Accuracy of concurrent visual and cytology screening in detecting cervical cancer precursors in rural India.

The high burden of cervical cancer and inadequate/suboptimal cytology screening in developing countries led to the evaluation of visual screening tests, like visual inspection with acetic acid (VIA) and Lugol's iodine (VILI). We describe the performance of VIA, VILI and cytology, carried out in a multinational project called "Screening Technologies to Advance Rapid Testing" in 5,519 women aged 30-49 years, in detecting cervical intraepithelial neoplasia (CIN). VIA, VILI and cytology were positive in 16.9%, 15.6% and 6.1% women, respectively. We found 57 cases of CIN2, 55 of CIN3 and 12 of cervical cancer; 90% of CIN3 and 43% CIN2 cases were positive for p16 overexpression and high-risk HPV infection, indicating a high validity of histological diagnosis. The sensitivity of VIA, VILI and cytology to detect high-grade CIN were 64.5%, 64.5% and 67.7%, respectively; specificities were 84.2%, 85.5% and 95.4%. A high proportion of p16 positive CIN 3 (93.8%) and 2 (76.9%) were positive on cytology compared with visual tests (68.8% and 53.8%, respectively) indicating a higher sensitivity of cytology to detect p16 positive high-grade CIN. However, the immediate availability of the results from the visual tests permits diagnosis and/or treatment to be performed in the same sitting, which can potentially reduce loss to follow-up when women must be recalled following positive cytology. Organizing visual screening services in low-resource countries may facilitate the gradual building of an infrastructure committed to screening allowing the eventual introduction of more sensitive, highly objective, reproducible and affordable human papillomavirus screening tests in future.

Case report of renal cell carcinoma diagnosed in voided urine confirmed by CD10 immunocytochemistry.

BACKGROUND: Preoperative diagnosis of renal cell carcinoma (RCC) by exfoliative urine cytology is difficult, as infiltration of RCC into the pelvicalyceal system is uncommon. The exfoliation of RCC cells in urine is a rare phenomenon and when it does occur, it is likely to be missed. Cytologic examination of the urine coupled with ancillary immunocytochemistry can clinch the diagnosis leading to appropriate clinical management. CASE: A 50-year-old man presented with complaints of hematuria and abdominal pain of 6 months' duration. Ultrasonographic examination of the abdomen and pelvis showed a well-defined mass lesion in the upper pole of the left kidney, suggestive of neoplastic etiology. In the given clinical context of renal mass, urine cytology was suggestive of RCC and biopsy confirmation was suggested. One cytology smear subjected to immunocytochemistry with anti-CD10 antibody which showed strong diffuse cytoplasmic positivity in these cells confirmed the diagnosis of RCC. Subsequently, fine needle aspiration cytology of the kidney mass was reported as RCC. CONCLUSIONS: RCC has distinct cytologic features that facilitate a diagnosis in urine in an appropriate clinical and radiological context. Their recognition in the urine smear is important to avoid costly and invasive modalities like image-guided needle biopsy.

Diagnosis of hidradenoma papilliferum of the vulva on cytologic smears: a case report.

BACKGROUND: Hidradenoma papilliferum is a rare benign tumor of the vulva that occurs mainly in women in the skin of the anogenital region. Clinically it may mimic a carcinoma, so the correct cytologic interpretation of the tumor cells with subsequent careful histologic confirmation is essential for good management. CASE: A 44-year-old woman who had a history of hysterectomy 14 years earlier presented with a small, exophytic growth on the labia majora. Fine needle aspiration cytology and vulvar scraping of the lesion were reported as a malignant tumor, and the case was referred to our center for further management. Repeat vulvar smears showed mainly groups and clusters of benign-looking glandular cells and were interpreted as a benign tumor. Subsequent histology confirmed the diagnosis of hidradenoma papilliferum. CONCLUSION: This case highlights the cytologic features of hidradenoma papilliferum on cytologic smears. The clinical presentation should not mislead the cytologic interpretation of the tumor cells.

Evaluation of atypical squamous cells on conventional cytology smears: An experience from a screening program practiced in limited resource settings.

