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Peroxisome proliferator-activated receptor-γ (PPAR-γ) is a ligand-activated transcription factor, implicated in various aspects of cancer biology, such as differentiation, proliferation, invasion and angiogenesis. The present study aimed to evaluate the clinical significance of PPAR-γ in mobile tongue squamous cell carcinoma (SCC).PPAR-γ protein expression was assessed immunohistochemically on 49 mobile tongue SCC tissue samples obtained from an equal number of patients. PPAR-γ expression and intensity of immunostaining were statistically analyzed in relation with clinicopathological characteristics, mitotic index and patients' survival.Elevated PPAR-γ expression was more frequently observed in patients with reduced depth of invasion (P = 0.0111). Moderate/intense PPAR-γ staining intensity was more frequently observed in patients with no evidence of muscular infiltration (P = 0.0229) and reduced depth of invasion (P = 0.0176). Mobile tongue SCC patients presenting enhanced PPAR-γ expression had significantly longer overall and disease-free survival times compared to those with low PPAR-γ expression (log-rank test, P = 0.0162 and P = 0.0114, respectively).PPAR-γ immunoreactivity in mobile tongue SCC was correlated with clinicopathological characteristics crucial for patients' management and prognosis. PPAR-γ may be considered as a useful prognostic marker in mobile tongue SCC and a potential therapeutic target for tongue cancer chemoprevention and treatment.

Myxofibrosarcoma (MFS) is a well-established nosologic entity different from the myxoid variant of malignant fibrous histiocytoma. In an attempt to better define the representative cytologic criteria of MFS, we undertook a review and a reanalysis of a series of 14 cytology samples in 12 patients whose tumors were diagnosed as MFS. Using FNA technique and reviewing the original diagnoses, 11 cases were diagnosed as malignant and three as benign tumors. The cytologic diagnosis of MFS was accurate in seven cases (2 primary tumors, 4 recurrences, and 1 metastasis). Four cases were classified malignant myxoid sarcoma (1 primary and 3 recurrences), whereas three cases (2 primary and 1 recurrence) were false-negative. The smears were cell-rich in 12 cases and cell-poor in two cases. They were constantly composed of isolated and regular small spindle-shaped and stellated cells with elongated nuclei containing small inconspicuous nucleoli. Cytoplasm was pale with elongated processes. Clusters of wavy spindle-shaped cells, round cells without specific pattern, moderate cytonuclear atypia, and abundant myxoid background as well as curvilinear vascular structures were always seen. In the vast majority of cases, the cytologic distinction of MFS from other low-grade myxoid lesions is difficult. Entities such as myxoid MFH, myxoid liposarcoma (MLP), myxoid DFSP, and myxoma should be considered in the differential diagnosis. The cytological misdiagnosis is of limited clinical consequence because FNA findings suggestive of a myxoid tumor will be indicative for a surgical removal followed by the histopathological analysis. Diagn. Cytopathol. 2010. © 2009 Wiley-Liss, Inc.

Objectives To retrospectively report the clinical and therapeutic features of children with nasopharyngeal carcinoma (NPC) treated by chemotherapy and doses adapted of radiotherapy. Patients and Methods From 1978 to 2005, 34 children were treated for NPC. All histologic and/or cytologic samples and CT scans were reviewed. Cervical nodal irradiation was reduced (

Whereas neuroblastoma (NB) with MYCN amplification presents a poor prognosis, no single marker allows to reliably predict outcome in tumours without MYCN amplification. We report here an extensive analysis of 147 NB samples at diagnosis, without MYCN amplification, by chromosomal comparative genomic hybridisation (CGH), providing a comprehensive overview of their genomic imbalances. Comparative genomic hybridisation profiles showed gains or losses of entire chromosomes (type 1) in 71 cases, whereas partial chromosome gains or losses (type 2), including gain involving 17q were observed in 68 cases. Atypical profiles were present in eight cases. A type 1 profile was observed more frequently in localised disease (P

Background Neuroblastoma is a heterogeneous pediatric disease. Most patients with localized disease usually have a favorable prognosis, but patients with advanced disease have a poor prognosis despite combination chemotherapy. Treatment failure may be attributable to resistance to cytotoxic drugs. Procedure Using quantitative RT-PCR, we investigated the clinical significance of the level of mRNA expression of multidrug resistance genes (MDR1, MRP1, MRP5, LRP) in a series of 29 advanced neuroblastoma samples. Results At the end of induction chemotherapy, 48% of patients achieved a clinical complete response, 28% achieved a partial response or stable disease, and 24% presented progressive disease. MDR1 mRNA overexpression (i.e., mRNA level >2 copies of MDR1 gene) was observed in 74% of samples, and MRP1, MRP5, LRP overexpression was observed less frequently (30, 33, and 33% of samples, respectively). None of these parameters were predictive of response, relapse, or survival. However, clinical response to treatment was highly predictive of relapse-free survival and overall survival. Conclusions High expression of these multidrug resistance genes in advanced neuroblastoma is not the main parameter of response to cytotoxic drugs; clinical response to treatment remains the most important parameter in predicting the prognosis of patients with advanced neuroblastoma, until other relevant laboratory parameters have been identified. Pediatr Blood Cancer 2007;48:311–317. © 2006 Wiley-Liss, Inc.

To determine diagnostic cytomorphologic features of rhabdomyosarcoma (RMS) on fine-needle aspiration (FNA) material, the cytologic material and corresponding histologic slides of 180 tumors obtained from 109 patients were reviewed. Fifty eight (32.2%) tumors were primary, 34 (18.9%) recurrent, and 88 (48.9%) metastatic. A review of original cytology reports revealed that 176 of 180 (97.8%) tumors were either diagnosed accurately or as round cell sarcoma, while 3 (1.7%) were reported as suspicious. In one case (0.5%), the material was unsatisfactory. No false negative samples were seen. When FNA morphology was correlated with different histological subtypes, the alveolar subtype RMSs were more cellular than the nonalveolar ones (91.4% vs. 64.9%). Similarly, alveolar subtype RMSs compared with nonalveolar ones exhibited more rhabdomyoblastic cells (77.1% vs. 52.7%), alveolar structures (67.6% vs. 10.8%), giant, multinucleated cells (22.9% vs. 6.7%), mitotic figures (57.1% vs. 18.9%), and cyto-nuclear atypia (77.1% vs. 43.2%). Inversely, spindle-shaped cells were more frequently seen in nonalveolar versus alveolar RMSs (37.8% vs. 20.9%). Diagn. Cytopathol. 2007;35:482–487. © 2007 Wiley-Liss, Inc.

