Additional slides from residual ThinPrep Pap tests: Of potential diagnostic benefit in equivocal cases?

The aim of this study was to further evaluate the diagnostic significance of additional slides prepared from residual ThinPrep (TP) Pap Tests. Up to 10 repeat slides were prepared from 105 residual TP cervical samples. All additional slides were evaluated for the presence of diagnostic elements which were not found on the primary TP slide. After the evaluation of the repeat slides, an upgraded diagnosis was noted in 15 cases (14.3%). The reclassified cases included: three negative cases reclassified as two ASC-US and as one LSIL, seven cases of ASC-US reclassified as six LSIL and as one HSIL, and five cases of LSIL reclassified as HSIL. The highest rate (7/15 cases, 46.7%) of cases with an upgraded diagnosis was noted in the ASC-US diagnostic category. Our results suggest that repeat processing of residual TP cervical samples may represent an adjunctive diagnostic tool for a more accurate classification of ASC-US cases. Nevertheless, the practical value of this approach seems to be limited by its significant cost and its uncertain effectiveness.

Repeat processing of residual ThinPrep Pap tests: sampling of the vial may not be invariably homogeneous.

OBJECTIVE: To re-evaluate the reproducibility of additional slides prepared from residual cervical ThinPrep (TP) samples. STUDY DESIGN: Sixty paired specimens (conventional smears and direct-to-vial TP) were studied. Up to 10 additional slides were prepared from each TP vial. All slides were reviewed for adequacy of material, presence of abnormal cells and presence of normal flora or other pathogens. The additional TP slides were further evaluated for the presence of diagnostic elements which were not found on the conventional smear and primary TP slide. RESULTS: Abnormal cells found on the primary TP slide were also identified on all additional slides in 48/50 cases (96%) with squamous cell lesions. The distribution of material on TP slides was evaluated as homogenous in 51 cases (85%) and as non-homogenous in 9 (15%). Using the primary slides (conventional smear and TP) as a reference, additional diagnostic cells upgrading the cytologic diagnosis were found on the repeat slides in 7 cases (11.7%) and fungi consistent with Candida in 3 (5%). CONCLUSION: Repeat processing of residual cervical TP samples may not be an invariably reproducible procedure and the first slide may not be necessarily representative of the specimen as a whole. Nevertheless, both primary and repeat TP slides seem to be extremely effective in detecting a lesion (regardless of grade) in abnormal cases. The exact impact of non-homogeneous sampling of the vial on the diagnostic accuracy of the TP method should be further investigated.

p53 protein expression and oestrogen and progesterone receptor status in invasive ductal breast carcinomas.

p53 protein expression and oestrogen and progesterone receptor status in invasive ductal breast carcinomas The p53 protein expression and oestrogen and progesterone receptors status was investigated in correlation to the grade of malignancy of primary breast carcinomas. Our material constituted imprints from surgical biopsies of 75 invasive ductal breast cancer cases. The p53 protein expression was investigated immunocytologically using the monoclonal antibody p53 DO-7 (DAKO). A biochemical DCC method was applied for the detection of oestrogen and progesterone receptors for all tumours. Fifty-one percent of breast cancer cases were p53 protein positive. A statistically significant association of p53 protein expression and high tumour grade was found (chi2=23.72, d.f.=2, P < 0.001). A statistically significant association was also found between oestrogen and progesterone receptor positive cases and the grade of malignancy (P < 0.001). A negative association between p53 protein expression and oestrogen (ER) and progesterone receptors (PgR) positivity was found. From our results it appears that it is possible to distinguish from grade II tumours two subgroups of cases, one with low malignancy potential and p53 (-), ER (+), PgR (+), and another subgroup with high malignancy potential and phenotype p53 (+), ER (-), PgR (-). The last subset of patients could actually benefit from adjuvant therapy.

Histological diagnosis of cutaneous "warty" carcinoma on a pre-existing HPV lesion.

A middle-aged man with an 8-year history of a fungating tumour mass on his thigh was histologically diagnosed as having an invasive "warty" carcinoma at the location of a pre-existing human papillomavirus (HPV) lesion. The tumour surface had a verruciform appearance with papillae containing fibrovascular cores. Many of the malignant cells displayed changes consistent with koilocytotic atypia. We noticed a greater degree of nuclear atypia in comparison with a verrucous carcinoma. Focally, some neoplastic cells demonstrated features of basaloid differentiation. Under in situ hybridization conditions, only HPV16 DNA, which is commonly associated with genital neoplasia, was detected selectively in rather superficial areas, corresponding to morphological evidence of HPV infection (i.e. koilocytotic atypia). It is worth noting that cutaneous location of such a warty carcinoma is very rare.

p53 protein expression and proliferative activity in ductal breast carcinomas.

