Biomedical subjects
Publications and source records attributed to R Montironi.
A case of sarcomatoid carcinoma of the anorectal junction in a 71-year-old woman showing neuroendocrine and rhabdomyoblastic features is described. The tumor was diffusely infiltrative and metastases to regional lymph nodes were found at laparotomy. The patient died 6 months after surgery with widespread metastases.
OBJECTIVE: To examine the potential of different constructs of Bayesian belief networks (BBN) to manage uncertainty in breast cancer grading. STUDY DESIGN: We developed four networks, two based on Bloom-Richardson's and two on Helpap's grading systems. The function of the networks was based either on an expert's experience or frequency counts derived from subjective grading of a large number of samples. The four BBNs were tested on 20 specimens, and the resulting final beliefs were compared with the subjective gradings. RESULTS: The BBNs showed agreement with the subjective gradings in 60-85% of cases. Different constructs of BBNs, however, differed in their performance. The mean beliefs in frequency-based networks were slightly higher than in experience-based networks. In addition, as compared with the Bloom-Richardson-based networks, the Helpap-based BBNs resulted in higher maximum beliefs but produced a larger fraction of discrepancies with the subjectively graded cases. Depending on the type of network, 65-90% of the BBN grades were associated with high beliefs. CONCLUSION: The results suggest that for reliable results, grading systems with more than three or four variables may be necessary. When based on relevant information, BBNs seem to have the potential to become a valuable method of assisting the pathologist in breast cancer grading.
OBJECTIVE: To develop procedures for the segmentation of cribriform prostatic glands. STUDY DESIGN: A knowledge-guided procedure following a model-based reasoning process was developed in the context of a set of interacting expert systems for machine vision in histometry. RESULTS: With 78 entities in the knowledge file, fully automated, correct segmentation of approximately 70-80% of cribriform glands was attained--i.e., outlining of histologic components agreed with visual assessment. Measurement of gland size, shape, lumen area, number of lumina per gland, epithelial layer thickness, degree of cribriformity and determination of completeness of lining of a gland by a basal cell layer could be taken from the correctly segmented images. CONCLUSION: The automated procedure allows a histometric characterization of premalignant and malignant prostatic lesions. Extension of system capabilities to the utilization of spectral information is expected to allow an increase in the correct segmentation rate.
Bayesian belief networks (BBNs) are a novel tool for representing knowledge about diagnostic decision making and for obtaining a numerical measure of certainty in the final diagnosis. Belief networks have been applied to the pathological assessment of breast, prostate and skin lesions and have been shown to provide consistency in the grading of microscopic features and improve diagnosis. These applications are reviewed in the current paper. The application of BBNs has been further facilitated through the use of standardised imagery which is stored digitally and used to enter evidence into a BBN. It is predicted that the further development of BBNs with improved logical capabilities represent the key to improved decision making in pathology.
The participants agreed that high grade prostatic intraepithelial neoplasia was the most likely precursor of prostate cancer. Consensus was reached regarding grading, suggesting that PIN be classified as low grade and high grade, noting that high grade PIN is the clinically significant end of the morphologic continuum. Grade 1 PIN is now considered low grade, and grades 2 and 3 are considered high grade. All participants agreed that urologists should be informed when high grade PIN is identified in isolation in tissue specimens, but consensus was not reached regarding the value of reporting low grade PIN. No therapy was recommended for patients with high grade PIN, although repeat biopsy and follow-up is of value. No consensus was reached regarding the biologic potential of the lesion known as atypical adenomatous hyperplasia. Further investigation is needed to determine the diagnostic utility of this finding in prostatic specimens.
