Imprint Specimens Research Articles

CELLULAR COMPOSITION OF THE LYMPHOID TISSUE OF THE CECAL IMMUNE FORMATIONS IN DUCKS

Observing the quantitative and qualitative composition of immunocompetent cell populations of the lymphoid tissue of the immunogenic organs allows to determine the immune status of the organism in a certain age period. The object of research is determing the cellular composition of the lymphoid tissue of the cecal Peyer’s patches and cecal (apical) diverticula in ducks in age-concerned aspect. Material simples for research were selected from broiler ducks of the Blagovarsky cross. Cytological testss were performed on imprint specimens. Immunoblasts, lymphocytes, proplasmocytes, and plasmocytes, monocytes, and macrophages are distinguished among the cells of the lymphoid tissue of the cecal Peyer’s patches and the cecal diverticula. The content of these cells is not the same. Population of lymphocytes in imprint specimens is the biggest. It consists of subpopulations of small, medium, and large lymphocytes, the ratio is uneven. The largest is a subpopulation of small lymphocytes, and the smallest is a subpopulation of large ones. The total content of lymphocyte in cecal Peyer’s patches and cecal diverticula decreases with age of the subject ducks. The content of small and medium-sized lymphocytes in the cecal diverticula and small lymphocytes in the cecal Peyer’s patches as well decreases. Simultaneously, the content of large lymphocytes in the cecal diverticula, large and medium lymphocytes in the cecal Peyer’s patches increases. The immunoblasts content in the lymphoid tissue of the studied immune formations decreases with age of ducks, while the quantity of macrophages and monocytes conversely increases. Proplasmocytes and plasmocytes are detected in the lymphoid tissue of cecal Peyer’s patches and cecal diverticula from the age of 10 days in ducks. Their content increases significantly with the poultry age. Reticular cells observation if complicated due to their location under a dense layer of lymphoid cells. Fibroblasts, M-cells, erythrocytes, and granulocytes in imprint specimens are detected in trace amounts. The established changes in the cellular composition of the lymphoid tissue of the cecal Peyer’s patches and the cecal diverticula in ducks in the age-related aspect confirm the occurring immune reactions within them. Consideration of these changes will improve the effectiveness of anti-epizootic measures

A case of anaplastic large cell lymphoma with Charcot-Leyden crystals with tumor cells in an imprint specimen

A case of anaplastic large cell lymphoma with Charcot-Leyden crystals with tumor cells in an imprint specimen

Filter paper-assisted cell transfer (FaCT) technique: A novel cell-sampling technique for intraoperative diagnosis of central nervous system tumors.

Intraoperative diagnosis of central nervous system (CNS) tumors provides critical guidance to surgeons in the determination of surgical resection margins and treatment. The techniques and preparations used for the intraoperative diagnosis of CNS tumors include frozen sectioning and cytologic methods (squash smear and touch imprint). Cytologic specimens, which do not have freezing artifacts, are important as an adjuvant tool to frozen sections. However, if the amount of submitted tissue samples is limited, then it is difficult to prepare both frozen sections and squash smears or touch imprint specimens from a single sample at the same time. Therefore, the objective of this study was to derive cells directly from filter paper on which tumor samples are placed. The authors established the filter paper-assisted cell transfer (FaCT) smear technique, in which tumor cells are transferred onto a glass slide directly from the filter paper sample spot after the biopsy is removed. Cell yields and diagnostic accuracy of the FaCT smears were assessed in 40 CNS tumors. FaCT smears had ample cell numbers and well preserved cell morphology sufficient for cytologic diagnosis, even if the submitted tissues were minimal. The overall diagnostic concordance rates between frozen sections and FaCT smears were 90% and 87.5%, respectively (no significant differences). When combining FaCT smears with frozen sections, the diagnostic concordance rate rose to 92.5%. The current results suggest that the FaCT smear technique is a simple and effective processing method that has significant value for intraoperative diagnosis of CNS tumors. Cancer Cytopathol 2017;125:277-282. © 2016 American Cancer Society.

Pleomorphic rhabdomyosarcoma arising in the anterior mediastinum: A case report with cytological features of imprint and liquid-based cytology specimens.

