Malignant Nuclei Research Articles

Using classification and regression trees, liquid‐based cytology and nuclear morphometry for the discrimination of endometrial lesions

'The objective of this study is to investigate the potential of classification and regression trees (CARTs) in discriminating benign from malignant endometrial nuclei and lesions. The study was performed on 222 histologically confirmed liquid based cytological smears, specifically: 117 benign cases, 62 malignant cases and 43 hyperplasias with or without atypia. About 100 nuclei were measured from each case using an image analysis system; in total, we collected 22783 nuclei. The nuclei from 50% of the cases (the training set) were used to construct a CART model that was used for knowledge extraction. The nuclei from the remaining 50% of cases (test set) were used to evaluate the stability and performance of the CART on unknown data. Based on the results of the CART for nuclei classification, we propose two classification methods to discriminate benign from malignant cases. The CART model had an overall accuracy for the classification of endometrial nuclei equal to 85%, specificity 90.68%, and sensitivity 72.05%. Both methods for case classification had similar performance: overall accuracy in the range 94-95%, specificity 95%, and sensitivity 91-94%. The results of the proposed system outperform the standard cytological diagnosis of endometrial lesions. This study highlights interesting diagnostic features of endometrial nuclear morphology and provides a new classification approach for endometrial nuclei and cases. The proposed method can be a useful tool for the everyday practice of the cytological laboratory.

Argyrophilic nucleolar organizer regions of thyroid lesions on fi ne needle aspiration smears

Background: Argyrophilic nucleolar organizer region is higher in malignant nuclei as compared to reactive and benign nuclei. The main objective of this study was to evaluate the role of Argyrophilic nucleolar organizer region and its extent in differentiating benign and malignant lesions, particularly with respect to follicular neoplasms of the thyroid. Materials and Methods: This was a prospective, cross sectional study which included 110 patients presenting with thyroid swelling. The fi ne needle aspiration smears were studied by conventional methods and silver staining for Argyrophilic nucleolar organizer region. In 50 cases, diagnosis was confi rmed by histopathology. The Argyrophilic nucleolar organizer region count was done and the mean Argyrophilic nucleolar organizer region count per nucleus was calculated along with pattern of distribution. Results: Mean Argyrophilic nucleolar organizer region count was higher in neoplastic as compared to non-neoplastic and infl ammatory lesions. Papillary carcinoma had the highest mean Argyrophilic nucleolar organizer region count (5.40 ± 0.86) and the lowest count was seen in Hashimoto’s thyroiditis (2.10 ± 0.36). Higher mean Argyrophilic nucleolar organizer region count was recorded in follicular carcinoma (4.84 ± 1.42) as compared to follicular adenoma (3.5 ± 0.27) which was found to be statistically signifi cant (P<0.05). Argyrophilic nucleolar organizer region dots were arranged in clusters in colloid goiter whereas scattered discrete black dots were seen in neoplastic lesions. Conclusion: Argyrophilic nucleolar organizer region study of thyroid can be used as an additional diagnostic adjunct with cytomorphological features to differentiate benign and malignant follicular neoplasms. Journal of Pathology of Nepal (2013) Vol. 3, No.1, Issue 5, 361-366 DOI: http://dx.doi.org/10.3126/jpn.v3i5.7859

The Sheep in Wolf’s Clothing: Vegetable and Fruit Particles Mimicking Cells and Microorganisms in Cytology Specimens

