Background: Current data regarding infertility suggests that male factor contributes up to 30% of the total cases of infertility. Semen analysis reveals the presence of spermatozoa as well as a number of non-sperm cells, presently being mentioned in routine semen report as "round cells" without further differentiating them into leucocytes or immature germ cells. Aim: The aim of this work was to study a simple, cost-effective, and convenient method for differentiating the round cells in semen into immature germ cells and leucocytes and correlating them with total sperm counts and motility. Materials and Methods: Semen samples from 120 males, who had come for investigation for infertility, were collected, semen parameters recorded, and stained smears studied for different round cells. Statistical analysis of the data was done to correlate total sperm counts and sperm motility with the occurrence of immature germ cells and leucocytes. The average shedding of immature germ cells in different groups with normal and low sperm counts was compared. The clinical significance of "round cells" in semen and their differentiation into leucocytes and immature germ cells are discussed. Conclusions: Round cells in semen can be differentiated into immature germ cells and leucocytes using simple staining methods. The differential counts mentioned in a semen report give valuable and clinically relevant information. In this study, we observed a negative correlation between total count and immature germ cells, as well as sperm motility and shedding of immature germ cells. The latter was statistically significant with a P value 0.000.

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Background: Endoscopic evaluation is critical in assessing the cause of obstructive jaundice. Cytological techniques including bile aspiration and biliary brushings have become the initial diagnostic modality. Aim: The aim of this study is to evaluate the role of endoscopic biliary tract cytology as a diagnostic tool in the evaluation of extrahepatic cholestatic jaundice. Materials and Methods: A total of 56 biliary tract specimens including 34 bile aspirations and 22 biliary brushings from 41 consecutive patients who had presented with obstructive jaundice and underwent endoscopic retrograde cholangiopancreatography (ERCP) were assessed by cytological examination. The smears prepared were analyzed for standard cytological features. Results: Cytologic diagnosis was adenocarcinoma in 13 (31.7%) cases, atypical in 2 (4.9%), reactive in 3 (7.3%) and benign changes in 19 (46.3%) cases. 4 (9.8%) cases were non-diagnostic. Serum bilirubin was significantly elevated in the malignant group. Biliary stricture was the most common finding on ERCP (68.3%). On cytological examination, presence of solitary, intact atypical cells, enlarged nuclei, irregular nuclear membrane, coarse chromatin and nucleoli were important cytologic criteria for differentiating malignant from benign biliary specimens. Conclusions: Regular use of bile cytology and brushings during ERCP evaluation of extrahepatic cholestatic jaundice is invaluable in obtaining a morphologic diagnosis. A systematic approach, use of strict cytomorphologic criteria and inclusion of significant atypia as malignant diagnosis may improve the sensitivity.

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Background: Spontaneously exfoliated benign-appearing endometrial cells (BEC) on a Papanicolaou smear might indicate endometrial pathology in postmenopausal women, necessitating further investigation. A cut-off age of 40 years was included in the Bethesda System 2001 based on studies of clinical significance of endometrial cells in Pap smears in Western countries. Aims: The purpose of this study was to determine the significance of age subgroup for women with a cytological diagnosis of BEC, regardless of menopausal status, in a retrospective cohort of Turkish women. Materials and Methods: Between October 2006 and November 2011, 41 patients with a BEC diagnosis and 64 patients with a cytological diagnosis of normal smear (NS) were enrolled; regardless of menopausal status, these women were 40 years and older and for whom follow-up endometrial biopsies had been performed. Results: On subsequent histopathologic evaluation, no malignant lesion was detected in women aged 40-50 years compared to three endometrioid-type adenocarcinomas in women older than 50 years with cytological diagnosis of BEC. There was a significant difference between women older than 50 years with cytologic diagnosis of BEC and NS in relation to premalignant lesions on histopathologic evaluation; however, this was not the case for women aged 40-50 years. Conclusions: According to our study, reporting BEC for women aged between 40 and 50 years has minor clinical significance but is significant for women older than 50 years, regardless of menopausal status. Larger sample size would be appropriate to confirm the results of the current study.

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Myoepithelial carcinoma (MC) is rare malignant salivary gland neoplasm and its cytologic features have been rarely described in the literature. Furthermore, MC shows varied cell types and patterns leading to the wide range of differential diagnosis on cytology. Histopathology and immunohistochemistry (IHC) are necessary to make a definite diagnosis. A 37-year-old female presented with painless, progressive swelling in the infra-auricular region since 2 years. Fine-needle aspiration cytology was performed and cytological possibilities of cellular pleomorphic adenoma and myoepithelial cell neoplasm were rendered and patient was advised excision and histopathologic examination for final diagnosis and subtyping. Final diagnosis of MC was made on hematoxylin and eosin sections and IHC. MC is rare malignant salivary gland tumor showing a clinic-pathologic diversity. The cytological features of MC are diverse and may lack overt feature of malignancy. Pathologists should be aware of this entity while evaluating cytological smears of salivary gland mass.

