Detection of bladder cancer by multitarget multicolour FISH: comparative analysis on archival cytology and paraffin-embedded tissue.

Detection of bladder cancer by multitarget multicolour FISH: comparative analysis on archival cytology and paraffin-embedded tissue We have evaluated the possibility of using the same specimen for both cytological diagnosis and multitarget multicolour FISH (MtMcFISH) analysis in order to determine whether the routinely processed specimens used for diagnosis were also suitable for this ancillary procedure. For this purpose 18 positive samples (11 voided urine and seven bladder washings) were selected, together with a representative section of the corresponding immediately previous or subsequent histological specimens. Two negative cytology slides were added as negative controls. FISH analysis revealed a normal pattern for each probe in the two negative controls and an abnormal pattern in the 18 positive cases. In the latter the same FISH alterations were found in the cytology samples and in the corresponding histological sections, and superimposable cytological/histological features were observed in two cases where two different histology samples were analyzed. The results clearly show that MtMcFISH may be successfully applied to destained routinely processed cytology slides.

Assessment of nodal involvement and survival analysis in breast cancer patients using image cytometric data: statistical, neural network and fuzzy approaches.

Accurate and reliable decision making in breast cancer prognosis can help in the planning of suitable surgery and therapy and, generally, optimise patient management through the different stages of the disease. In recent years, several prognostic factors have been used as indicators of disease progression in breast cancer. In this paper we investigate a fuzzy method, namely fuzzy k-nearest neighbour technique for breast cancer prognosis, and for determining the significance of prognostic markers and subsets of the markers, which include histology type, tumour grade, DNA ploidy, S-phase fraction, G0G1/G2M ratio, and minimum (start) and maximum (end) nuclear pleomorphism indices. We also compare the method with (a) logistic regression as a statistical method, and (b) multilayer feed forward backpropagation neural networks as an artificial neural network tool, the latter two techniques having been widely used for cancer prognosis. Nodal involvement and survival analyses in breast cancer are carried out for 100 women who were clinically diagnosed with breast disease in the form of carcinoma and benign conditions, and seven prognostic markers collected for each patient. For nodal involvement analysis, node positive and negative patients are predicted whereas survival analysis is carried out for two categories: whether a patient is alive or dead within 5 years of diagnosis. The results obtained show that the fuzzy method yields the highest predictive accuracy of 88% for both nodal involvement and survival analyses obtained from the subsets of [tumour grade, S-phase fraction, minimum (start) nuclear pleomorphism index] and [tumour histology type, DNA ploidy, S-phase fraction, G0G1/G2M ratio], respectively. We believe that this technique has produced more reliable prognostic factor models than those obtained using either the statistical or artificial neural networks-based methods.

Performance of the AutoPap Primary Screening System in the detection of high-risk cases in cervicovaginal smears.

OBJECTIVE: To assess the performance of the AutoPap Primary Screening System (APSS) (TriPath Imaging, Inc., Burlington, North Carolina, U.S.A.) for the detection of high grade cervical squamous intraepithelial lesions and invasive cervical cancer. STUDY DESIGN: A total of 14,779 consecutive conventional Pap smears were processed by the APSS. All slides designated as "Review" by the device were manually screened according to the Bethesda System. The ranking scores obtained from the device were compared with the cytologic interpretations in all cases and with the final histologic diagnoses in the cases with cytologic severe abnormalities. RESULTS: The device classified 10,349 slides as Review (78%) and 2,912 (22%) as "No Further Review." In the 78% Review cases, the samples were ranked in descending order of potential abnormality, broken into quintiles. The correlation between the slide quintile ranks and the manual cytologic diagnosis indicated that 90% of abnormal smears were categorized by the device as in the first and second quintile rank, and the correlation between the rank report of the device and the histologic diagnosis showed that all cases of HSIL or invasive carcinoma were in the top two ranks. No significant abnormalities were observed in any of the smears categorized as No Further Review. CONCLUSION: This study confirmed the effectiveness of APSS for the detection of Pap smears with severe abnormalities.

