Fractal dimension of cell clusters in effusion cytology

In this study, the fractal dimension (FD) of the ball‐like tight clusters of known metastatic adenocarcinoma cells is compared with the tight cell clusters of the benign cells in the effusion fluid to find out the role of FD in differentiating malignant from benign cluster in effusion. A total of 68 and 72 images of cluster of cells from 12 cases of benign and 13 cases of metastatic pleural and peritoneal effusion were studied for FD. The mean FD of the cluster of cells of benign and malignant effusion was 1.4801 ± 0.23260 and 1.7175 ± 0.09006, respectively. Mann–Whitney U‐test showed significant difference of FD between the cell clusters of malignant and benign effusion cases (P < 0.0001). Cytological features along with the measurement of FD of cell clusters in effusion cytology may be helpful in differentiating benign from malignant cases. Diagn. Cytopathol. 2010;38:866–868. © 2010 Wiley‐Liss, Inc.     

Fractal Analysis of Contours of Breast Masses in Mammograms

Fractal analysis has been shown to be useful in image processing for characterizing shape and gray-scale complexity. Breast masses present shape and gray-scale characteristics that vary between benign masses and malignant tumors in mammograms. Limited studies have been conducted on the application of fractal analysis specifically for classifying breast masses based on shape. The fractal dimension of the contour of a mass may be computed either directly from the 2-dimensional (2D) contour or from a 1-dimensional (1D) signature derived from the contour. We present a study of four methods to compute the fractal dimension of the contours of breast masses, including the ruler method and the box counting method applied to 1D and 2D representations of the contours. The methods were applied to a data set of 111 contours of breast masses. Receiver operating characteristics (ROC) analysis was performed to assess and compare the performance of fractal dimension and four previously developed shape factors in the classification of breast masses as benign or malignant. Fractal dimension was observed to complement the other shape factors, in particular fractional concavity, in the representation of the complexity of the contours. The combination of fractal dimension with fractional concavity yielded the highest area (A ( z )) under the ROC curve of 0.93; the two measures, on their own, resulted in A ( z ) values of 0.89 and 0.88, respectively.     

Image analysis of irregularity of cluster shape in cytological diagnosis of breast tumors: cluster analysis with 2D-fractal dimension

To establish diagnostic criteria using comparison of cell cluster shapes, between benign and malignant tumors, breast tumors demonstrating weak cellular atypia in low grade invasive ductal carcinoma (IDC) were compared. Fine-needle aspiration (FNA) specimens of breast tumors were obtained from 37 patients. Among these, 16 were histologically diagnosed as IDC low-grade and the other 21 as benign fibroadenoma (FA). For evaluation, we examined 740 clusters from these 37 FNA specimens. Nine image morphometric parameters were studied, including the cluster area, circumference, maximal length, maximal breadth, ratio of length to breadth, cluster roundness, cluster size, and the edge and distribution image fractal dimensions for cluster analysis. We evaluated the irregularity in cell cluster shape using fractal dimension analysis, and determined the correlation to cluster size. The irregularity in the IDC cluster shape was higher than that in the FA cluster shape. However, six cases (28.5%) of 21 FA clusters showed high fractal dimensions similar to those for IDC. The clusters were classified by cluster analysis into three types: IDC clusters, FA with irregular cluster shape, and FA with no irregular clusters. The average cell cluster area of the FA with irregular shape was found to be about three times larger than that of IDC clusters. When the differential diagnosis between IDC and FA is difficult, it is important to focus on irregularities in the shape and on overall size of the cell clusters. For accurate diagnosis, the cell cluster shape is as important as the individual cellular atypia.     

