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.

Assessment of HER-2/neu status in breast cancer using fluorescence in situ hybridization & immunohistochemistry: Experience of a tertiary cancer referral centre in India.

BACKGROUND & OBJECTIVES: Determination of HER2 status in breast cancer has become important to identify potential candidates for anti-HER2 therapy. In this study we compared fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) for the determination of HER2 status in breast cancer patients referred to a tertiary care referral centre. METHODS: A total of 200 cases of invasive breast cancer were evaluated for HER2 status using IHC and FISH and results were compared. RESULTS: The IHC 3+ (93.9%) and IHC negative (85.9%) cases showed good concordance with the corresponding FISH results; while 66.6 per cent of IHC 2+ cases showed gene amplification by FISH. In addition, hormone receptor expression and HER2 gene status showed a statistically significant inverse association (P<0.05). INTERPRETATION & CONCLUSION: These findings reaffirm IHC as a prudent first-step to screen tissue samples for HER2 status and to determine suitability for technically demanding FISH test and the dual coloured FISH as a gold standard for determination of HER2/neu status in IHC equivocal cases of breast carcinoma.

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.

Diagnostic dilemmas in fine needle aspiration cytology of an ectopic kidney: a case report.

BACKGROUND: Congenital anomalies are easily diagnosed on radiology and rarely have an unusual presentation requiring an invasive diagnostic procedure. Fine needle aspiration cytology (FNAC), though a well-established diagnostic technique for abdominal and retroperitoneal lesions, is fraught with several challenges and pitfalls. CASE: We report a case of a 39-year-old woman with an ectopic kidney presenting as an abdominal mass. On FNAC the cellular aspirate was misinterpreted as a paraganglioma or metastatic carcinoma. CONCLUSION: This case highlights the fact that on FNAC, normal cellular components, especially from the kidney, provide a significant pitfall for overdiagnosis, potentially resulting in unnecessary surgical explorations.

Presence of a micropapillary pattern in mucinous carcinomas of the breast and its impact on the clinical behavior.

Infiltrating micropapillary carcinomas (IMPC) of breast are highly angioinvasive tumors with poor prognosis. This study is based on the observation that a similar micropapillary pattern is also observed in mucinous carcinomas of breast. About 102 mucinous carcinomas were evaluated for the presence and impact of this micropapillary pattern on the clinical behavior. Of these, 68 were mucinous carcinomas with a micropapillary pattern (MUMPC), 20 had MUMPC mixed with an infiltrating duct carcinoma component, two were solid variants of papillary carcinoma with mucin (SVPCMU), five had collision of the MUMPC and SVPCMU patterns and seven were mucinous carcinomas with signet ring cells (MUS). The factors negatively affecting overall survival (OAS) and disease-free survival (DFS) included the histological type of mucinous carcinoma, nodal metastases, an irregular tumor border, <50% mucin and an IMPC type of local recurrence or metastases. In the multivariate analysis, the histologic type of mucinous carcinoma and an irregular tumor border were most significant for OAS and DFS. Thus, 86% of mucinous carcinomas in this study were mucinous variants of the angioinvasive infiltrating micropapillary carcinomas. These tumors can produce IMPC type of metastases and thus should be treated aggressively.

Epithelioid sarcoma of the foot with subsequent lesion in hand: metastatic lesion or second primary?

Epithelioid sarcomas (ES) are rare tumors of soft tissue that have a propensity to occur in the extremities. Epithelioid sarcomas are known to metastatise to draining lymph nodes and commonly to the lungs. Herein, a case of epithelioid sarcoma which recurred in an unusual site namely the distal phalanx of left middle finger, six months post amputation of the primary lesion in the left foot is being reported. The ipsilateral inguinal lymph node showed metastatic deposits. The tumor at both these sites had similar histology and an identical immunohistochemical (IHC) pattern showing reactivity to cytokeratin (CK), epithelial membrane antigen (EMA), vimentin (Vim) and CD34. This case is presented to record an unusual occurrence of ES in the distal phalanx of middle finger with an ES of foot. The metastasis of ES to the distal acral bones has not been documented till date.

Changes in the tumor grade and biological markers in locally advanced breast cancer after chemotherapy--implications for a pathologist.