Purpose Phosphorylated epidermal growth factor receptor (pEGFR) activates several signaling pathways, resulting in tumor-promoting cellular activities, and has been implicated in malignant transformation and disease progression. The present study evaluated the clinical significance of pEGFR protein expression in mobile tongue squamous cell carcinoma (SCC). Materials and Methods The present cohort study included 48 patients with mobile tongue SCC. We evaluated whether pEGFR immunohistochemical protein expression is associated with clinical variables and patient outcome. Results Of the 48 patients included in the present cohort study, 25 were men and 23 were women. The median patient age was 60 years (interquartile range 53 to 72). pEGFR protein expression was significantly increased in well-differentiated tumors compared with poorly differentiated tumors ( P = .001). Elevated pEGFR protein expression was significantly more frequently observed in mobile tongue SCC cases with a well-defined tumor shape and an earlier disease stage ( P = .010 and P = .019, respectively). Patients with mobile tongue SCC presenting with elevated pEGFR expression had longer overall and disease-free survival times compared with those with low pEGFR expression ( P = .015 and P = .006, respectively; log-rank test). On multivariate analysis, pEGFR expression proved to be an independent prognostic factor of both overall and disease-free survival ( P = .008 and P = .044, respectively; Cox regression analysis). Conclusions The results of the present study support evidence that the pEGFR signaling pathway might be implicated in the malignant transformation of mobile tongue SCC. Additional studies are recommended to validate whether pEGFR could be used as a potential biomarker and therapeutic target in mobile tongue SCC.

Schwannoma accounts for one of the most common benign mesenchymal neoplasms of soft tissues. Although it is well defined in the cytology literature, particular histologic subtypes such as “ancient,” cellular and epitheliod variants could be a source of diagnostic difficulties. We have reviewed cytology aspirates and corresponding histologic sections from 34 schwannomas diagnosed at Institut Curie. Histologically, 24 cases were classic, 5 were “ancient,” 4 were cellular, and 1 was epithelioid schwannomas. No example of melanotic schwannoma was recorded. Original cytologic diagnosis was schwannoma in 13 (38.2%) cases, benign soft tissue tumor in 11 (32.4%), pleomorphic adenoma in 2 (6%) cases, angioma in 1 (2.9%) case, nodular fasciitis in 1 (2.9%) case, suspicious in 3 (8.8%) cases, and not satisfactory in 3 (8.8%) cases. There were no major differences between classical, “ancient,” cellular, and epithelioid variants on cytology smears. Myxoid stroma, mast cells, and intranuclear inclusions were limited to classical subtype. Similarly, cyto-nuclear atypia was more frequent in classical subtype than in other subtypes. Schwannoma should be differentiated from well-differentiated malignant peripheral nerve sheath tumor, neurofibroma, and pleomorphic adenoma, in the last instance particularly for head and neck lesions. Diagn. Cytopathol. 2006;34:517–522. © 2006 Wiley-Liss, Inc.

Resume La cytologie des sereuses est deja assez complexe et souvent la cause d'embuches. C'est pour cela que l'on doit savoir detecter les anomalies cellulaires liees a l'action des radiations ionisantes ainsi qu'a la chimiotherapie. Dans cet article, sont decrits les modifications rencontrees au cours des reactions aigues et chroniques apres irradiation ainsi que les effets directs et indirects de la chimiotherapie sur les cellules mesotheliales. Enfin, pour chaque versant de la pathologie des diagnostics differentiels sont proposes.

Recent biological studies have classified breast carcinomas into HER2-overexpressing, estrogen receptor-positive/luminal, basal- and normal-like groups. According to this new biological classification, the objectives of our study were to assess the clinical, morphologic and immunophenotypic characteristics of adenoid cystic carcinoma of the breast in order to classify this subtype of breast carcinoma. A total of 18 cases of adenoid cystic carcinoma were identified from the Institut Curie files. Clinical information was available for 16 patients with a median follow-up of 6.5 years. Morphologically, all tumors were graded according to the system defined by Kleer and Oberman (histologic and nuclear grade). Immunophenotype was assessed with anti-ER, PR, HER-2, KIT, basal (CK5/6) and luminal cytokeratins (CK8/18) and p63 antibodies. One out of 18 tumors was nuclear grade 1 (16%), nine were nuclear grade 2 (50%) and eight were nuclear grade 3 (44%). All cases were estrogen receptor, progesterone receptor and HER-2 negative. Epithelial cells were strongly positive around glandular lumina with one or both cytokeratins, identifying the coexistence of CK5/6+ cells, CK5/6 and CK8/18+ cells, CK8/18+ cells and p63+ cells. All cases (100%) were also KIT positive. In all, 15 patients were treated by surgery. Nine of them received adjuvant radiotherapy. Follow-up was available for 16 patients. In all, 14 patients were alive. Two of them, initially treated by surgery only, presented a local recurrence. Two patients died (one of them treated by radiation therapy only died from her disease). Our study shows that adenoid cystic carcinoma of the breast is a special, estrogen receptor, progesterone receptor, HER-2 negative and highly KIT-positive, basal-like breast carcinoma, associated with an excellent prognosis. This highly specific immunophenotype could be useful to differentiate adenoid cystic carcinoma of the breast from other subtypes of breast carcinoma such as cribriform carcinoma.

Tumor infiltrating lymphocytes (TIL) display activation markers and their presence is often associated with a favorable outcome. The role of tumor antigens in T cell recruitment into tumors is unclear. In an attempt to address this issue, we purified lymphocytes from breast tumor or nontumor, mammary tissue from patients, and normal mammary tissue from healthy individuals. In all groups, including healthy individuals, the majority of cells displayed an effector/memory (CD45RA lo /CD27 1/2 ) phenotype and quite surprisingly the early and transient activation marker CD69, thus, questioning the tumor antigen specificity of TIL. Because the human repertoire is diverse, the T cells found in the tumors could recognize both self/tumor and environmental antigens through cross-reactivity. To test this hypothesis, we used two anti-male HY monospecific TCR transgenic mouse models. We found an infiltration of HY negative tumors by the CD4 1 and CD8 1 monoclonal T cells after priming with HY positive cells in the periphery. Thus, the presence of activated effector/memory T lymphocytes in tumors can be independent of reactivity against tumor antigens. These results suggest that to find activated effector T cells in a tissue does not always mean that a specific immune response is taking place. ' 2005 Wiley-Liss, Inc.

