Dipeptidyl aminopeptidase IV staining of cytologic preparations to distinguish benign from malignant thyroid diseases.

Staining for dipeptidyl aminopeptidase IV (DAP IV [EC: 3.4.14.5]) activity was applied to aspiration biopsy specimens or imprint preparations of surgical biopsy specimens from thyroid tumors. Material was obtained from 55 patients with histologically proven thyroid diseases: 9 with papillary carcinoma, 5 with follicular carcinoma, 11 with follicular adenoma, 13 with adenomatous goiter, 8 with Hashimoto's thyroiditis, and 9 with other benign conditions. Most tumor cells, follicular lumina in cell clusters, and intranuclear inclusions were strongly positive for DAP IV in all examples of papillary or follicular carcinoma. In contrast, only a few epithelial cells were labeled for DAP IV in follicular adenoma and adenomatous goiter. Some Hürthle cells in Hashimoto's thyroiditis also were positive for DAP IV. When a DAP IV scoring system based on the percentage of positive cells and staining intensity was used, all benign tissues except one (from a follicular adenoma) were found to have extremely low scores. These results indicate that staining for DAP IV activity is a simple but useful tool to aid in distinguishing benign from malignant thyroid neoplasms.     

identification of Cryptococcus neoformans in cytologic preparations of cerebrospinal fluid.

Review of routine Papanicolaou-stained cerebrospinal fluid preparations from 13 patients who had meningeal cryptococcosis documented by other methods demonstrated the yeast in 11 cases. Special stains greatly facilitated the detection of the organisms in two samples and discriminated them from artifacts. An increased number of cells was present in nine cases. Correlation with the clinical data revealed that every patient but one had a malignant lymphoma, most commonly Hodgkin's disease. The exception was a patient who had disseminated carcinoma of the breast treated with adrenal corticosteroids. The clinical history and the cellularity of the smear should alert one to the possibility of cryptococcosis.     

Prognostic factors in breast cancer: Use of cytologic preparations

There is increasing evidence that certain morphological features and biological markers found in breast tumors may provide prognostic information by predicting the risk of recurrence and metastasis in early breast cancer. This information may also be important in choosing therapeutic options in patients with advanced disease. Prognostic testing is commonly performed on surgically excised lesions. However, there are clinical conditions in which a surgical specimen may not be suitable or available for such analysis. In these circumstances, fine-needle aspiration biopsy and imprint preparation provide an attractive sample for prognostic testings. This review summarizes the current approach to the use of cytologic preparation for assessment of established and newly recognized prognostic factors. © 1995 Wiley-Liss, Inc.     

Comparison of three commonly used cytologic preparations in effusion immunocytochemistry.

Discrepant results in effusion immunocytochemistry are often the result of specimen processing. Smears, cytospins, cell blocks, and monolayer preparations have all been used in various published studies; thus, there is no consistency in the immunostaining process for cytology to compare with the surgical pathology "gold standard" results. We sought to evaluate optimal specimen preparation for the immunostaining of effusion samples. Fourteen reactive and 15 malignant effusion samples (various epithelial/mesothelial neoplasms) were each prepared in three forms: air-dried cytospins (postfixed in ethanol), formalin-fixed, paraffin-embedded cell blocks, and liquid-based thin-layer (ThinPrep, CYTYC, Boxborough, MA) processing. All slides were immunostained with antibodies commonly used in effusion cytology: HBME-1, calretinin, E-cadherin, BerEP4, B72.3, LeuM1, and CA19-9. Cytospin and ThinPrep samples performed in a similar manner: high background staining was encountered in 66% of cases, most evident in three-dimensional clusters of cells. In addition, membrane staining patterns were difficult to interpret. Cell blocks provided the best milieu for morphologic interpretation, with less background staining (only 17% of cases) and results that most closely approximated those reported in the surgical pathology literature. The cost per test for cell block immunocytochemistry was also the most economical for our laboratory.     

Identification of "papillary-like" clusters on endometrial cytologic preparations

The presence of papillary cell clusters is taken as a serious indication in the diagnosis of well-differentiated endometrial adenocarcinoma in Japan. However, papillary-like clusters have been observed in both normal and benign samples. Therefore, overdiagnosis may occur in the observation of cytopreparations. The present study attempts to prepare a histological sample from an Auto Cyto Fix (ACF) sample prepared by the automatic fixation apparatus (ACF1000) using a membrane filter method after cytological observation and to examine its usefulness. In an ACF sample, a papillary-like cluster was observed and the coverslip was then removed, the circumference of the membrane filter was cut off, and the target cell cluster was smeared, embedded, and sliced. In the cases in which a papillary-like cluster was observed, even if differential diagnosis of the derivation was difficult due to a resemblance between the respective morphological findings, it was easily made by histological observation of an ACF sample.     