There is insurgence of literature evaluating prognostic and predictive factors in breast carcinomas treated with chemotherapy, with a parallel need to develop guidelines for the pathologist interpreting such excisions. Prechemotherapy gun biopsy and postchemotherapy excision specimens from 78 women with locally advanced breast cancer were analyzed for histological changes in the tumor, changes in the tumor grade, hormone receptors, cerb2, and bcl2 and their impact on disease-free survival (DFS). An unusually prominent granulomatous response to tumor was seen in three cases. The tumor grade changed in five patients, estrogen receptor (ER) expression was altered in 10 cases, progesterone receptor detection changed in 16 cases, cerb2 in one case and bcl2 in 16 cases. Fixation of the gun biopsy in Bouin's fluid and severe damage of nuclei after chemotherapy were the reasons for shift in the expression of hormone receptors. A low-grade tumor was associated with better response to chemotherapy. In the Kaplan-Meier analysis the ER expression and a low-grade tumor (grade I and II) significantly affected DFS. None of the factors evaluated impacted the overall survival of patients. To conclude there is a change in the tumor grade, bcl2, cerb2 and hormone receptors after chemotherapy. A pathologist interpreting specimens of breast cancer after chemotherapy must always record the postchemotherapy grade as it is an indicator of better response to chemotherapy and survival.

Cytologic findings in infiltrating micropapillary carcinoma and mucinous carcinomas with micropapillary pattern.

OBJECTIVE: To examine the observation that some mucinous carcinomas have a micropapillary pattern and are mucinous variants of the highly angioinvasive infiltrating micropapillary carcinomas (IMPC). STUDY DESIGN: We evaluated cytologic findings of 13 IMPC and 55 mucinous carcinomas for comparative features. RESULTS: In mucinous carcinomas, 37 of 50 (74%) had a micropapillary pattern. This group included 27 cases with pure mucinous micropapillary morphology (MUMPC), 8 MUMPC associated with a ductal carcinoma of the IMPC type (MUIDC) and 2 cases of mixed mucinous carcinomas with an MUMPC and a solid variant ofpapillary carcinoma (SVPC) component. On cytology both IMPC and mucinous carcinomas with micropapillary pattern demonstrated the micropapillary pattern, that is, angulated clusters or abortive papillae and ball-like clusters. However, the IMPC smears revealed numerous singly scattered tumor cells and larger fragments with shrub-like branching and the MUMPC had psammoma bodies. The mixed MUMPC and SVPC showed the classic cytologic features of MUMPC admixed with abundant singly dispersed tumor cells in the background representing the SVPC component. CONCLUSION: Although IMPC and the MUMPC share the micropapillary pattern on histologic examination, mucin alters the appearances in aspirates. Recognition of this morphologic spectrum will help in understanding the behavior of these tumors.

Proximal-type epithelioid sarcoma--a rare, aggressive subtype of epithelioid sarcoma presenting as a recurrent perineal mass in a middle-aged male.

BACKGROUND: Epithelioid sarcoma (ES) is an uncommon soft tissue sarcoma. Lately, subtypes of ES, including proximal-type ES have been recognized, with relatively few reports on such cases. CASE PRESENTATION: A 47-year-old male presented with a perineal soft tissue mass that was excised elsewhere and the biopsy was submitted for a review diagnosis. On histology, a multi nodular tumor was seen comprising sheets of oval to polygonal cells with moderate amount of cytoplasm. Interspersed were larger, rhabdoid cells with abundant eosinophilic cytoplasm and prominent nucleoli. Focal necrosis was noted. A wide panel of immunohistochemical (IHC) markers was performed to rule out a range of differential diagnoses, including a poorly differentiated carcinoma, a melanoma and a variety of sarcomas with epithelioid differentiation. On IHC, the tumor cells showed a polyphenotypic expression, including positivity for epithelial markers i.e cytokeratin (CK), CK7, EMA and mesenchymal markers like vimentin and CD 34. Desmin was focally positive. CK20, CEA, S-100, HMB-45, SMA, LCA and CD31 were negative. A diagnosis of a proximal-type ES was formed. Six moths later, despite adjuvant chemo and radiotherapy (CT and RT), the patient continued to have the lesion and was referred again. In addition to the earlier histological features, sections from the persistent tumor mass showed an increased number of larger cells along with multinucleated tumor giant cells. CONCLUSION: The value of identifying this uncommon tumor from a list of differential diagnoses is in view of its aggressive behavior, as seen in our case. A wide excision with clear margins is imperative with options of post-operative CT/RT in individual cases during a close follow-up.