We examined 21 brains from individuals more than 65 years of age by MRI and neuropathological methods to study the frequency and morphology of white matter changes. There were 16 brains from neurologically normal subjects (Group 1) while the remaining 5 (Group 2) had neurological disturbances. In Group 1 MRI showed high signal areas in the periventricular white matter in 12 brains and in the deep white matter in 9. All had focal areas, with confluent zones in 4; 3 cystic infarcts were also detected. Neuropathology in this Group showed periventricular changes of variable extent in all cases, vacuolated myelin around the perivascular spaces in 14 and degenerate myelin in 4. Macroscopic inspection showed 3 cystic lacunar infarcts, while areas of recent infarction were present on histology in 2. Four of the Group 2 brains had periventricular MRI changes; high signal areas in deep white matter were focal in 2 and confluent in 1. Cystic infarcts were detected in 3 cases. Neuropathology showed periventricular changes in all the brains; in 4 myelin around the perivascular spaces was vacuolated while degenerate myelin was demonstrated in 1. There were also old (1) and recent (2) lacunar infarcts. High signal areas in the white matter thus have different histological backgrounds but only in a minority of cases do they seem to be of pathological significance and, as a rule, they are not related to the presence of neurological disturbances. Correlative MRI-neuropathological studies are helpful for characterising abnormalities detected by techniques, like MRI, which are sensitive but not very specific.
Mucinous ovarian tumors are still a subject of controversy because they can show either intestinal or endocervical differentiation. Morphologic distinction between borderline and malignant tumors is sometimes difficult, and their clinical behavior has not been definitively ascertained. We selected 10 mucinous cystadenomas (MCAs), 32 intestinal mucinous borderline tumors (IMBTs), and 15 well-differentiated mucinous carcinomas (MCCs), all with goblet cells, at least focally. In all cases, we studied the clinicopathologic features, mucin content, intermediate filament expression, and some nuclear quantitative features, namely, the volume-corrected mitotic index (M/Vi), percentage of nucleolated nuclei, mean number of nucleoli per nucleus, percentage of nucleoli touching the nuclear membrane, and mean nuclear area. The quantitative nuclear study included cytometric DNA analysis and the results were expressed as relative mean ploidy value (RMPV) and as diploid-tetraploid or aneuploid histograms. The results of the quantitative study were evaluated statistically. All patients had stage IA tumors, had received surgical therapy only, and were alive and well after a follow-up of more than 5 years. Light microscopic examination revealed that destructive stromal invasion was not present in any MCCs and that IMBTs and MCCs were easily recognizable using the Hart and Norris criteria, later expanded by Hart. Mucin histochemistry and intermediate filament immunohistochemistry failed to detect substantial differences between the diagnostic categories. DNA analysis demonstrated an increase in aneuploid tumors going from IMBTs to MCCs, but these differences were not statistically significant. On the other hand, nuclear quantitative morphology showed significant differences among the three groups of tumors for all features considered. Forward stepwise discriminant analysis highlighted that MCAs, IMBTs, and MCCs were contiguous but different categories. These data support the separation of IMBTs and MCCs into morphologically different categories as underlined by the results of quantitative nuclear morphologic analysis. The favorable outcome of all patients confirms the excellent prognosis of stage I IMBTs and suggests that well-differentiated MCCs without destructive stromal invasion at stage IA could be assimilated, in terms of prognosis and therapy, into stage I IMBTs.
