Romanowsky staining in cytopathology: history, advantages and limitations.

If the entire discipline of diagnostic cytopathology could be distilled into a single theme, it would be the Papanicolaou stain. Yet it was the Romanowsky stain upon which the discipline of cytopathology was founded. Both stains are used today in the cytopathology laboratory, each for a different and complementary purpose. We trace the history of cytopathological stains and discuss the advantages and limitations of Romanowsky-type stains for cytological evaluation. We also provide suggestions for the advantageous use of Romanowsky-type stains in cytopathology.

Endobronchial ultrasound-guided transbronchial needle aspiration cytology: a state of the art review.

Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is a recently developed, accurate, safe and cost-effective technique that allows sampling of mediastinal lymph nodes and peribronchial lesions including pulmonary and mediastinal lesions. Its major indications are the nodal staging of non-small cell carcinomas of the lung, their restaging after chemotherapy and/or radiation, the diagnosis of sarcoidosis and of metastases from extrathoracic malignancies, and the diagnosis of mediastinal lymphadenopathy and masses of unknown aetiology. From our experience at the University of Minnesota and a comprehensive review of the literature, we discuss technical aspects of the procedure, its advantages and limitations in comparison with other methods of sampling mediastinal lymph nodes, focusing on the role of the cytopathologist in ensuring the effectiveness of the procedure. An algorithmic approach to the cytological diagnosis, starting with the determination of the adequacy of the sample, is also presented.

Beta 1-integrins regulate the formation and adhesion of ovarian carcinoma multicellular spheroids.

Ovarian carcinoma multicellular spheroids are an in vitro model of micrometastasis whose adhesive abilities have not been elucidated. In this study, we identified adhesion molecules that mediate the formation of ovarian carcinoma spheroids and their subsequent adhesion to extracellular matrix proteins. The NIH:OVCAR5, but not the SKOV3, ovarian carcinoma cell line formed spheroids similar to multicellular aggregates isolated from patient ascitic fluid. NIH:OVCAR5 spheroid formation was augmented by a beta 1-integrin-stimulating monoclonal antibody or exogenous fibronectin, but was inhibited by blocking monoclonal antibodies against the alpha 5- or beta 1-integrin subunits. By immunohistochemical staining, alpha 2-, alpha 3-, alpha 5-, alpha 6-, and beta 1-integrin subunits, CD44, and fibronectin were detected in NIH:OVCAR5 spheroids. NIH:OVCAR5 spheroids adhered to fibronectin, laminin, and type IV collagen, and this adhesion was partially inhibited by blocking antibodies against the alpha 5-, alpha 6-, and alpha 2-integrin subunits, respectively. A blocking monoclonal antibody against the beta 1-integrin subunit completely inhibited adhesion of the spheroids to all three proteins. These results suggest that interactions between the alpha 5 beta 1-integrin and fibronectin mediate the formation of ovarian carcinoma spheroids and that their adhesion to extracellular matrix proteins at sites of secondary tumor growth may be mediated by a complex interaction between multiple integrins and their ligands.

Fine-needle aspiration diagnosis of unusual cutaneous neoplasms of the scalp in HIV-infected patients: a report of two cases and review of the literature.

We report on two unusual, non-AIDS-defining scalp neoplasms, Merkel-cell carcinoma (MCC) and malignant melanoma, in 2 men with acquired immunodeficiency syndrome (AIDS). In the first patient, metastatic MCC was initially diagnosed by fine-needle aspiration (FNA) of a posterior cervical lymph node, based on the cytomorphology and the characteristic immunohistochemical and ultrastructural features. No skin lesion was initially apparent, but a 0.3-mm scalp primary was found during the ensuing neck dissection. In the second patient, recurrent and metastatic malignant melanoma from a Breslow 1.3-mm scalp primary was diagnosed by FNA. Both patients developed generalized disease in a relatively short time, despite their small primaries. These cases illustrate the occurrence of Merkel-cell carcinoma and melanoma in AIDS patients, and stress the need to consider these unusual cutaneous neoplasms when evaluating lymph node FNA samples from HIV-positive patients, especially since both may present as metastases from clinically occult primaries.

Signet ring cell lobular carcinoma of the breast presenting in a cervicovaginal smear. A case report.

BACKGROUND: The presence of extragenital malignant cells in cervicovaginal smears is a rare and usually late event in a patient with a long history of cancer. This, to the best of our knowledge, is the first case of breast cancer initially diagnosed on a Pap smear. CASE: A 50-year-old woman presented with abdominal distension and weight gain. A Pap smear showed numerous signet ring cells and was diagnosed as signet ring cell adenocarcinoma, most likely metastatic from the breast or stomach. Subsequent evaluation revealed bilateral adnexal masses and inguinal lymphadenopathy, leading to hysterectomy and bilateral salpingo-oophorectomy. The ovaries, corpus and cervix were involved by signet ring cell carcinoma. The metastatic tumor proved to be positive for CK7, CEA, estrogen receptor and progesterone receptors and negative for CK20. Despite the absence of a discrete palpable breast mass, a mammogram was recommended based on these results, and an ensuing breast biopsy showed the presence of an infiltrating lobular carcinoma. CONCLUSION: Lobular carcinoma of the breast may present in a cervicovaginal smear. Correct interpretation of signet ring cells as metastatic, most likely from the breast or stomach, is helpful in guiding management.