The immunocytochemical expression of p53 protein and Ki-67 labelling index in tumour cells of 100 ductal breast carcinomas of different histological grade and stage was evaluated in cytological material. In order to investigate p53 expression and Ki-67 expression an avidin-extravidin immunocytochemical technique was applied to imprints. Monoclonal antibody (MoAb) DO-p53 and proliferating cell monoclonal antibody were used as primary antibodies. A statistically significant difference was observed between p53 protein expression and grade of malignancy and clinical stage (P = 0.001, P < 0.001, respectively). A statistically significant difference was also observed between Ki-67 LI and histological grade and stage of the tumours (P < 0.001, P < 0.001 correspondingly). A correlation was observed between p53 protein expression and Ki-67 LI (P < 0.001). The immunocytochemical study of p53 protein and Ki-67 expression in cytological material represents a simple method which can be applied in routine cytological laboratories for the investigation of potential malignancy of ductal breast cancer.

p53 protein expression and proliferative activity in imprints of superficial transitional cell carcinoma of the bladder.

OBJECTIVE: To investigate p53 protein expression and proliferative activity in imprints of tumor biopsies from superficial transitional cell carcinoma of the bladder in relation to the histologic grade of malignancy and recurrence status. STUDY DESIGN: The study group consisted of 70 cases of superficial transitional cell carcinoma of the bladder. In order to investigate p53 protein expression and Ki-67 expression, an immunocytochemical avidin-extravidin complex technique was performed using monoclonal antibodies p53 D0-7 and proliferating cells correspondingly. RESULTS: Thirty-seven percent of superficial transitional cell carcinoma cases showed positive expression of p53 protein. No correlation was found between p53 protein expression and grade of malignancy (P = .45). p53 Protein expression was statistically correlated with a high Ki-67 labeling index (LI) (P < .001) and recurrence status (P < .001). Forty-seven percent of cases showed a Ki-67 LI > 25%. No correlation was found between a high Ki-67 LI and grade of malignancy (P = .703). A significant difference in high Ki-67 LI between recurrent and nonrecurrent tumors of the same grade (P < .001) and between recurrent and nonrecurrent tumors was found independently of grade (P < .001). CONCLUSION: These results on cytologic material could provide useful information on the biologic behavior of superficial transitional cell carcinoma of the bladder at the time of diagnosis.

DNA ploidy and p53 protein expression in superficial transitional cell carcinoma of the bladder.

Superficial transitional cell carcinoma of the bladder (STCCB) is a heterogeneous group of neoplasias with an unpredictable clinical course. In recent years many techniques have been used in order to predict the behaviour of these tumours at individual patient level. The aim of this study was to investigate in imprints from tumour biopsies the DNA ploidy and p53 protein expression in a group of 80 STCCB (pTa-pT1) patients in relation to histological grade and recurrence status. The DNA content was studied in Feulgen-stained imprints by the image analysis technique using a SAMBA 2005 analyser. In order to investigate p53 protein expression an avidin-extravidin immunocytochemical technique was used. According to our measurements a strong correlation was observed between recurrence status and DNA ploidy status (P < 0.001). No statistical difference was found in DNA ploidy status and grade of malignancy (P = 0.68). A statistically significant difference was found in p53 protein expression between recurrent and nonrecurrent tumours (P < 0.001). No statistically significant difference was found among tumours of grade I, grade II and grade III (P = 0.42). These results could provide useful information on the potential behaviour of STCCB.

Learning vector quantizer in the investigation of thyroid lesions.

OBJECTIVE: To investigate the capability of the learning vector quantizer (LVQ) in the discrimination of benign from malignant thyroid lesions. STUDY DESIGN: The study was performed on May-Grünwald-Giemsa-stained smears taken by fine needle aspiration (FNA). Using a custom image analysis system, 25 features that describe the size, shape and texture of approximately 100 nuclei were measured from each case. Statistical features were extracted from each case, and a linear regression analysis was performed to detect the statistically significant features. The cases were distributed by category, as follows: 100 cases of goiter and follicular adenomas, 11 cases of follicular carcinoma, 35 cases of papillary carcinoma, 24 cases of oncocytic adenoma, 8 cases of oncocytic carcinoma and 20 cases of Hashimoto thyroiditis. About 30% of the cases from each class were used for training two LVQ classifiers. The remaining 139 cases, out of a total of 198, were used as the test set. A classifier was used to discriminate into four classes and a second into two classes. RESULTS: The application of LVQ neural networks allows good discrimination between benign and malignant lesions (O.A. = 97.8). However, reliable discrimination of the cytologic types of the lesions was not obtained. CONCLUSION: These results indicate that the use of neural networks combined with image morphometry may offer useful information on the potential for malignancy of thyroid lesions and may improve the diagnostic accuracy of FNA of the thyroid gland, especially in cases of follicular neoplasms classified as suspicious for malignancy and in cases of oncocytic tumors.

