, 118 benign, 10 atypia or follicular lesion of undetermined significance, 25 follicular neoplasms, 6 suspicious for malignancy, and 34 malignant thyroid FNA cases based on routine conventional smears. Cellularity of conventional smears was significantly higher than paired LBC smears (paired t test, P < 0.005). Comparison of overall diagnostic efficacy showed that LBC and CS were equivalent in 59% of cases; CS was superior to LBC in 37% cases and LBC was better than CS in 4% cases only. LBC smears showed higher unsatisfactory rate as compared to CS (18% vs. 3.5%). In category 2, the two techniques were equivalent in 87% of cases. In categories 4 and 6, CS were superior to LBC. Conclusions: In thyroid FNA cases, conventional smears are superior to LBC preparation in terms of cellularity and diagnostic efficacy and hence, LBC preparations should not replace conventional smears for making a routine cytomorphological diagnosis.
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Histopathological review of diagnostic categories of the milan system for reporting salivary gland cytopathology – An institutional experience of 6 years
Irem S Isgor, Selim Yigit Ercetin, Necati Enver, Leyla Cinel
October-December 2021, 38(4):203-209
Introduction: Salivary gland malignancies account for 2 to 4% of head and neck cancers. Fine needle aspiration cytology (FNAC) is used in preoperative diagnosis of salivary gland lesions. Although FNAC is a highly reliable technique for preoperative diagnosis, there were no consensus on salivary gland cytopathology reporting. Recently, an international group has recommended a classification system for salivary gland FNAC reporting titled “Milan System for Reporting Salivary Gland Cytopathology” (MSRSGC). In this study, we aimed to evaluate the usability of the Milan System, its ability to determine the risk of malignancy for each category, with comparisons of inital cytologic and final histopathological diagnosis. Materials and Methods: We performed a retrospective analysis of salivary gland lesion FNAC in our department from 2013 to 2019. A total of 578 FNACs were performed in 514 patients. Of these, 85 cases had surgical follow-up (parotid gland, n = 73, submandibular gland, n = 12). The cytological samples were categorized according to the MSRSGC into six categories by two pathologists. The risk of malignancy (ROM) and diagnostic accuracy values were calculated for each diagnostic categories. Results: A total of 85 aspirates of the patients with follow-up, the MSRSGC diagnostic categories were as follows: non-diagnostic in 7 aspirates (8.2%), non-neoplastic in 3 (3.5%), atypia of undetermined significance (AUS) in 9 (10.5%), benign neoplasm in 43 (50.5%), salivary gland neoplasm of undetermined malignant potential in 7 (8.2%), suspicious for malignancy in 10 (11.7%), and malignant in 6 (7%). The ROM for each category was 28, 5%, 0%, 33%, 0%, 28.5%, 90%, and 100%, respectively. Conclusion: FNAC plays a critical role in the evaluation of patients with salivary gland lesions. The MSRSGC helps in the standardization of the process of diagnosis and clinical management of salivary gland lesions, especially of AUS and SUMP categories that are indeterminate categories in nature.
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Factors affecting the histopathological outcomes of atypical glandular cells on pap test
Esra Keles, Ugur K Ozturk, Cihat M Alinca, Burak Giray, Canan Kabaca, Handan Cetiner
October-December 2021, 38(4):210-215
Background: Glandular cell abnormalities may indicate the presence of pre-malignant or malignant lesions. Aim: This study aimed to investigate the relationship between atypical glandular cells (AGC) and patients' demographics, histopathological outcomes, Human Papillomavirus (HPV) test results. Material and Methods: Between January 2015 and December 2019, women with AGC on Pap tests were retrieved from the hospital electronic database. The patients with AGC on cervicovaginal smears who underwent further pathological, laboratory, and imaging diagnostic testing and who were followed up at least 1-year were included in the study, while those who had a history of cervical dysplasia or cancer, lost during follow-up, or had missing data were excluded. Results: Of 85,692 Pap smears, 114 (0.13%) were diagnosed with AGC, of those 88 cases were eligible for final analysis. Gynecological malignancies were detected in 13 (14.8%) patients; including 6 (6.8%) endometrioid endometrial cancers, 3 (3.4%) non-endometrioid endometrial cancers, 2 (2.3%) cervical adenocarcinomas, 1 (1.1%) cervical squamous cell carcinoma, and 1 (1.1%) high-grade tubal serous cancer. Multivariate analysis revealed that presence of concomitant abnormal squamous lesion (P = 0.002), being 50 years and older (P = 0.028), HPV positivity (P < 0.001), and menopause (P = 0.023) were risk factors for significant pathology. Conclusion: The diagnosis of AGC may be related to the preneoplastic/neoplastic processes. A further comprehensive histopathological examination is required in women with AGC, aged 50 years and older, postmenopausal, HPV-positivity and concomitant squamous cell abnormality Clinicians should consider ovarian pathologies when there is no pathological finding on endometrial or cervical histopathological examination.