Role of DNA flow cytometry and image cytometry on effusion fluid

The objective of the study was to assess the value of DNA flow cytometry (FCM) and image cytometry (ICM) as an adjunct to routine diagnostic cytology. In this prospective study, 100 consecutive effusion fluids were studied for routine cytology, DNA FCM, and in selected cases, ICM. One half of the centrifuged fluid sample was used for routine cytology and the remaining portion was used for DNA FCM. Nuclear area, nuclear diameter, nuclear perimeter, nuclear convex perimeter, nuclear roundess, and nuclear convex area were measured on at least 100 cells by ICM in cytologically malignant or DNA aneuploid cases along with control cases. Clinical follow-up was done in all cases. There were 22 cytologically malignant cases and 78 cytologically benign cases. Among the 22 cytologically malignant cases, there were 11 aneuploid and diploid cases each by DNA FCM. Out of 78 cytologically benign cases, six (7.7%) were aneuploid by DNA FCM. Smears of these cases showed predominantly reactive mesothelial cells, but the DNA histograms showed hypodiploid (one), hyperdiploid (three), tetraploid (one), and hypertetraploid (one) aneuploidy. Follow-up of these cases showed clinical or histologic features of malignancy except in one case of tetraploid aneuploidy, which did not show any features of malignancy and responded well to antitubercular therapy. Therefore, out of 27 malignant effusions, DNA FCM picked up 16 cases and routine cytology detected 22 cases. Sensitivity and specificity of DNA FCM were thus 59.25% and 98.63%, respectively. There was a statistically significant difference (Student's unpaired t-test, P < 0.05) between cytologically malignant cases and control benign cases in all the nuclear morphometric parameters except for nuclear roundness. There was, however, no statistically significant difference of nuclear morphometric parameters between cytologically benign vs. DNA aneuploid cases and control benign cases. DNA FCM is a useful adjunct for routine diagnostic cytology. Visual diagnostic cytology and morphometric digital microscopy miss some cases of malignancy which can be detected by DNA flow cytometry. Diagn. Cytopathol. 2000;22:81-85.     

Cell cannibalism: diagnostic marker of malignancy

In this study, we tried to find out the cytological relevance of cannibalism as a dependable feature of malignancy in effusion and urine cytology. We randomly selected a total of 40 cases consisting of 10 each of malignant effusion, benign effusion, malignant urine samples, and benign urine samples. These smears were assessed for the presence of cell cannibalism. The number of cannibalistic cells/100 tumor cells was counted. The cannibalistic cells were seen more commonly in malignant effusion cases (3.4/100 cells) compared with malignant urine cases (2/100 cells). There was not a single cannibalistic cell in benign conditions. The finding of an increased number of cannibalistic cell was highly significant in malignant versus benign samples (P > 0.0000, Student's t-test). The present study highlights the significance of cannibalism in malignant urine and effusion cytology. We suggest that cell cannibalism is a dependable cytological feature of malignancy.     

Micronucleus assay in buccal smears of breast carcinoma patients

The aim of this study is to compare micronucleus assay in buccal smear of breast carcinoma patients versus normal benign cases as control group. In this prospective study, we selected a total 32 patients of carcinoma of breast and 49 patients of benign breast lesions diagnosed in fine needle aspiration cytology (FNAC). Acridine orange stain was done on buccal smears of these cases and micronucleus (MN) scoring was performed in 40× 0bjective in a fluorescent microscope. The MN score was expressed as positivity per 1,000 cells. The MN scoring in buccal smear was compared in malignant and benign breast cases. In fluorescence microscope, the micronucleus was detected as round orange shaped small intracytoplasmic structure around the nucleus. The mean MN scores in buccal smears of benign and carcinoma cases were 0.5014 ± 0.45768 and 2.1938 ± 1.08656 cases respectively. Independent sample Student's t test showed significantly high MN score in buccal smear of the cancer patients (P < 0.001). Micronucleated cells are significantly increased in buccal cells of the breast carcinoma cases. The increased number of MN in buccal smears raises the possibility that the genetic damage in breast cancer patients is generalized. In future, MN scoring could be used as biomonitoring of DNA damage and in early detection of high risk cases of carcinoma of breast.     

Expression of Sialyl-Tn in fine-needle aspirates from mammographically detected breast lesions: A marker of malignancy?