We herein report the cytological features of a very rare case of pleomorphic rhabdomyosarcoma arising in the anterior mediastinum on imprint and liquid-based cytology (LBC) specimens. A 58-year-old man had an approximately 10-cm tumor in the anterior mediastinum as shown on computed tomography. Thymectomy with complete resection of the left lung was performed. The fresh cut surface of the tumor was used to prepare imprint and LBC specimens. The imprint specimens showed four types of tumor cells dispersed on a background of hemorrhage, necrosis, and mucus. On the other hand, only two types of tumor cells (spindle-shaped and spiderweb cells) were scattered or present in clusters in the LBC specimens. Immunocytologically, both of these cell types were positive for desmin and myoglobin, negative for pan-keratin and epithelial membrane antigen. Cytological and immunocytological features are useful for the correct diagnosis of pleomorphic rhabdomyosarcoma, and LBC specimens show clearer results than do imprint specimens. Diagn. Cytopathol. 2017;45:333-338. © 2016 Wiley Periodicals, Inc.

Identification of intranuclear inclusions is useful for the cytological diagnosis of ovarian clear cell carcinoma.

The aim of this study was to clarify the diagnostic significance of the presence of intranuclear inclusions in clear cell carcinoma (CCC). We analyzed 98 imprint specimens and 53 ascites specimens from 98 ovarian carcinoma cases [28 CCCs, 37 serous carcinomas (SCs), 22 endometrioid carcinomas (ECs), and 11 mucinous carcinomas (MCs)]. We examined (1) frequency of intranuclear inclusion-positive cases of each ovarian carcinoma subtype, using imprint specimens, (2) frequency of intranuclear inclusion-positive cells of each ovarian carcinoma subtype, using imprint specimens, (3) frequency of intranuclear inclusion-positive cases of each ovarian carcinoma subtype, using ascites specimens, and (4) sensitivity and specificity of the presence of intranuclear inclusions for the cytological diagnosis of CCC. (1) The frequency of intranuclear inclusion-positive cases in CCC (96.4%) was significantly higher than in SC (13.5%), EC (13.6%), and MC (18.2%) (P < 0.001). Two or more intranuclear inclusions in a single nucleus were observed only in CCC. (2) The frequency of intranuclear inclusion-positive cells in CCC (median, 0.41%) was significantly higher than in non-CCC subtypes (0.010%) (P < 0.001). (3) Using ascites specimens, the frequency of intranuclear inclusion-positive cases in CCC (78.6%) was significantly higher than in SC (10.3%), EC (0%), and MC (0%) (P < 0.001). (4) The sensitivity of intranuclear inclusions was 96.4%, and the specificity was 85.7%. The identification of intranuclear inclusions, in particular a high frequency and multiple intranuclear inclusions in a single nucleus, is useful for the cytological diagnosis of CCC. Furthermore, these results may be applicable to ascites cytology.

Utility of intraoperative cytology of resection margins in biliary tract and pancreas tumors.

Intraoperative diagnosis of resection margins in the biliary tract and pancreas tumors is important in deciding extent of resection; however, diagnosis based solely on frozen sections is sometimes difficult. Therefore, we investigated the usefulness of intraoperative cytology (IC) combined with frozen section (FS) histology. We present the results with a discussion of the value of this combination and its associated problems. We examined 80 bile duct resection margin specimens from 42 patients, and 34 pancreatic resection margin specimens from 29 patients, who underwent intraoperative diagnosis of resection margins during surgery for biliary tract or pancreatic tumors between October 2012 and January 2014. IC was performed on imprint specimens prepared from surfaces of margins being examined. The results were compared with FS and final histology of operative materials. In IC, excluding cases with insufficient material, the results for bile duct margins; sensitivity was 96.7%, specificity 100% and accuracy 98.7%. The results for pancreatic margins; sensitivity was 100%, specificity 92.9%, and accuracy 93.3%. In FS, the results for bile duct margins; sensitivity was 96.8%, specificity 100%, and accuracy 98.8%. The results for pancreatic margins; sensitivity was 66.7%, specificity 100%, and accuracy 97.1%. IC is quick, highly accurate and very easy to perform. This study even included a specimen for which only IC led to an accurate diagnosis. IC used in combination with FS can achieve intraoperative diagnosis with high overall accuracy.