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Signet ring cell carcinoma (SRCC) of the breast is a rare tumor and it is classified by World Health Organization in 2003 classification under 'mucin producing carcinomas'. Pure form of SRCC breast is an extremely rare entity and very few cases have been reported in literature so far. We present a case of pure primary SRCC of the breast in a 70-year-old female, which was diagnosed on fine needle aspiration cytology. Cytological features generally show cellular smears with tumor cells showing eccentrically placed large, irregular nuclei showing indentations at places with cytoplasmic vacuoles. This case is being presented in view of its characteristic cytological features and its rarity.

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Background: Ayre spatula for cervical smear collection is being used despite the suggestion that different modified spatulas provide more satisfactory sampling. Aims: To see whether the cytological pickup improves with the use of long tipped spatula. Setting and Design: Rurally based University Hospital; crossover study. Materials and Methods: Pap smear using Ayre spatula in 500 and with plastic narrow long tip (Szalay) spatula in 500 clinic attending women was taken and analyzed. Crossover smears were taken with modified spatula in 163 and using Ayre spatula in 187 women after 2 weeks of initial smears. The same pathologist made cytological reporting for all smears and was unaware of the type of spatula used. Results: Smears from Ayre spatula had significantly higher reports of inadequate smears (94 of 500 vs. 68 of 500 for Ayre and Szalay, respectively; P = 0.032) and it remained so even after crossover (94 of 187 vs. 70 of 163 for Ayre and Szalay, respectively; P = 0.2). Cellular quality appeared better with smears taken using Szalay spatula, but the overall abnormal smear detection rate remained similar with either collection tool (χ² = 1.5; P = 0.2). Conclusions: Proportion of satisfactory smears is higher when long tip plastic spatula is used for collection of sample.

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Background: Oral habits such as alcohol consumption and tobacco chewing are considered to be initiators of dysplastic changes in the oral mucosa. Aim: The aim of this study was to determine and compare the alteration in apparently normal buccal mucosal cells due to effect of alcohol and tobacco by assessing silver-stained nucleolar organiser regions (AgNORs) and micronuclei. Materials and Methods: The study comprised a total of 100 subjects which were divided into four groups with 25 subjects having alcohol consuming habit, 25 subjects were tobacco consumer, 25 were both alcohol and tobacco consumer and 25 formed control group who neither consumed alcohol nor tobacco. Two cytological smears were taken from each subject with the help of cytological brush. The smear was then wet fixed and stained with AgNOR and acridine orange staining technique and assessed for nucleolar organiser region and micronuclei count respectively. 500 cells per slide were counted to note the changes. Results: Mann-Whitney test was applied to assess the variation in the number of AgNORs and micronuclei count between different groups. Cytological changes in each group revealed the increase in mean AgNORs and micronuclei count in subjects with combined alcohol and tobacco consumption when compared with individual groups. Conclusions: Tobacco and alcohol consumption produce alteration in apparently normal buccal mucosal cells, which may cumulatively lead to carcinomatous changes. Result of these changes may be used as educational tool in cessation of habits.

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Granular cell tumors (GCTs) are uncommon soft tissue tumors, which are difficult to diagnose merely on clinical examination. Being an effective first-line investigation, the fine-needle aspiration cytology (FNAC) plays a significant role in its pre-operative recognition. However, as the tumor is likely to mimic certain other lesions, a cytopathologist needs to be aware of its characteristic cytomorphology. We report two cases of GCT who presented with subcutaneous swellings in the left lower back and the right-sided anterior abdominal wall for 6 and 2 months, respectively. Both the patients had a clinical diagnosis of lipoma/neurofibroma. FNAC was done in both. In the first case a cytodiagnosis of xanthogranuloma was suggested and GCT in the second. Subsequent histologic examination of both showed features of GCT. FNAC would aid in presumptive diagnosis of GCT.