AIMS: To investigate the effect of combination endocrine treatment (CET) or luteinising hormone releasing hormone agonist and flutamide on non-neoplastic prostate, prostatic intraepithelial neoplasia, and prostatic adenocarcinoma. METHODS: The morphology, including the mitotic activity, of 12 radical prostatectomies from patients with prostatic adenocarcinoma pretreated for three months with CET was evaluated in haematoxylin and eosin stained sections and compared with an untreated age and stage matched control group. RESULTS: A differential effect on the non-neoplastic prostate was observed. In fact, the transition zone of the treated prostate showed simplification of the glandular lobules: the ducts and acini were small without undulations of the epithelial border and with a prominent basal cell layer. Within the peripheral zone there was inconspicuous branching of the ducts and acini which looked dilatated and lined by flattened atrophic epithelium. Prostatic intraepithelial neoplasia occurred in scattered ducts and acini in the peripheral zone of 10 of the 12 patients. The epithelial cell lining showed a prominent basal cell layer. A certain degree of secretory cell type stratification was always present. However, crowding was less evident than in the untreated prostate because of cytoplasmic clearing and enlargement as a result of coalescence of vacuoles. The treated adenocarcinomas had neoplastic acini which looked small and shrunken, and areas of individual infiltrating tumour cells separated by abundant interglandular connective tissue. The secretory cells of the nonneoplastic, prostatic intraepithelial neoplasia, and prostatic adenocarcinoma lesions had inconspicuous nucleoli, nuclear shrinkage, chromatin condensation, and cytoplasmic clearing. Apoptotic bodies were easily identifiable in all the cell layers. The lumina were rich in macrophages, sloughed secretory cells with degenerative features, and apoptotic bodies. Mitoses were not observed in any of the treated non-neoplastic prostate, prostatic intraepithelial neoplasia, or prostatic adenocarcinomas, whereas the mitotic frequency increased from non-neoplastic prostate through prostatic intraepithelial neoplasia up to prostatic adenocarcinomas in the untreated specimens. CONCLUSIONS: CET before radical prostatectomy causes regressive epithelial changes together with enhanced apoptosis and blocked mitotic activity.
Surrogate endpoint biomarkers (SEBs) are needed in clinical chemoprevention trials to avoid the excessively long study periods and high costs associated with the use of cancer incidence reduction as an endpoint, particularly with relatively slow-growing tumors such as prostatic adenocarcinoma. SEBs should be directly associated with the evolution of neoplasia, and develop with high frequency in abnormal cells of susceptible individuals. If SEBs can be modified by a particular intervention regimen in short-term studies, the rationale for carrying out long-term studies may be strengthened. The consensus panel identified a small and manageable group of biomarkers measured in tissue or serum as the most promising in prostate cancer chemoprevention, including (1) prostate specific antigen (PSA); (2) morphometric markers, such as nuclear size and roundness; (3) proliferation markers, such as MIB-1 and PCNA; (4) nuclear DNA content (ploidy); (5) oncogene c-erbB-2 (HER-2/neu) expression; (6) angiogenesis; and (7) high-grade prostatic intraepithelial neoplasia (PIN). Information regarding many of these and other biomarkers is limited, calling for further investigation. Also, these factors, chosen chiefly for their proven or proposed utility as prognostic factors, may be less useful as SEBs. It was agreed that concurrent study of numerous markers rather than single markers allows comparison of their relative utility, including assessment of ease of quantitation and the sensitivity, specificity, and positive and negative predictive value.
This paper evaluates the use of quantitative methods to accurately assess cell proliferation and death in untreated and treated prostate lesions. The analysis of proliferating cell nuclear antigen (PCNA)-stained nuclei allow precise evaluation of the proliferating cells and exact identification of their location in the progression of untreated prostatic intraepithelial neoplasia (PIN) to prostatic adenocarcinoma (PAC). The evaluation of the frequency and location of apoptotic bodies (ABs) gives accurate information on the apoptotic phenomenon in PIN compared to normal prostate (NP) and PAC. In fact, the frequency of ABs increases from NP to PIN to PAC and parallels that observed with PCNA. However, the AB-related values were approximately one-eighth to one-tenth of those obtained with PCNA immunostaining. Combination endocrine therapy (CET) decreases the proliferative activity and enhances the apoptosis phenomenon in NP, PIN, and PAC. This might indicate that CET could induce a certain degree of regression not only of PAC, but also of PIN.