A case of blastic plasmacytoid dendritic cell neoplasm: Cytomorphological findings of the touch imprint specimen of lymph node

The patient is a 75-year-old man with axillary lymphadenopathy presenting an indurated papule on his buttock. Touch imprint cytology of the biopsied axillary lymph node revealed the monotonous appearance of medium-sized tumor cells. The nuclei had a slightly irregular contour, finely dispersed chromatin, and a conspicuous nucleolus. Some tumor cells had intracytoplasmic microvacuoles. Immunohistochemistry of the imprint specimens showed that the tumor cells were positive for CD56 and CD123. Histological diagnosis of the lesion was blastic plasmacytoid dendritic cell neoplasm (BPDCN). Epstein-Barr virus-encoded RNAs were not detected in the tumor cells. Neither immunoglobulin heavy chain genes nor T- cell receptor genes was clonally rearranged. BPDCN should be strongly considered during the differential diagnosis of CD56-positive neoplasms of the skin. We demonstrated a possible contribution of the cytomorphological and immunohistochemical findings of the touch imprint specimens to the diagnosis of BPDCN.

Role of Intra-Operative Touch Imprint Cytology in the Treatment of Breast Cancer

Objectives : A prospective study was carried out to evaluate the role of intra-operative touch imprint cytology (TIC) in the assessment of sentinel lymph node (SLN) involvement for staging and treatment of early-stage, clinically node-negative breast carcinoma.Methods : Forty-five patients with early-stage, clinically node-negative breast cancer underwent a SLN biopsy with intra-operative TIC. The SLN was bisected if its width was less than 4 mm or sliced every 2 mm if it was more than 4 mm. The imprint specimens were stained with haematoxylin and eosin (H&E). Rapid immunochemistry (IH) was performed in case of equivocal cytological result. Permanent sections were evaluated with H&E and IH staining. The results of TIC were compared to histopathological results.Results : The sensitivity, specificity and overall accuracy of TIC on a node basis were 65.5%, 96.3%, 85.5%, respectively. When calculated according to the size of SLN metastasis, the sensitivity of TIC for overt metastasis was 84.6%, while it was 62.5% for micrometastasis and 37.5% for sub-micrometastasis. The mean size of nodal metastasis was 5.08 mm and 1.25 mm for true positive and false negative results, respectively (P = 0.0236). Because of intra-operative TIC, 76.5% of the patients who needed further axillary lymph node dissection (ALND) could undergo this during the same operating time.Conclusions : TIC is a rapid and reliable method for the intra-operative assessment of metastatic sentinel node involvement in patients with early-stage, clinically node-negative breast carcinoma. Despite a low sensitivity comparable to frozen section (FS) in detecting micro-and sub-micrometastases, the technique offers the advantage of full tissue preservation for subsequent histological analysis.

Automated Microscopy to Evaluate Surgical Specimens Via Touch Prep in Breast Cancer

Breast conservation therapy is the standard treatment for breast cancer; however, 20-50% of operations have a positive margin leading to secondary procedures. The standard of care to evaluate surgical margins is based on permanent section. Imprint cytology (touch prep) has been used to evaluate surgical samples, but conventional techniques require an experienced cytopathologist for correct interpretation. An automated image screening process has been developed to discern cancer cells from normal epithelial cells. This technique is based on cellularity of the imprint specimen and does not require expertise in cytopathology. A rapid immunofluorescent staining technique coupled with automated microscopy was used to classify specimens as cancer vs. noncancer based on the density of epithelial cells captured on touch prep of tumor cross-sections. The results of the automated analysis vs. a manual screen of ten 20x fields were compared to the pathology interpretation on permanent section. A total of 34 consecutive cases were analyzed: 10 normal cases, and 24 cancer cases. The cross-section specimens for invasive cancer were correctly classified in at least 65% of the cases by using manual microscopy and at least 83% by using automated microscopy. The manual and automated microscopy correlated well for measurements of epithelial cell density (R(2)=0.64); however, the automated microscopy was more accurate. This preliminary study using an automated system for intraoperative interpretation does not require a cytopathologist and shows that rapid, low-resolution imaging can correctly identify cancer cells for invasive carcinoma in surgical specimens. Therefore, automated determination of cellularity in touch prep is a promising technique for future margin interpretation of breast conservation therapy.