Exact Touch: a new device for cervical cytology. Comparison with Ayre spatula plus Cytobrush.

Exact Touch is a new plastic device for the collection of cells from the ectocervix and endocervix. A total of 189 consecutive women were evaluated, 94 with the Ayre/cytobrush and 95 with Exact Touch. Sampling was performed by only one clinician, and the slides were analysed by only one cytotechnologist, who had no information about the sampling method. Our results showed that more endocervical and metaplastic cells were collected by Exact Touch than by Ayre/cytobrush.

TP53 mutations and mdm2 protein overexpression in cholangiocarcinomas.

Tumor suppressor protein p53 is a positive regulator of MDM2 gene expression and the mdm2 protein can bind to p53, preventing the transactivation of p53 responsive genes, thus mimicking TP53 mutation. The authors looked for alterations that could affect, directly and indirectly, p53 function in 13 patients with extrahepatic cholangiocarcinoma. Molecular analysis by single strand conformation polymorphism and DNA sequencing revealed that TP53 gene mutations occurred in only 2 of 13 cholangiocarcinomas. High levels of mdm2 protein were found, by immunohistochemical staining, in 61% of the cholangiocarcinomas and in almost all specimens (70%) displaying stabilized p53 protein in the absence and in the presence of TP53 mutations. The finding of co-overexpressed mdm2 and p53 proteins in cholangiocarcinomas indicates that they can upregulate the expression of mdm2 protein to a level sufficient for binding and accumulating p53 in a presumably inactive complexed form. The presence of TP53 mutations or upregulation of MDM2 gene expression in 9 of the 13 cholangiocarcinomas strongly supports that the impairment of the p53 pathway is an important and specific step in cholangiocarcinoma pathogenesis. At variance with other authors, no alteration of p16ink4/CDKN2 gene was observed in all 13 cholangiocarcinomas.

Fluctuation of HER2 expression in breast carcinomas during the menstrual cycle.

The hormonal milieu at time of tumor surgery seems to have a significant impact on survival in premenopausal breast cancer patients. Indeed, surgery performed during the follicular phase of the menstrual cycle was suggested to correlate with a poor prognosis. To investigate the relationship between prognosis and menstrual cycle at time of surgery, we analyzed the expression of some markers associated with tumor aggressiveness, such as the hormone receptors, HER2, p53, Bcl2, and cathepsin D in breast carcinomas obtained from 198 premenopausal women who underwent surgery during different phases of the menstrual cycle. HER2 overexpression was found to fluctuate in hormone receptor-positive tumors. In actual fact, 20% of the tumors removed during the follicular phase scored HER2-positive, versus 8% of those removed during the luteal phase. Similarly, a number of hormone receptor-positive tumor specimens, obtained from the same patients during follicular and luteal phases, were scored HER2-positive when the sample was removed during the follicular phase and HER2-negative when removed in the luteal phase. Southern blot analysis of the HER2 gene indicated that, in hormone receptor-positive cases, the overexpression of HER2 is often not associated with gene amplification. The finding that overexpression of the HER2 gene, associated with tumor aggressiveness, can fluctuate according to the hormonal milieu may explain the increased survival of patients operated during the luteal phase. It is also relevant to the selection and treatment of patients most likely to benefit from anti-HER2 antibody therapy.

c-erbB2/neu gene and chromosome 17 analysis in breast cancer by FISH on archival cytological fine-needle aspirates.