Langerhans cell histiocytosis of the skull on cytologic squash preparations

We present a case in which a primary cytodiagnosis of Langerhans cell histiocytosis (LCH) of the skull was made using squash preparations. The patient, a 25-year-old male, presented with raised intracranial pressure and decreased visual acuity. Magnetic resonance imaging revealed a large skull lesion with osteolytic features in the left frontal bone. The patient underwent surgical resection by the extended basal frontal epidural approach. The squash preparation smears were cellular and demonstrated a mixed population of small, mature lymphocytes, eosinophils, and a high histiocytes content. The histiocytes occurred as isolated or loosely cohesive and clustered. They possessed abundant cytoplasm with rounded cell shape and had characteristic nuclear features, composed of fine chromatin and delicate nuclear membranes. The cytologic features of these histiocytes were consistent with Langerhans cells (LCs). A final impression of LCH of the skull was rendered. Subsequent histopathology confirmed the diagnosis. LCs reacted with both S-100 protein and CD1a immunohistochemically. The demonstration of Birbeck granules on electron microscopic study was also noted. Whenever squash preparation yields a mixed population of mature lymphocytes, eosinophils, and histiocytes, the cytologists should be aware of and consider LCH as a diagnostic possibility.     

Obtaining preparations for the differential staining of chromosomes

A study of correlation between preservation of the three-dimensional structure of chromosomes and their ability to undergo differential staining showed that chromosomes shaped like a convex half-cylinder give a regular pattern of longitudinal differentiation, but those shaped like concave half-cylinders do not. It was found that if the chromosomes are exposed for a brief period to the action of water during the staining procedure their outlines are changed and they become shaped like concave half-cylinders. Another method of obtaining chromosome preparations — ruling out possible exposure to water and increasing both the percentage of metaphase plates with differential staining of the chromosomes and also the quality of the staining — is thus suggested.Laboratory of General Cytogenetics, Institute of Medical Genetics, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. A. Kraevskii.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 78, No. 8, pp. 120–122, August, 1974.     

Immunohistochemical analysis of gel-transferred cells in cytologic preparations following smear division

Fine-needle aspirations on solid tumors are used increasingly as a means of obtaining a primary diagnosis. In many cases, a panel of immunostains performed on these aspirates is necessary to further characterize the cytologic interpretation. The amount of material obtained through aspiration, however, is often quite limited and is present on few glass slides. Previous studies have demonstrated the success of dividing cytologic smear preparations into smaller parts that could then be used for a panel of immunohistochemical stains. These results, however, did not compare the immunoreactivities of various antibodies before and after tissue transfer on cytologic preparations. In the present study, 41 immunohistochemical stains that employed 16 antibodies on 15 tumor preparations were performed following smear partition using the tissue-transfer technique. The percentage of cells that stained positive after transfer was determined and was correlated quantitatively to the untransferred controls. Specific immunoreactivity was demonstrated in 30 of 38 cases (79%) but was significantly decreased or lost in 8 of 38 cases (21%), which included antibodies for S-100, estrogen and progesterone receptors, chromogranin, neuron-specific enolase, and cytokeratin. Morphology was well preserved following tissue transfer, although limited cytoplasmic damage was seen in up to 25% of tumor cells. Immunopositive samples were found to be easily interpretable. Because sporadic cases fail to show immunohistochemical staining reactions following cytologic smear division and transfer, negative immunohistochemical stains in such preparations should be approached with caution. Diagn. Cytopathol. 1998;18:377–380. © 1998 Wiley-Liss, Inc.     

ThinPrep vs. conventional smear cytologic preparations in analyzing fine-needle aspiration specimens from palpable breast masses.