Malignant transformation is frequently associated with abnormal expression of cell surface carbohydrates. Sialyl-Tn (STn) is a core carbohydrate antigen of tumor-associated mucin formed by the premature 2-6 sialylation of N-acetylgalactosamine. In an attempt to verify whether this antigen is restricted to malignant cells, we studied 30 cases of fine-needle aspiration (FNA) cytology from mammographically detected breast lesions. The rationale for choosing this material was the acknowledged difficulty in diagnosing cytologically small breast lesions, especially epithelial intraductal proliferations. The cases were divided in benign lesions (two fibroadenomas and ten ductal hyperplasias) and malignant lesions (16 ductal carcinomas). Ten of sixteen malignant cases (62.5%) were positive for STn. Five of fourteen benign cases (35.7%) were also positive for STn (two fibroadenomas and three ductal hyperplasias). The most consistent positive results in benign lesions resulted from cases that displayed apocrine metaplasia, although positivity has also been observed in ductal cells without metaplasia. We did not find statistical significant differences among STn expression in benign and malignant breast lesions detected by FNA (P = 0.14). Thus, we conclude that STn is neither specific nor sensitive for detection of malignancy in FNA from mammographically detected breast lesions. Diagn. Cytopathol. 1998;18:325–329. © 1998 Wiley-Liss, Inc.     

The estimation of mean nuclear volume in the diagnosis of breast carcinoma.

Sixty-two cases of breast pathology were randomly selected from the files of the Dunedin Public Hospital for evaluation of mean epithelial nuclear volume. The cases were comprised of both benign and malignant ductal epithelial disease, diagnosed in cytological smears or in histological sections. Nuclear volume in histological preparations was estimated by the stereological technique of point-line intercept measurements to derive volume-weighted mean volumes (vV). An index of the nuclear volume (Vi) of cytology smears was calculated from measurements of nuclear areas by either image analysis Vi(e) or point-line intercepts Vi(p). By all methods of analysis a clear distinction of nuclear volume was found between the benign [means for the cytology were 148 microns 3 (Vi(e)) or 246 microns 3 (Vi(p)), and 203 microns 3 (Vv) for the histology specimens] and malignant diseased cases [means for cytology: 524 microns 3 (Vi(e)) or 886 microns 3 (Vi(p)), and 587 microns 3 (vV) for the histology specimens].     

Utility of CK7 and CK20 immunohistochemistry in the detection of synchronous breast and colon carcinoma in a pleural effusion: a case report and supporting survey of archival material

We present a case of synchronous breast and colon carcinoma in a pleural effusion, to our knowledge the first such reported case in the English-language literature. The patient was a 55-yr-old white female with known metastatic breast and colon carcinoma who developed a malignant pleural effusion which demonstrated two strikingly different populations of malignant cells by immunohistochemical study of cell block material. One cell population demonstrated a cytokeratin (CK)7+/CK20-/ER+ phenotype, while the other demonstrated a CK7-/CK20+/ER- phenotype, consistent with breast and colon origin, respectively. An immunohistochemical survey of archival breast and colon primary and metastatic carcinomas confirmed the established CK7+/CK20- phenotype of breast and CK7-/CK20+ phenotype of colon primary carcinomas, and the maintenance of this phenotype in metastases thereof. A survey of benign and malignant mesothelial lesions confirmed the absence of staining for estrogen receptor, but showed 6/10 cases weakly positive for CK20, which has not been described in other published series. This unusual case graphically illustrates the utility of cytokeratin subset immunohistochemistry in effusion cytology.     

Are epithelial cells in fat or connective tissue a reliable indicator of tumor invasion in fine-needle aspiration of the breast?