Fibromatosis-like carcinoma-an unusual phenotype of a metaplastic breast tumor associated with a micropapilloma.

BACKGROUND: Fibromatosis-like metaplastic carcinoma is a newly described metaplastic breast tumor, literature on which is still evolving. CASE PRESENTATION: A 77-year-old lady presented with a 2 x 2 cm mass with irregular margins in the upper and outer quadrant of left breast. Fine needle aspiration cytology (FNAC) from the lump was inconclusive. A lumpectomy was performed and sent for frozen section, which revealed presence of spindle cells showing mild atypia in a sclerotic stroma. The tumor cells revealed prominent infiltration into the adjacent fat. A differential diagnosis of a low-grade sarcoma vs. a metaplastic carcinoma, favoring the former, was offered. Final histology sections revealed an infiltrating tumor with predominant spindle cells in a collagenous background, simulating a fibromatosis. Adjacent to the tumor were foci of benign ductal hyperplasia and a micropapilloma. Immunohistochemistry (IHC) showed diffuse co-expression of epithelial markers i.e. cytokeratins (CK, HMWCK, CK7) and EMA along with a mesenchymal marker i.e. vimentin in the tumor cells. Myoepithelial markers (SMA and p63) showed focal positivity. A diagnosis of a low-grade fibromatosis-like carcinoma breast associated with a micropapilloma was formed. CONCLUSION: Fibromatosis-like carcinoma is a rare form of a metaplastic breast tumor. This diagnosis requires an index of suspicion while dealing with spindle cell breast tumors. The importance of making this diagnosis to facilitate an intra operative surgical planning is marred by diagnostic difficulties. In such cases, IHC is imperative in forming an objective diagnosis.

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.

Comparative analysis of routine histology, immunohistochemistry, reverse transcriptase polymerase chain reaction, and fluorescence in situ hybridization in diagnosis of Ewing family of tumors.

CONTEXT: The Ewing family of tumors are often difficult to distinguish from other malignant small round cell tumors, but more than 90% have EWS-FLI1 chimeric transcript, which acts as a potential molecular diagnostic marker. OBJECTIVE: To do a comparative analysis of 32 cases with EWS-FLI1: Ewing family of tumors (n = 30), desmoplastic small round cell tumor (n = 1), and undifferentiated sarcoma (n = 1). DESIGN: The initial diagnosis was made on core biopsy (n = 22) and open biopsy (n = 4) specimens by using morphology and immunohistochemistry and on fine-needle aspiration cytology ([FNAC], n = 6) specimens. EWS-FLI1 was detected by reverse transcriptase polymerase chain reaction on all 32 fresh FNAC samples and by fluorescence in situ hybridization on 16 paraffin blocks. RESULTS: The 19 male and 13 female patients had bone (n = 19) or soft tissue (n = 13) tumors. Histologic groups were typical Ewing sarcoma (n = 15), atypical Ewing sarcoma (n = 4), Askin Rosai tumors (n = 5), desmoplastic small round cell tumor (n = 1), undifferentiated sarcoma (n = 1), and cases diagnosed as malignant small round cell tumors on FNAC (n = 6). All tumors except desmoplastic small round cell tumor and undifferentiated sarcoma were CD99 positive. EWS-FLI1 by reverse transcriptase polymerase chain reaction was noted in 15 cases of typical Ewing sarcoma, 4 cases of atypical Ewing sarcoma, 5 cases of Askin Rosai tumor, and no cases of desmoplastic small round cell tumor or undifferentiated sarcoma. With use of fluorescence in situ hybridization, EWS break was detected in 10 of 11 paraffin blocks used and was negative in desmoplastic small round cell tumor. CONCLUSIONS: The excellent correlation of routine histologic findings in Ewing family of tumors with results on immunohistochemistry and fluorescence in situ hybridization on archival material and reverse transcriptase polymerase chain reaction on fresh FNAC specimens underscores that the traditional observation on routine histologic examination is a time-tested tool. The diagnosis of Ewing family of tumors can be validated on archival material or fresh biopsy samples, including those obtained by FNAC.

An analysis of 46 static telecytology cases over a period of two years.