BACKGROUND Large retrospective archival studies of S-phase fraction (SPF) measured by DNA flow cytometry in patients with breast carcinoma have shown its long-term prognostic relevance. However, contradictory results have also been reported, some of them being related to the various methods of SPF calculation using different commercially available software. METHODS DNA flow cytometric list mode data, initially computed with Cellfit software, were blindly reanalyzed using Modfit and Multicycle software. The data, acquired prospectively between 1990 and 2003 from cytologic fine-needle aspiration biopsy samples of 397 patients with breast carcinoma, were compared with patient outcome with a median follow-up of 99 months (8.2 yrs). RESULTS Measurement of SPF was successful in 321 (81.7%), 362 (92.1%), and 335 cases (85.2%) by means of the Cellfit, Modfit, and Multicycle software programs, respectively. In 306 cases (77.9%), SPF values were obtained using all 3 methods. Comparisons between SPF measurements showed a good agreement between Modfit and Multicycle computations. In the series of 306 patients, SPF median values of 2.5%, 4.3%, and 5.45% for Cellfit, Modfit, and Multicycle, respectively, were also found to be statistically different. Regardless of the software used, high SPF defined as above the median value was an independent factor of prognosis in a multivariate analysis including all traditional clinicopathologic parameters. It is noteworthy that this was also observed in the subgroups of patients either treated by primary surgery for an early tumor (n = 133) or by neoadjuvant chemotherapy for a locally advanced breast carcinoma (n = 173). CONCLUSIONS The data in the current study supported the prognostic relevance of SPF measurement in predicting the long-term overall survival of patients with early-stage or locally advanced invasive breast carcinoma. Cancer (Cancer Cytopathol) 2005. © 2005 American Cancer Society.

The preoperative cytological diagnosis of malignant phyllodes tumor (MPT) is challenging due to the heterogeneity of its clinical, radiological, and morphological presentation. To better define the cytopathological characteristics of MPT, we reviewed 22 examples seen at the Institut Curie. The original cytologic diagnosis was benign breast tumor in four cases (18.2%), suspicious in seven cases (31.8%) (low-grade phyllodes tumor in six cases needing histological evaluation, suspicious of sarcoma in one case), and malignant in 11 cases (50%). Smears were composed of different proportions of clusters of epithelial cells (68.2%), phyllodes fragments (31.8%), spindle cells within stromal tissue (31.8%), isolated spindle-shaped or round cells (45.5%), and bipolar naked nuclei (22.7%). Giant cells and mitotic figures were also occasionally seen. The cytological findings on smears were correlated with histopathological observations. One of the difficulties to reach an accurate cytological diagnosis for MPT is the frequent overwhelming of clearly malignant sarcomatous cell by the presence of largely predominant clusters of epithelial cell. Diagn. Cytopathol. 2004;31:71–76. © 2004 Wiley-Liss, Inc.

The cell proliferation rate has been correlated to the response of breast carcinomas to preoperative chemotherapy (CT) and to disease outcome. However, this parameter is not yet used to select which tumours should be treated with preoperative CT. Furthermore, there is no consensus in the method used to evaluate cell proliferation. In poor prognosis breast carcinomas (PPBCs) treated by intensive preoperative CT, we compared the predictive value of S phase fraction (SPF), mitotic index (MI) and Ki67. We also evaluated the prognostic significance of the variation of the MI after CT. A series of 55 T2-T4N0N1M0 breast carcinomas were treated with 4 cycles of cyclophosphamide, 5-fluorouracil (5-FU) and doxorubicin. SPF was determined by flow cytometry on pretherapeutic needle aspiration products. NU and Ki67 were evaluated on pre-therapeutic biopsy samples and on the tumours after CT. Fifteen patients (27%) had a pathological complete response (pCR), whereas 40 (73%) had residual disease. All three proliferative markers were found to have predictive value, but this value was higher for MI than for SPF (P = 0.04) and Ki67 (P = 0.03): the rate of pCR was 50% in cases with MI > 17/3.3 mm(2), but was only 70% in cases with MI under this threshold (P = 0.0003). A significant decrease of MI (mean 10.97) was observed after CT (P 0.001). Furthermore, we observed that even for patients with residual tumour, the variation of MI after CT was a prognostic parameter and overall survival. The sequential analysis of MI in breast cancers treated by preoperative CT thus provides a surrogate for predicting long-term outcome. (C) 2004 Elsevier Ltd. All rights reserved.

We have reviewed cytopathology and the corresponding histopathology material of 86 liposarcomas (55 patients) seen at Institut Curie. The liposarcomas (LS) were well differentiated in 14 cases (9 pure, 2 dedifferentiated, 3 sclerosing), 64 myxoid, and 8 pleomorphic. Twenty-four tumors were primary, 34 recurrent, and 28 secondary. Smears in LS were composed in different proportions of round, spindle cells, lipoblasts, and myxoid and vascular arborizing structures. Pure well-differentiated LS were frequently composed of lipoblasts, and round or spindle cells were occasionally seen. Dedifferentiated and sclerosing liposarcomas were composed of spindle or round cells, but lipoblasts were also occasionally present. Myxoid or vascular arborizing structures were absent. Myxoid LS (including round and spindle cell LS) frequently showed a myxoid background and less frequently vascular arborizing structures. Tumor cells were round or spindle. Lipoblasts were also seen. Pleomorphic LS were composed of an admixture of all cellular and stromal elements. Well-differentiated LS should be distinguished from hibernoma and spindle cell lipoma, and myxoid LS from myxoma, myxoid chondrosarcoma, chordoma, myxoid leiomyosarcoma, and myxoid malignant fibrous histiocytoma. The demonstration of the specific translocation t(12;16)(q13;p11) of myxoid LS is very helpful to establish the diagnosis. Pleomorphic LS should be differentiated from other high-grade sarcomas, whenever possible. Diagn. Cytopathol. 2004;30:307–312. © 2004 Wiley-Liss, Inc.

Histone synthesis and chromatin assembly are mainly associated with DNA replication and are thus intimately involved in cell cycle regulation. The expression of key components involved in these events in human cells was studied in relation to cell-proliferative status. Among several chromatin assembly factors, chromatin assembly factor (CAF)-1 stood out as the most discriminating marker of the proliferative state. We show, using both immunofluorescence and Western blot analysis, that the expression of both CAF-1 large subunits, p150 and p60, is massively down-regulated during quiescence in several cell lines. Upon exit from the quiescent state, the CAF-1 subunits are re-expressed early, before DNA replication. The amounts of either total or chromatin-associated pools of CAF-1 proteins correlate directly with cell proliferation. Regulation of CAF-1 expression is partly controlled at the RNA level, as shown by quantitative reverse transcription-PCR and Northern blot experiments. Biological material from benign and malignant human breast tumors analyzed by immunocytochemistry and immunohistochemistry exhibits a strong positive correlation between CAF-1 p60 expression and the following proliferation markers: S-phase fraction (r = 0.84, P < 0.0001); Ki-67 (r = 0.94, P < 0.0001); and proliferating cell nuclear antigen (r = 0.95, P = 0.0001). We discuss the advantages of using CAF-1 to assess cell proliferation. High CAF-1 p60 levels are also shown to be associated with various prognostic factors. Our data highlight the precise association of CAF-1 expression with the proliferative state and validate the use of this factor as a useful proliferation marker and prognostic indicator in malignant and benign breast lesions.