Background: Microorganisms and contaminants, including food contaminants, can be difficult to distinguish in various cytologic specimens. Vegetable and fruit contaminants can be found in specimens from the gastrointestinal tract, as in anal Pap smears, or aspirated into the respiratory tract, as in bronchiolar-alveolar lavage. Some of these materials mimic microorganisms, normal human cells, and even malignant cells, making correct diagnosis of the cytology specimen a challenge. A catalogue of the cytologic appearance of these contaminants will increase awareness of these diagnostic stumbling blocks. Methods: Commonly eaten fruits and vegetables were selected. In each preparation, a small amount of the edible portions of the fruit or uncooked vegetable were ground with mortar and pestle, and smeared onto glass slides. The slides were fixed in 100% alcohol for Papanicolaou (PAP) stain or air-dried for May-Grunswald-Giemsa (MGG) stain. Results: All vegetable cells contained nuclei, cytoplasm, and cell walls. Fruit cells contained nuclei and cytoplasm, but variably contained cell walls. Many of the deeply stained nuclei resembled overly stained malignant nuclei or dysplastic cells. Vegetable contaminants can resemble anucleated to intermediate squamous cells, respiratory columnar cells, viral inclusions such as cytomegalovirus or Molluscum contagiosum, fungal elements, and even organisms such as nematodes, Stronglyoides, and Toxoplasma. Fruit contaminants can mimic similar infectious parasites or fungal spores, and contribute to a seemingly necrotic background. Conclusion: The distinctive morphologic pattern of fruit and vegetable cells make identification of these mimickers of pathologic processes possible, and differentiates them from human cells.

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.

Expression of glutathione-S-transferases isoenzymes and p53 in exfoliated human bladder cancer cells

Objectives This study investigates the usefulness of glutathione-S-transferases (GST) isoenzymes and p53 immunostaining as a marker of malignancy in urinary cytology, and evaluates their potential effect in increasing diagnostic accuracy in a series of urine cytologic samples. They are also correlated with cytopathology diagnosis and histopathologic diagnosis. Materials and methods In this study, the slides from 124 bladder carcinoma patients prepared by the cytocentrifugation method were observed. The cytomorphologic properties of these cancer cells were determined. Moreover, the immunocytochemical distributions of GST alpha (GSTA), pi (GSTP), mu (GSTM4), theta (GSTT1) isoenzymes and p53 protein were studied for the patients. Results The urothelial cancer cells had small cytoplasm and rough nuclear membrane. The chromatin granules were heterogeneously distributed in each malignant cell's nucleus. There was a pleomorphism of the malignant cells' nuclei. According to immunocytopathologic observations, the urothelial cancer cells had stronger staining intensity than the benign cells had in 48% of cases for GSTA, 46% of cases for GSTP, 38% of cases for GSTM4, and 42% of cases for GSTT1. For all papillary cases, the malignant cells were stained negative, while the benign cells were positive. For 83% of patients, the malignant cells were stained positive for p53. There was a significant difference in GSTA ( P = 0.006), GSTT1 ( P = 0.004), GSTP ( P = 0.000) and p53 ( P = 0.000) expressions for benign cells whereas, a non-statistical difference in the malignant cells for GSTA, GSTT1, GSTP, GSTM4, and p53 expressions ( P > 0.05). Conclusions GST isoenzymes and p53 immunostaining were not found to be markers of malignancy in urinary cytology.

Contribution of Nuclear Morphometry by Confocal Laser Scanning Microscopy to the Diagnosis of Malignant Bile Duct Strictures

To determine the clinical utility of nuclear morphometry by confocal laser scanning microscopy for the diagnosis of malignant biliary strictures.The study included 51 patients with bile duct strictures who underwent endoscopic retrograde cholangiopancreatography (ERCP). Based on the initial workup, 6 patients were diagnosed with benign strictures, and 12 patients had malignant strictures, while in the remaining 33 cases the diagnoses were inconsistent, due mainly to inadequate samples. Smears from ERCP brushings were stained for DNA with propidium iodide. Nuclear morphometry was assessed on images acquired by a confocal laser scanning microscope. Three parameters-nuclear volume, nuclear shape and nuclear staining intensity-were calculated. Based on these features, a distinctive nuclear morphometric pattern was attributed to the malignant nuclei, and its predictive value was assessed prospectively in the 33 undiagnosed cases.After an overall median follow-up period of 8 months, 19 patients were diagnosed with malignant strictures, and 14 patients were considered to have benign strictures. With respect to the prediction of malignancy, the sensitivity of the described method was 78%, the specificity was 63%, the positive predictive value was 64%, and the negative predictive value was 80%.Nuclear morphometry may provide significant information for the diagnosis of malignant bile duct strictures when conventional cytology fails to.