BACKGROUND: The Bethesda system (TBS) 2001 has subdivided the category of atypical squamous cells (ASC) into: ASC-US (undetermined significance) and ASC-H (cannot exclude high-grade squamous intraepithelial lesion (HSIL)). The present study is an analysis of ASC-US and ASC-H cases diagnosed in a screening program practiced in limited resource settings. METHODS: During the period January 2005 to December 2008, a total of 9190 smears were received, of which 568 were unsatisfactory. Cases initially diagnosed as ASC-US (n=74) and ASC-H (n=29) on conventional cytology smears were reviewed. Biopsy and human papilloma virus (HPV) results were available in limited cases. RESULTS: On review, diagnosis of ASC-US was retained in 49 (66.2%) of the 74 initially diagnosed ASC-US cases. Remaining 12 cases were re-labeled as negative for intraepithelial lesion or malignancy (NILM), nine as low-grade squamous intraepithelial lesion (LSIL), three as ASC-H and one case as squamous carcinoma (SCC). Similarly, on review, diagnosis of ASC-H cases was retained in 17 of the 29 initially diagnosed ASC-H cases. Seven cases were re-labeled as NILM, three as HSIL and one case each as ASC-US and SCC. Overall, 8622 cases (96.6%) were diagnosed as NILM, 72 (0.83%) as LSIL, 121 (1.40%) as HSIL, 23 (0.26%) as SCC, 50 (0.57%) as ASC-US cases, 20 (0.23%) as ASC-H, five (0.05%) as atypical glandular cells (AGC) and two cases as adenocarcinomas. Out of 50 ASC-US cases, biopsy in 23 cases showed presence of CIN 1 in 16 cases (69.5%) and CIN 2 in one case (4.34%), while the remaining six cases were negative for CIN/malignancy. The remaining 20 cases with unavailable biopsy results were HPV-positive. Out of 20 ASC-H cases, biopsy in 15 revealed CIN 2 and above in 11 cases (73.3%). Three cases (20%) revealed CIN 1. CONCLUSIONS: Critical review is helpful in further reducing the number of ASC cases. The percentage of cases with CIN 2 and above is higher with ASC-H cases. The reason for relative increase in HSILs in the present study included referral bias in the screening program.

Has urine cytology a role to play in the era of fluorescence in situ hybridization?

OBJECTIVE: To determine the efficacy of urine cytology as a diagnostic tool and in follow-up of patients with urinary bladder cancer. STUDY DESIGN: From 951 cases, 1,831 urine specimens were collected in a 5-year period (2000-2004). Six hundred fifty-two cases were suspected to have primary bladder cancer and formed the basis of the study. The final diagnosis was based on histology. When histology was not available, clinical or radiologic findings formed the basis for the final diagnosis. RESULTS: We had 173 false negative and 6 false positive cases in our series, giving an overall sensitivity of 82% and specificity of 96%. The main reason for false negativity was screening error, noticed in low grade urothelial carcinoma. False positivity was due to inflammatory atypia and polyomaviral cytopathic changes. CONCLUSION: The sensitivity and specificity in our study compared well with those in the world literature. Urine cytology can be successfully used as a primary diagnostic tool and also as a follow-up modality as the cystoscopically occult malignancy can be detected easily.

"Signet-ring" cells--a caveat in the diagnosis of a diffuse peritoneal mesothelioma occurring in a lady presenting with recurrent ascites: an unusual case report.

A diffuse peritoneal mesothelioma is a rare tumor. Exfoliative cytology forms the first step in the diagnosis of mesothelioma, since most of these cases presented with effusion. Despite well established cytomorphological features, a challenge exists in differentiating mesothelial cells, including reactive and malignant types from carcinoma cells and macrophages. Presence of "signet-ring" cells increases the diagnostic challenge as these can be forms of benign and malignant cells. Ancillary techniques like immunohistochemical (IHC) markers and ultrastructural analysis form useful adjunct in substantiating exact diagnosis. We report an unusual case study of a diffuse peritoneal mesothelioma in a 57-years-old lady, with no history of asbestos exposure, presenting with recurrent ascites, diagnosed on ascitic fluid cytology and on histology as an adenocarcinoma, based upon the presence of "signet-ring" cells. On review, clinicopathological correlation with IHC was helpful in forming correct diagnosis.

Pulmonary non-Hodgkin's lymphoma (NHL) of diffuse large B-cell type with simultaneous humeral involvement in a young lady: an uncommon presentation with cytologic implications.