Dermatofibrosarcoma protuberans (DFSP) is a nodular cutaneous mesenchymal tumor of intermediate malignancy. Studies on fine-needle aspiration of DFSP are extremely rare; to our knowledge, only 33 cases have been reported. We have reviewed 14 examples of DFSP in 13 patients. Ten primary tumors were aspirated before surgical biopsy and four recurrent lesions (all from superficial lesions) were also investigated by fine-needle aspiration. All smears were surprisingly homogeneous and composed of isolated spindle cells in all cases (one unsatisfactory smear is excluded). Tissue fragments with a stroriform pattern were seen in 11 cases, fibrillary stromal fragments in 10 cases, naked nuclei in 8 cases, slight to moderate cytonuclear atypia in 5 cases. Mitotic figures, myxoid background, mast cells, and dispersed adipocytes were rare. Giant cells, necrosis, or marked cytonuclear atypia were not seen. DFSP shares morphological characteristics of some low-grade spindle-cell neoplasms. It should be differentiated from other benign low- and intermediate-grade spindle neoplasm such as low-grade fibrosarcoma, fibromyxosarcoma, low-grade malignant peripheral nerve sheath tumor, benign peripheral nerve sheath tumor, nodular fasciitis, and fibrous histiocytoma.

Clinical management of in situ carcinoma of the breast is different from invasive carcinomas. Thus, it is important to find cytomorphologic criteria to distinguish between these two entities. The current study is designed to assess whether, by applying strict cytologic criteria, the status of stromal invasion can be predicted. In this retrospective study, 223 consecutive nonpalpable tumors sampled by ultrasound-guided breast fine-needle aspirates with diagnosis of in situ and invasive carcinoma with histologic follow-up evaluation were retrieved. Ten cytologic parameters were evaluated, which included cellular clustering, eosinophilic differentiation, necrosis, tubular structures, dirty background, nuclear anisonucleosis, cellular pleomorphism, cribriform pattern, tubular structures, and stromal infiltration. Among all the parameters examined, stromal infiltration was the most powerful predictor of status of invasion. Stromal infiltration was significantly higher in invasive (88%) than in situ (11%), P < 0.0001. In contrast, cribriform pattern (16% vs 36%) and necrosis (19% vs 59%) were more frequently seen in situ than in invasive carcinomas with P-values of 0.0008 and

To characterize the cytological features of angiosarcomas, we reviewed the fine-needle aspiration material and corresponding histologic sections of 29 tumors in 23 patients. Histologically, 24 tumors were of the classic type, and 5 were epithelioid angiosarcomas. The original corresponding cytologic diagnoses were as follows: angiosarcoma, 17 cases; sarcoma not otherwise specified, 8 cases; and rhabdomyosarcoma, 1 case. Three samples were cell-poor and were considered suspicious of malignancy. The review of cytology samples showed that smears were cell-rich in 17 tumors and cell-poor in 12 tumors. A hemorrhagic background was present in 9 cases. Tumor cells were polymorphous, including spindle-shaped, round to oval, and polygonal epithelioid cells and giant cells in different proportions. Erythrophagocytosis was seen in 12 tumors. Smears of classic angiosarcomas were polymorphous and lacking specific characteristics, whereas smears of epithelioid tumors were morphologically similar and composed of round to oval and polygonal, epithelial cells frequently arranged in clusters, and showing erythrophagocytosis. The wide spectrum of cellular components of angiosarcomas accounts for the difficulty in establishing accurate tumor typing, particularly with cell-poor samples and low-grade classic angiosarcoma. Entities to consider in the differential diagnosis are carcinoma, epithelioid sarcoma, pleomorphic rhabdomyosarcoma, and malignant melanoma.

To better define the cytological features of various leiomyosarcoma (LMS) variants, we reviewed the fine-needle aspiration material and the corresponding histologic sections of 96 tumors in 68 patients. Histological variants of LMS were as follows: 80 (83.3%) were of the classical/usual, seven (7.3%) were epithelioid, and nine (9.4%) were myxoid. Review of original cytology reports showed that 23 (24%) tumors were diagnosed as LMS and 69 (71.8%) as other types of malignancies. Two (2.1%) cases were reported as suspicious and two (2.1%) were unsatisfactory. The classical variants of LMS were characterized cytologically by various proportions of spindle-shaped, cohesive, small- or large-sized cells arranged in parallel alignment. Large spindle, round, binucleated, giant cells with intracytoplasmic granulations were frequently seen. Blunt-ended nuclei, intranuclear inclusions and mitotic figures were occasionally seen, as well as stromal fragments. The epithelioid tumors were composed of an admixture of small and large, spindle-shaped and round cells, also arranged in parallel alignment. Tumor cells with granular cytoplasm, blunt-ended nuclei, intranuclear inclusions, mitotic figures, fibrous or myxoid stroma were not observed. The myxoid tumors disclosed large amounts of background myxoid matrix containing large spindle-shaped and giant cells. Entities such as leiomyoma, malignant peripheral nerve sheath tumor, monophasic synovial sarcoma, and malignant fibrous histiocytoma should be considered in the differential diagnosis of LMS of the classical type. Epithelioid leiomyoma may share similar cytological features with epithelioid LMS. The cytological features of the myxoid variant of LMS can be easily confused with other types of benign and malignant mesenchymal tumors depicting degenerative myxoid changes and/or a myxoid matrix component.

Synovial sarcoma (SS) is a high-grade malignant soft tissue tumor that manifests different phenotypic subtypes that may render their cytologic evaluation challenging. Although several cytologic studies of SS have been published, correlative studies of cytologic and corresponding histologic features are limited. To better define the cytological features of various SS forms, we reviewed the cytologic and the corresponding histologic material of 56 tumors from 36 patients. Classical patterns were defined as dispersed or small clusters of cells with bland chromatin, inconspicuous nucleoli, oval to spindle-shaped cytoplasm and branching tumor tissue fragments, vessel stalks, acinar structures in scant mucin background, seen in all 53 (94.7%) cellular cases. Epithelial, squamous, round cells, mast cells, necrosis, comma-like nuclei, marked nuclear atypia, secretory mucin, and rosette-like structures were also occasionally observed. Comparing the histological subtype we noted that epithelial cells and secretory mucin were restricted to biphasic SS, round cells to poorly differentiated SS, and comma-like nuclei to monophasic fibrous SS. We conclude that the classical pattern is highly suggestive of SS of all three monophasic, biphasic, or poorly differentiated subtypes. These characteristics, along with molecular genetic studies, may improve the cytologic diagnosis of SS.