Alcohol-fixed, toluidine-blue-stained smears from 24 astrocytomas (12 low grade and 12 high grade or anaplastic) were included in the study. In each case 50 nuclei from representative areas of the tumor were selected for analysis; quantitative features pertaining to both the entire nucleus and the nucleoli were computed. Nuclear features were nuclear area and total optical density. Nucleolar features were number of nucleoli per nucleus, nucleolar area, variance of the nucleolar area, and mean and variance of the distance of nucleoli from the nuclear membrane. The results showed distinct changes in a number of nuclear and nucleolar features from low to high grade astrocytomas. Features expressing the most pronounced nuclear changes were area, grey level nonuniformity and run percentage. Changes were also found in the following nucleolar features: number of nucleoli, nucleolar area, variance of nucleolar area, nucleolar location and variance of nucleolar location. Linear discriminant analysis was carried out to determine a direction in feature space along which astrocytomas of low and high grade might be ranked. The nuclear area, number of low gray value pixels and a run length feature provided a useful linear combination. The study showed that one can derive a set of objective criteria from morphometric measurements that allows an ordering of astrocytoma cases along an axis and that might be used for continuous grading.
The diagnosis and grading of prostatic intraepithelial neoplasia (PIN) are affected by uncertainties that arise from the fact that almost all our knowledge of PIN histopathology is not expressed in numeric form but rather in descriptive linguistic terms, words and concepts. A Bayesian belief network (BBN) was used to reduce the problem of uncertainty in diagnostic clue assessment while considering the dependencies between elements in the reasoning sequence. A shallow network was developed with an open-tree topology, with a root node containing the diagnostic alternatives and seven first-level descendant nodes for the diagnostic clues. One of these nodes was based on tissue architecture and the others on cell features. The results obtained with prototypes of relative likelihood ratios showed that beliefs for the diagnostic alternatives are very high. The network can grade and differentiate PIN lesions from other prostate lesions with certainty. A number of diagnostic clues greater than seven did not significantly improve network performance, whereas a reduced number of clues resulted in decreased beliefs. A BBN for PIN diagnosis and grading offers a descriptive classifier that is readily implemented and allows the use of linguistic, fuzzy variables. A BBN allows the accumulation of evidence presented by diagnostic clues, each offering only weak evidence.
The aim of our study was to assess the frequency and location of apoptotic bodies (ABs) in haematoxylin and eosin-stained sections of prostatic intraepithelial neoplasia (PIN) and then to compare the patterns with those in benign prostatic hyperplasia (BPH) and prostatic invasive adenocarcinoma (PAC). ABs were identified in all epithelial cell layers of the ducts, acini and tumour islands, as well as in the lumina contained in such structures. In the epithelial cell layers, ABs were found in general in the intercellular space and occasionally in the cytoplasm of epithelial cells. The frequency of ABs increased from BPH through PIN up to PAC. The proportions of ABs in PIN lesions of low grade (PINlow) and high grade (PINhigh) were greater than in BPH, the values decreasing from the nuclei in the basal position towards those in the luminal layer. In PINlow, the mean category values were 0.85% (standard error, SE, 0.311%) in the basal, 0.623% (SE 0.065%) in the intermediate and 0.474% (SE 0.138%) in the luminal position. In PINhigh, the mean category values were 1.006% (SE 0.16%) in the basal position, 0.713% (SE 0.182%) in the intermediate and 0.618% (SE 0.172%) in the luminal position. The proportions of ABs in adenocarcinoma with cribriform pattern decreased from the basal towards the luminal layer, as for PIN: 1.806% (SE 0.346%) in the basal position, 1.15% (SE 0.172%) in the intermediate and 0.886% (SE 0.137%) in the luminal position. In the solid/trabecular adenocarcinomas, the mean category value in the cell layer adjacent to the stroma was 2.154% (SE 0.203%), whereas in the other cell layers it was 2.052% (SE 0.239%). In small and large acinar adenocarcinomas, the proportions of positive nuclei were 1.022% (SE 0.1%) and 0.922% (SE 0.163%), respectively. The evaluation of the frequency and location of ABs gives accurate information on cell death in PIN in comparison with BPH and PAC.