Limited data exist concerning the cellular features of the ThinPrep (Cytyc Corp., Boxborough, MA) technique in the analysis of breast fine-needle aspiration specimens. Therefore, we analyzed a series of 75 surgically excised palpable breast masses and compared ThinPrep and conventional smear fine-needle aspiration preparations. Each mass was aspirated twice. The first sample was used for two alcohol-fixed conventional smears, and the second sample was rinsed into CytoLyt (Cytyc Corp., Boxborough, MA) solution for processing into a ThinPrep slide. The paired slides were separated and independently analyzed for adequacy, overall cellularity, single epithelial cells (absent, rare, moderate, or numerous), epithelial architecture (sheets or three-dimensional clusters), myoepithelial cells and stripped bipolar nuclei (present or absent), and nuclear detail (poor, satisfactory, or excellent). Each sample was classified as negative, negative consistent with fibroadenoma, atypical favoring benign, atypical favoring malignant, or positive for malignant cells. The 75 breast masses included 32 carcinomas and 43 benign lesions. Four conventional smears and one ThinPrep were unsatisfactory. Significantly, more conventional smears were limited by drying artifact (9 vs. 0). ThinPrep aspirates of carcinomas had better nuclear detail (P = 0.03) and greater cellularity (P = 0.05). ThinPrep aspirates of benign masses had greater epithelial cellularity (P = 0.007) and better nuclear detail (P < 0.001), and more specimens had myoepithelial cells (P = 0.007). The ThinPrep interpretation classified 29 of 32 carcinomas (91%) as positive and three as atypical favoring malignant (sensitivity = 100%). The conventional smear interpretation classified 28 of 31 carcinomas (90%) as positive and three as atypical favoring malignant (sensitivity = 100%). The ThinPrep interpretation classified 42 benign lesions as negative (23 cases), negative consistent with fibroadenoma (8 cases), atypical favoring benign (10 cases), and atypical favoring malignant (1 case) (specificity = 74%). The conventional smear interpretation classified 40 benign lesions as negative (25 cases), negative consistent with fibroadenoma (12 cases), and atypical favoring benign (3 cases) (specificity = 93%). ThinPrep was less specific, but the difference was not statistically significant (P = 0.065). In summary, ThinPrep aspirates had greater cellularity and better nuclear detail than conventional smears, and were just as sensitive in identifying the carcinomas. The difference in specificity between the two techniques was not statistically significant (P = 0.065). Diagn. Cytopathol. 1999;21:137-141.     

Antineuron specific enolase staining reactions in sarcomas and carcinomas: its lack of neuroendocrine specificity.

A commercially available polyclonal antiserum (Dakopatts) raised against bovine neuron specific enolase (NSE) was reacted with 197 sarcomas, 32 carcinomas, 11 carcinoid tumours and 20 malignant melanomas to assess its specificity for neuroendocrine tumours. All the tumours had been fixed in formalin and embedded in paraffin. Positive tumour cells were found in two of 11 squamous cell carcinomas, one of 11 adenocarcinomas, 10 of 10 oat cell carcinomas, 11 of 11 carcinoid tumours, 16 of 20 malignant melanomas, four of seven clear cell sarcomas, nine of 25 leiomyosarcomas, four of 22 rhabdomyosarcomas, one of seven angiosarcomas and one of 20 synovial sarcomas.     

Ultrasound-guided fine-needle aspiration biopsy of the thyroid: role of on-site assessment and multiple cytologic preparations

Several studies have shown that ultrasound guidance can serve as a valuable aid in improving the diagnostic yield of fine-needle aspiration (FNA) biopsy of thyroid nodules. In this study, we evaluated the combined impact of ultrasound-guidance, rapid on-site evaluation of FNA specimens, and different cytologic preparations (fresh and alcohol-fixed smears, Millipore filter) and staining methods (Diff-Quik and Papanicolaou stains) on the diagnostic yield of thyroid FNA. Ultrasound-guided FNA was performed on 282 patients (313 cases) between November 1997 and April 1999. The diagnostic categories included: benign (198 cases, 63.2%); indeterminate (42 cases, 13.4%); suspicious for follicular variant of papillary carcinoma (26 cases, 8.3%), malignant (32 cases, 10.1%); and nondiagnostic (15 cases, 5%). The nondiagnostic cases also included 6 cystic lesions without any solid component and 3 thyroid-bed aspirations. After excluding these, the nondiagnostic rate was only 2%. Histological follow-up was available in 77 (77/313) cases. The concordance rate between cytological and histological diagnosis was 100% in malignant, 67% in suspicious, and 56% in indeterminate cases. All cases with histologic follow-up were selected to evaluate the independent diagnostic efficacy of each aforementioned cytologic staining method. A definite diagnosis could be made solely on the basis of air-dried, Diff-Quik-stained preparations in 50 (65%), alcohol-fixed, Papanicolaou stained smears in 68 (88%), and Millipore filter preparations in 70 (91%) cases. We conclude that ultrasound-guided FNA combined with on-site evaluation and different cytologic preparations can significantly improve the diagnostic accuracy of thyroid FNA specimens.     