6021 Background: Afatinib, a pan-HER irreversible tyrosine kinase inhibitor, demonstrated limited antitumor activity compared to methotrexate in unselected recurrent and/or metastatic HNSCC patients (LUX-HN1, Machiels et al, Lancet Oncol 2015). The UNICANCER (GEP 11) PREDICTOR study’s objective was to identify predictive and pharmacodynamic biomarkers of biological activity and efficacy of afatinib (EUDRACT N° 2010-024046-29). Methods: This open-label, randomized, multicentric, controlled, phase II study included untreated patients with operable T2-4N0-2M0 HNSCC of the oral cavity, pharynx and larynx, with a PS < 2, adequate organ function and LVEF > 50%. Patients were randomized (2:1) to: oral afatinib (A) 40mg/day (d) for 14-28d or no treatment (NT). Patients had pre-treatment tumor biopsies, tumor imaging, and PET CT scan, with a 2nd tumor imaging before surgery and a PET scan at D15. Adverse events were classified by NCI CTCAE criteria. Based on the biological primary endpoint of tumor reduction the sample size was designed to identify biomarkers associated with a 20% difference between the study arms. Results: 61 patients were included (A: 41/NT: 20). 2 patients in the NT arm were not analyzed (consent withdrawal, no surgery). 7 patients in arm A received < 14d of treatment, including 6 patients with unacceptable toxicity. Afatinib-related toxicities were: grade (G)1 37%, G2 41%, G3 7%, G4 5%, and G5 0%. G≥3 toxicities were mainly gastrointestinal. Partial responses (RECIST1.1) were observed in 3 patients (7.3%) in arm A versus none in the NT arm (p = 0.018). Progressive disease was not observed in arm A versus 3 (16.6%) in the NT arm. Partial responses on PET CT scan by PERCIST were observed in 15/31 evaluable patients (48%) in arm A versus 1/15 (6.7%) in the NT arm (p = 0.005). Conclusions: Afatinib given to HNSCC patients in the preoperative setting is safe and is associated with improved response according to RECIST1.1 and PERCIST compared to no treatment. Clinical trial information: NCT01415674.

Microfluidic systems bear promise to provide new powerful tools for the molecular characterization of cancer cells, in particular for the routine detection of multiple cancer biomarkers using a minute amount of the sample. However, taking miniaturized cell-based assays into the clinics requires the implementation and validation of complex biological protocols on chip, as well as the development of disposable microdevices produced at a low cost. Based on a recently developed microfluidic chip made of Cyclic Olefin Copolymer for cell immobilization with minimal dead volume and controlled shear stress, we developed a protocol performed entirely in the liquid phase, allowing the immobilization and fixation of cells and their quantitative characterization by fluorescence in situ hybridization. We demonstrated first in cell lines and then in two clinical case studies the potential of this method to perform quantitative copy number measurement and clinical scoring of the amplification of the ERBB2 gene, a decisive biomarker for the prescription of HER2+ related targeted therapies. This validation was performed in a blind protocol in two clinical case studies, in reference to the gold standard and clinically used method based on glass slides. We obtained a comparable reproducibility and a minor difference in apparent amplification, which can be corrected by internal calibration. The method thus reaches the standard of robustness needed for clinical use. The protocol can be fully automated, and its consumption of samples and DNA probes is reduced as compared to glass slide protocols by a factor of at least 10. The total duration of the assay is divided by two.

Background The B-subunit of Shiga toxin (STxB) specifically binds to the glycosphingolipid Gb3 that is highly expressed on a number of human tumors and has been shown to target tumor cells in mouse models and ex vivo on primary colon carcinoma specimen. Methods Using a novel ex vivo STxB labeling (ESL) method we studied Gb3 expression in cytological specimens of primary human breast tumors from 107 patients, and in synchronous lymph node metastases from 20 patients. Fluorescent STxB was incubated with fine-needle aspiration (FNA) specimens, and Gb3 expression was evaluated by fluorescence microscopy. Furthermore, 11 patient-derived human breast cancer xenografts (HBCx) were evaluated for expression of Gb3 by ESL and FACS. In addition, the biodistribution of fluorescent STxB conjugate was studied after intravenous injection in a Gb3 positive HBCx model. Results Gb3 expression was detected in 62 of 107 patients (57.9%), mainly in epithelial tumor cells. Gb3 positivity correlated with estrogen receptor expression (p ≤ 0.01), whereas absence of Gb3 expression in primary tumors was correlated with the presence of lymph node metastases (p ≤ 0.03). 65% of lymph node metastases were Gb3 positive and in 40% of tested patients, we observed a statistically significant increase of metastatic Gb3 expression (p ≤ 0.04). Using concordant ESL and flow cytometry analysis, 6 out of 11 HBCx samples were scored positive. Intravenous injections of fluorescent STxB into HBC xenografted mice showed preferential STxB accumulation in epithelial cells and cells with endothelial morphology of the tumor. Conclusion The enhanced expression of Gb3 in primary breast carcinomas and its lymph node metastases indicate that the development of STxB-based therapeutic strategies is of interest in this pathology. Gb3 expressing HBCx can be used as a model for preclinical studies with STxB conjugates. Finally, the ESL technique on FNA represents a rapid and cost effective method for the stratification of patients in future clinical trials.

The diagnosis of a soft tissue tumor (STT) is a complex process that requires a multidisciplinary team consisting of a medical oncologist, a radiologist, a pathologist, and a molecular biologist. The combination of this information gives highly accurate results. Fine-needle aspiration in this setting is one of the most performed techniques for obtaining, in a noninvasive way, optimal material for morphological, immunohistochemical, and molecular purposes. On the other hand, an accurate tumor diagnosis is a dynamic process, as evidenced by the constant modifications of international putative classifications, which are more and more based on molecular aberrations (2013 World Health Organization classification). In the future, STT classification will certainly be a hybrid of morphology and molecular data and, consequently, will be very "useful" for cytology.

Objectives: To describe cytology patterns in low-grade adenosquamous carcinomas (LGASCs) of the breast. Study Design: Low-grade adenosquamous carcinomas of the breast are a recently described rare variant of primary metaplastic carcinomas characterized by clinical indolence, slow evolution and excellent survival. To date, only 7 cases of LGASC were studied cytologically, and it was demonstrated that LGASC identification was difficult because its cellular components exhibited unspecific and nonsuspicious features. They consisted of irregularly clustered cells without prominent cytonuclear atypia, mitosis or necrosis. The presence of metaplastic cells or keratin debris was helpful in accurate tumor typing. We report here 3 additional cases of LGASC that were initially studied by fine-needle aspiration. Results: We have also encountered diagnostic difficulties and misdiagnosed tumors, since 2 cases were underdiagnosed as ‘suspicious' and only 1 was accurately diagnosed as malignancy. Conclusion: The review of our cases and the literature confirms that, despite its putative metaplastic origin, LGASC is an entity which is difficult to diagnose using classical cytological methods. Moreover, core-needle biopsy as well as frozen sections may also misdiagnose LGASC as a benign breast lesion.