Morphometry of Isolated Tumor Cells in Breast Cancer Sentinel Lymph Nodes: Metastases or Displacement?

Iatrogenic displacement and mechanical transport of epithelial cells to the sentinel node (SN) has been suggested to result in false-positive findings in breast cancer patients, but little biologic evidence has yet been presented for this hypothesis. As malignant nuclei are larger than benign ones, nuclear morphometry of SN isolated tumor cells (ITC) could provide relevant information with regard to the malignant origin-or-not of epithelial cells in the SN. In patients with primary invasive breast cancer and SN ITC with (N=16) or without (N=45) non-SN involvement, nuclear morphometry was performed on the primary tumor as well as on the ITC in the SN. Nuclear size in the primary tumor was compared with that in the corresponding ITC. Patients with SN micrometastases (N=30) and SN macrometastases (N=30) served as controls. Nuclear size of ITC was significantly smaller compared with nuclear size of the corresponding primary tumor (P<0.0001). In contrast, there were no differences in nuclear size between SN micrometastases and macrometastases on the one hand and their corresponding primary tumors on the other. In addition, a subgroup of cases (10/61, 16%) with benign morphometric features of SN ITC nuclei (small and isomorph) could be discerned that had no non-SN metastases. In conclusion, nuclei of SN ITC are significantly smaller compared with the corresponding primary tumor and are often not associated with non-SN involvement. This supports the hypothesis that some of these deposits could represent benign epithelium or degenerated malignant cells lacking outgrowth potential.

Refractometry of melanocyte cell nuclei using optical scatter images recorded by digital Fourier microscopy

The cell nucleus is the dominant optical scatterer in the cell. Neoplastic cells are characterized by cell nucleus polymorphism and polychromism-i.e., the nuclei exhibits an increase in the distribution of both size and refractive index. The relative size parameter, and its distribution, is proportional to the product of the nucleus size and its relative refractive index and is a useful discriminant between normal and abnormal (cancerous) cells. We demonstrate a recently introduced holographic technique, digital Fourier microscopy (DFM), to provide a sensitive measure of this relative size parameter. Fourier holograms were recorded and optical scatter of individual scatterers were extracted and modeled with Mie theory to determine the relative size parameter. The relative size parameter of individual melanocyte cell nuclei were found to be 16.5+/-0.2, which gives a cell nucleus refractive index of 1.38+/-0.01 and is in good agreement with previously reported data. The relative size parameters of individual malignant melanocyte cell nuclei are expected to be greater than 16.5.

A Novel Camptothecin Derivative Incorporated in Nano-Carrier Induced Distinguished Improvement in Solubility, Stability and Anti-tumor Activity Both In Vitro and In Vivo