The detection of specific genetic alterations in breast cancer is useful for diagnosing, predicting prognosis and planning preoperative treatment. c-erbB2/neu overexpression is usually detected by immunocytochemistry (ICC), although this technique is neither completely reproducible nor highly reliable, owing to specimen and methodologic variability and antibody sensitivity. Here, we combine two well-established techniques, fine-needle aspiration (FNA) and fluorescence in situ hybridization (FISH), to detect c-erbB2/neu amplification in patients candidate to primary chemotherapy and, in part, previously analysed for c-erbB2/neu overexpression. Sixty smears from FNA were used to simultaneously detect c-erbB2/neu and chromosome 17 centromere. FISH was successful in 58 cases and detected 24 amplified cases, three of which were negative by immunophenotyping, 28 negative cases, with evidence of two normal c-erbB2/neu/signals, two cases with deletion of c-erbB2/neu, and four cases with polysomy, thus providing more reliable and informative results than ICC. This study underlines the advantages offered by the FNA and FISH combination which are two rapid, reliable, simple and informative techniques, to analyse one of the most important genetic markers for predicting prognosis and chemotherapy planning for breast carcinoma in particular in the light of the recently proposed trials of primary chemotherapy.

Percutaneous intraductal sampling for cyto-histologic diagnosis of biliary duct strictures.

BACKGROUND: Percutaneous transhepatic biliary drainage (PTBD) allows ductal material to be collected for cyto-histologic examination. We evaluated the data from a large series of patients with a PTBD in whom endobiliary cyto-histologic sampling techniques were employed in order to define a strategy for their use in the diagnostic work-up. PATIENTS AND METHODS: Ductal samples for cyto-histologic examination were obtained from 409 consecutive patients with a PTBD for stenosing lesions of the biliary tree. Bile aspirate cytology was performed for all patients and ductal biopsy specimens were obtained, generally after negative cytology, from 49 of them (11.9%), all candidates for a therapeutic procedure. The cyto-histologic results of intraductal sampling were compared with pathologic surgical data in 210 patients and with clinical-radiologic follow-up in 199. RESULTS: Overall, 22 out of the 409 patients had a final diagnosis of benign stenosis and 177 had samples positive for neoplastic disease. The sensitivity of bile cytology was 43.8% while ductal biopsies showed a sensitivity of 60.4%. The combination of the two sampling techniques achieved a sensitivity of 65.1%. For both sampling methods the specificity was 100%. Hilar metastases from neoplastic lesions of the GI tract and primary lesions of the biliary ducts showed the highest sensitivity. CONCLUSION: Cyto-histologic assessment of stenosing lesions of the biliary ducts is mandatory when highly sophisticated interventions (e.g. wide hepatic resection or liver transplantation) or non-surgical treatments are envisaged. The collection of cyto-histologic samples from bile ducts, in patients with a percutaneous bile drainage, is an easy, safe and valuable method to obtain the diagnosis. In view of the absence of false positive results in our series and in others, intraductal biopsy serves no purpose when positive exfoliative cytology is positive for malignancy. In the presence of negative cytology it is felt that an intraductal biopsy should be mandatory when the choice of a therapeutic program depends on the result of the cyto-histologic diagnosis.

PCR analysis of IgH and BCL2 gene rearrangement in the diagnosis of follicular lymphoma in lymph node fine-needle aspiration. A critical appraisal.

In order to improve the cytomorphologic diagnosis of malignant lymphoma on lymph node fine-needle aspiration (FNA), and to make a confident discrimination between low-grade follicular non-Hodgkin's lymphoma (NHL) and lymphoid hyperplasia, polymerase chain reaction (PCR) analysis was performed of the Ig CDR3 region and BCL2 breakpoint region in 25 nonselected cases of malignant lymphoma (17 NHL and 8 Hodgkin's disease [HD]) with histologic control, and 22 cases of lymph nodal hyperplasia with histologic and/or clinical control. Among lymphomas, IgH monoclonality was detected in 7 (77%) of 9 NHLs and BCL2 rearrangement in 3 (17.6%) of 17 NHLs, all of which were follicular centroblastic-centrocytic (FCBCC). Three BCL2/JH negative FCBCC cases were monoclonal for CDR3. Neither IgH monoclonality nor BCL2 rearrangement were found in HD. Among cytologically diagnosed lymphoid hyperplasias, one IgH polyclonal case was considered false-negative, being histologically diagnosed as lymphoplasmacytic NHL on the subsequent excisional biopsy. Another 4 cases (2 BCL2 rearranged and 2 monoclonal for IgH) were considered false-positive on the basis of histologic features or clinical control. These data indicate that the combined PCR analysis of IgH and BCL2 rearrangements can confirm a cytologic diagnosis of lymphoma in FNAs while, due to the occurrence of both false-positive and false-negative results, it is of limited value in the distinction between follicular lymphoma and lymphoid hyperplasia without morphologic or clinical support.