The diagnosis of breast carcinoma tumor invasion by fine-needle aspiration (FNA) cytology continues to be controversial. To assess the reliability of predicting tumor invasion by FNA, we examined the cytologic smears of 183 FNAs of benign and malignant solid epithelial lesions of the breast for which histologic follow-up was available. The study group consisted of 94 invasive carcinomas, eight pure ductal carcinomas in situ (DCIS), and 81 benign lesions (fibroadenoma, fibrocystic changes, papilloma, adenosis). Epithelial cellularity, presence of epithelial cells in dispersed fat droplets and presence of epithelium within intact fragments of fibrofatty connective tissue were tabulated. Epithelial cellularity in dispersed fat was semiquantitatively scored. The cytologic diagnosis of the epithelial cells in all cases was recorded as benign, malignant, or indeterminant for malignancy. Findings showed that 95.5% of invasive carcinomas, 100% of DCIS, and 68.1% of benign lesions contained epithelial cells in dispersed fat; 80.8% of invasive carcinomas, 66.7% of DCIS, and 60.7% of benign lesions contained epithelial cells in intact fibrofatty connective tissue. Corrected score of epithelium within fat was 0.781 for invasive carcinoma, 0.727 for DCIS, and 0.562 for benign lesions. The difference in values for all parameters was not statistically significant between invasive carcinoma and DCIS, but reached significance between invasive carcinoma and benign lesions. Eighteen cases (7/94 invasive carcinoma, 5/8 DCIS, 6/81 benign lesions) contained atypical epithelial cells indeterminant for malignancy, all of which had epithelial cells present in dispersed fat when dispersed fat was present on the slides, indicating that this criterion was not helpful in discriminating between a benign and malignant diagnosis. We conclude that the presence of epithelial cells either admixed within dispersed fatty droplets or seemingly within fragments of fibrofatty connective tissue is not a reliable indicator of tumor invasion in FNA of the breast, and is frequently found in both benign and malignant breast lesions. The presence of epithelial cells in intact or dispersed fat is most likely a mechanical artifact of aspiration and/or smear preparation. Diagn. Cytopathol. 16:137–142, 1997. © 1997 Wiley-Liss, Inc.     

Role of morphometry in evaluation of cytologically borderline breast lesions: a study of 70 cases

Fine needle aspiration, an accepted minimally invasive diagnostic procedure for breast masses, leads to an indeterminate (borderline) diagnosis in a significant proportion of cases. This study was aimed at evaluation of the utility of image analysis and morphometry in the cytologic assessment of borderline breast cases. A 'study' group of 45 cases (including fibroadenoma, fibrocystic change and ductal carcinoma) and a 'test' group of 25 cytologically borderline cases were included. Nuclei from each of the groups (nuclei of monolayer sheet of fibroadenoma, naked nuclei of fibroadenoma, ductal cells nuclei of fibrocystic change and nuclei of carcinoma) were manually traced and analyzed using self-written image analysis software. Discriminant analysis was done on the 'study' group and the resultant discriminant functions were applied to the 'test' group. The percentage of nuclei correctly classified of the study as well as test group was noted. On step-wise discriminant analysis, various morphometric, textural parameters and color information parameters were selected. Of the original 'study' group, 98.49% of nuclei could be correctly classified. No overlapping of benign and malignant nuclei was observed in the 'study' group. When the resulting functions were tested on the 'test' group, all the cases were correctly classified as benign or malignant (compared with the final histologic diagnosis). This study suggests that application of computerized image analysis to cytologically borderline breast cases can help in the classification of these cases as benign or malignant. This may prevent unnecessary surgery in many of these cases.     

Fractal patterns for cells in culture

The present study aims to understand the growth of human malignant tumours in vitro, using the geometry of fractals as a method of analysis. The fractal dimensions of HN-5 and MDCK cell growth patterns have been measured. The first results may suggest the possibility of distinct growth processes characterized by different (time-dependent) effective fractal dimensions for MDCK and HN-5 cells. If this is true, the fractal dimension may yet prove to be a useful discriminant for comparing different diagnostic categories.     