An oil/water nanoemulsion was developed in the present study to enhance the solubility, stability and anti-tumor activity of a novel 10-methoxy-9-nitrocamptothecin (MONCPT). MONCPT nanoemulsion was prepared using Lipoid E80 and cremophor EL as main emulsifiers by microfluidization. The droplet size of the nanoemulsion was measured by dynamic light scattering. In vitro drug release was monitored by membrane dialysis. Kinetics of MONCPT transformed into carboxylic salt was performed in phosphate buffer at different pH. Hemolysis of MONCPT nanoemulsion was conducted in rabbit erythrocytes. Solubilization character of MONCPT in nanoemulsion was experimented using Nile red as a solvatochromic probe. In vitro cytotoxicity of the nanoemulsion was measured in A549 and S180 cells using Sulforhodamine B protein stain method, and suppression rate of tumor growth was investigated in S180-bearing mice. The cell cycle effects of MONCPT nanoemulsion on S180 cells were analyzed by flow cytometry. Distribution of the nanoemulsion in A549 cells and S180-bearing mice were also investigated by fluorescence image. MONCPT is incorporated in the nanoemulsion in form of lactone with concentration of 489 microg/ml, more than 200 folds higher than that in water. Experiments using Nile red as a solvatochromic probe indicated that more MONCPT might be located in the interfacial surfactant layer of the nanoemulsion than that in discrete oil droplet or continuous aqueous phase. Nanoemulsion could release MONCPT in a sustained way, and it was further shown to notably postpone the hydrolysis of MONCPT with longer hydrolysis half-life time (11.38 h) in nanoemulsion at pH 7.4 than that of MONCPT solution (4.03 h). No obvious hemolysis was caused by MOCPT nanoemulsion in rabbit erythrocytes. MONCPT nanoemulsion showed a marked increase in cytotoxic activity, 23.6 folds and 28.6 folds in S180 cells and A549 cells respectively via arresting the cell at G2 phase, compared to that induced by MONCPT injection. It correlated well to the in vivo anti-tumor activity of MONCPT nanoemulsion with suppression rate of 93.6%, while that of MONCPT injection was only 24.2% at the same dosage. Moreover, nanoemulsion exhibited enhanced capability of delivering drug into malignant cell's nucleus in vitro and induced drug accumulation in tumor in S180-bearing mice using in vivo imaging. The nanoemulsion prepared exhibited an improved MONCPT solubility, stability and anti-tumor activity, providing a promising carrier for cancer chemotherapy using MONCPT.

Osteoclast nuclei of myeloma patients show chromosome translocations specific for the myeloma cell clone: a new type of cancer–host partnership?

A major clinical manifestation of bone cancers is bone destruction. It is widely accepted that this destruction is not caused by the malignant cells themselves, but by osteoclasts, multinucleated cells of monocytic origin that are considered to be the only cells able to degrade bone. The present study demonstrates that bone-resorbing osteoclasts from myeloma patients contain nuclei with translocated chromosomes of myeloma B-cell clone origin, in addition to nuclei without these translocations, by using combined FISH and immunohistochemistry on bone sections. These nuclei of malignant origin are transcriptionally active and appear fully integrated amongst the other nuclei. The contribution of malignant nuclei to the osteoclast population analysed in this study was greater than 30%. Osteoclast-myeloma clone hybrids contained more nuclei than normal osteoclasts and their occurrence correlated with the proximity of myeloma cells. Similar hybrid cells were generated in myeloma cell-osteoclast co-cultures, as revealed by tracing myeloma nuclei using translocations, bromo-deoxyuridine, or the Y chromosome of male myeloma cells in female osteoclasts. These observations indicate that hybrid cells can originate through fusion between myeloma cells and osteoclasts. In conclusion, malignant cells contribute significantly to the formation of bone-resorbing osteoclasts in multiple myeloma. Osteoclast-myeloma clone hybrids reflect a previously unrecognized mechanism of bone destruction in which malignant cells participate directly. The possibility that malignant cells corrupt host cells by the transfer of malignant DNA may have been underestimated to date in cancer research.

NON-DIFFERENTIABLE DEFORMATIONS OF ℝn

Many problems of physics or biology involve very irregular objects like the rugged surface of a malignant cell nucleus or the structure of space-time at the atomic scale. We define and study non-differentiable deformations of the classical Cartesian space ℝn which can be viewed as the basic bricks to construct irregular objects. They are obtained by taking the topological product of n-graphs of nowhere differentiable real valued functions. Our point of view is to replace the study of a non-differentiable function by the dynamical study of a one-parameter family of smooth regularization of this function. In particular, this allows us to construct a one-parameter family of smooth coordinates systems on non-differentiable deformations of ℝn, which depend on the smoothing parameter via an explicit differential equation called a scale law. Deformations of ℝn are examples of a new class of geometrical objects called scale manifolds which are defined in this paper. As an application, we derive rigorously the main results of the scale-relativity theory developed by Nottale in the framework of a scale space-time manifold.