Fine needle aspiration (FNA) appearances of malignant myoepithelioma of the parotid gland.

We describe the FNA features of five cases of malignant parotid myoepithelioma, the majority of which were thought clinically to be recurrent pleomorphic adenomas. A major finding was cell shape variation: round-oval, polygonal or spindle-shaped, with basophilic cytoplasm. Many were epithelial and plasmacytoid-like and had nuclear grooves, pseudoinclusions, and multinucleation. The true myoepithelial nature of the neoplastic cells was identified in all cases, but only two showed obvious cytological malignant features, both initially and on re-examination. FNA of malignant myoepithelioma may thus show overt features of malignancy, or may lack atypia and malignancy can only be identified on histology. The correct diagnosis can be predicted in FNA samples in certain cases, both in terms of typing and malignancy, whilst sometimes only the myoepithelial nature of the lesion can be assessed.

Diagnostic assessment of enlarged superficial lymph nodes by fine needle aspiration.

Two hundred eighty-five consecutive outpatients with enlarged superficial lymph node either clinically suspicious (152) or with a previous diagnosis of a malignant tumor (133) underwent fine needle aspiration (FNA) followed by excisional biopsy. Cytologic and/or cytologic-immunophenotypic diagnoses made on direct smears were compared with subsequent histologic findings. The comparison demonstrated (1) a high rate of conclusive cytologic diagnoses in the assessment of metastatic malignancies, with an overall accuracy rate of 99.1% and a typing accuracy rate of 96.5%; (2) a high rate of conclusive diagnoses in the assessment of high grade non-Hodgkin's lymphomas and Hodgkin's disease, with the exception of the lymphocytic predominance variant of the latter; and (3) significant limitations in the assessment of low grade non-Hodgkin's lymphomas because of the high rate of false-negative diagnoses in cases with a substantial nonmalignant cell component. This was particularly evident in follicular centroblastic-centrocytic lymphomas. Immunocytochemistry appeared to be of limited value in the distinction between centroblastic-centrocytic follicular lymphomas and reactive follicular hyperplasia. The results confirmed the diagnostic value of fine needle aspiration as the first step in the workup of patients with nodal enlargement suspicious for malignancy. In the area of low grade non-Hodgkin's lymphomas, morphologic and immunocytochemical methods need to be supplemented by molecular techniques in order to achieve conclusive diagnoses.

Fine-needle aspiration of parotid masses.

BACKGROUND: There is controversy concerning the utility of fine-needle aspiration in diagnosing parotid masses. Even studies on large series of patients have compared aspiration findings with the histology in much fewer cases. METHODS: Preoperative fine-needle aspiration findings were compared with the histopathologic diagnoses from surgically resected specimens in 246 patients presenting with and treated for parotid mass from 1980-1990. RESULTS: Of 173 benign tumors, 159 (91.9%) were diagnosed correctly and 110 of 144 (> 60%) were typed. Of 36 malignant tumors, malignancy was recognized in 22 cases (61.1%). There were nine false-negatives, and in five cases, the specimen was unsatisfactory. The four cases of metastatic disease were correctly typed. Only two of seven lymphomas (28.6%) were identified. The cytologic and histologic diagnoses were concordant in all cases of nonneoplastic disease. Overall accuracy was 87%. CONCLUSIONS: Fine-needle aspiration speeds up the diagnostic process and, with close cooperation between clinician and pathologist, the technique is a valuable adjunct to preoperative assessment in patients with parotid masses.