Immunocytochemical differentiation of canine mesenchymal tumors in cytologic imprint preparations

Immunocytochemical techniques are a potentially valuable diagnostic tool to support cytologic diagnosis in dogs. However, detailed studies of staining patterns and intensity in cytologic specimens of mesenchymal tumor types are lacking. The aim of this study was to evaluate commercially available antibodies against human proteins for use in the characterization of canine tumors of mesenchymal origin in cytologic samples. Immunocytochemical staining was performed on air-dried imprint specimens of biopsies obtained from 103 mesenchymal neoplasms and 14 metastatic lesions from 98 dogs. All specimens were stained with anti-cytokeratin AE1/AE3 and vimentin. Based on the histologic diagnosis, tumors of muscle, endothelial, histiocytic, and melanocytic origin also were stained with cell-specific antibodies. Staining intensity was subjectively graded and the percentage of positive tumor cells was estimated. All mesenchymal tumors and metastases, with the exception of mesotheliomas, were vimentin-positive and cytokeratin-negative; mesotheliomas (n=6) were positive for both vimentin and cytokeratin. Tumors of muscle (n=5), endothelial (n=15), and histiocytic (n=18) origin stained moderately to strongly positive in a majority of tumor cells with desmin, von Willebrand factor, and lysozyme, respectively. Malignant melanomas (n=15) had variable staining and a variable percentage of positive cells with Melan-A and S100. Our results indicate that immunocytochemical staining of canine cytologic specimens is a reliable and sensitive technique that may be of benefit for the differentiation of poorly differentiated mesenchymal tumors and metastases. Additional study is needed to assess the specificity of immunocytochemical stains in mesenchymal tumors.

Topoisomerase II α gene amplification in gastric carcinomas: correlation with the HER2 gene. An immunohistochemical, immunoblotting, and multicolor fluorescence in situ hybridization study

Topoisomerase IIalpha (topoIIalpha) is an enzyme required for DNA replication and a molecular target for drugs called anthracyclines. The topoIIalpha gene (TOP2A) is located close to the HER-2/neu oncogene (HER2). We assessed gastric cancers to (1) clarify the relationship between gene amplification and protein overexpression of topoIIalpha and HER2; (2) evaluate the correlation between gene amplification and protein overexpression of topoIIalpha; and (3) examine the relationship between the results of immunohistochemistry and Western blot analysis for topoIIalpha. In a combined analysis of immunohistochemistry and fluorescence in situ hybridization on 552 formalin-fixed and paraffin-embedded gastric cancer tissues, 38 cases were found to have HER2 amplification. Further examination by fluorescence in situ hybridization revealed amplification of TOP2A in 13 of the 38 cases. No aberrations in the TOP2A gene were observed in cases without HER2 overexpression, except for one containing a gene deletion. The TopoIIalpha protein-labeling index was not correlated with TOP2A amplification. Fluorescence in situ hybridization was performed on nuclear imprint specimens obtained from 9 cases using simultaneous probes for TOP2A, HER2, and centromere 17. Of these 9 cases, 3 displayed coamplification of TOP2A and HER2, and only 1 of the 3 cases revealed a high expression of topoIIalpha in Western blot. Although patients having gastric adenocarcinoma with TOP2A amplification could be considered suitable for clinical trials, information involving anthracycline therapy is not firmly understood in regards to the status of TOP2A amplification or protein overexpression. Therefore, results of the current study will provide further insight for the clinical application of anthracycline in gastric cancers.

A case of polymorphous low-grade adenocarcinoma in the palate with similar features to adenoid cystic carcinoma

Background: We report a case of polymorphous low-grade adenocarcinoma (PLGA) of the palate showing features similar to adenoid cystic carcinoma (ACC) in smear cytology.Case and Conclusion: Imprint specimens from the cut surface of biopsied tissue in a 78-year-old woman with a painless mass in the palate contained many neoplastic cells arranged in tight clusters, and mucous ball-like structures, pseudopapillary, tubular, and solid patterns were observed. Cells forming these clusters varied in size and had abundant eosinophilic, granular cytoplasm. Nuclei were round to oval and had a relatively smooth configuration. Although ACC was suspected cytologically, PLGA was confirmed by histopathological section. As opposed to imprint specimens obtained from resected tumors, no indian file arrays or concentric targetoids were detected in the imprint from the biopsied specimen, which thus led to diagnostic failure.