Discrimination between benign and malignant prostate tissue using chromatin texture analysis in 3‐D by confocal laser scanning microscopy

Analysis of chromatin texture may improve both the diagnosis and the assessment of the prognosis of prostate cancer. Confocal laser scanning microscopy (CLSM) allows performing measurements in nuclei reconstructed in 3-D. The aim of this study was to evaluate the clinical usefulness of 3-D texture analysis of prostate tissue. Image stacks of eight prostate cancer sections were obtained by CLSM of both benign and malignant areas. Texture feature values were computed for individual nuclei. The discriminative power of the texture features was established by receiver operating characteristic (ROC) analysis and linear discriminant analysis (LDA). Texture features were identified that could discriminate between benign and malignant nuclei. LDA correctly classified 89% of the nuclei of the pooled set of benign and malignant nuclei. 3-D nuclear texture features allow discrimination of most benign and malignant prostate nuclei. We estimate that the classification rates can be increased by improving the image quality.

The potential of feature selection by statistical techniques and the use of statistical classifiers in the discrimination of benign from malignant gastric lesions

The objective of this study was the investigation of the potential value of morphometry, feature selection and statistical classifiers techniques, such as neural networks, for the classification of benign from malignant gastric nuclei and cases. One hundred and twenty gastric smears, routinely processed and stained by Papanicolaou technique, were analyzed by a customized image analysis system. Data from half of the cases were selected to form the training set, while the remaining data formed the test set. A feature selection technique was applied in order to identify the most important nuclear features, which were used in a second stage by statistical classifiers to classify a nucleus as benign or malignant. Using the classifier results for the nuclear classification, a method to classify each individual patient was developed. The performance of the proposed method was validated through the test set. The technique described in this report produces significant results at the nuclear and patient level and promises to be a powerful assistance tool for everyday cytological laboratory routine.

Appendiceal Mass in a Middle-Aged Woman

Appendiceal Mass in a Middle-Aged Woman

Development of 3D Chromatin Texture Analysis Using Confocal Laser Scanning Microscopy

Introduction: Analysis of nuclear texture features as a measure of nuclear chromatin changes has been proven to be useful when measured on thin (5–6 μm) tissue sections using conventional 2D bright field microscopy. The drawback of this approach is that most nuclei are not intact because of those thin sections. Confocal laser scanning microscopy (CLSM) allows measurements of texture in 3D reconstructed nuclei. The aim of this study was to develop 3D texture features that quantitatively describe changes in chromatin architecture associated with malignancy using CLSM images. Methods: Thirty-five features thoughtfully chosen from 4 categories of 3D texture features (discrete texture features, Markovian features, fractal features, grey value distribution features) were selected and tested for invariance properties (rotation and scaling) using artificial images with a known grey value distribution. The discriminative power of the 3D texture features was tested on artificially constructed benign and malignant 3D nuclei with increasing nucleolar size and advancing chromatin margination towards the periphery of the nucleus. As a clinical proof of principle, the discriminative power of the texture features was assessed on 10 benign and 10 malignant human prostate nuclei, evaluating also whether there was more texture information in 3D whole nuclei compared to a single 2D plane from the middle of the nucleus. Results: All texture features showed the expected invariance properties. Almost all features were sensitive to variations in the nucleolar size and to the degree of margination of chromatin. Fourteen texture features from different categories had high discriminative power for separating the benign and malignant nuclei. The discrete texture features performed less than expected. There was more information on nuclear texture in 3D than in 2D. Conclusion: A set of 35 3D nuclear texture features was used successfully to assess nuclear chromatin patterns in 3D images obtained by confocal laser scanning microscopy, and as a proof of principle we showed that these features may be clinically useful for analysis of prostate neoplasia.