A bronchogenic carcinoma, almost invariably, presents as a lung mass. Primary pulmonary lymphomas are rare. We report an unusual case of a pulmonary non-Hodgkin's lymphoma (NHL) with simultaneous involvement of the right humerus in a 37 year old lady. Bronchial lavage smears showed atypical cells with irregular nuclear membranes raising a suspicion of a hematolymphoid tumor, over a small cell carcinoma that was the closest differential diagnosis. Biopsy from the lung mass and from the lesion in the humerus showed an identical malignant round cell tumor with prominent apoptosis. On immunohistochemistry (IHC), tumor cells were diffusely positive for leukocyte common antigen (LCA), CD20 and MIB1 (70%), while negative for cytokeratin (CK), epithelial membrane antigen (EMA) synaptophysin, chromogranin, neuron specific enolase (NSE), CD3, and CD10. Diagnosis of a pulmonary NHL of diffuse large B-cell type with involvement of the humerus was formed. The case is presented to create an index of suspicion for the possibility of a NHL on respiratory samples, while dealing with small round cells with irregular nuclear membranes. IHC is necessary to confirm he diagnosis. A simultaneous association in the humerus in our case makes it unusual.

Cytodiagnostic problems in cervicovaginal smears from symptomatic breast cancer patients on tamoxifen therapy.

OBJECTIVE: To evaluate the effect of tamoxifen on cervicovaginal epithelium, identify tamoxifen-related changes that mimic cancer and detennine the morphologic features differentiating the 2 changes. STUDY DESIGN: Cervicovaginal smears from 153 conventionally treated primary breast cancer patients presenting with gynecologic symptoms were studied. RESULTS: All 153 patients presented with menorrhagia or irregular periods. Of 4 patients with a cytodiagnosis of atypical glandular changes, 2 had negative histology; 1 each had a uterine leiomyoma and endometrial hyperplasia. Of the 6 cases reported as adenocarcinoma, 3 were histologically confirmed, and the others were false positives. Conversely, 1 false negative case histologically was an endometrioid carcinoma. CONCLUSION: Our study revealed that reactive glandular cells are a cause of false positive diagnoses. Tamoxifen-associated cellular changes can mimic morphologic features of cancer. To avoid diagnostic errors, cervicovaginal smears should be repeated after discontinuing tamoxifen treatment. Clinical correlation is mandatory. Regular follow-up with cervicovaginal smears from patients on tamoxifen treatment is recommended.

Utility of the thromboplastin-plasma cell-block technique for fine-needle aspiration and serous effusions.

(I) To assess the feasibility of thromboplastin-plasma (TP) method for cell block, (II) to concentrate the minimal cellular material from effusions and needle-rinses by block preparation and improve visual details, (III) to compare conventional cytological smears with cell blocks for final assessment, and (IV) to assess utility of immunocytochemistry (ICC) for diagnostic accuracy. Seventy cell blocks were prepared by TP technique using surplus fluid from 38 serous effusions, and for 32 ultrasonography-guided fine-needle aspiration cytology (FNAC) material, rinses of syringes and needles were collected in normal saline after conventional cytological smears. Then, cell blocks were compared with conventional smears for adequacy, morphologic preservation, and ICC. Absolute concordance seen in 66 cases (94%) between the smears and cell blocks. Advantages with the blocks were cellular concentration in a limited field and better cellular preservation with architectural pattern. Quality of ICC was comparable to that of standard controls. Diagnostic discrepancy was seen in two cases where cell blocks were positive but smears were negative. Two cell blocks were nonrepresentative. Cell block serves as a useful adjunct to traditional cytological smears. TP method is simple, cost effective, and reproducible. It is easy when compared with agar-embedding technique. Ancillary techniques like ICC can be performed successfully.

Papanicolaou stain: Is it economical to switch to rapid, economical, acetic acid, papanicolaou stain?

OBJECTIVE: To standardize an inexpensive and rapid Papanicolaou staining technique with limited ethanol usage. STUDY DESIGN: Smears from 200 patients were collected (2 per patient) and fixed in methanol. Half were subjected to conventional Papanicolaou and half to stain ing with rapid, economical, acetic acid Papanicolaou (REAP) stain. In REAP, pre-OG6 and post-OG6 and post-EA36 ethanol baths were replaced by 1% acetic acid and Scott's tap water with tap water. Hematoxylin was preheated to 60 degrees C. Final dehydration was with methanol. REAP smears were compared with Papanicolaou smears for optimal cytoplasmic and nuclear staining, stain preservation, cost and turnaround time. RESULTS: With the REAP method, cytoplasmic and nuclear staining was optimal in 181 and 192 cases, respectively. The staining time was considerably reduced, to 3 minutes, and the cost per smear was reduced to one fourth. The staining quality remained good in all the smears for > 2 years. CONCLUSION: REAP is a rapid, cost-effective alternative to Papanicolaou stain. Though low stain penetration in large cell clusters is a limitation, final interpretation was not compromised.