The role of fine needle aspiration in the assessment of renal masses.

An investigation of the role of fine needle aspiration in the assessment of renal masses was carried out on 132 consecutive patients, 11 of whom were children. Selection was based on the presence of a solid or mixed solid and cystic renal mass that could not be defined by radiology, either in symptomatic patients or in patients to be submitted to embolization of the renal artery and at high surgical risk. Histologic and clinical data showed 49% of the cases to have a malignant, predominantly (45%) primary disease of the kidney and the remainder to have a nonneoplastic lesion. In five cases, the primary lesion was in the adrenal gland (three neuroblastomas and two pheochromocytomas). Sensitivity, specificity and predictive values for positive results were, respectively, 0.93, 0.96 and 0.935 because of a false-positive diagnosis in a case of multilocular cystic nephroma. Furthermore, in 43 of 65 cases (66%), consisting of 33 renal cell carcinomas, 1 transitional cell carcinoma, 3 Wilm's tumors, 1 neuroblastoma, 2 pheochromocytomas and 3 metastatic lesions, the histologic type could be ascertained on the tissue yielded by the fine needle aspiration. The findings stress the usefulness of this method for the clarification of radiologically not unequivocal space-occupying lesions of the renal area, especially when, in addition to the smears, histologic sections of paraffin-embedded tissues are available. Ultrastructural and immunohistochemical studies are cost-effective mainly in pediatric patients.

Sensitivity enhancement of the cytologic detection of cancer cells in effusions by monoclonal antibodies.

Cells from 229 pleural and peritoneal spontaneous fluids and 51 peritoneal lavage fluids from patients with neoplastic and nonneoplastic diseases were studied by indirect immunofluorescence with two monoclonal antibodies; MBr1, prepared against breast carcinoma, and MOv2, prepared against ovarian carcinoma. The results were correlated with those obtained by conventional cytologic methods. A cytologic diagnosis of metastatic carcinoma was established in about 50% of the fluids examined. Sixty percent of the cytologically malignant fluids contained tumor cells reactive with at least one of the two monoclonal antibodies tested. The specificity of the labeling was confirmed by immunoelectron microscopy. In addition, 16 fluids with a negative cytologic diagnosis contained cells strongly immunopositive with MBr1 and/or MOv2. Reactive mesothelial cells were consistently negative. These results suggest that antibodies MBr1 and MOv2 are able to identify cancer cells that do not fully meet conventional morphologic criteria for malignancy. The two reagents, when used in support of cytologic analysis, may substantially reduce the number of false negative cytologic diagnoses of fluids from patients with breast and ovarian carcinomas.

Histologic evidence for an association of cervical intraepithelial neoplasia with human papilloma virus infection.

All the cytologic and histologic material pertaining to 100 patients who underwent cervical conization for advanced cervical intraepithelial neoplasia (CIN) was reviewed. The revision of the histology of the biopsies and cones showed in 56 cases the association of CIN with viral cytopathic effects (VCE) attributable to human pappiloma virus (HPV) and in 52 the coexistence of a predominantly flat condyloma. The comparison of the two groups of CIN, with and without VCE, showed that in the first the association had favored in 20% of the cases the histologic overestimation of the severity of the lesion. Of the patients with CIN III, 46% showed additional changes due to VCE. The mean age of the patients with CIN and VCE was 39.8 years and that of the patients with CIN was 48.6 (p less than 0.0001). The exocervix was significantly more often involved by CIN + VCE than by CIN alone (p less than 0.00001). Follow-up studies revealed in both groups the same percentage of residual disease and, preliminarily, a trend to a better control of CIN with VCE. New disease developed more often in the group of patients with CIN without VCE. Cytologic sensitivity for VCE in cervical smears was high (95%) in the cases of CIN II and somewhat lower (81%) in those with CIN III. Cytologic follow-up showed the persistence of VCE in 17% of the patients treated surgically for CIN and VCE. The morphologic and clinical features displayed by CIN associated with VCE warrant its recognition as a distinct variant of CIN.

