Surgical treatment of aggressive vertebral hemangioma causing progressive paraparesis.

Vertebral hemangioma is a benign vascular lesion that may onset with neurologic symptoms due to spinal cord compression by epidural extension. Surgical procedure, embolization and radiotherapy are the gold standard for the treatment of this disease. We present a case of a 84 years old woman admitted at our department with worsening paraparesis and urinary retention. Her magnetic resonance images (MRI) showed a lesion involving both anterior and posterior vertebral element of D5, with extension into epidural space and spinal cord compression. The patient was operated for laminectomy and epidural lesion removal. Histological examination confirmed the diagnosis of cavernous hemangioma.

Direct demonstration of cytokeratin filaments by electron microscopy in K562 cell lines in liquid culture.

Our previous study suggested the presence of cytokeratins in the supernatant of human K562 erythroleukemic cell line. In this study we confirm, by using an electron microscopy technique, that K562 cells contain typical intermediate tonofilaments with the characteristics of cytokeratins. After variable intervals of liquid culture, K562 cells have been examined for their clonogenic ability by immunostaining and ultrastructural study. K562 cells showed variable amounts of medium-sized filaments (intermediate filaments) of 10 nm mean size, having the submicroscopic pattern consistent with tonofilaments, arranged as electron-dense curve-shaped bundles, in perinuclear position independently from the different growth phases. A pool of monoclonal antibodies against cytokeratins confirmed the presence of cytokeratins in immunostaining, while the cytofluorimetric analysis showed an unexpected positivity of the CD34 antigen, associated with the typical adhesion molecule VLA5. In conclusion, the immunostaining with monoclonal antibodies and the ultrastructural findings suggest the expression of epithelial features in human K562 leukemic cells.

Standardized mitotic counts in breast cancer. Evaluation of the method.

Twenty-one pathologists and technicians participated in a study evaluating the variation present in mitotic counts for prognostication of breast cancer. The participants counted the mitotic figures in 20 breast cancer samples from ten high power fields (mitotic activity index, MAI, giving the results in mitotic figures per 10 fields) and also made a correction for field size and area fraction of the neoplastic epithelium to get the standardized mitotic index (volume fraction corrected mitotic index, or M/VV index, giving the result in mitotic figures per square mm of neoplastic epithelium). The difference in variation between the two methods was not big, but the standardized mitotic index (SMI) showed consistently smaller variation among all participants and different subgroups. Experienced pathologists had the highest variation in mitotic counts, and specially trained technicians, the lowest. The efficiency of the mitotic counts in grading (the grading efficiency) was used to evaluate the mitotic counts. In groups without special training for mitotic counts the mean grading efficiency was lower (experienced and training pathologists both on average had the potential to grade 88% of the cases correctly) than in the group specially trained for the purpose (trained technicians had the potential to grade 95% of the cases correctly). Among the specially trained technicians, the grading efficiency was of the same magnitude as the grading efficiency achieved in determining the S-Phase fraction of cells from paraffin embedded breast cancers by flow cytometry in different laboratories. The results suggest that special training is helpful in making mitotic counts more reproducible, and that in trained hands, the mitotic counts give results comparable to more sophisticated methods of determining proliferative activity in breast cancer.

Primary myelofibrosis: a detailed statistical analysis of the clinicopathological variables influencing survival.

In the present study we analyzed the prognostic significance of several clinical, hematological, and histological parameters recorded at diagnosis in a consecutive series of 72 patients with primary myelofibrosis (PMF). Univariate analysis showed that the most significant indicators of poor survival were the following: age greater than 60, splenomegaly, anemia (hemoglobin > 10 g/dl), leukopenia (WBC < 4 x 10(9)/l or leukocytosis > 14 x 10(9)/l), and any of these histological features: adipose tissue and megakaryocyte reduction, prominent osteoblastic rims along the trabecular bone, presence of peritrabecular megakaryocytes (Mk), absence of normal or giant Mk. The multivariate analysis showed that only the level of hemoglobin and the presence of both normal Mk and fever independently influenced the prognosis. These parameters were used to set up a prognostic scoring system, allowing a feasible prognosis to be made for each patient at the time of diagnosis and identifying those patients in urgent need of new therapeutic approaches.