Racial Differences in Androgen Receptor Protein Expression in Men With Clinically Localized Prostate Cancer

Black American men experience disproportionate mortality from prostate cancer (CaP) compared with white American men. Differences in outcome may stem from differences within the androgen axis. Since serum testosterone levels appear to be similar by race in men with CaP, we measured and compared androgen receptor (AR) protein expression in malignant and benign prostate tissue from black and white men who underwent radical prostatectomy for clinically localized CaP. Archived radical prostatectomy specimens obtained from 25 white and 25 black men had AR protein antigen retrieved and immunostained. AR protein expression from CaP and benign tissue was assessed by 2 methods. Automated digital color video image analysis was used to measure the percent area immunostained for AR protein and the intensity of expression (mean optical density). Visual scoring was performed to compare results with automated values. In black compared with white men malignant nuclei were 27% more likely to immunostain for AR (p = 0.005) and in immunopositive nuclei AR protein expression was 81% greater (p = 0.002). Visual scoring of malignant nuclei revealed that AR immunostaining was significantly increased in black vs white men (171 +/- 40 vs 149 +/- 37, p = 0.048). In immunopositive benign nuclei AR protein expression was 22% greater in black than in white men (p = 0.027). Visual scoring of benign nuclei revealed 20% increased immunostaining in black vs white men, although this difference did not attain statistical significance (p = 0.065). Racial differences in AR protein expression were not explained by age, pathological grade or stage, although serum prostate specific antigen levels were higher in black men (9.7 +/- 7.5 vs 15.5 +/- 12.2 ng/ml, p = 0.049). AR protein expression was 22% higher in the benign prostate and 81% higher in the CaP of black African compared with white men. CaP may occur at a younger age and progress more rapidly in black than in white men due to racial differences in androgenic stimulation of the prostate.

FGF8 isoform b expression in human prostate cancer.

Overexpression of fibroblast growth factor 8 (FGF8) mRNA has been previously described in prostate cancer. Of its four isoforms, FGF8b is thought to be the most important in carcinogenesis. We hypothesised that immunodetection of FGF8b in archival prostate cancer specimens is of potential prognostic value. Using a selected cohort of prostate tumours from transurethral (n=30) and radical prostatectomies (n=59), an optimised protocol for FGF8b immunoreactivity was used to corroborate expression with clinical parameters. No expression was observed in benign prostates (n=10). In prostate cancer, immunoreactivity was localised to the malignant epithelium with weak signals in the adjacent stroma. Expression of FGF8b in stage T1 and T2 cancers were 40 and 67%, respectively. In contrast, FGF8b expression was present in 94% of T3 and 100% of T4 cancers. By histological grade, FGF8b was found in 41% of low-grade cancers (Gleason score 4–6), 60% of intermediate-grade cancers (Gleason score 7 and 92% of high-grade cancers (Gleason score 8–10). The intensity of expression was significantly associated with stage (P=0.0004) and grade (P<0.0001) of disease. We further hypothesised that FGF8b overexpression resulted from enhanced transcription and translation rather than from abnormalities involving the FGF8 gene locus. This was tested by means of fluorescent in situ hybridisation in 20 cancer specimens to map the FGF8 gene locus. FGF8 gene copy number in benign and malignant nuclei was found to be similar (2.33±0.57 and 2.0±0.81, respectively P=0.51). Based on these findings, we propose a multicentre study on cohorts of patients to further evaluate FGF8b as a potential prognostic marker in prostate cancer.

Mucinous tumors of the ovary: a clinicopathologic analysis of 75 borderline tumors (of intestinal type) and carcinomas.