Inflammatory myofibroblastic tumours (IMTs) were initially considered to be benign reactive processes, but cases with an unfavourable outcome have been reported. Moreover, clonal genetic alterations have recently been published in some cases, suggesting that IMT may represent a malignant neoplastic entity. This paper reports a case of IMT that developed in the mammary gland, an unusual site. The histological picture was characterized by a proliferation of spindle cells with little cellular atypia and rare mitoses, associated with a polymorphous inflammatory infiltrate. Their immunophenotype, characterized by the expression of vimentin, smooth muscle actin, and cytokeratins, corresponded to that of myofibroblasts. Cytogenetic analysis revealed the clonal nature of the lesion. The modal karyotype was 48, X, ins(2;X)(q34;p21.2p22.2), +7, del(9)(p23), +19. Including the present observation, a 9p deletion has now been found in three cases of IMT. These observations show that IMT may be a clonal neoplasm, even in sites different from deep soft tissues. Copyright © 2001 John Wiley & Sons, Ltd.

BACKGROUND Because false-positive cytologic diagnoses in breast tumors are rare, few cases have been reported, although their consequences may be highly detrimental to the patient. The authors report the Institut Curie's experience, by using a multidisciplinary approach. METHODS Of 9334 benign breast tumors examined preoperatively for cytologic diagnosis by fine-needle sampling (FNS), the 23 (0.25%) FNS cases considered to be false-positive were retrospectively reviewed and analyzed. RESULTS Tumors were situated close to the nipple in 7 cases and away from the nipple in 16 cases. Tumor stage was T0 for 1 case, T1 for 18 cases, and T2 for 4 cases. Radiologically, six tumors were classified as malignant, seven as indeterminate or suspicious, and nine as benign. Three of six tumors studied by flow cytometry were DNA aneuploid. Based on a multidisciplinary clinicopathologic review, 20 FNS cases were finally classified as false-positive, and the remaining 3 tumors with malignant FNS and subsequent benign histology were classified as true-positive, because local and/or metastatic progression was observed in the short term. CONCLUSIONS The authors' review suggests two categories of false-positive cases: the first in which cytologic benign patterns are overdiagnosed, and the second in which atypical morphologic criteria were present. Nevertheless, as shown by the malignant course in three cases, patients with malignant preoperative FNS and corresponding benign histology always require close clinical follow-up. Finally, surgical overtreatment rate could be decreased if all radiologically benign tumors with positive/suspicious FNS were subject to intraoperative frozen section examination. Cancer (Cancer Cytopathol) 2001;93:132–139. © 2001 American Cancer Society.

Fine-needle samplings of nine examples of primary neuroendocrine carcinoma of salivary glands were evaluated for their cytologic characteristics and were correlated with the corresponding histological sections. Consistent cytological findings were dispersed or loose clusters of poorly differentiated small- to intermediate-sized cells and occasional smudged nuclei. Mild to moderate nuclear pleomorphism, scant or absent cytoplasm, and nuclear molding were also observed. Rosette-like patterns and multinucleated cells were occasionally seen. Immunostaining of one recent case showed positivity for chromogranin and keratin. The differential diagnosis of primary and metastatic tumors with neuroendocrine features of the salivary glands is discussed.

Background Desmoplastic small round cell tumor (DSRCT) is a recently described neoplasm. This entity is well defined, with distinct clinical, pathologic and immunohistochemical features. Molecular studies have shown a specific reciprocal translocation t(11;22)(p13;q12). To our knowledge, no report of DSRCT with molecular confirmation on cytologic material has been reported before. Case Fine needle aspiration (FNA) was performed on an intraabdominal mass in a 37-year-old man. A May-Grunwald-Giemsa-stained preparation showed clusters of small round tumor cells associated with desmoplastic stromal cells, highly suggestive of DSRCT. FNA of a supraclavicular node showed cytologic features similar to those of the primary abdominal mass, including a prominent desmoplastic reaction of the stroma. Immunocytochemical studies showed myogenic and epithelial differentiation. Molecular analysis was performed on FNA, revealing the EWS/ WT1 chimeric transcript and thus confirming the cytologic diagnosis. Conclusion Cytomorphologically, a definitive diagnosis of DSRCT may be difficult, as this tumor bears considerable resemblance to other small round cell tumors. The diagnosis can be confirmed by ancillary techniques, such as immunocytochemistry, and particularly by molecular analysis, which may also be performed on cytologic material.

BACKGROUND Metastatic tumors to the salivary glands are common, but documentation in the cytologic literature has been limited. The current study presents the authors' experience with fine-needle aspiration (FNA) in metastatic nonlymphomatous tumors to the salivary glands. METHODS From a retrospective review of 1675 salivary gland lesions (1535 patients), the authors collected 40 salivary gland lesions (39 patients who had histories of extrasalivary cancer) that were diagnosed cytologically as metastases to the salivary glands and were correlated histologically. RESULTS FNAs of 34 parotid gland and 6 submandibular gland tumors were performed. The cytologic diagnoses of metastases of squamous cell carcinoma (15 cases), melanoma (12 cases), carcinoma (5 cases), rhabdomyosarcoma (3 cases), and retinoblastoma (2 cases) were confirmed (95%) histologically. Two (5%) FNAs were false-negative. CONCLUSIONS In patients who had a history of extrasalivary cancer, cytology examination was very helpful and sufficient for adequate patient management. Cancer (Cancer Cytopathol) 2000;90:350–356. © 2000 American Cancer Society.

The rates of cell proliferation and programmed cell death (apoptosis) reflect tumor cell dynamics and are considered to directly influence biological progression and tumor response to therapy. Bax and Bcl-2 are members of a gene family that influence apoptosis and have been used as surrogate markers in the evaluation of this process. Sixty-three fine-needle tumor samples from an equal number of patients with breast carcinomas were analyzed for Bax, Bcl-2, and DNA content by flow cytometry. The results were correlated with classical clinicopathological parameters. Bax values varied widely among tumors and showed no significant correlation with any of the clinicopathological parameters analyzed. Bcl-2 levels ranged from 4% to 91%, correlated positively with estrogen (P = 0.0004) and progesterone (P = 0.0045) receptor positivity, and were more associated with low S-phase tumor values. In contrast, high S-phase values correlated with estrogen receptor negativity, high grade, and DNA aneuploidy. The study results indicate that Bcl-2 and S-phase analysis of fine-needle samples of breast carcinomas provide a convenient tool for the assessment of these tumors.