The expression and location of proliferating cell nuclear antigen (PCNA) immunostaining in epithelial, endothelial and stromal nuclei were assessed in prostatic intra-epithelial neoplasia (PIN). It was then compared with patterns in benign lesions and in invasive adenocarcinomas of the prostate. The PCNA-positive nuclei showed homogeneous or granular types of staining, or a mixture of both, and a gradation in the intensity of staining. Nuclei with granular and mixed patterns appeared lighter brown than those with a homogeneous pattern, which are darker and more often noted in PIN and invasive adenocarcinomas than in benign lesions. For epithelial PCNA-stained nuclei, the proportions in the two grades of PIN were greater than in benign prostatic hyperplasia (mean 3.16%, SE 0.31%) and prostatic atrophic ducts and acini (mean 0.56%, SE 0.09%), the values decreasing from the nuclei in the basal position towards those in the luminal layer. In grade 1, the category mean values were 9.51% (SE 1.14%) in the basal, 7.02% (SE 1.27%) in the intermediate and 6.02% (SE 0.90%) in the luminal position. In grade 2, the category mean values were 13.81% (SE 1.42%) in the basal position, 10.99% (SE 1.17%) in the intermediate and 7.91% (SE 1.43%) in the luminal position. In small and large acinar adenocarcinomas, the proportions of positive nuclei were 8.66% (SE 0.30%) and 9.06% (SE 0.30%), respectively. The category mean values in the cribriform adenocarcinomas were 14.40% (SE 0.61%) in the basal position, 11.84% (SE 1.30%) in the intermediate and 9.26% (SE 0.66%) in the luminal position. As in PIN, the proportions of immunostained nuclei in the adenocarcinoma with cribriform pattern decreased from the basal towards the luminal layer. In the solid/trabecular adenocarcinomas, the category mean value in the cell layer adjacent to the stroma was 17.60% (SE 2.92%), whereas in the other cell layers it was lower than that in the cells adjacent the stroma (mean 13.88%, SE 1.71%). For capillary endothelial and stromal cells, the percentages of PCNA-stained nuclei were much lower than those in the epithelial component. The lowest mean values were obtained in benign lesions, whereas the highest were in invasive adenocarcinomas, the percentages in PIN being intermediate.
Expression and location of Proliferating Cell Nuclear Antigen immunostaining in epithelial nuclei were assessed on histological sections from 32 cases of invasive adenocarcinoma of the prostate gland: 20 untreated and 12 treated with combination endocrine therapy or CET. The PCNA-positive nuclei showed homogeneous or granular types of staining or a mixture of both, and a gradation in the intensity of staining. Nuclei with homogeneous patterns appeared darker brown than the lighter granular and mixed patterns. Darker nuclei were more frequently noted, mainly among the epithelial cells adjacent to the stroma, in the untreated cases. In contrast, nuclei with pyknotic chromatin, unstained and corresponding to apoptotic bodies, were more frequently seen in the treated patients. For the untreated invasive adenocarcinomas, the mean proportion of PCNA-stained epithelial nuclei in the 10 cases with an acinar pattern (small and large) was 8.86% (SE 0.23%). The mean value in the 5 cases with a cribriform pattern was 11.76% (SE 0.52%), that is, greater than in the acinar pattern, and decreased from the nuclei in the basal position, or adjacent to the stroma, toward the lumen: 14.40% (SE 0.61%) in the basal position, 11.84% (SE 1.30%) in the intermediate and 9.26% (SE 0.66%) in the lumenal. In the 5 cases with a solid/trabecular pattern, the proportion of PCNA-positive nuclei was 15.74% (SE 2.30%), that is, higher than in all the other patterns, and decreased from the cell layer adjacent to the stroma (17.60%, SE 2.92%) toward the other layers (13.88%, SE 1.71%).(ABSTRACT TRUNCATED AT 250 WORDS)
AIMS: To analyse the cell cycle and DNA histogram components in data from DNA static cytometry and, in particular, to investigate the influence of the length of time the slides are exposed to the light of the cytophotometer in evaluating the G0/G1 peak. METHODS: DNA static cytometry was performed on 18 Feulgen stained imprints and six histological sections taken from six breast carcinomas. The total optical density values obtained were analysed using software commercially available as Multicycle. DNA flow cytometry was performed on the same cases. RESULTS: The proportions of nuclei related to the cell cycle components from DNA static cytometric data, obtained from Feulgen stained cytological smears, were almost identical with those obtained from DNA flow cytometric data. Moreover, additional information was obtained from the DNA static cytometry frequency histogram and the proportions of nuclei below the diploid G0/G1 peak and above the G2 phase. Discrepancies between DNA static cytometry and DNA flow cytometry were seen in the large coefficients of variation of the G0/G1 peaks obtained with the former method of analysis, even though a better correspondence was found when the exposure time of the slides to the light of the cytophometer was conspicuously shortened. The information obtained from histological sections seemed to be similar to that obtained from DNA flow cytometry when a single cell population was present; a single cell population was detected in two out of the three cases in which two distinct populations had been present in DNA flow cytometry. CONCLUSIONS: The computer analysis of DNA static cytometric data obtained from Feulgen stained cytological specimens provides the type of information on the cell cycle which is usually obtainable only from DNA flow cytometry. Correspondence with the DNA data from histological sections, however, was poor.