Condylomata of the uterine cervix and koilocytosis of cervical intraepithelial neoplasia.

In 202 women with koilocytotic atypia in cervical smears, 136 had predominantly small condylomata of the uterine cervix, and 66 had cervical intraepithelial neoplasia (CIN) of varying degree either with koilocytosis of the neoplasia or associated with condylomata. Koilocytosis correlated well with the histological diagnosis of condylomata, but occasionally it obscured the cytological evidence of CIN. Human papilloma virus particles were found in the cells of condylomata in 10 cases and in those of CIN II with koilocytosis in two cases of 21 examined ultrastructurally. There was evidence that the condyloma of the uterine cervix is a well-defined morphological entity and also that cytopathie changes similar to those seen in condylomata are present in some cases of CIN.

[Cytological changes caused by intrauterine devices]. / Alterazioni citologiche da dispositivi intrauterini.

PIP: As part of a mass screening program, 100 cervical-endocervical smears of IUD wearers were examined and compared with the smears of 100 women in a control group. A positive relationship between the use of IUDs and the cellular changes in the glandular epithelium, both of endocervical and endometrial origin, could be demonstrated. The most striking abnormalities occurred in the cytoplasms; however, the nuclei also showed some degree of variation in size and shape, thus giving rise to problems concerning the differential diagnosis with atypias due to cervical intraepithelial neoplasia in various stages as well as to adenocarcinoma and its precursors. (author's)^ieng

Cytopathologic manifestation of uterine leiomyosarcoma. Case report.

The presence of malignant tumor cells in a vaginal and cervical smear of a 53-year-old female with vaginal bleeding and with subsequent negative hsitology of the scraping material could be later correlated with a leiomyosarcoma of the myometrium that produced deep local invasion and pulmonary metastases. The findings that favor the cytologic diagnosis of this neoplasm are the presence of isolated cells or the side-by-side arrangement of the tumor cells, the elongated shape of the cytoplasm, and the ovoidal cigar-shaped nuclei with sparse, coarse chromocenters. Cytologic differential diagnosis of other malignant neoplasms capable of cellular exfoliation into the vagina is discussed.

The cytologic diagnosis of gastric carcinoma related to the histologic type.

In gastric smears obtained by direct vision fiberoptic brush technique from 78 patients with carcinoma of the stomach, an attempt was made to recognize cytologically the histologic type of the tumor with reference to Lauren's classification. The cytologic diagnosis of intestinal type carcinoma was made in 36/45 positive cases on the basis of an abundant cellularity and the presence of rather large pleomorphic cohesive cells often arranged in sheets with a moderately increased N/C ratio. One case of intramucous and two of "early" invasive carcinoma revealed malignant cells which did not differ from those of the advanced cases. In these cases as well as in some of the advanced ones, atypical epithelial cells were found in addition to the malignant ones; these cells could have derived from the histologic areas of atypical hyperplasia of the gastric mucosa surrounding the carcinoma. 14/15 cases of advanced diffuse carcinoma of the stomach could be cytologically identified on the basis of a scanty cellularity and the presence of rather small, monomorphic poorly differentiated cells with a high N/C ratio. The cytologic diagnosis of the mixed-type carcinoma was made in 2/5 positive cases on the basis of the presence of an admixture of both cell types described above. In two cases of the mixed-type carcinoma, only intestinal type cells were found. In the smears of nine cases of intestinal type carcinoma, one of which was intramucous, and of one case of mixed-type carcinoma, the tumor cells could not be specified. 13/78 cases (16.7%) showed negative cytology. The overall accuracy rate was 83.3 per cent. The statistical analysis of a number of cytologic parameters indicated that morphologic differences between Type I and Type D carcinomas of the stomach do exist and that they can be evaluated for differential diagnostic purposes.