Multivariate quantitative histopathological assessment of prognostic factors in ductal infiltrating breast carcinoma.

The aim of this study was to evaluate the power of quantitative pathology in improving the accuracy of prognosis in ductal infiltrating breast carcinoma. Ninety tumours were studied, with a diameter < or = 2.5 cm; positive (metastatic) axillary lymph nodes were found in 48 of the 90 cases; no patient had systemic metastases. Surviving patients had a mean follow-up of 106 months (minimum 69 months). At the end of the study, 45 patients were alive, 45 deceased. The histopathologic study of tumour nuclei and nucleoli included: a) geometric features, i.e., the mean and standard deviation of nuclear area, perimeter, diameter and form factor, nucleolar area and nucleolar/nuclear ratio, measured with a Kontron-IBAS automatic image analyser; b) immunohistochemical features, i.e., the percentage of PCNA (Proliferating Cell Nuclear Antigen)-positive nuclei, the expression (positive or negative) of vimentin (as a marker of tumour dedifferentiation), and the nuclear protein p53. Statistically significant differences were found between surviving and decreased patients for most features; multivariate analysis gave 92.2% accuracy in predicting outcome. Among the classes defined with multivariate analysis, it was possible to distinguish subsets of patients with bad prognosis by studying the expression of vimentin and p53, although these markers were positive in a small number of cases: 5 out of 6 patients with vimentin positivity and 4 out of 8 with p53 positivity died within 24 months of the diagnosis.

Evolutionary somatic cell changes in cervical tumour progression quantitatively evaluated with morphological, histochemical and kinetic parameters.

The somatic cell changes which characterise malignancy evolution in human cervical preneoplastic and neoplastic lesions have been assessed on histological sections by means of a computerised image analyser. Many features have been simultaneously measured on each cell of the lesions studied, and the following results have been obtained: Some features, mainly kinetic, show continuously increasing values which express changes correlated to the increasing malignancy; other features, especially related to nuclear atypia, cellular heterogeneity and the degree of aneuploidy, have values dropping at the level of early stromal infiltration, which can be morphometrically characterised as composed of relatively homogeneous phenotypes; these features seem to express the degree of genetic instability and relate to the evolutionary somatic cell changes; tumour progression evolves through sequential discontinuous steps, each of them characterised by specific phenotypical features of the neoplastic cell population; the neoplastic cells in the foci of early stromal infiltration and vascular invasion, phenotypically more homogeneous than the parent cell populations of carcinoma in situ and infiltrating carcinoma, seem to possess a greater genetic stability.

Quantitative analysis of quadriceps muscle biopsy in systemic sclerosis.

The lesser diameter of the fibers, separately of type 1 and type 2 fibers, and the capillary density evaluated as number of capillaries/area and number of capillaries/fiber were quantitatively estimated in muscle biopsies from 12 patients with systemic sclerosis. Nine patients with polymyositis or rheumatoid arthritis and six patients with muscle dystrophy and neurogenic atrophy served as controls. The results showed that patients with systemic sclerosis had low values of the lesser diameter of the fibers. This reduction was most obvious in type 2 fibers. The capillaries were also significantly reduced when compared with the control groups. When considering the capillaries/fiber ratio, seven out of eight patients with values lower than 0.7 belonged to the systemic sclerosis group. In systemic sclerosis the capillary density values were not significantly correlated with those of the lesser diameters of the fibers.

Cytometric evidence that cervical intraepithelial neoplasia I and II are dysplasias rather than true neoplasias. An image analysis study of factors involved in the progression of cervical lesions.