Cytologic diagnosis ofLawsonia intracellularisproliferative ileitis in a Japanese snow macaque (Macaca fuscata)

Diffuse ileal thickening and ileocecocolic lymphadenomegaly were observed during exploratory laparotomy in a 2-year-old male Japanese snow macaque (Macaca fuscata) that had flu-like signs and diarrhea. Cytologic examination of ileal biopsy imprints revealed many mature, mildly karyolytic neutrophils and fewer well-differentiated lymphocytes, eosinophils, macrophages, and plasma cells in a background containing amorphous, necrotic material. Tightly cohesive sheets of moderately pleomorphic epithelial cells also were seen. The cytologic diagnosis was chronic, active, mixed inflammation with atypical epithelial cells and necrosis. Histologically, the mucosal and crypt epithelium was moderately hyperplastic with a loss of goblet cells, increased mitoses, and frequent crypt abscesses. Within the lamina propria and extending into the submucosa was a marked neutrophilic infiltrate, with low numbers of lymphocytes, histiocytes, plasma cells, and eosinophils. The histologic diagnosis was chronic, diffuse, marked suppurative and lymphocytic ileitis. Warthin-Starry silver staining of the ileal biopsy and imprint specimens demonstrated numerous pleomorphic, curved bacilli consistent with Lawsonia intracellularis. Polymerase chain reaction (PCR) and immunohistochemistry confirmed the identity of the infectious agent. L intracellularis infection may be underdiagnosed because silver stain is required to visualize the organism with light microscopy and because the pathognomonic crypt hyperplasia may be complicated by secondary pathologic changes. Application of silver stain to cytologic specimens should be considered when distal intestinal lesions associated with hyperplastic epithelium, with or without inflammation, hemorrhage, or necrosis, are identified in animals with clinical signs of enteritis, especially in frequently affected species or in stressed or young animals.

Immunocytochemical study of Ki-67 as a prognostic marker in canine mammary neoplasia.

Canine mammary tumors are challenging for clinicians and pathologists because of complex histologic classification, low specificity of cytologic diagnosis, and unpredictable biological behavior. In histologic specimens, expression of tumor proliferation marker Ki-67, a nuclear nonhistone protein, has been shown to have prognostic value for canine mammary tumors and to correlate with malignancy and low survival rates. The objective of this study was to measure the proliferation index of canine mammary tumors by immunochemical detection of Ki-67 in cytologic specimens and to determine its relationship to clinical and pathologic variables and patient outcome. Spontaneous mammary tumors from 31 female dogs were surgically excised. Imprint specimens for cytologic evaluation were wet-fixed in ethanol; histologic specimens were prepared routinely. Immunostaining was performed with the PH 177 monoclonal antibody against Ki-67; proliferation index was graded from negative to +++. Dogs were followed for 18 months. Multivariate logistic regression analysis was used to determine correlations between immunocytochemical results, tumor and clinical variables, and patient outcome. Ki-67 proliferation indices in cytologic specimens were significantly lower for nonmalignant tumors than for malignant tumors. High index values of Ki-67 were positively correlated with metastasis, death from neoplasia, low disease-free survival rates, and low overall survival rate. With the exception of 4 specimens for which cellularity was insufficient, positive expression of Ki-67 in cytologic specimens correlated with that of histologic specimens. The prognostic value of the Ki-67 index in canine mammary tumors by using wet-fixed cytology imprint specimens was similar to that observed previously for histologic specimens. Immunocytochemical detection of Ki-67 could improve the accuracy and value of cytology by providing safe and rapid information about malignancy and patient outcome.

