Cytology

Cytomorphological spectrum of Hashimoto's thyroiditis and its correlation with hormonal profile and hematological parameters Archana Shetty, Vijaya Chowdappa Journal of Cytology 2019 36(3):137-141 Introduction: Hashimoto's thyroiditis (HT) is a well-known autoimmune disorder of the thyroid diagnosed on fine needle aspiration cytology (FNAC) and a common cause of hypothyroidism in women. Often serological and hematological parameters are additional investigations aiding the diagnosis of this entity. Aim: To grade HT based on cytomorphology and to correlate the cytological grades with thyroid hormone status and basic hematological parameters. Materials and Methods: During a period of 2.5 years, 1762 patients underwent FNAC of thyroid at our tertiary healthcare center. Cytological evidence of lymphocytic thyroiditis was seen in 102 cases, of which 58 cases in addition had thyroid hormone levels and hematological parameters for correlation. Results: Of the 58 cases, 55 were females. Majority of the patients had grade II thyroiditis (56.9%), followed by grade I (34.5%) and grade III (8.6%). Elevated thyroid-stimulating hormone was seen in 74.2% of cases, with 39.7% of patients presenting with subclinical hypothyroidism and 18.9% being euthyroid. Mean hemoglobin was low in all grades, more so in hypothyroid state, while other hematological parameters were normal when correlated with grade and hormonal status without any significant P value. Conclusion: Cytomorphological grading of HT can explain the pathogenesis of this autoimmune disease. Subclinical hypothyroidism was significantly observed. There was no significant statistical correlation of cytological grades with thyroid status. In this study, most of the hypothyroid cases had low hemoglobin levels while other basic hematological parameters did not show any statistically significant correlation with the thyroid hormonal status.

Cytological correlates of axillary nodal involvement in invasive ductal carcinoma of breast Anamika Sinha, Satyajit Singh Gill Journal of Cytology 2019 36(3):142-145 Context: Fine needle aspiration cytology (FNAC) plays an important role in the diagnosis of breast carcinoma. However, its role as a prognostic tool needs to be explored. This can be achieved by studying its correlation with an established prognostic marker such as axillary nodal metastasis. Aims: This study was undertaken to correlate the cytological features of invasive ductal carcinoma (IDC) of breast with axillary lymph node status. Settings and Design: Tertiary care hospital, retrospective analytical study. Materials and Methods: The study group included 150 cases of IDC of breast diagnosed on FNAC, who had subsequently undergone modified radical mastectomy. Cytologic grades were assigned as per Robinson's grading system. Histopathological sections of axillary lymph nodes were assessed for metastasis. Statistical Analysis Used: The cytologic grade and each feature of the cytologic grade were correlated with the lymph node metastasis using χ2 test. Values of P < 0.05 were considered significant. Result: A statistically significant correlation was noted between cytologic grade of tumor and axillary lymph node metastasis (P < 0.05). In addition, a positive correlation was found between two of the individual features of cytologic grade, namely, nuclear size and cell uniformity with axillary lymph node metastasis (P < 0.05). Conclusion: Robinson's cytologic grade of breast carcinoma correlates well with the presence of axillary lymph node metastasis and hence can be used as a prognostic tool. As there is an increasing trend toward conservative approach to management of breast carcinoma, patients receive preoperative neoadjuvant therapy which may alter the nodal status on the resected specimen. Hence, a high cytological grade of primary tumor, as assessed on FNAC before initiation of therapy, should alert the treating team of the possibility of axillary lymph nodal metastasis.

Performance of a convolutional neural network in screening liquid based cervical cytology smears Parikshit Sanyal, Sanghita Barui, Prabal Deb, Harish Chander Sharma Journal of Cytology 2019 36(3):146-151 Context: Cervical cancer is the second most common cancer in women. The liquid based cervical cytology (LBCC) is a useful tool of choice for screening cervical cancer. Aims: To train a convolutional neural network (CNN) to identify abnormal foci from LBCC smears. Settings and Design: We have chosen retrospective study design from archived smears of patients undergoing screening from cervical cancer by LBCC smears. Materials and Methods: 2816 images, each of 256 × 256 pixels, were prepared from microphotographs of these LBCC smears, which included 816 “abnormal” foci (low grade or high grade squamous intraepithelial lesion) and 2000 'normal' foci (benign epithelial cells and reactive changes). The images were split into three sets, Training, Testing, and Evaluation. A convolutional neural network (CNN) was developed with the python programming language. The CNN was trained with the Training dataset; performance was assayed concurrently with the Testing dataset. Two CNN models were developed, after 20 and 10 epochs of training, respectively. The models were then run on the Evaluation dataset. Statistical Analysis Used: A contingency table was prepared from the original image labels and the labels predicted by the CNN. Results: Combined assessment of both models yielded a sensitivity of 95.63% in detecting abnormal foci, with 79.85% specificity. The negative predictive value was high (99.19%), suggesting potential utility in screening. False positives due to overlapping cells, neutrophils, and debris was the principal difficulty met during evaluation. Conclusions: The CNN shows promise as a screening tool; however, for its use in confirmatory diagnosis, further training with a more diverse dataset will be required.