Image analysis was performed on 40 Feulgen-stained histologic samples and 48 Feulgen-stained cytologic preparations representing normal squamous epithelium and all grades of cervical lesions (from mild dysplasia to invasive carcinoma) in order to characterize the evolutionary progressive changes in cervical epithelial proliferative disease toward malignancy. Quantitative studies included the analysis of proliferative features, differentiation features, nuclear morphology and DNA content. The data obtained on the histologic sections showed that the various features, to a different extent, detected a gradual increase in phenotypic cellular disarrangements related to the progression of the cervical lesions toward malignancy--that is, the modifications to nuclear area, perimeter, DNA content, percentage of nuclei with nucleoli, nuclear/cytoplasmic ratio and percentage of cells with no membrane positivity for soybean agglutinin lectin were progressively greater, moving from normal epithelium and mild dysplasia toward infiltrating carcinoma. In particular, all the morphologic and histochemical features appeared to parallel a diploid reduction and the appearance of aneuploidy. The simultaneous evaluation of proliferation- and differentiation-related features, together with those of nuclear DNA content, showed two main successive preneoplastic lesions: one characterized by an increase in cell turnover without alterations in its organization and another by a true neoplastic disorder. The data obtained on sequential cytologic examinations showed that individual cell changes are detectable and seem basically to be characterized by the appearance of clusters of cells with somatic characteristics not observed in previous cytologic checks. From the results of our study, the cervical intraepithelial neoplasia (CIN) concept appears to be inaccurate. In fact, only CIN III (severe dysplasia/carcinoma in situ) lesions have the morphologic and proliferative alterations of true neoplasia. In contrast, CIN I and some cases of CIN II lesions lack these characteristics and seem to be properly classified as dysplasia, thus avoiding the term neoplasia, implicit in CIN. Moreover, the multivariate study of data sets of features related to the progressive somatic changes, both in histologically and cytologically studied cases, allows us to detect the steps of progression; they are marked by the appearance of cell clusters with qualitatively different phenotypic characters when compared to the cell populations from which they presumably arise. These results seem to provide a further argument against the CIN theory, which stresses the concept that progression is related only to a gradual numerical increase in an initially established phenotype with the characteristics of malignancy.

Well differentiated follicular neoplasms of the thyroid: reproducibility and validity of a 'decision tree' classification based on nucleolar and karyometric features.

This study was conducted on fine-needle aspirates of well differentiated follicular neoplasms of the thyroid. A 'decision tree' classification based on the percentage of nucleolated nuclei, percentage of nuclei with two or more nucleoli and mean major nuclear diameter was adopted. We observed that the reproducibility and the validity of the follicular adenoma vs follicular carcinoma discrimination are greater than in the subjective evaluation. Moreover, similar classification results were obtained when measurements were performed either with a fully automated image analysis system or with semiautomatic or manual instrumentation. As for reproducibility of the inter-instrument comparisons, the k statistic values ranged from 0.85 to 1.00 (mean value 0.90, that is, an 'almost perfect' degree of agreement); in the subjective evaluations, the inter-observer comparisons showed values ranging from 0.20 to 0.56 (mean value 0.37, that is, 'fair'). In the decision tree classification, feature value thresholds were selected in order to have specificity of 100% and the predictive value of a positive result (carcinoma) of 100%; accuracy was 87% (range 86-89%), sensitivity 74% (71-79%), the predictive value of a negative result (adenoma) 79% (78-82%). In the subjective evaluation the values were as follows: accuracy 67% (64-71%), sensitivity 57% (50-64%), specificity 77% (71-79%), predictive value of a negative result (adenoma) 64% (61-69%), predictive value of a positive result (carcinoma) 71% (67-75%). The conclusion is that, by using a routine microscope equipped with a micrometer, the preoperative diagnosis of follicular carcinoma from smears can be formulated with a high degree of certainty.

Quantitative analysis of prostatic intraepithelial neoplasia on tissue sections.