Amyloidoma of bone, a plasma cell/plasmacytoid neoplasm. Report of three cases and review of the literature.

Tumoral amyloidosis (amyloidoma) of bone is a rare condition characterized by the massive destructive deposition of AL amyloid in bones. We report three cases. The patients ranged in age from 45 to 78 years and had tumors located in the lumbar spine, scapula, and humeral head measuring 6.5 to 18 cm. The radiologic diagnosis was chondrosarcoma in two cases. Microscopically, there were large, rounded deposits of amorphous eosinophilic material surrounded by numerous giant cells and a sparse lymphoplasmacytic infiltrate. The deposits proved to be composed of AL amyloid showing potassium permanganate resistant congophilia. Immunohistochemistry showed immunoglobulin IgG lambda, IgG kappa, and IgM lambda monoclonality of the plasma cell and (in one case) lymphoid infiltrate. The tumors were classified by morphology and immunohistochemistry as solitary plasmacytomas of bone (two cases) and plasmacytoid lymphoma (one case). During the relatively short follow-up period, one patient progressed to symptomatic generalized amyloidosis and died, one patient died of recurrent tumor, and one patient is alive with no evidence of disease. An extensive review of the world literature showed 34 well-documented similar cases, occurring most often in the spine and skull, causing neurologic symptoms, tending to occur in middle-aged men and frequently progressing to generalized disease. Most if not all AL amyloidomas of bone represent solitary plasmacytomas of bone or plasmacytoid lymphomas.

Differential diagnosis of Hürthle cell neoplasms on fine needle aspirates. Can we do any better with morphometry?

OBJECTIVE: To determine the value of computerized interactive morphometry in the preoperative prediction of malignancy in fine needle aspirates of Hürthle cell neoplasms. STUDY DESIGN: Alcohol-fixed, Papanicolaou-stained fine needle aspiration smears of histologically proven Hürthle cell adenomas (HCA) (n = 10) and Hürthle cell carcinomas (HCC) (n = 9) were studied by interactive computerized morphometry. The measured features included the areas, perimeters and shape factors of individual cells, nuclei and nucleoli; the nucleocytoplasmic and nucleolonuclear ratios; and the eccentricities of nuclei and nucleoli. RESULTS: Only nucleolar features showed statistically significant differences between HCA and HCC. These features were the nucleolar area and its standard deviation, the nucleolar form factor and circularity, and the nucleolonuclear ratio. The most effective, albeit imperfect, discrimination was achieved by the nucleolar form factor. CONCLUSION: Nucleolar features, such as size, variation in size and roundness, may be more effective than cellular or nuclear features in differentiating between HCA and HCC in fine needle aspiration smears.

Lymphoid monoclonal antibodies reactive with lung tumors. Diagnostic applications.

In the course of investigating 30 monoclonal antibodies (MAbs) for their potential reactivity with 25 lung tumors of different histologic types, we found that three MAbs commonly used for their specificities for lymphoid markers were highly reactive with non-small-cell carcinomas (NSCLC) and totally nonreactive with small-cell carcinomas (SCLC). Immunostaining was performed by the standard streptavidin-biotin-peroxidase method after microwave antigen retrieval on formalin-fixed, paraffin-embedded tissue sections. LN2 (CD74), LN3 (HLA-DR), and BLA-36, which are commonly used for the identification of B-lymphocytes, strongly immunostained 19 of 25 squamous and adenocarcinomas and none of 34 small-cell carcinomas and carcinoids. Moreover, in combined tumors, these MAbs selectively stained the adenocarcinoma cells but not the adjacent small-cell carcinoma cells. A cocktail mixture of LN2, LN3, and BLA-36 assayed on 24 additional lung tumors produced similar results with even stronger and sharper stainings. Other lymphoid MAbs showed some selective staining but to a lesser degree. Among nonlymphoid MAbs, the results were as expected, with MAbs for cytokeratin (B72.3) and epithelial membrane antigen staining NSCLC but also some SCLC. The MAbs for chromogranin and neuron-specific enolase were not entirely specific, whereas some nerve-cell adhesion molecule MAbs showed good specificity for SCLC. In a field with few specific MAbs, the newly discovered ability of these lymphoid MAbs to discriminate between SCLC and NSCLC may prove useful in the immunohistochemical diagnosis of lung tumors.