BACKGROUND Fine-needle sampling, although a practical and noninvasive method of tissue acquisition, has rarely been used for HER-2/neu fluorescent in situ hybridization (FISH). To assess HER-2/neu gene amplification in mammary carcinoma, FISH signals on cytology and corresponding tissue biopsies were detected visually and measured by image analysis. The results were correlated with patient and tumor characteristics. METHODS In situ HER-2/neu DNA probe hybridization was performed on 61 cytology specimens and on 47 corresponding frozen sections of breast carcinomas. Tumors were classified by visual evaluation as unamplified, moderately amplified, or highly amplified. Multiparametric image analysis was performed using the Discovery automated image analyzer (Becton Dickinson, Leiden, Netherlands). The integrated fluorescence ratio (IFR) was calculated for each sample as the integrated FISH fluorescence of the tumor cells divided by the integrated FISH fluorescence of internal control cells containing two spots. The percentage positive nuclear area (PPN), calculated as the area of FISH fluorescence divided by the area of nuclear DNA fluorescence, and the PPR, ratio of the PPN of the tumor cells divided by the control cells, were also calculated for each sample. RESULTS Visual analysis yielded 46 unamplified and 15 (24.6%) amplified (seven moderately amplified and eight highly-amplified) tumors. Strong (P < 0.001) correlation between results on cytological and histological materials was obtained. The FISH spots on the cytological preparations were more easily visualized and scored than those on the corresponding tissue sections. Visual HER-2/neu signal scoring was strongly correlated with IFR (P = 0.0001) and PPR (P = 0.0001). Within the tumors classified as highly amplified by visual examination, quantitation of the degree of amplification fluorescence signal was possible using image analysis. CONCLUSIONS Cytologic specimens were a suitable and representative source of materials for detection and quantitation of HER-2/neu gene amplification by FISH and image analysis. Cancer (Cancer Cytopathol) 1999;87:312–8. © 1999 American Cancer Society.

Background Restricted presentation of malignancy at both lacrimal and parotid salivary glands are unusual occurrences that may pose clinical and diagnostic difficulties. Methods We applied certain clinicopathological criteria to distinguish between lacrimal and parotid gland primaries in a series of 11 patients with synchronous and/or metachronous tumors of similar histology at both sites. Results Of eight patients with primary lacrimal carcinomas, seven had metachronous metastases to parotid glands. All three patients with primary parotid carcinomas had concurrent metastases to lacrimal glands. Patients with primary lacrimal tumors metastasizing to the parotid gland appear to have better clinical outcome in comparison with those with parotid primary with secondary lacrimal metastases. Conclusion Careful examination of parotid glands in the assessment of any lacrimal gland tumor with high-grade adenocarcinoma morphology is recommended to rule out the possibility of a parotid primary. © 1999 John Wiley & Sons, Inc. Head Neck 21: 512–516, 1999.

Fine-needle sampling (FNS) of five cases of epithelial-myoepithelial carcinoma, three primary tumors, and two local recurrences, was performed preoperatively in five patients. Cytologic diagnoses of malignancy were established in all tumors (three were reported as adenoid cystic carcinoma, two as adenocarcinoma not otherwise specified). Material for cytologic evaluation was satisfactory in all cases. Adenoid cystic carcinoma, polymorphous low-grade adenocarcinoma, papillary cystadenocarcinoma and cellular type of pleomorphic adenoma are the main differential diagnoses.

BACKGROUND Automated image cytometry can allow concurrent quantification of several parameters in each individual cell within a population, opening new possibilities for diagnosis and prognosis. In this study, the authors investigated the capacity of this method for performing a bivariate analysis of DNA ploidy and synthesis in fine-needle samplings obtained without aspiration from breast tumors. METHODS Samplings from 25 unselected cases of ductal infiltrative breast adenocarcinoma and 2 cases of fibroadenoma were analyzed. For each case, 3-5 slides (containing approximately 1000 cells each) were quantified to assess experimental precision. Ploidy was determined by fluorescent staining of DNA using 4,6-diamidino-2-phenylindole (DAPI). Contaminating lymphocytes were taken as internal controls to calculate DNA indices. DNA synthesis was analyzed by immunofluorescent detection of 5-bromodeoxyuridine (BrdU) incorporation. Measurements were compared with flow cytometric data obtained from the same patients. RESULTS Relative error in determination of DNA indices was generally below 5%. Determination of proliferation indices were more variable, with a mean relative error of 25%. Two different populations of BrdU positive cells were detected systematically, one in the diploid and another in the aneuploid fraction. For both cytometric methods, DNA indices were similar in all 27 cases, whereas BrdU labeling indices showed no significant correlation in 13 cases. The remaining cases were not comparable due to lack of flow cytometric data. Labeling indices obtained by image cytometry did not reveal any significant correlation with Scarff-Bloom-Richardson grading or clinical staging. CONCLUSIONS Automated image cytometry allows concurrent measurement of ploidy and cell proliferation within individual breast carcinoma cells. Statistical reliability can be reached with a relative small number of cells (1000), which is crucial for samples in which the cell number is too low for flow cytometry analysis. Visual control for artifact elimination and better characterization of cell populations makes this a powerful tool for tumor cell investigation. Automated image cytometry allows the obtainment of valuable prognostic parameters of traditional flow cytometry with the relatively small number of cells obtained in aspiration procedures. Cancer (Cancer Cytopathol) 1998;84:309-316. © 1998 American Cancer Society.

BACKGROUND In contrast to most major salivary gland malignancies, mucoepidermoid carcinoma has been rarely documented and tentatively accepted as carcinoma ex pleomorphic adenoma. The authors present the cytologic and clinical findings for six examples of these rare tumors. METHODS The six examples were identified during an extensive review of the authors' salivary gland files, and all were high grade mucoepidermoid carcinomas on histologic evaluation. RESULTS Overall, mucoepidermoid carcinomas cytologically presented as nonspecific high grade malignancies and consisted of highly malignant cells, in clusters or isolated, that were rarely associated with intracellular and extracellular mucin. In only two cases, a background of cellular and stromal elements consistent with pleomorphic adenoma was identified. CONCLUSIONS Although it is nearly impossible cytologically to distinguish these lesions from other high grade primary or metastatic carcinomas (such as poorly differentiated squamous cell carcinomas, adenosquamous carcinomas, and salivary duct carcinomas), this limitation is not dramatic, as a cytologic diagnosis of high grade malignancy per se allows for proper preoperative patient management. Cancer (Cancer Cytopathol) 1998;84:231-234. © 1998 American Cancer Society.