Image cytometric DNA analysis and proliferating cell nuclear antigen (PCNA) expression in transitional cell carcinoma of the bladder.

The proliferation rate as determined by Proliferating Cell Nuclear Antigen (PCNA) immunostaining and the DNA ploidy status as measured by static cytometry were studied in 70 transitional cell carcinomas of the bladder (TCCB) in relation to grade, stage, and recurrence. The follow-up period was 2 years. A significant difference was observed in PCNA expression among grades I, II, and III (P < 0.02), between superficial (pTa-pT1) and invasive (pT2-pT4) tumors (p < 0. 04), between recurring and non-recurring tumors (p < 0.001), and between tumors of the same grade with and without recurrence (p < 0. 05). A significant difference was also found in the ploidy pattern among grades I, II, and III (p = 0.002), and between superficial and invasive (p = 0.02) and recurring and non-recurring tumors (p < 0. 01). Finally, the recurrence status seems to be strongly influenced by the proliferation rate and ploidy of TCCB (p < 0.001). Our results suggest that the above-studied parameters may offer useful information on the biological behavior of TCCB.

Changes of chromosomes 1, 3, 6, and 11 in metastatic effusions arising from breast and ovarian cancer.

This cytogenetic study deals with cell material obtained from 15 pleural fluids from 11 patients with breast cancer and 27 ascitic fluids from 16 patients with ovarian cancer; in addition, 8 pleural, 5 ascitic, and 1 pericardial fluid from patients with tuberculosis, liver cirrhosis, and heart insufficiency, were studied. Using mainly direct methods, as well as short-term cell cultures, the chromosome spreads were GTG-banded. Cancerous biopsies showed a plethora of numerical and structural chromosome anomalies and exhibited broad aneuploidy. Chromosomes participating more often in numerical and structural aberrations were 1, 3, 6, 7, 8, 9, 11, 12, and 17. This study provides further cytogenetic evidence for the involvement of these chromosomes in breast and ovarian malignancy.

p53 protein expression in benign and malignant breast lesions.

OBJECTIVE: To investigate p53 protein expression in imprints from benign and ductal breast carcinoma cases in relation to the histologic grade of malignancy and clinical stage. STUDY DESIGN: The study group consisted of 60 cases of primary ductal breast carcinomas and 20 benign lesions. For the demonstration of p53 protein expression, an immunocytochemical avidin-extravidin complex technique was applied. Monoclonal antibody p53 was used as the primary antibody, diaminobenzidine as the chromogen and hematoxylin as the counterstain. RESULTS: Forty-five percent of breast cancer cases showed positive expression of p53. A statistically significant difference in p53 protein expression was observed between grade 1, 2 and 3 carcinomas and stage I, II and III cases. All benign lesions were negative for p53 protein expression. CONCLUSION: Immunocytochemical p53 protein expression in cytologic material is a simple method that can be applied in routine cytologic laboratories for the identification of genetic alterations in primary ductal breast cancer.

Evaluation of cytological morphologic criteria and AgNOR expression in male breast lesions.

Fine needle aspiration (FNA) breast smears from 81 male patients have been examined in our laboratory between 1988 and 1994. The cytological criteria for diagnosing male breast lesions and the expression of nuclear organizer regions were evaluated. Of the 81 cases examined cytologically, 50 cases were proven cytologically and histologically to be inactive gynaecomastia, two cases showed florid gynaecomastia and there were 10 adenocarcinomas; in the 17 cases in which no cytological evidence of gynaecomastia or malignancy was found, the histological diagnosis was gynaecomastia in 13 and there was one case of mastopathy; in two cases suspicious of malignancy on cytology the histological examination proved to be florid gynaecomastia in one case and the other showed an adenocarcinoma. The absolute specificity of FNA in this study was 74.28%, the complete specificity 98.5%, the absolute sensitivity 90.9% and the complete sensitivity 100%. The overall accuracy was 97.5%, the positive predictive value 91.66% and the negative predictive value 98.5%. In all cases of male breast carcinoma, AgNOR mean value was > or = 3; thus, it appears that AgNOR mean value 3 could be used as a cut-off value between benign and malignant male breast lesions. Our experience suggests that FNA is an acceptable procedure for the investigation of male breast lesions.