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The effect of the extent of neuroendocrine differentiation on cytopathological findings in primary neuroendocrine neoplasms of the breast
Burcu Guzelbey, Ezgi Hacihasanoglu, Canan Kelten Talu, Yasemin Cakir, Mehmet A Nazli
October-December 2021, 38(4):216-224
Objective: This study aimed to describe the cytological features of neuroendocrine breast tumors and to show the effect of the extent of neuroendocrine differentiation on cytological features. Methods: Breast tumor excision materials showing immunostaining with neuroendocrine markers (Synaptophysin or Chromogranin A) were determined and divided into two groups: cases with focal (10%–50% of tumor cells) staining and cases with diffuse (>50% of tumor cells) staining. A group of cases without neuroendocrine features/staining was used as control group. Fine needle aspiration biopsy specimens of the tumor mass or metastatic lymph nodes were examined and compared. Results: Twenty cases with neuroendocrine differentiation were included. Eleven cases were in the diffuse group, nine cases were in the focal group. Clean background, high cellularity, loosely cohesive cell groups with monotonous appearance, and naked nuclei were more common in the diffuse group. On the contrary, tight cohesive cell groups, the proportion of large cells, nuclear pleomorphism, and nucleolar prominence were higher in the group with focal staining. Plasmocytoid appearance, isolated cell groups, and binucleation were in similar distribution in both groups. Although round-oval nuclei were dominant in both groups, round nuclei were observed to be slightly more in the diffuse group. Only two cases in diffuse group showed cytoplasmic granularity and one case in focal group showed necrosis and mitosis. In the control group, tight cohesive groups, large cell size, pleomorphism, prominent nucleoli, and coarse chromatin were more commonly encountered. Conclusions: Clean background, hypercellularity, loss of cohesion, naked nuclei, monotonous cells with round nucleus, and granular cytoplasm were more prominent in cases showing diffuse staining with neuroendocrine markers. Suspecting neuroendocrine differentiation in tumors that show focal staining with neuroendocrine markers can be challenging in cytological preparations.
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Genotoxicity in oral mucosal epithelial cells of petrol station attendants: A micronucleus study
Shweta Rehani, Naresh Raj, Prabhakar Jeergal, Mohit Sharma, Kundendu Arya Bishen, Ruchi Nagpal
October-December 2021, 38(4):225-230
Introduction: Occupational exposure to petrol derivatives possesses an increased risk of various cancers including that of the oral mucosa. Scientific studies have shown the correlation of micronuclei assay (MN) with the cytogenotoxic changes in petrol station attendants. However, very few have reported the use of MN assay as a promising tool for assessing the impact of smoking in these workers. Aim: To explore the cytogenotoxic damage in exfoliated buccal cells obtained from petrol station attendants and control subjects using the MN assay along with additional effects due to smoking. Materials and Methods: The study comprised 60 males who were divided into Group I–IV with each having 15 subjects. These subjects were categorized as exposed smokers, exposed nonsmokers, unexposed smoker group, and unexposed nonsmokers. The MN and additional nuclear abnormalities (karyorrhexis [KH], binucleation [BN], pyknosis [P], and karyolysis [KL]) were calculated in PAP-stained slides. Results: Statistically higher mean frequencies of overall nuclear anomalies were observed in petrol pump workers in comparison with the control group. Petrol pump smokers carry the highest nuclear anomalies followed by non-exposed smokers than exposed non-smokers and the count was the least among unexposed non-smoker workers. Discussion and Conclusion: The present study indicated that the petrol pump workers are under higher cytogenotoxic damage. Also, smoking added to the frequency of damage. Thus, MN and other nuclear anomalies are in-vitro reliable biomarker assays available and should be routinely employed as a screening tool in their periodic medical evaluation.
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