With the exception of benign cystadenomas, mucinous ovarian tumors are rare and heterogeneous neoplasms. They have been classified as either borderline tumors or carcinomas for almost 30 years. Subsequently, the borderline tumors have been subclassified into endocervical-like and intestinal types. The diagnostic criteria for distinguishing borderline tumors of the intestinal type from mucinous carcinomas have varied, making difficult the interpretation of prognostic information. More recently, a further subdivision of the former tumors into forms with only epithelial atypia and variants with focal intraepithelial carcinoma has been proposed. Consequently, in this study of 41 mucinous borderline tumors of intestinal type and 34 mucinous carcinomas, the former were also subdivided into 30 cases with mild to moderate atypia only and 11 with areas of intraepithelial carcinoma. All 30 purely borderline tumors were stage I tumors, and all 15 with follow-up information (including one case with microinvasion) were clinically benign. All 11 mucinous borderline tumors that had foci of intraepithelial carcinoma were also stage I neoplasms, and none of the eight patients with follow-up data (including one with microinvasive carcinoma) recurred. Thirty-four invasive carcinomas were subclassified into 15 expansile and 19 infiltrative subtypes. All 15 carcinomas with only expansile invasion were stage I; none of the 11 with follow-up data recurred. Three of nine patients with stage I infiltrative carcinomas with follow-up information had a fatal recurrence. Eight of the remaining 10 infiltrative carcinomas had extended beyond the ovary at the time of diagnosis (stages II and III); of the six patients with follow-up data, four died of tumor and two were alive with disease. In stage I carcinomas nuclear grade and tumor rupture correlated with unfavorable prognosis, but less than infiltrative invasion. However, all three fatal tumors were infiltrative carcinomas that had ruptured, and two contained grade 3 malignant nuclei. Combination of infiltrative invasion, high nuclear grade, and tumor rupture is a strong predictor of recurrence for stage I mucinous ovarian tumors. Among the 19 infiltrative tumors, 13 contained foci of anaplastic carcinoma. Of the seven patients with stage I tumors and follow-up data, only one patient whose tumor had ruptured intraoperatively had a fatal recurrence. The presence of anaplastic components in stage Ia (intact) carcinomas did not have an adverse effect in their outcome, even when the undifferentiated carcinomatous elements appeared in the form of mural nodules.

Predictive value of topoisomerase IIα immunostaining in urothelial bladder carcinoma

Aims—The nuclear enzyme DNA topoisomerase II has been shown to be required for chromatin condensation and chromosomal segregation during mitosis; its isoform topo IIα is linked with active cell proliferation...

DNA Topoisomerase II-Alpha Immunoreactivity as a Marker of Tumor Aggressiveness in Invasive Breast Cancer

Objective: The nuclear enzyme DNA topoisomerase (topo) II breaks and rejoins DNA strands; its isoform topo IIα is associated with active cell proliferation of mammalian cells. The aim of this study was to examine the relationship between the expression of topo IIα and biological behavior markers in breast cancer. Methods: Formalin-fixed, paraffin-embedded tissue from 88 samples of infiltrating breast cancer was immunohistochemically stained for topo IIα. For each case, a topo IIα index was determined by image analysis. Similar indexes were available for Ki-67 protein, a known cell proliferation marker, and p53, bcl-2 and c-erbB-2 oncoproteins. Each case had been staged and graded and the patients had been followed up for a mean period of 61.62 months. Results: Elevated topo IIα immunopositivity (in >10% of malignant nuclei) was detected in 22 tumors, and this immunostatus was statistically associated with poor nuclear differentiation, absence of steroid hormone receptors, high Ki-67 immunoexpression, p53 protein accumulation and c-erbB-2 protein overexpression. Topo IIα expression was not linked with disease extent (stage or lymph node status). Neither proliferation marker (topo IIα or Ki-67) had any significant influence on the patients’ recurrence-free survival. Conclusion: From the above results, we conclude that topo IIα overexpression appears to be linked with cellular dedifferentiation and potentially aggressive tumor phenotype in invasive breast cancer.