The changes in nuclear morphology (karyometry) and DNA content in prostatic intraepithelial neoplasia (PIN) were analyzed on tissue sections. The cases of PIN were subdivided into PIN 1 and PIN 2 based on the degree of proliferation and the anaplasia of the secretory cells lining the ducts and acini. Cases of nodular hyperplasia (NH) and adenocarcinoma were also studied for comparative purposes. Karyometric analysis showed a progression of most values from NH to PIN to carcinoma. The DNA analysis showed a decrease in the frequency of nuclei in the diploid range and an increase in the percentage of nuclei in the other ploidy regions (especially between 2c and 4c and in the tetraploid range) from NH to PIN to carcinoma. Forward stepwise discriminant analysis showed similarities between NH and PIN 1 and between PIN 2 and carcinoma. These findings suggest that the evolution towards adenocarcinoma is characterized by progressive morphologic derangements of the nuclei and by the transformation of the diploid DNA content into a nondiploid one, with the changes taking place at the level of PIN 2.

Morphological and quantitative analyses of intraductal dysplasia of the prostate.

The aim of our study was to characterize quantitatively the cellular and architectural features of intraductal dysplasia (ID) (subdivided into two grades), in comparison with benign prostatic hyperplasia (BPH) and adenocarcinoma (AC). The research was carried out on histological sections where the mean nuclear and cellular area, the mean nuclear cytoplasmic (N/C) ratio and parameters related to nuclear crowding and stratification were evaluated. The mean nuclear area and the mean N/C ratio steadily increased from BPH to ID and to AC. As for the mean cellular area, the values decreased from BPH to ID grade 1: in the two grades of ID the values remained about the same; in the AC, the mean cellular area was intermediate between ID and PBH. The mean crowding index showed increase in ID grade I, in comparison with BPH, and then a decline from ID grade 2 to AC. The stratification related features (mean epithelial height, mean nuclear stratification height, stratification index) showed ID values greater than in BPH, while in AC smaller than in BPH; in ID grades 1 and 2 the values were similar. The results clearly indicate that mean nuclear area helps to discriminate BPH and AC. A partial separation between the two grades of ID is possible by combining the mean nuclear area and the mean nuclear crowding index in a bivariate analysis.

[Alclometasone dipropionate (Legederm) for the treatment of steroid-sensitive dermatoses in the elderly]. / Alclometasone dipropionato (Legederm) nel trattamento delle dermatosi steroido-sensibili dell'anziano.

This is a randomized single blind parallel comparison of alclometasone dipropionate cream 0.1% vs hydrocortisone butyrrate in 39 geriatric patients (greater than 60 years old) with steroid sensible skin diseases. The regimen consisted of dosing patients with two applications of the two drugs every day for 4 weeks. Follow-up evaluations have been done weekly, and subjective and objective clinical symptoms, adherence to the study protocol, evolution of the disease, onset of adverse reactions including atrophy have been recorded. Furthermore possible side-effects on hypophysis-adrenal axis have been monitored in baseline conditions, after 7 days and at the end of therapy. In most patients bioptic skin patterns for histomorphometric examination have been drawn before and after therapy to be evaluated by a blind examinator. Alclometasone has reduced initial skin lesions by 82.2%, the extent as hydrocortisone butyrrate. In five patients the complete clearance of the disease has been obtained. Study drugs have been tolerated well by all patients, nor clinical signs of atrophy have been observed. Fluctuations of blood cortisol levels ranged between the normal values. In patients treated with alclometasone histomorphometry revealed a better skin trophism than in patients treated with hydrocortisone butyrrate.

The number of nucleoli in benign and malignant thyroid lesions: a useful diagnostic sign in cytological preparations.

The slides of fine needle aspiration cytology specimens from 99 cases of cold thyroid nodules with known histology were reviewed and the number of nucleoli per nucleus counted and correlated with the different histopathological groups. Significant differences were observed between benign and malignant thyroid lesions in the number of nucleoli in the cytological material. Lower values were present in nodular goitres and follicular adenomas compared to carcinomas. In benign lesions the majority of nuclei contained one nucleolus and nuclei with two, three or more nucleoli were less frequent than in follicular, papillary, medullary and anaplastic carcinomas. Only one case of follicular adenoma had cells containing three or more nucleoli compared to more than half the cases of follicular carcinoma.