Cytology of lleal conduit urine in bladder cancer patients: diagnostic utility and pitfalls.

OBJECTIVE: To study the cytomorphology of urine obtained from the ileal conduit and to determine its utility and identify the pitfalls. STUDY DESIGN: Urine specimens from 469 cases of suspected or proven bladder cancer received over a period of 5 years were analyzed in the cytology laboratory. In 35 cases, total bladder resection was followed by ileal conduit reconstruction. The follow-up cytologic analysis of these 35 ileal conduit cases formed the basis of this study. RESULTS: There was absence of urothelial cells in all but 2 cases. The smear predominantly showed small, scattered intestinal mucosal cells with pyknotic nuclei, extensive karyorrhexis and numerous bacteria. In 2 cases, cytology proved superior to endoscopy and radiology in detecting recurrent disease. We had 2 false negative cases, and the negativity was attributed to sampling errors. There was 1 false positive case in which 3-dimensional clusters of intestinal columnar cells were erroneously diagnosed as adenocarcinoma. CONCLUSION: Urine obtained from ileal conduit specimens shows a smear picture that is different from that of specimens from the bladder. Thus, it is imperative to understand the difference between the cytomorphology of bladder urine and ileal conduit urine, to minimize the pitfalls and increase diagnostic utility.

Collection devices for cervicovaginal cytology: a comparison.

OBJECTIVE: To compare cervicovaginal smears obtained by a cotton-tipped swab with those obtained by cervix brush and modified Ayre spatula. STUDY DESIGN: A combined cervicovaginal smear was collected from 100 women using 3 different collection devices: cotton-tipped swab, cervix brush, and modified Ayre spatula. In each patient a set of 3 smears was collected by the same cytotechnologist using all 3 devices in random order. Smears were evaluated using parameters mandatory for an optimal smear: evenly dispersed, well-preserved, adequate cells from the transformation zone. The cost and availability of the collection devices were also considered. RESULTS: The swab was the most effective device in obtaining thin, evenly spread, adequate, well-preserved smears as against the cervix brush and Ayre spatula. The pickup of abnormal cells was similar with the cotton-tipped swab and cervix brush, while the Ayre spatula failed to yield high grade squamous intraepithelial lesions, atypical glandular cells of undetermined significance and adenocarcinoma cells. The cotton-tipped swab proved to be the most cost effective. CONCLUSION: A properly prepared cotton-tipped swab is an inexpensive, readily available, nontraumatic collection device that yields smear of optimal quality.

Concurrent evaluation of visual, cytological and HPV testing as screening methods for the early detection of cervical neoplasia in Mumbai, India.

OBJECTIVE: Naked eye visual inspection with acetic acid (VIA), magnified VIA (VIAM), visual inspection with Lugol's iodine (VILI), cytology and human papillomavirus (HPV) testing were evaluated as screening methods for the detection of high-grade squamous intraepithelial lesions (HSIL) of the uterine cervix in a cross-sectional study in Mumbai, India. METHODS: Cytology, HPV testing, VIA, VIAM and VILI were carried out concurrently for 4039 women aged 30-65 years. All women were investigated with colposcopy and biopsies were taken from 939 women who had colposcopic abnormalities. The reference standard for final disease status was histology or negative colposcopy. The presence of HSIL was confirmed in 57 women (1.4%). The test characteristics for each method were calculated using standard formulae. RESULTS: The sensitivities of cytology, HPV testing, VIA, VIAM and VILI were 57.4%, 62.0%, 59.7%, 64.9%, and 75.4%, respectively (differences were not statistically significant). The specificities were 98.6%, 93.5%, 88.4%, 86.3%, and 84.3%, respectively. Adding a visual test to cytology or HPV testing in parallel combination resulted in a substantial increase in sensitivity, with a moderate decrease in specificity. The parallel combination of VILI and HPV testing resulted in a sensitivity of 92.0% and a specificity of 79.9%. CONCLUSION: As a single test, cytology had the best balance of sensitivity and specificity. Visual tests are promising in low-resource settings, such as India. The use of both VIA and VILI may be considered where good quality cytology or HPV testing are not feasible. The sensitivity of cytology and HPV testing increased significantly when combined with VIA or VILI.