Utility of reflex Gomori methenamine silver staining for Pneumocystis jirovecii on bronchoalveolar lavage cytologic specimens: a review

Pneumocystis jiroveci (Pj; formerly Pneumocystis carinii) is an opportunistic pathogen causing life-threatening pneumonia (Pneumocystis pneumonia) in immunosuppressed individuals. Its diagnosis is dependent on identification in bronchoalveolar lavage (BAL) specimens. Gomori's methenamine silver nitrate (GMS) stain has been advocated to highlight the organisms in BAL specimens. This study was performed to determine the utility of reflex GMS staining on all BAL specimens for identifying Pj.All BAL specimens from years 2000 to 2004 were processed as cytospins and stained with Papanicolaou (Pap) and GMS stains. A total of 2,984 BAL specimens were identified. A total of 116 (3.9% of total BAL) BAL specimens were diagnostic of Pj. The diagnostic specimens were grouped as follows: 103 (88.8% of total positive cases) Pj identified with both Pap and GMS staining; 11 (9.5% of total positive cases) Pj identified only with Pap staining; and 2 (1.7% of total positive cases) Pj identified only with GMS staining.In conclusion, the prevalence of Pj in BAL specimens is 3.9%, which can be attributed to improved management of immunocompromised patients. Performing reflex GMS staining on all BAL specimens does not improve the diagnostic identification of Pj since the majority (98.3%) of diagnoses can be rendered on Pap stained slides. A cost analysis for GMS staining on 2,879 GMS-negative BAL specimens was estimated at $143,950. Thus, from diagnostic and cost benefit perspectives, GMS staining can be recommended only on cases where Pap stain is negative, and the clinical presentation is consistent with Pneumocystis pneumonia.     

Differentiation of herpes simplex virus type 1 and type 2 by immunofluorescence: discriminative staining by labelled IgG preparations.

While evaluating herpes simplex virus (HSV) typing by indirect immunofluorescence staining, an undesired specific staining pattern turned out to be a reliable marker for herpes simplex type 1. Cells infected with herpes simplex type 1 displayed clear staining with a FITC-conjugated antiglobulin preparation, also in the absence of herpes simplex-specific antibodies. Using the same conjugate, herpes simplex type-2-infected cells exhibited no fluorescence. The particular type of staining observed was influenced by neither the anatomical site of origin of the virus isolate nor the cell type used for virus preparation. Herpes simplex type 1-specific fluorescence was only obtained with the use of FITC-conjugates possessing anti-IgG activity. Both reliability and specificity of this discriminating procedure as a diagnostic tool has been established by typing 282 virus isolates over a period of 4 years.     

Intercellular adhesion molecule expression in ductal carcinoma of the breast: correlation of immunohistochemical staining with cytologic smear pattern

Recent studies suggest that altered expression of intercellular adhesion molecules (ICAM) in ductal carcinoma of the breast is associated with a higher incidence of metastases and decreased patient survival. In addition, the presence of significant cellular dyscohesion in cytologic smear preparations has been found to correlate with the presence of regional and distant metastases in a subset of patients. In this study, we correlate the smear pattern in preparations taken directly from surgically excised breast tumors with their immunohistochemical staining pattern, using antibodies directed against a panel of ICAM. We found excellent correlation, as all three tumors with an extremely high degree of tumor cell cohesion showed strong staining with all ICAM antibodies in the vast majority (>/=90%) of tumor cells in corresponding tissue sections. In contrast, five tumors displaying a largely dyscohesive smear pattern demonstrated decreased staining (相似文献   

Immunohistochemical staining of cytologic smears with MIB-1 helps distinguish low-grade from high-grade neuroendocrine neoplasms

Neuroendocrine neoplasms (NENs) of the lung and gastrointestinal tract constitute a pathologic and biologic spectrum of tumors. Accurate cytologic diagnosis of a neuroendocrine neoplasm is important since definitive treatment frequently is based on low- and high-grade categories without histologic sampling. In many instances, however, low- and high-grade NENs share cytologic features, hindering a precise classification. Since the histologic diagnostic criteria for separation of low- from high-grade categories can be based on the proliferation rate, we proposed to evaluate the usefulness of the immunocytochemical stain for the proliferation marker MIB-1 in the grading of NENs. Cytologic preparations of 63 NENs were retrieved from the files of Memorial Sloan-Kettering Cancer Center, New York, NY. One representative alcohol-fixed slide from each case was destained and restained immunocytochemically for MIB-1. When MIB-1 immunoreactivity was considered, all low-grade NENs showed immunoreactivity in fewer than 25% of the neoplastic cells, and all high-grade NENs demonstrated immunoreactivity in more than 50% of neoplastic cells. Our study demonstrates that MIB-1 dramatically stratifies NENs as low-grade or high-grade. Therefore, the proliferation index also correlates with grade of NEN in cytology specimens.