Diagnostic value of imprint cytology during image-guided core biopsy in improving breast health care

The aim of this study was to investigate the accuracy of imprint cytology (IC) of breast core biopsy under ultrasound guidance and to assess the value of a rapid on-site preliminary diagnosis of breast lesions. A total of 437 breast core needle biopsies under ultrasound guidance with touch imprint cytology, histology, and final diagnosis were reviewed. These cases were collected from archived files at our institution. Of 437 core biopsies, IC classified 241 (55%) as benign; 22 (5%) as probably benign; 28 (6%) as probably malignant; 107 (25%) as malignant; and 39 (9%) as inadequate for IC diagnosis. Histological classifications for the 437 cases were: 285 (65%) benign; 132 (30%) malignant; 16 (4%) atypical hyperplasia; and 4 (1%) inadequate specimen. The overall sensitivity and specificity indices of IC were 95% and 96%, respectively, for benign and probably benign lesions vs malignant and probably malignant breast lesions. The overall positive and negative predictive values were 91% and 97%, respectively. The overall accuracy was 95% (379 of 398 cases, excluding specimens inadequate for IC diagnosis). IC of ultrasound-guided core needle biopsy provides a rapid and reliable preliminary diagnosis for breast lesions; it also serves as a means to verify the adequacy of biopsy specimens and to optimize the biopsy procedure. Use of IC may reduce anxiety in patients with benign lesions and expedite the diagnosis and assessment of treatment options in patients with breast cancer.     

Feature Extraction from a Signature Based on the Turning Angle Function for the Classification of Breast Tumors

Malignant breast tumors and benign masses appear in mammograms with different shape characteristics: the former usually have rough, spiculated, or microlobulated contours, whereas the latter commonly have smooth, round, oval, or macrolobulated contours. Features that characterize shape roughness and complexity can assist in distinguishing between malignant tumors and benign masses. Signatures of contours may be used to analyze their shapes. We propose to use a signature based on the turning angle function of contours of breast masses to derive features that capture the characteristics of shape roughness as described above. We propose methods to derive an index of the presence of convex regions (XR ( TA )), an index of the presence of concave regions (VR ( TA )), an index of convexity (CX ( TA )), and two measures of fractal dimension (FD ( TA ) and FDd ( TA )) from the turning angle function. The methods were tested with a set of 111 contours of 65 benign masses and 46 malignant tumors with different parameters. The best classification accuracies in discriminating between benign masses and malignant tumors, obtained for XR ( TA ), VR ( TA ), CX ( TA ), FD ( TA ), and FDd ( TA ) in terms of the area under the receiver operating characteristics curve, were 0.92, 0.92, 0.93, 0.93, and, 0.92, respectively.     

Fractal geometric analysis of material from molar and non-molar pregnancies

Histological sections from 25 non-molar pregnancies, nine partial hydatidiform moles, and 16 complete hydatidiform moles were examined (diagnosis was taken as the consensus of seven experienced histopathologists) and the fractal dimension was measured using a box-counting method implemented on a microcomputer-based image analysis system. The fractal dimensions of the different diagnostic categories were normally distributed with a mean of 1.50 for non-molar pregnancies, 1.44 for partial moles, and 1.42 for complete moles. All the measured fractal dimensions were greater than the topological dimension (1), demonstrating that the specimens had a fractal element to their structure. There was a significant difference between the fractal dimensions of non-molar pregnancies and complete moles (P=0.0005), but not between partial moles and non-molar pregnancies (P=0.0823) or complete and partial moles (P=0.4400). Using the fractal dimension to predict the histopathological diagnosis assigned 56 per cent of the cases to the correct category with a kappa statistic of 0.26, so the fractal dimension, used alone, is not a useful morphometric discriminant in the diagnosis of molar and non-molar pregnancy.     