Fine-needle samples of 75 non-Hodgkin's lymphomas were investigated for apoptosis immediately and after 24 h of culture after in vitro irradiation (2 Gy, 10 Gy, and nonirradiated controls). Apoptotic cells were simultaneously quantified by fluorescence microscopic enumeration of apoptotic cells using Hoechst 33342 staining, and by flow cytometric detection of sub-G1 peak cells. The nonirradiated controls showed a similar mean percent apoptotic cells using both methods, analyzed immediately (9% by morphology vs. 10% by flow) or after 24 h of culture (40% by morphology vs. 41% by flow). In the irradiated samples, the mean percent apoptotic cells quantified by morphology was higher than by flow cytometry (64% by morphology vs. 55% by flow after 2 Gy irradiation, and 71% vs. 58% after 10 Gy). The results of the two methods were correlated, although large differences were seen between the techniques in individual tumors. In our system, flow cytometric sub-G1 peak analysis appears to underestimate apoptosis. Of these two methods, we find the Hoechst morphology method to be more reliable for quantitation of apoptosis utilizing fresh fine-needle sample material, in that discrimination of apoptotic cells from debris is easier and that both early and late apoptotic cells are detectable.

Fine-needle sampling (FNS) of 44 salivary squamous cell carcinomas, including 34 primary tumors arising from parotid gland, 6 local recurrences, and 4 lymph node cervical metastases, was performed in 44 patients. Cytologic diagnoses were concordant with histological diagnoses in 36 (81.8%) cases, whereas 5 (11.4%) cases were classified as malignant (carcinoma or adenocarcinoma), 1 (2.3%) case as suspicious, and 2 (4.5%) cases were false-negative (necrosis). No samples were unsatisfactory for cytologic evaluation. Diagn. Cytopathol. 1998;18:174–178. © 1998 Wiley-Liss, Inc.

BACKGROUND The value of ultrasound-guided fine-needle aspiration cytology (FNAC) in the diagnosis of nonpalpable breast lesions remains a subject of debate. To determine the accuracy of this procedure in these lesions, a comparative analysis with open surgical biopsy in 188 patients was performed. METHODS Six hundred and fifty-four nonpalpable breast lesions detected by ultrasound in 585 patients were subjected to FNAC. Of these, 198 lesions (30.3%) from 188 patients were surgically excised. RESULTS Histologically, 105 lesions (53%) were malignant and 93 (47%) were benign. Among carcinomas, 65 cases (61.9%) were concordant, whereas 21 cases (20%) were suspicious, 12 cases (11.4%) were false-negative (benign), and 7 cases (6.7%) were unsatisfactory for diagnosis. Among benign lesions, 80 cases (86%) were concordant, whereas 5 cases (5.4%) were suspicious, and 8 cases (8.6%) were unsatisfactory. No false-positive diagnoses were made. The sensitivity of the FNAC method was 87.8%, the specificity was 94.5%, and the positive predictive value was 94.6%. CONCLUSIONS These results indicate that, with the exception of random microcalcified lesions, ultrasound-guided FNAC is a valuable and reliable technique in the diagnosis of nonpalpable breast lesions. Cancer (Cancer Cytopathol) 1998;84: 36-41. © 1998 American Cancer Society.

Fine-needle sampling (FNS) of 22 acinic cell carcinomas, including 17 primary tumors, 4 local recurrences, and 1 lymph node metastasis was performed preoperatively in 17 patients. Cytologic diagnoses were concordant with histology in 3 (13.7%) cases, whereas 15 (68.2%) cases were cytologically classified as malignant, 2 (9.1%) as suspicious, and 1 (4.5%) as benign (pleomorphic adenoma). The material was unsatisfactory for cytologic evaluation in 1 (4.5%) case. Preoperative FNS technique is, therefore, useful in acinic cell carcinoma with a concordant malignant/suspicious rate of 91%.

Background It has been suggested that metastasizing pleomorphic adenomas may represent unrecognized malignancy. Methods The cytologic and clinical characteristics of two metastasizing pleomorphic adenomas diagnosed by fine-needle sampling are reported. Results Both showed malignant evolution: the primary tumors arose from the palate and the parotid salivary glands, respectively. Metastases occurred simultaneously with local recurrence in the first patient and after a second local recurrence in the second patient. Both patients were treated by surgery and radiotherapy but died of disseminated disease 8 and 4 years after initial diagnosis. No histologic evidence of malignancy was observed on cytology smears or histology sections in either case. Conclusion Clinically, our cases support the hypothesis that metastasizing pleomorphic adenomas represent unrecognized malignancy, because the biologic course of two tumors led to unequivocally lethal outcome.

Fine-needle sampling (FNS) of 50 mucoepidermoid carcinomas, including 44 primary tumors, five local recurrences, and one lymph node metastasis, was performed preoperatively in 44 patients. Concordant cytologic diagnoses were established in only 19 (38%) tumors, whereas 15 (30%) were classified as carcinoma, five (10%) as suspicious, and six (12%) as benign tumors. The material was insufficient for cytologic evaluation in five (10%) cases. The tumors were classified histologically as high-, intermediate-, and low-grade in 15, 13, and 22 cases, respectively. The quality of diagnosis did not vary between high- and intermediate-grade, but was lower in low-grade tumors: Malignancy was diagnosed or suspected in 13 (87%) high-grade tumors, 11 (85%) intermediate-grade tumors, and 15 (68%) low-grade tumors. In conclusion, FNS is an accurate technique in high- or intermediate-grade mucoepidermoid carcinomas, but quite unsatisfactory in low-grade tumors. Diagn. Cytopathol. 17:92–98, 1997. © 1997 Wiley-Liss, Inc.

DNA quantitation using fluorescent dyes is of interest in image cytometry in that it is nondestructive in its mode of staining and is compatible with techniques such as FISH and immunofluorescence, allowing multicolor analysis of a wide range of cellular markers of interest. Optimal preparation techniques were sought using human peripheral blood lymphocytes and fine needle samples of breast carcinomas. Unfixed (air-dried only), ethanol, ethanol/acetic acid, and paraformaldehyde/ethanol fixations were tested. Unfixed or fixed cells were placed on slides as a drop or by cytocentrifugation, stained with 4′,6-diamidino-2-phenylindole dihydrochloride and DNA content was measured using image analysis. Histogram quality was evaluated using G0G1 peak coefficient of variation, and compared to those generated by flow cytometry. Drop preparations of ethanol/acetic acid fixed and cytospin preparations of paraformaldehyde/ethanol fixed cells appeared to give the best histograms for image analysis, which were inferior in quality to those generated by flow cytometry. Comparison of breast carcinoma histograms generated by flow and image showed the same DNA aneuploid populations but with slightly higher DNA indices measured by image analysis. Cytometry 26:123–129, 1997. © 1997 Wiley-Liss, Inc.