Touch imprint cytology and immunohistochemistry for the assessment of sentinel lymph nodes in patients with breast cancer

Aims: Accurate evaluation of sentinel nodes is of clinical importance to avoid further surgery for axillary node dissection. A prospective study was carried out to investigate the feasibility and accuracy of touch imprint cytology (TIC) and touch imprint immunohistochemistry (TIHC).Methods: Two hundred and five sentinel nodes from consecutive 118 patients with primary breast cancer were studied after successful identification of sentinel nodes. Sentinel nodes were sectioned at 2 mm intervals and imprint specimens prepared from all cut surfaces were subjected to Papanicolaou staining and immunohistochemical staining using anti-cytokeratin antibody.Results: Forty-nine sentinel nodes from 40 patients were positive by permanent section. The sensitivity of TIC was 84% (41/49) per sentinel node and 83% (33/40) on a per patient basis. The sensitivity of TIHC was 86% (42/49) per sentinel node and 83% (33/40) on a per patient basis. When the results of TIC and TIHC were combined, the sensitivity was 88% (43/49) per sentinel node and 85% (34/40) on a per patient basis. Among the 156 negative sentinel nodes, four sentinel nodes from four different patients were consistently positive by TIC and TIHC, but only one patient out of 78 node-negative patients was upstaged.Conclusions: Touch imprint cytology is sufficiently sensitive for intraoperative evaluation of sentinel nodes. A slight improvement in the sensitivity is expected when immunohistochemistry is used. The combination of these methods provides better sensitivity than either method alone.

Cytologic Diagnosis of Estrogen and Progesterone Receptors in Breast Imprints

To investigate estrogen receptor (ER) and progesterone receptor (PR) levels in imprint specimens obtained at breast surgery and to compare their correlation with that of standard methods. Imprint specimens for cytology were obtained from 101 mass-forming lesions in 66 patients, and specimens were frozen in liquid nitrogen for later assay. The imprint specimens were immunocytochemically (ICC) stained by monoclonal antibody to ER or PR; diaminobenzidine-stained cell nuclei in clusters were regarded as positive. Tissue specimens were assayed by the standard method of dextran-coated charcoal assay (DCC) and enzyme immunoassay. Forty-five primary breast cancer lesions, 2 contralateral breast cancer, 49 dissected nodes and 5 benign breast lesions were collected. The correlation between DCC and ICC was 81% (82/101) for ER and 74% (66/101) for PR. That between EIA and ICC was 88% (88/99) for ER and 80% (79/100) for PR, higher than that between DCC and ICC for ER and PR. ICC assessment of ER or PR on imprint cytology is a promising clinical test with an acceptable correlation.

Detection of chromosome 11q13 breakpoints by interphase fluorescence in situ hybridization. A useful ancillary method for the diagnosis of mantle cell lymphoma.

We assessed cytologic specimens from 11 mantle cell lymphomas (MCLs) and 32 other B-cell non-Hodgkin lymphomas (NHLs) for 11q13 breakpoints using a 2-color fluorescence in situ hybridization (FISH) assay that uses an 11q13 probe centered on the CCND1 gene and a centromeric chromosome 11 probe (CEP11). The number of nuclei in 200 cells were counted, and results were expressed as an 11q13/CEP11 ratio. All MCLs showed a high percentage of interphase nuclei with 3 or more 11q13 signals (mean, 74.8%; range 57%-90%). In contrast, in other B-cell NHLs the mean percentage of cells with 3 or more 11q13 signals was 9.2%. All MCLs had an elevated 11q13/CEP11 ratio (mean, 1.38). The mean ratio for other B-cell NHLs was 0.99. Two non-MCL cases, 1 large B-cell and 1 B-cell unclassified NHL, had high 11q13/CEP11 ratios of 1.15 and 1.30, respectively. Conventional cytogenetic analysis performed on the former case revealed a t(5;11)(q31;q13). Interphase FISH analysis using 11q13 and CEP11 probes is a convenient ancillary method for assisting in the diagnosis of MCL. This commercially available assay is simple to use on cytology or imprint specimens, and results can be obtained within 24 hours.