The aim of our study was to qualitatively and quantitatively investigate the capillary architecture on lectin Ulex Europaeus agglutinin I-stained histological section in prostatic intra-epithelial neoplasia. The capillaries appeared as small, short or elongated vessels with either a smooth or undulated external contour and either virtual or visible lumen, sometimes with a clearly identifiable endothelial nucleus/i. In the benign prostatic hyperplasia and prostatic intra-epithelial neoplasia categories, the capillaries appeared located in close contact with (i.e. touching) or in proximity to the basement membrane of ducts and acini. In the invasive adenocarcinoma category, on the contrary, the capillaries in general appeared interspersed within the tumour stroma and septa. Our quantitative studies of the capillary architecture showed that, going from benign prostatic hyperplasia through prostatic intra-epithelial neoplasia up to invasive adenocarcinoma, an increasing proportion of capillaries becomes shorter, with open lumen and undulated external contour and with a greater number of endothelial cells. The highest proportion of touching capillaries was seen in benign prostatic hyperplasia, while the lowest was in invasive adenocarcinoma, being intermediate in prostatic intra-epithelial neoplasia. When the prostatic intra-epithelial neoplasia samples were divided into low-grade and high-grade, the feature values in the low-grade approached those in benign prostatic hyperplasia, whereas in the high-grade they were close to invasive adenocarcinoma. Half of the benign prostatic hyperplasia samples were taken from total prostatectomies because of the preoperative diagnosis of prostatic adenocarcinoma. The feature values in this subcategory were close to those of prostatic intra-epithelial neoplasia of low grade.
The clinicopathologic features of 44 serous borderline tumors (SBT) of the ovary were evaluated. Nineteen were Stages II and III, and 9 had invasive peritoneal implants. All 19 patients received chemotherapy and 4, who had invasive implants, died of disease after 3, 4.3, 8, and 9 years. The other 25 patients were free of tumor 1-14 years (mean, 5.3 years) after surgery. Coexpression of low molecular weight keratins (AE1, CAM 5.2) and vimentin was found in all tumors and their implants. No significant differences were found between SBT with different volume-corrected mitotic indices (M/Vi) with respect to gross features, presence or absence of implants, stage, and survival. Cytometric DNA analysis also was performed on the primary ovarian tumors and the implants. Twenty-one primary tumors had diploid or tetraploid histograms, and 23 had aneuploid histograms. DNA ploidy of the primary ovarian tumors did not correlate with gross features, the presence or absence of implants, M/Vi, stage, and survival. The data from this study confirm that most SBT are clinically benign, but SBT with invasive implants may behave aggressively. Expression of intermediate filaments, M/Vi, and DNA ploidy evaluation of the primary ovarian tumors seem to be of no value in predicting prognosis. However, four of seven patients with aneuploid DNA implants died of tumor.