Quantitative evaluation of morphological parameters for infiltrating ductal breast cancer prognosis.

The aim of our study was to determine the parameters intrinsic to ductal infiltrating carcinoma. 42 cases of about the same diameter (greater than 1.5 and less than 2.5 cm) with a minimum follow-up of more than seven years were selected. The nuclear and nucleolar shape parameters of one hundred cells for each case and the DNA content of 100 nuclei were measured by static cytometry. By means of multivariate discriminant analysis, the nucleolus/nucleus ratio and the DNA content proved to be the two best prognostic indicators based only on the examination of the parameters of the primary neoplasia. We were able to correctly classify 90% of the cases, and in particular to assign 100% of living cases and 81% of deceased ones to their exact categories.

Sources and nature of variation in DNA analysis of follicular thyroid adenoma.

We investigated the sources and nature of variation that may occur in the DNA analysis of thyroid adenomas from cytological and histological samples. Imprints and smears gave identical results. However, the nuclear area was higher in smears where the optical density of the nuclei was lower. In measuring imprints, the interactive selection of nuclei was preferred to the automatic, because the risk of measuring nuclear fragments or undesired objects was thus avoided. The reproducibility and the variation of the DNA measurements depended on the degree of observer training in quantitative pathology, the method of field selection, and the type of instrumentation. Biological variation in the spatial distribution of nuclei with different ploidy values in some adenomas seemed to hide the influence of section thickness on measurements. Our data seem to suggest that it is best to apply a constant section thickness and 5 micron sections seem acceptable.

Multiparametric quantitation of the progression of uterine cervix preneoplasia towards neoplasia.

The aim of the research was to quantify the malignancy progression of preneoplastic lesions towards carcinoma of the uterine cervix. The study has shown that the modifications to nuclear area, perimeter, DNA content, percentage of nuclei with nucleoli, nuclear/cytoplasmic ratio and percentage of SOYA BEAN AGGLUTININ stained cells are progressively greater moving from normal epithelium and mild dysplasia towards infiltrating carcinoma. In particular all the morphologic and histochemical parameters have shown an increasing derangement which appears to parallel the diploid reduction and the appearance of aneuploidy. In order to identify the changes in the stroma, the modifications to the capillaries were investigated: in mild dysplasia only the vessel density increases; the capillary area, perimeter and diameter increase in moderate and severe dysplasia, and in CIS; the values of these features are similar in the carcinoma groups. The capillary density increases mainly in the invasive carcinoma. The data obtained in the study have shown that: 1) the process of malignancy progression is characterised by a gradual and continuous derangement of cell characteristics and modifications to capillaries of the stroma; 2) the mild dysplastic changes have the characteristics of hyperplastic lesions; 3) CIS, qualitatively similar to severe dysplasia from which it differs quantitatively, has features of neoplastic proliferation; 4) the appearance of invasion is associated with DNA parameter changes which may indicate a ploidy reduction.

Quantitative evaluation of recurrent meningiomas.

Clinical, histological and karyometric parameters, nuclear DNA content and the number of nucleolar organizer regions were investigated in 9 recurrent meningiomas and 10 meningiomas which had not recurred within a 10-year period. There were no significant differences between the two groups as to age, sex, site of the tumours and most of the histological parameters scored. Recurrent tumours showed a higher number of mitotic figures and the nucleolus was visible in most of the cells. Cell density, nuclear area, perimeter and nuclear DNA content had values with no statistically significant differences between the two groups. However, significant differences were found in the distribution of the nuclei in the different ploidy regions. Most of the nuclei in the non recurrent cases were in the diploid range, whereas in recurrent tumours there was a reduction in the number of diploid cells associated with an increase in 2c--4c and 4c components. Recurrent tumours also showed a higher number of nucleolar organizer regions positively stained using an argyrophil method. The mitotic count and the nucleolar organizer regions appeared to be the best predictors for recurrence.