A comparative study of three variations of the learning vector quantizer in the discrimination of benign from malignant gastric cells.

A prospective study was undertaken to investigate the potential value of morphometry and artificial neural networks (ANN) for the discrimination of benign and malignant gastric lesions. Two thousand five hundred cells from 23 cases of cancer, 19 cases of gastritis and 58 cases of ulcer were selected as a training set, and an additional 8524 cells from an equal number of cases of cancer, gastritis and ulcer were used as a test set. Images of routine processed gastric smears stained by the Papanicolaou technique were processed by a custom image analysis system. The application of the learning vector quantization (LVQ) classifier enabled correct classification of > 97% of benign cells and > 95% of malignant cells, obtaining an overall accuracy of > 97%. This study presents the capabilities of ANN, and also indicates that ANN and image morphometry may offer useful information on the potential of malignancy in gastric cells.

P53 protein expression and DNA ploidy in common epithelial tumors of the ovary.

OBJECTIVE: To investigate p53 protein expression and DNA content in imprints from surgical biopsies of common epithelial tumors of the ovary. STUDY DESIGN: The study was based on 60 cases of epithelial tumors of the ovary (15 benign, 3 border-line and 42 malignant). For the demonstration of p53 protein, immunocytochemical staining with the avidin-extravidin technique was performed using monoclonal antibody p53 DO-7. DNA content was measured by image cytometry after Feulgen staining. RESULTS: There was a strong correlation between p53 expression and aneuploidy, with the difference between diploid and aneuploid tumors statistically significant (P < .001). A correlation was found between DNA ploidy, histologic grade and clinical stage (P < .001 and P < .05), respectively. There was no correlation between DNA ploidy and histologic type (P = .89). No correlation was observed between p53 protein expression and grade or clinical stage of the tumors. Nevertheless, a correlation of p53 expression between early (I, II) and advanced stages (III, IV) (P < .05) was observed. All benign and borderline tumors were diploid and did not express p53 protein. CONCLUSION: The results of the present study and the data in the literature stress the value of p53 expression and DNA ploidy in assessing the malignant potential of common epithelial ovarian cancers. However, the clinical application of these data requires further study.

Application of the learning vector quantizer to the classification of breast lesions.

OBJECTIVE: To investigate the potential of the learning vector quantization (LVQ) neural network for the discrimination of benign from malignant breast lesions. STUDY DESIGN: Using a custom image analysis system on Giemsa-stained smears, 25 parameters describing the size, shape and texture of the cell nucleus were measured. Three thousand nuclei from a total of 9,356 were selected as a training set for the neural network, and the whole data set was used for testing. An additional 238 cells from 16 cases without final cytologic diagnoses were evaluated by the system. The total number of cells (9,594) was collected from 100 patients (68 carcinomas and 32 benign lesions). RESULTS: Cytologic examination of the cases gave two false positive and two false negative results. However, in eight cases of ductal breast carcinoma and in eight cases of benign lesions, histologic confirmation was necessary in order to confirm the cytologic diagnosis. Application of the LVQ permitted correct classification of 87.41% of the cells. Classification at the patient level by using a hypothesis test for proportion with a hypothesis value equal to 50% permitted the correct diagnosis in 98% of patients. CONCLUSION: These results indicate that the use of neural networks combined with image morphometry and statistical techniques may offer useful information about the potential for malignancy, improving the diagnostic accuracy of fine needle aspiration of breast lesions.

DNA content and p53 protein expression in ductal breast cancer.

The DNA content of 85 ductal breast cancers of different histological grades was evaluated using static cytometry and correlated with immunocytochemical expression of p53 protein in tumour cells in cytological material. A statistically significant difference was observed between p53 protein expression and grade of malignancy (P < 0.001). The percentage of euploid tumours significantly decreased from grade I through grade II to grade III tumours (P < 0.001). Clonal DNA heterogeneity was observed in 26.6% of cases analysed and was correlated with p53 protein expression (P < 0.001). These changes probably reflect genomic alterations which may affect potential malignancy of breast cancer.

DNA ploidy and pS2 protein expression in breast cancer.