We analysed 46 telecytology cases sent from two rural hospitals about 500 km from a tertiary cancer centre. The cases were submitted for second opinion over a period of two years and evaluated using a static store and forward telecytology approach. A total of 715 digital images were studied (average 15 per case). Forty-one of the 46 cases (89%) were reported within 3 days and 54% of cases were reported within one working day. The aspiration smears and images were found to be of diagnosable quality in 89 and 93% of the cases, respectively. The diagnostic concordance was assessed by comparing the telecytology diagnosis, glass slide diagnosis and final histopathology diagnosis (when available). A clinically useful diagnosis was rendered in 91% cases with 74% complete concordance. Five out of 46 cases (11%) were deferred for glass slide review. Store and forward telecytology using the Internet is a rapid and effective method of providing expert diagnosis in cytology.

Primary angiosarcoma of the breast: observations in Asian Indian women.

BACKGROUND: Primary angiosarcomas of breast are rare tumors, with a fatal outcome. MATERIAL AND METHODS: We studied histological prognostic factors and c-kit expression by immunohistochemistry (IHC) in 12 angiosarcomas accessioned at a cancer referral center in India. RESULTS: All patients had primary angiosarcoma; no case of secondary angiosarcoma was accessioned during the study period. Median age of patients was 24.5 years. Nine patients had intermediate grade tumors, one a well differentiated tumor and three patients had high-grade tumors. Interesting cases encountered included an epithelioid angiosarcoma and an angiosarcoma arising on the background of a biphasic tumor. Eight patients had lumpectomy, four mastectomy and two patients were given radiotherapy. Of the nine patients (seven type I/II and two high grade) with follow up, eight patients developed disseminated metastases within a year of presentation. The patient with well-differentiated angiosarcoma also died of metastasis albeit after a longer time. On IHC c-kit staining was weakly seen in two cases. CONCLUSION: Primary angiosarcoma was fatal in young Indian women even in lower grade tumors. The low expression of c-kit on IHC suggests that targeting this protein for therapy may not be successful in treating these tumors.

Cytomorphologic patterns in papillary cystic variant of acinic cell carcinoma of the salivary gland.

OBJECTIVE: To study the cytomorphologic profile of the papillary and cystic variant of acinic cell carcinoma (ACC-PCV) of the salivary glands. STUDY DESIGN: We studied 6 aspirates from 5 cases of ACC-PCV. RESULTS: All the cases had varied cytomorphologic features depending upon the degree of vacuolation of cells. However, common to all was a papillary pattern and a cystic fluid background with or without mucin blobs; that led to misdiagnosing the tumor as mucoepidermoid carcinoma on 2 occasions. The smears showed papillary fragments, sheets or clusters of vacuolated/histiocyte like cells and granular cells. The histiocytelike cells resembled macrophages, with finely vacuolated cytoplasm and an eccentrically placed nucleus with frequent binucleation. Vascular cores were seen in a few aspirates. The granular cells were similar to those seen in the usual acinic cell carcinoma but were smaller. The tumor did not show any acinar pattern and lacked naked nuclei in the background. In 4 aspirates finely distributed, brown hemosiderin pigment was detected in the vacuolated cells. CONCLUSION: ACC-PCV is papillary and cystic and hence is often not recognized as acinic cell carcinoma. However, papillary fragments of vacuolated cells or histiocytelike cells and granular cells are clues to the diagnosis.

Pure malignant myoepitheliomas of the breast: an immunohistochemical study.

Myoepithelioma of breast are extremely rare. We report two cases of pure malignant myoepithelioma of the breast, utilising light microscopic and immunohistochemical methods for diagnosis. Both the cases presented as breast lump. Hematoxylin and Eosin (H&E) stained microscopic sections revealed a predominantly spindle cell tumor. Immunohistochemical work up was done. Case number one expressed positivity for vimentin, Smooth Muscle Actin (SMA), S-100 and CD10. Case number two expressed positivity for Vimentin, CD10 and p63. This led to the diagnoses of malignant myoepithelioma in both of them. Documentation of such cases prospectively and from archival material, using immunohistochemistry, is of extreme importance to assess the prevalence, various phenotypic patterns, long-term biological behaviour and to establish management protocols for malignant myoepithelioma.

Case report: metastatic adamantinoma of the tibia--an unusual presentation.