Evaluation of p16/Ki-67 dual staining compared with HPV genotyping in anal cytology with diagnosis of ASC-US for detection of high-grade anal intraepithelial lesions Maxime Pichon, Marie Joly, Frédérique Lebreton, Medhi Benchaïb, Yahia Mekki, Mojgan Devouassoux-Shisheboran Journal of Cytology 2019 36(3):152-156 Introduction: Human Papillomavirus (HPV) infection is the main risk factor for anogenital cancer. The objective of this study was to compare p16/Ki-67 dual staining to HPV genotyping in anal cytology samples with an atypical squamous cell of undetermined significance (ASC-US) for the identification of high-grade squamous intraepithelial lesion (HSIL). Methods: Anal cytology samples with an ASC-US result (n = 111) were collected from patients of a university hospital (Lyon, France) from 2014 to 2015. Cases with remaining squamous cells (n = 82) were stained using p16/Ki-67 dual staining (CINtec-Plus kit) and analyzed for HPV screening (CLART2-PCR kit) using a composite endpoint of biopsy and cytology results on follow-up specimens. Results: Detection of HSIL on follow-up specimens (5/22 biopsies; 1/29 cytology samples) was obtained in two out of six cases with p16/Ki-17 versus. five out of six with HPV genotyping alone. Sensitivity and specificity to detect HSIL for p16/Ki-67 was 33% (95% confidence interval [CI] [4; 77]) and 49% (95%CI [34; 64]) versus. 83% (95%CI [36; 99.6]) and 13% (95%CI [5; 27]) for HPV genotyping. Conclusion: Herein, HPV genotyping was more sensitive but less specific than p16/Ki-67 staining for the detection of subsequent HSIL in ASC-US anal cytology. A larger study is required to evaluate the combination of these biomarkers for triage.

Evaluation of imprint smears of bronchoscopic biopsy in lung tumors: A cytohistological correlation Abhishek Chowdhury Journal of Cytology 2019 36(3):157-159 Background: There are several methods for obtaining samples in patients of lung tumors, of which bronchoscopic biopsy is the most common. In most of Indian scenario, however, histopathology diagnosis is time taking. Aims and Objectives: To evaluate imprint cytology as a tool for rapid diagnosis of lung carcinoma and its histopathological correlation. Study Type: Prospective study on accuracy of a diagnostic test. Materials and Methods: A total of 175 cases were included in the study, and all of them were subjected to brochoscopic biopsy. Imprint smears were prepared from all the bronchoscopy specimens obtained from 175 cases. Imprint smears were stained with Leishman-Giemsa cocktail and Pap stain, and histopathology sections were stained with hematoxlin and eosin. Histopathological findings were confirmed by immunohistochemistry. Results: Sensitivity and specificity of imprint cytology was 84.9% and 72.4%, respectively when compared to histopathology as standard. Conclusion: Imprint cytology can be used as a preliminary tool for diagnosis in lung tumor bronchoscopic biopsies.