The DNA content of ductal breast carcinomas of varying histological grade was measured using static image cytometry and correlated with pS2 expression in the tumour cells. Our study was performed on imprint of surgical biopsies of 60 women with ductal breast cancer. A statistically significant difference was observed between pS2+ expression and grade of malignancy (P < 0.001). The percentage of euploid tumours significantly decreased from grade I to grade II to grade III (P = 0.01). The percentage of aneuploid tumours increased from pS2+ to pS2- breast tumours (P < 0.001). These findings may be indicative of pS2 and DNA ploidy alterations and tumour aggressiveness.

Expression of pS2 protein and estrogen and progesterone receptor status in breast cancer.

OBJECTIVE: To investigate pS2 protein expression and estrogen (ER) and progesterone receptor (PR) status of imprints from surgical biopsies of breast cancer cases in relation to the histologic grade of malignancy. STUDY DESIGN: The study group consisted of 50 cases of primary breast carcinomas. For the demonstration of pS2 protein expression an immunocyto-chemical avidin-biotin complex technique was applied. Monoclonal antibody pS2 was used as the primary antibody, diaminobenzidine as the chromogene and hematoxylin as the counterstain. For the evaluation of ERs and PRs, a biochemical method was applied. RESULTS: Sixty-two percent of breast cancer cases showed positive expression of pS2. Of the 31 pS2+ cases, 74% had positive ERs and PRs. A statistically significant difference was observed between pS2 protein expression, ER+, PR+ and histologic grade of malignancy (P < .001). CONCLUSION: Two groups of breast cancer cases can be distinguished: one group that is ER+, PR+ and pS2+, with low malignancy potential, and another group that is ER-, PR- and pS2-, with high malignancy potential.

DNA content and proliferation activity in superficial transitional cell carcinoma of the bladder.

The identification of new prognostic parameters in Superficial Transitional Cell Carcinoma of the Bladder (STCCB) is important since conventional methods are often insufficient for prognostic purposes. We studied the proliferation activity and the DNA ploidy status of 60 pTa and pT1 Superficial Transitional Cell Carcinoma of the Bladder in relation to grade and recurrence rate. The proliferative activity was investigated by measuring the PCNA expression in paraffin embedded tissue sections. The DNA content was studied in Feulgen stained imprints by image analysis technique using a SAMBA 2005 analyser. According to our measurements a statistically significant difference was found in PCNA expression among tumors grade I, grade II, grade III (F = 5.43, p < 0.001), between tumors of the same grade with, and without recurrence (p < 0.001); and between recurrent and non-recurrent tumors (T58 = -6.03, p < 0.001). A statistically significant difference was also observed concerning the DNA-index among grade I, grade II and grade III (F = 4.81, p < 0.01), and between recurrent and non-recurrent tumors concerning the DNA DNA ploidy status (DNA-euploid vs. DNA-aneuploid tumors) (X2 = 24.96, p < 0.001). The recurrence status is also strongly influenced by the proliferation rate and the DNA ploidy status of tumors (X23 = 41.19, p < 0.001). No cases recurrence were found in the group of DNA-euploid tumors with PCNA. expression lower than 30%, in contrast a very high percentage of recurrence in patients with DNA-aneuploid tumors with PCNA expression higher than 30%. Although a small proportion of cases could not be included in me previous categories, STCCB may be classified in to main groups concerning the risk of recurrence. In keeping with this view of proliferation rate and DNA ploidy status could provide useful information, on the potential malignancy of Superficial Transitional Cell Carcinoma of the Bladder. However further studies are required to establish the clinical utility of these parameters.

DNA ploidy and vimentin expression in primary breast cancer.

The DNA content of 50 breast cancers of varying tumour type, grade and stage was measured using static image cytometry, and correlated with vimentin expression in the tumour cells. A tendency to increased vimentin expression and aneuploidy was observed in high grade and late stage tumours. A statistically significant difference was observed in DNA index and ploidy balance between grade 1 and grades 2 and 3 carcinomas (P < 0.05) and between grade I and stage II carcinomas (P < 0.05). There was a significant difference in the expression of vimentin between grades 1, 2, 3 (P < 0.001), and stages I, II, and III ductal carcinomas (P < 0.05). No significant difference was observed in the proliferation index and the degree of hyperploidy (P > 0.05) Clonal heterogeneity was observed in 25% of breast carcinomas, and was associated with increased vimentin expression. These changes may be indicative of genomic alteration and tumour aggressiveness.