Multifocality of transitional cell carcinoma results from genetic instability of entire transitional epithelium

Objectives. Multifocality of transitional cell carcinoma (TCC) has been attributed to seeding of exfoliated tumor cells or to a general sensitivity of the entire urothelium to carcinogenic stimuli. By contrast, TCC has been shown to evolve as a consequence of genetic defects and chromosomal instability. We analyzed chromosomal patterns, total DNA content, and p53 and Ki67 expression in malignant and normal transitional cells to evaluate their relationship to the development of multifocal TCC. Methods. Included in the study were 47 patients, 16 women and 31 men, with a mean age of 70.04 years (range 37 to 83). Of 47 patients, 45 had TCC of the urinary bladder and 7 of those had synchronous ureteral involvement. Two patients had ureteral TCC and a history of TCC of the bladder. Using fluorescence in situ hybridization, numerical aberrations of chromosomes 7, 9, and 17 were detected in imprint specimens of histologically verified tumor and “normal” urothelium and were compared with static ploidy and p53 and Ki67 expression. Results. Chromosome 7 was altered in 93.6%, chromosome 9 in 63.8% (including monosomy), and chromosome 17 in 87.2% of the 47 analyzed tumor and normal imprints. Differences between tumor and normal epithelium were observed in aberrational frequencies (number of cells showing chromosomal aberrations calculated on 200 cells counted, given in percentages). DNA content was aneuploid in all tumor specimens, but diploid in 20 (42.5%) of 47 normal specimens, according to lower aberration frequencies in these patients. p53 detection was positive in 82.9% of the tumor specimens and 76.6% of the normal specimens. Ki67 was positive in 87.2% of the tumor imprints and in 72.3% of the normal specimens. Conclusions. These data suggest a general genetic instability as a reason for multifocality in the entire transitional epithelium.

Imprint cytology of large b-cell lymphoma with high content of epithelioid cells. A report of two cases

An association of epithelioid cell reaction in follicular center cell lymphomas seems to be a rare phenomenon. The cytological features of this type of B-cell lymphoma from two patients are presented. In both cases, immunohistochemistry applied to paraffin sections revealed reactivity of tumor cells for CD10, 20 and 79a. The first patient, a 77-year-old male, presented with bilateral inguinal bulky masses. The second patient, a 76-year-old female, revealed left supraclavicular bulky masses. Imprint cytological specimens of these two cases showed large lymphoid cells interspersed with epithelioid histiocytes, which were present in small aggregated clusters or in isolated forms. Most tumor cells showed centroblastic morphology, whereas the minority of them looked like immunoblasts. Occasionally, a monotonous proliferation of these large atypical blastic cells was also detected. On the other hand, in some parts of the imprint specimens, numerous epithelioid cells obscured the underlining tumor cells. Reactive cells other than epithelioid cells were not prominent. These cytologic features closely resembled the histologic patterns of this rare follicular center cell lymphoma. Careful examination of the cytological specimens is needed to detect the monotonous proliferation of tumor cells to establish the diagnosis of malignant lymphoma.

Fluorescence in Situ Hybridization Identifies More Aggressive Types of Primarily Noninvasive (Stage pTa) Bladder Cancer

Purpose We evaluated the genetic changes in cytological specimens of bladder cancer by fluorescence in situ hybridization, and related them to stage and grade of the tumor, ploidy, p53 and Ki-67 expression, and clinical outcome to determine a simple method to identify tumors with a poorer prognosis. Materials and Methods Using fluorescence in situ hybridization the numerical aberrations of chromosomes 7, 9 and 17 in barbotages and imprints of 50 patients with transitional cell cancer of the bladder were determined. Of the patients 29 had a primary stage pTa tumor, while 21 with stage pT1 or greater disease formed the control group. Data were compared to ploidy status, and Ki-67 and p53 immunoreactivity. Results Repeated monosomy 9 and haploid or diploid status on static ploidy determination were found in patients with primary stage pTa tumors without recurrence. Immunoreactivity of p53 was negative in all of these patients, while there was a low percentage of positive staining for Ki-67. Patients with recurrent and progressive disease had a high incidence of trisomy 7 and 17, aneuploid status and high positivity for both immunological markers. For chromosomes 7 and 17, and ploidy status bivariate analysis showed a significant difference. Conclusions The evaluation of chromosomal aberrations in barbotage and imprint specimens clearly establishes a relationship between chromosomal defects and aggressiveness of the tumor. The majority of nonaggressive stage pTa transitional cell carcinomas can be distinguished from potentially lethal cases by fluorescence in situ hybridization at a diagnostic point when the grading is not yet prognostic.