Milan system for reporting salivary gland cytopathology: An experience with the implication for risk of malignancy Chayanika Kala, Sanjay Kala, Lubna Khan Journal of Cytology 2019 36(3):160-164 Introduction: Fine-needle aspiration cytology (FNAC) is a well-established technique for evaluation of salivary gland lesions, but because of the heterogenicity and morphological overlap between spectrum of lesion, there are a few challenges in its wide use. Recently, “The Milan system for reporting salivary gland cytopathology” (MSRSGC) was introduced, providing guide for diagnosis and management according to the risk of malignancy (ROM) in different categories. The current study was conducted retrospectively to reclassify the salivary gland lesions from previous diagnosis and to evaluate the ROM in different categories. Material and Methods: Clinical data, FNAC specimen, histological, and clinical follow-up of cases were retrieved, cytological features were re-evaluated, and cases were reclassified as follows: Category 1: Non-diagnostic (ND); Category 2: Non-neoplastic (NN); Category 3: Atypia of undetermined significance (AUS); Category 4a: Neoplasm: benign (NB), Category 4b: Neoplasm: salivary gland neoplasm of uncertain malignant potential (SUMP); Category 5: suspicious of malignancy (SM); and Category 6: Malignant (M). Result: Total 293 cases were evaluated cytologically, and histological follow-up was available in 172 cases. The distribution of cases into different categories was as follows ND (6.1%), NN (38.2%), AUS (2.7%), NB (33.4%), SUMP (2.0%), SM (2.4%), and M (15%). Overall, ROM reported were 25%, 5%, 20%, 4.4%, 33.3%, 85.7%, and 97.5%, respectively for each category. Overall, sensitivity was 83.33%, specificity was 98.31%, positive predictive value was 95.74%, and negative predictive value was 92.80%. Conclusion: MSRSGC is a recently proposed six category scheme, which places salivary gland FNAC into well-defined categories that limit the possibilities of false negative and false positive cases.

von Hansemann cells from fresh urine sediment samples in the diagnosis of malakoplakia Fernando Guerra, Adriana E Rocher, Anabela Angeleri, María Juarez, Gisela Coliva, Luis A Palaoro Journal of Cytology 2019 36(3):165-168 Background: Malakoplakia is characterized by the presence of plaques with macrophages containing inclusion bodies. The diagnosis of this disease is carried out by biopsy of the lesion. The objective of this paper was to assess the value of fresh urine sediment in the diagnosis of malakoplakia. Materials and Methods: Five suspected cases of malakoplakia that showed macrophages with inclusions called bodies of Michaelis-Gutmann (von Hansemann cells) in unstained urine sediment were processed with Papanicolaou, Giemsa, and periodic acid-Schiff (PAS) stains. Four of the five patients had a history of cystitis and had developed antibiotic resistance. The other patient had the characteristics cells in a routine urinalysis. Results: Papanicolaou stain revealed intracytoplasmic eosinophilic or basophilic bodies, single or multiple in macrophages. Such bodies were stained deep red with PAS technique. Giemsa stain showed these bodies with a faint basophilic coloration, sometimes with a central core. Bladder biopsies established the definitive diagnosis, showing bodies within and outside macrophages, with a concentric “birds-eye” or “owl-eye” (targetoid) appearance. Conclusions: Finding of von Hansemann cells in fresh urine sediment of patients with cystitis and a history of resistance to antibiotic scan leads to the diagnosis of malakoplakia. Giemsa stain can show in some cases the characteristic central core of Michaelis-Gutmann bodies. Malakoplakia is probably the result of an acquired defect in macrophage function causing impairment of bactericidal activity. A correct diagnosis is important because the spread to ureters with bilateral stenosis and obstruction can lead to kidney failure.

A quest for accuracy: Evaluation of the Paris system in diagnosis of urothelial carcinomas Sharada Rai, Bhagat S Lali, Chaithra G Venkataramana, Cheryl S Philipose, Ranjitha Rao, G G Laxman Prabhu Journal of Cytology 2019 36(3):169-173 Introduction: Urine cytology is an important screening tool of patients for urothelial carcinoma (UC) and follow-up of patients with treated disease. Ease of procurement, cost-effectiveness, and lower turnaround time are the major advantages. Objective: To compare current system of reporting (CSR) at our institute with The Paris System (TPS) and analyze utility of urine cytology based on TPS reporting in correlation with urine culture and histopathology. Materials and Methods: One-year retrospective study of 90 cases was undertaken wherein cases presenting with painless hematuria and clinically suspicious of UC were included. Urine cytology slides were reviewed and reported with TPS guidelines. These findings were correlated with histopathological diagnosis and urine culture as indicated. Statistical analysis was done using SPSS 17 software. Results: With TPS guidelines, 11.1% and 5.6% cases were reported as high-grade UC (HGUC) and low-grade urothelial neoplasm (LGUN), respectively. Suspicious for HGUC category included 17.8% of cases. The rate of reporting “atypical urothelial cells (AUC)” was significantly lower (11.1%) with TPS on comparison with CSR (16.7%). Histopathological correlation of positive predictive value for HGUC was better (100%) on using TPS when compared with CSR (64.3%). Among 11 cases with microbial growth on urine culture, 9.1% were reported as atypical. Sensitivity and accuracy of TPS in detecting UC were 83.3% and 86.52%, respectively. Both were higher when compared with CSR. Conclusion: In comparison to CSR, criteria of TPS limit the AUC category and enhance the sensitivity and accuracy of detecting HGUC. Adopting TPS for urinary cytology will ensure uniformity and accuracy of HGUC diagnosis.