Sclerosing papillary lesion of the breast: a diagnostic pitfall for malignancy in fine needle aspiration biopsy

Papillary neoplasms of breast constitute a group of lesions that show broad spectrum of morphological changes, ranging from benign to malignant and posing challenges at all diagnostic levels. Some benign papillary lesions may form well-defined solid masses with a dominant sclerosed architecture, known as complex sclerosing papillary lesion or simply sclerosing papilloma. The purpose of this study is to apply the previously published criteria for papillary lesions and to identify the cytomorphologic findings that lead to false-positive diagnosis of these cases. We reviewed the fine needle aspiration biopsies (FNAB) of six histologically proven sclerosing papilloma that were called suspicious or malignant on FNAB. The patient age ranged from 40 to 69, with a mean of (43 +/- 6) yr. Three patients presented with a palpable lump and two patients had history of fibrocystic disease. All six patients had abnormal screening mammograms. FNAB was performed using a 23-gauge syringe attached to a commercial holder. FNA smears were markedly hypercellular with large number of epithelial fragments and papillary clusters, discohesive single cells that are hyperchromatic with mild to moderate nuclear pleomorphism. Bipolar cells were present in all cases, varying from low to abundant. Intraoperative consultation was requested on four cases. Touch preparations were made on two cases and were reported as suspicious based on the cellularity and nuclear atypia. All surgically excised specimens showed sclerosing complex papillary proliferative lesions with epithelial hyperplasia. In conclusion, FNA cytology of this proliferative lesions may be highly cellular and may display cellular atypia similar to breast carcinoma and thus leads to false-positive interpretation.     

Metaplastic breast carcinoma on fine-needle cytology samples: a report of three cases

Metaplastic breast carcinoma (MBC) may have a varied presentation on fine-needle cytology samples. We herewith describe three cases of MBC found in our series. One of these cases showed a peculiar mixture of malignant ductal, apocrine type, and squamous epithelial cells with fascicles of spindle cells with variable degree of atypia and was diagnosed as metaplastic carcinoma of the carcino-sarcomatous type. The other two lesions were characterized by an abundant chondroid extracellular matrix to which were variably admixed carcinomatous and chondroid-type cells, with variable degree of atypia. Both these latter cases were defined as matrix-producing metaplastic carcinomas. Because of the various presentation of MBC on fine-needle cytology samples and the possible influence of needle "sampling" on the cytological specimen, the spectrum of differential diagnoses to be considered may encompass a number of benign and malignant entities, like keratinous subareolar cysts, malignant fibroepithelial lesions with myxo-chondroid stroma, and true sarcomas of the breast, with cartilaginous metaplasia. It is the Authors' feeling that, with optimal samples, the cytomorphological findings of this rare variant of breast carcinoma permit its accurate pre-operative diagnosis.     

Fibroadenoma mimicking papillary carcinoma on ThinPrep of fine-needle aspiration of the breast

OBJECTIVE: To compare and contrast benign and malignant lesions of the breast that have similar appearances on fine-needle aspiration cytology and that constitute diagnostic pitfalls. DESIGN: The cytology files (dated November 1995 through May 1998) of the Beth Israel Deaconess Medical Center were searched to identify cases of breast fine-needle aspiration biopsies that were highly cellular and composed of bland-appearing spindle/columnar cells and that could represent either epithelial or stromal cells; these cases were reported as indeterminate (atypical/suspicious) and had subsequent excisional biopsies taken. RESULTS: Four such cases were found. Two were fibroadenomas and 2 were papillary carcinomas. Their appearances were strikingly similar on aspiration cytology. All cases were prepared with the ThinPrep method. On microscopic examination, all 4 cases were hypercellular and had many single cells and clusters of columnar/elongate cells. Immunocytochemistry proved these cells to be of epithelial origin. At least occasional bipolar stromal cells were seen in the background. The only appreciable difference between the benign and malignant cases was more significant nuclear atypia, which was barely discernible, in the malignant cases. Immunocytochemistry for smooth muscle actin was helpful in 2 cases that had sufficient material. CONCLUSIONS: Some cases of fibroadenomas and papillary carcinomas can be very difficult, if not impossible, to distinguish on fine-needle aspiration cytology. Immunocytochemistry may be helpful if sufficient material is available. To avoid false-negative or false-positive diagnosis on cytology, it is best to report such cases as atypical or suspicious with final diagnosis pending excisional biopsy.