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Background: Thalassemia is a genetic disorder of hemoglobin synthesis, which requires regular blood transfusion therapy leading to iron overload in the body tissues. Transfusional hemosiderosis is a major cause of morbidity and mortality in these patients. Reliable methods for evaluation of iron overload are either invasive, costly or remotely available. Therefore, a simple technique of monitoring iron overload is desirable. Aim: To know whether iron can be demonstrated in exfoliated buccal cells of β-thalassemia major patients using Perls' Prussian blue method and to correlate it with serum ferritin levels. Materials and Methods: Smears were obtained from buccal mucosa of 60 randomly selected β-thalassemia major patients and 30 healthy subjects as controls. Smears were stained with Perls' Prussian blue method. Blood samples were taken for estimation of serum ferritin levels. Statistical Analysis: Chi-square, Mann-Whitney, and Spearman Rank's Correlation tests. Results: Perls' positivity was observed in 71.7% of thalassemic patients with a moderately positive correlation to serum ferritin levels. Conclusion: Oral exfoliative cytology can be a useful tool in demonstration of iron overload in thalassemic patients, however, further research in this field in the direction of quantification of these procedures is required, which can establish this non-invasive procedure as an ideal screening tool.

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Background: Solid pseudopapillary tumor of pancreas (SPTP) is a rare pancreatic tumor of uncertain histogenesis usually affecting young women. Though these tumors have characteristic cytomorphology, it is sometimes difficult to differentiate them from neuroendocrine tumors of the pancreas. We reviewed cases of SPTP to delineate the diagnostic cytological features and also observed utility of CD 99 (MIC 2) immunostaining to aid in the diagnosis of this tumor. Aims: This study was designed to demonstrate the utility of CD 99 immunostaining along with cytological features for making a pre-operative diagnosis and delineating it from the neuroendocrine tumor of pancreas which is a close mimic. Materials and Methods: Cytomorphological features of 11 cases of solid pseudopapillary neoplasm diagnosed by pre-operative fine-needle aspiration cytology (FNAC) at our institute were reviewed. Immunocytochemistry for CD 99 was also performed on the smears. Results: All the cases had cellular smears with monomorphic cells lying singly, as loosely cohesive clusters as well as forming delicate pseudopapillae. Presence of intra and extra-cellular basement membrane material, background foamy macrophages and nuclear grooves were the other salient features. Immunocytochemistry for CD 99 could be performed on eight cases and demonstrated typical paranuclear dot-like positivity. Conclusions: Pre-operative early diagnosis of SPTP can be made by FNAC which can further be aided by CD 99 immunocytochemistry.

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Castleman's disease (CD), a rare benign disease is characterized by lymphoid hyperplasia of uncertain etiology that may present as a nodal or extranodal mass. We describe herein a rare case of CD occurring in the soft-tissue of right arm in a 55-year-old woman. The present case is instructive in the differential diagnosis of primary soft-tissue tumors, for which the possibility of CD should be considered. On clinical examination as well as gross examination, this disease mimics lymphomas and tuberculosis. Since cytological appearances vary depending on the type and extent of hyperplasia, fine needle aspiration cytology findings may not always be conclusive in all cases. We studied aspirates which revealed reactive hyperplasia with increased vascularity. Excision confirmed the diagnosis.

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Extra-medullary leukemic infiltration of the breast by acute lymphoblastic leukemia (ALL) is very rare. We report two cases of ALL presenting as breast masses and diagnosed on fine-needle aspiration (FNA). Our first patient, a post-partum 30-year-old female, developed bilateral breast lumps in her last trimester of pregnancy and complained of easy fatigability. Our second patient, a 14-year-old girl, presented with a right-breast lump of 1-week duration. She had received treatment for ALL 1 year back and had been in complete remission for the last 1 year. FNA of the breast nodules done in both the cases revealed diffuse infiltration by lymphoblasts. Subsequent hematological investigations confirmed bone marrow involvement by ALL in the first case and extra-medullary relapse in the second case. Fine-needle aspiration cytology (FNAC) is an easy and cost effective method for the early diagnosis of metastatic leukemic infiltration, avoiding unnecessary excisional biopsies in such cases.