Intraoperative Scrape Cytology from Ovarian Masss Lesions: A Study of 81 Cases Amita Jain Gupta, Meeta Singh, Jenna B Bhattacharya, S Anusha, Shyama Jain, Nita Khurana Journal of Cytology 2019 36(3):174-179 Background: Intraoperative evaluation of an ovarian mass is of crucial importance in its further management, accomplished by frozen section (FS) and scrape smear (SS) examination. Aim: To evaluate utility of SS over FS and to study scrape cytological features of a variety of ovarian neoplasms. Materials and Methods: The study comprised ovarian tumors studied over a period of 1 year (2014–2015) that were submitted for intraoperative assessment. SS and FS were examined and evaluated independently. The results were compared with final pathological diagnosis in each case, and cases with discordant diagnoses were reviewed. All the SSs of ovarian tumors were re-evaluated with Giemsa-stained smears, and cytological features were described. Results: The results of SS and FS were 100% concordant. On histopathology, of 81 cases, 43 were epithelial [(categorized further as serous, mucinous, or malignant mixed Mullerian tumor (MMMT)] along with subcategorization of benign, borderline, and malignant), 16 were germ cell (categorized as teratoma: mature/immature and yolk sac tumor), 11 were sex cord stromal tumors (fibroma, granulosa cell tumor, Sertoli–Leydig cell tumor), 8 cases were hemorrhagic cysts (torsion, endometroid, corpus luteal cyst, etc.), and 3 were metastasis. There were 61 benign, 2 borderline, and 18 malignant cases on FS and scrape. Combining all the values, sensitivity and specificity were 76% and 98.21%, respectively. The diagnostic accuracy in diagnosing malignant lesions was 91%. Conclusion: Adequate knowledge on cytohistological correlation of ovarian scrape cytology may phase out the use of cryostat in intraoperative diagnosis of ovarian neoplasms, and thus be a boon for resource-deprived settings.

High-risk Human papillomavirus testing in young Japanese women with atypical squamous cells of undetermined significance Takashi Mitamura, Yosuke Konno, Satomi Kikawa, Yutaka Iwaki, Kurumi Iwaki, Fumie Tanuma, Soromon Kataoka Journal of Cytology 2019 36(3):180-183 Introduction: The mortality due to uterine cervical cancer has been gradually increasing in women under 40 years of age (U40) in Japan. We investigated the effect of high-risk human papillomavirus (HR-HPV) on U40 subjects without any overt cytological abnormalities. Materials and Methods: We retrospectively examined the clinical data, including the findings of a cobas 4800 HPV test that was approved in Japan in 2013 to triage women with atypical squamous cells of undetermined significance (ASC-US) and a histological examination in 589 Japanese women. Results: The overall prevalence rate of HR-HPV was 34.5%. Biopsy-confirmed cervical intraepithelial neoplasia (CIN) 2, or worse (CIN2+) was identified in 45.1% (23/51) of HR-HPV-positive women with ASC-US, who underwent colposcopy immediately. The mean period from the HPV test to the diagnosis of CIN2+ was 3.7 months. CIN2+ was more common (69.6%) in U40 patients. The rates of single or multiple infections of HPV-16, HPV-18, and 12 other HR-HPV (31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68) in CIN2+ U40 patients were 31.3%, 0%, and 81.3%, respectively. The relative risk for CIN 2+ among U40 women with HPV-16 was not significantly different from that of the patients with infection of any of the 12 other HR-HPVs. Conclusion: The results of this study suggest that the 12 other HR-HPVs have a potential to generate high-grade cervical lesions among young women, and the examination rate of colposcopy should be increased.