A 26-year-old female with a tibial lesion diagnosed as an adamantinoma was treated with intra-lesional curettage, bone grafting and intra-medullary nailing. Six years post-surgery, she presented with an asymptomatic primary site but with a metastatic lesion in the mid-shaft of the ipsilateral femur and lung metastases. The femoral lesion was treated with wide excision and reconstructed with an allograft and plate fixation. Pulmonary metastatectomy was carried out for the lung lesions. A follow-up CT scan of the chest at 1 year after the surgery for the metastatic lesions revealed fresh unresectable bilateral metastases. Although cases of local recurrences and pulmonary metastases in adamantinoma are reported, this case is unusual in presenting without a local recurrence but with simultaneous skeletal and pulmonary metastases.

Renal angiomyolipomas--a study of 18 cases.

Angiomyolipoma is a distinctive neoplasm composed of an intimate admixture of three components, viz. mature adipocytes, smooth muscle cells and blood vessels. This study was undertaken to better define the various morphological patterns of angiomyolipoma and their immunohistochemical profile. The paraffin blocks and slides of 18 cases of renal angiomyolipoma, accessioned over a period of 8 years from Tata Memorial Hospital, were reviewed. There were 2 men and 16 women in the age range of 17 to 68 years. Pre-operative fine needle aspiration cytology (FNAC) was performed in 6 cases, of which 5 were erroneously diagnosed as renal cell carcinomal sarcoma. Histologically, 14 cases revealed conventional histology. The remaining 4 cases were particularly misleading posing diagnostic problems due to variant patterns (leiomyomatous variant-3 cases and epithelioid variant- 1 case). Apart from the usual histology, necrosis, giant cells, and varying degrees of nuclear pleomorphism were present in three of the cases. Immunohistochemistry performed in 16 cases revealed immunoreactivity for HMB45 in all the 16 tumors (100%), estrogen receptor (ER) and progesterone receptor ((PR) in 2 cases (12.5%) and 6 cases (37.5%) respectively.

Desmoplastic round cell tumor of childhood: can cytology with immunocytochemistry serve as an alternative for tissue diagnosis?

There are limited reports on the cytology of desmoplastic small round cell tumors (DSRCT). Fine needle aspiration biopsy (FNAB) findings in seven aspirates from four cases of histologically and immunohistochemically confirmed cases were analyzed with the main intention of ascertaining if cytological diagnosis of DSRCT is possible. Also assessed were the immunocytochemistry(ICC) findings in these cases. The basic cytological impression was that of a cohesive small round cell tumor. Nuclei showed granular chromatin with grooves, nuclear molding and inconspicuous nucleoli. Stromal fragments were noted in all four cases. In two cases, awareness of cytological features in the appropriate clinical context led to a suggestion of the diagnosis of DSRCT on cytology itself. ICC on destained smears showed positivity for cytokeratin, epithelial membrane antigen (EMA), desmin and WT-1 in two cases. In conclusion, given the right clinical setting, a cytological diagnosis of DSRCT is plausible and in conjunction with ICC may help in documenting the polyphenotypic nature and thereby confirming the diagnosis.

Experience with telepathology at a tertiary cancer centre and a rural cancer hospital.

BACKGROUND: Telepathology allows quick and timely access to an expert opinion, no matter where the patient is located. We analysed the experience with the use of telepathology between a tertiary cancer centre and a rural cancer hospital. METHODS: Ninety-three cases were analysed in which static telepathology was used to obtain a consultation between Tata Memorial Centre and Nargis Dutt Memorial Cancer Hospital at Barshi, a rural area. RESULTS: Successful teleconsultation was achieved in all cases. A diagnosis was offered in 92 cases (98.9%) and was deferred in 1 case (1.1%). Complete concordance, clinically unimportant minor discrepancy and hedged diagnosis were obtained in 83 cases (90.2%). Major discrepancies were encountered in 9 cases (9.7%). The number of images per case ranged from 3 to 27 (average: 7 images). Images were of diagnosable quality in 89.2% of cases. Most of the cases (77.4%) were reported within 3 days; 32.2% were reported within 8 hours (a single working day) and 45.1% within 1-3 days. CONCLUSION: Telemedicine can be effectively used to bridge the gap between medically underprivileged, geographically distant rural areas and advanced centres using the static store and forward methodology.