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Background: Genital human papillomavirus (HPV) infection is a sexually transmitted disease that is caused by HPV. Some types of HPV, called high-risk (HR) types may cause cell changes that sometimes lead to cervical cancer. HPV screening has been proposed for symptomatic female population; however, Pap test is the main stay in low resource setting. Aim: To detect HR HPV 16 positivity in perimenopausal and postmenopausal women and its association with cytological entities diagnosed on Pap smear. Materials and Methods: Pap smears and cervical scrapes were collected from 230 women consisting of 120 perimenopausal women approaching menopause and 110 postmenopausal women with a cervix after cessation of menstruation and processed as per routine procedure for detection of HR-HPV 16 deoxyribonucleic acid (DNA). Cytologically abnormal HPV 16 negative cases were also tested for other HR-HPV types. Results: Among the perimenopausal women 12 (10%) cases were positive for HR-HPV 16 consisting of 6 (5%) abnormal cases and 108 (90%) were HPV 16 negative consisting of 5 (4.1%) abnormal cases. However, among 110 postmenopausal women 14 (12.7%) were positive for HPV 16 DNA consisting of 6 (5.4%) abnormal cases and 96 (87.2%) were HPV 16 negative consisting of 4 (3.6%) abnormal cases. HPV 16 negative abnormal cases (9) were positive for low risk-HPV 6/11 consisting of atypical squamous cells (3) and low-grade squamous intraepithelial lesions-HPV (6). Conclusions: There is not much variation in HPV 16 positive cases in peri and postmenopausal women. By combining HPV DNA testing with Pap smear more cases having potential for pre-cancer lesions may be detected; however, HPV test cannot replace the Pap smear in low resource setting.

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Extramedullary hematopoiesis (EMH) evidenced by erythropoietic cells and megakaryocytes is a characteristic feature of hepatoblastoma (HB). The typical cytomorphology, the presence of EMH and associated clinical and radiological findings offer a reliable diagnosis of hepatoblastoma by fine-needle aspiration cytology (FNAC). We describe the cytologic features of hepatoblastomas and discuss the differential diagnosis in two children, aged 53 days and 19 years. The usefulness of EMH in differentiating HB from other small round cell tumors and well differentiated hepatocellular carcinoma (HCC) on cytology is highlighted.

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Background: Neuropathology centers are expected to offer a prompt and accurate intraoperative diagnosis regarding tumor/lesion type and grade on fresh unfixed tissue. Level of diagnostic accuracy according to type and grade and also, the experience at a new center has not been reported before. Aims: The aim of this study is to review the agreement patterns according to tumor/lesion type and grade between intraoperative and final histopathologic diagnosis in central nervous system (CNS) lesion samples received by a newly established neuropathology center at a tertiary care neuropsychiatric hospital. Materials and Methods: Agreement between intraoperative and final histopathologic diagnosis was classified as: (I) Grade in agreement but type not in agreement; (II) grade not in agreement but type in agreement; (III) grade and type both not in agreement; (IV) grade and type both in agreement. Statistical Analysis: Confidence interval (CI) of agreements was calculated for various categories of neoplastic as well as non-neoplastic lesions. CI was also calculated for groups where n × p and n × (1 − p) were more than 5, i.e., fulfilled the requirement of the central limit theorem. Results: On retrospective analysis of 333 cases, 284 (85.3%) cases were categorized as neoplastic while 49 (14.7%) cases were categorized as non-neoplastic. Among the neoplastic lesions agreement was seen in 237 (83.5%) cases while 47 (16.5%) cases showed disagreement. Similarly in non-neoplastic category; 46 (93.9%) cases showed agreement while 3 (6.15%) cases showed disagreement. Of the non-neoplastic lesions, one case fell into the agreement category I, 2 in category III and 46 in IV. Among neoplastic lesions, there were 21 cases in agreement category I, 17 in II, 9 in III and 237 in IV. On analyzing the accuracy of intraoperative reporting according to tumor type, the break up was: - Astrocytic: 2 (I), 16 (II), 2 (III), 86 (IV); oligodendroglial: 8 (I), 1 (II); ependymal: 2 (III), 6 (IV); embryonal: 23 (IV); cranial and spinal nerve tumors: 2 (II), 21 (IV); choroid plexus tumors: 4 (IV); meningeal tumors: 3 (I), 1 (III), 49 (IV); metastatic tumors: 3 (I), 17 (IV); cysts (tumor-like conditions): 14 (IV); neuronal and mixed neuronal glial tumors: 1 (III); malignant lymphoma: 1 (III); sellar tumors: 17 (IV); and mixed gliomas: 5 (I). Conclusions: This study identifies problem areas of CNS intraoperative reporting, in a new center, with reference to tumor typing and grading. It may forewarn upcoming centers of neuropathology about the potential problem areas of intraoperative reporting.

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Phaeohypomycosis is a clinical syndrome caused by melanised or dematiaceous fungi characterized by the presence of brown mycelial structures in tissue section. These fungi are associated with a repertoire of the clinical manifestations that includes superficial and deep local infection to disseminated infection. Herein, we describe the clinical and fine-needle aspiration cytology and histopathologic features of a case of subcutaneous phaeohypomycosis.

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