Clinical significance of the histomorphometric evaluation of diabetic microangiopathy in the oral mucosa.

Several structural parameters of the capillary vessels were measured in the oral mucosa of patients with diabetes mellitus of type 1 (D.1) and of type 2 (D.2), and of control cases (C), by means of an image analyser in histological sections of routinely processed biopsies. The studied parameters included: a) capillary wall thickness; b) capillary diameter; c) the ratio of capillary wall thickness and diameter; d) capillary wall area; e) capillary area; f) the ratio of capillary wall area and capillary area; g) density of capillary vessels in the lamina propria; h) density of endothelial cells; i) endothelial nuclear area. Clinical and laboratory parameters were also evaluated (duration of the disease, systolic and diastolic blood pressure levels, glycemia, glycosylated haemoglobin, glycosylated albumin, fructosamines, apolipoproteins A1 and B), in order to assess whether a relationship exists with the morphometric parameters studied. Statistically significant differences, at the level of p less than 0.05, were found in the following morphometric parameters between controls and each group of diabetic patients: mean and standard deviation of capillary wall thickness, mean capillary wall area, mean ratio of the capillary wall area and capillary area. A reduction in the capillary density, i.e. the number of capillary vessels per mm2 of lamina propria, was also observed in diabetic patients with respect to the control group, although it was not statistically significant (C vs. D.1: p less than 0.21; C vs. D.2: p less than 0.10).(ABSTRACT TRUNCATED AT 250 WORDS)

Malignant glial tumours: prognostic value of quantitative microscopy.

Nuclear and cell density features have been measured in 22 cases of glioblastoma divided into two groups according to their survival periods, i.e. less than 12 months or more than 12. The results have demonstrated that the logarithmic transformation of the following features show up statistically significant differences (p less than 0.05): mean of the logarithm of nuclear area, standard deviation of the logarithm of perimeter and standard deviation of the logarithm of the roundness factor. The standard deviation of the roundness factor has been shown to be another parameter with statistically significant differences between the two groups. Forward stepwise discriminant analysis was adopted in order to identify the quantitative features which contributed most significantly to discriminating between the two groups. The results suggested a combination of two features, i.e. the standard deviation of the logarithm of the roundness factor and the mean of the logarithm of the roundness factor. The comparison between actual and predicted categories showed 68.18% agreement: 7 out of 22 cases were allocated incorrectly by the computer. However, when a classification probability threshold was adopted, the 7 incorrectly allocated cases assumed an "intermediate" position between the two groups, in agreement with their survival.

Discrimination between follicular adenoma and follicular carcinoma of the thyroid: preoperative validity of cytometry on aspiration smears.

The aim of our study was to evaluate the validity of the quantitative analysis of nuclear features on aspiration smears in the distinction between follicular adenoma and follicular carcinoma. Karyometric and DNA features, including the nucleolar count, were evaluated on the cytologic material of 28 cases; the values were in general slightly higher in the carcinomas than in the adenomas, with overlap between the two diagnostic groups. By plotting the two most discriminant and least correlated parameters against each other and by drawing two straight lines (one perpendicular to the axis representing the mean of the major nuclear diameter at the 9-microns point, the other perpendicular to the axis of the percentage of nucleolated cells at the 75% point), we obtained two regions: a rectangle delimited by the two axes and the two lines containing 86% of adenomas, and the remaining part of the plane containing 79% of carcinomas. The validity of this approach was tested in 32 new cases not included in the original bivariate classification: accuracy 87%; sensitivity 86%; specificity 88%; predictive value of adenoma 88%; predictive value of carcinoma 85%. In conclusion, a simple cytometric method, i.e., the combined evaluation of the mean of the major nuclear diameter with the percentage of nucleolated nuclei, allows a substantial distinction to be made between follicular adenoma and carcinoma.