Morphometry of gastric endocrine cells in hypergastrinemic patients treated with the somatostatin analogue octreotide.

The aim of the study was to evaluate whether treatment with 200 micrograms/d of the somatostatin analogue octreotide (SMS 201-995) for three months can influence the trophic action exerted by hypergastrinemia on endocrine cells of the oxyntic mucosa, a condition potentially leading to hyperplasia and carcinoid tumors. Endocrine cells were morphometrically investigated in Grimelius silver stained sections of endoscopic biopsies of oxyntic mucosa collected from 13 hypergastrinemic patients with Zollinger-Ellison syndrome (ZES) (n = 5), antral G cell hyperfunction (AGCH) (n = 4) and atrophic gastritis type A (AG-A) (n = 4) before and after 3 months treatment and 3 months after drug discontinuance. The treatment induced a reduction of the volume density (P < 0.015), profile cross sectional area (P < 0.05) and number of cell profiles per unit area (P < 0.015) of argyrophil cells. A rebound of all these parameters was observed 3 months after drug withdrawal with values usually exceeding those at the entry, except in cases of AG-A. The patients' plasma gastrin concentrations presented similar variations showing a significant relation with all morphometric parameters of argyrophil cells. Also, the cell content in alpha subunit of human chorionic gonadotropin was related to the plasma gastrin levels, a finding confirming the close gastrin dependence of the expression of this protein by oxyntic endocrine cells. No significant changes were observed in mucosal somatostatin D cells. These results indicate that variations in circulating gastrin levels are the most likely factor responsible for the hypotrophic effect of octreotide on oxyntic argyrophil cells (mostly corresponding to the ECL cells) of hypergastrinemic patients.

Morphometric assessment of nucleolar activity in liver cells during the evolutive pathway of chronic hepatitis C to cirrhosis.

Using a computer-assisted image analysis system, we performed a morphometric study of silver-stained nucleoli of hepatocytes in liver biopsy specimens from hepatitis C virus-positive patients with chronic persistent hepatitis (3 cases), chronic active hepatitis (4 cases), and cirrhosis (4 cases). The number and the total area of nucleoli, the average area of each nucleolus and the nuclear area were determined for each of 100 hepatocytes per case. A continuing increase in the area of both nucleoli and nuclei paralleled a progressive decrease in the number of nucleoli during the evolution of chronic hepatitis C to liver cirrhosis. These findings would indicate that the hepatitis C virus-induced liver damage causes reactive changes in surviving hepatocytes resulting in an increased nucleolar biosynthetic activity rather than in an increment of cell proliferation rate. Therefore, the liver response to hepatitis C virus injury seems to be mainly based on a condition of cell "hypertrophy", whereas we previously showed that a process also of compensatory hyperplasia occurs in chronic hepatitis B, possibly resulting from a different pathogenesis of the viral damage.

Quantitative analysis of nucleoli and nucleolar organizer regions in cultured primary human normal, reactive and malignant mesothelial cells.

The number and the size of silver-stained intranuclear granules, which correspond to the nucleolus and nucleolar organizer regions, have been determined by means of quantitative methods in cultured primary human mesothelial cells obtained from normal, reactive and malignant mesothelium. The mean values per nucleus of the number, the total area, the average area, and the relative area of the silver-stained granules and the mean nuclear area were determined for each of the three conditions. Normal, reactive and malignant mesothelial cells differed significantly in all the features. These findings at the optical level reflect the differing rate of the nucleolar biosynthetic activity related to the different biological properties of the three cell types, and the features can be useful morphometric descriptors in the diagnostic pathology of the mesothelium.

A model to assess the risk of recurrences of superficial bladder carcinoma. A preliminary report based on 27 patients with a two-year follow-up.

Being able to identify patients with superficial bladder carcinoma at high risk of tumour relapse would be helpful in reducing the high recurrence rate observed in these cases, because a more aggressive prophylactic treatment could be applied. We obtained a series of cystoscopic and histological findings from 27 patients with pTa and pT1 bladder carcinomas, of whom 19 recurred within 2 years following transurethral resection. Histological grade, shape and number of tumours were chosen as discriminating features between patients who relapsed and those who did not. These three variables were used to derive a discriminant function which classified correctly 23 out of the 27 patients on the basis of their actual situation of tumour relapse at 2 years. This method might therefore prove to be useful in predicting accurately the outcome of each patient, thus allowing us to follow an individualized protocol of surveillance and treatment based on a quantified risk for tumour recurrence.

Changes in nucleolar transcriptional activity in hepatitis B virus-associated chronic liver diseases. Preliminary results from a quantitative study of silver-stained nucleoli.

Modifications of gene expression may occur in hepatitis B virus (HBV)-related chronic liver diseases, possibly also involving ribosomal RNA (rRNA) genes contained in the nucleolus. Changes in the level of transcriptional activity of rRNA genes are reflected by variations in the number and/or size of the nucleoli. Therefore a quantitative analysis of the silver-stained nucleoli (AgNus) was performed in a small series of liver needle biopsies from patients with HBV+ chronic persistent hepatitis (CPH) (n = 3), HBV+ chronic active hepatitis (CAH) (n = 3) and HBV+ cirrhosis (CIR) (n = 3). In each case, 100 hepatocytes were selected. The number of the nucleoli (AgNuN), their total area (tAgNuA), the average area of each nucleolus (xAgNuA), the nuclear area (NA) and the percentage ratio of tAgNuA related to NA (rAgNuA) were determined for each hepatocyte nucleus. The pooled mean values of all the features were significantly different (p less than 0.001) among the case groups. The results point towards a remarkable increase of nucleolar activity in CAH in comparison with CPH, whereas an additional increment of this activity is associated with the progress from CAH to CIR.

Synovial sarcoma: a new approach to the histological classification and a comparison with malignant mesothelioma. Immunohistochemical study with anti-mesothelial cell serum.

Routine paraffin-embedded sections from synovial sarcomas in 70 patients were stained immunohistochemically with monoclonal anti-cytokeratin antibody and serum against mesothelial cells. Positive reactions occurred in the epithelioid cells of biphasic tumors only. On the ground of the reaction with anti-mesothelial serum a hypothetical scheme of cell differentiation in synovial sarcoma was set up.

A new insight into the histogenesis of 'mesodermomas'--malignant mesotheliomas.

A myogenic phenotype was induced in cultures of human mesothelial cells treated for 72 h with atrazine, a triazine derivative. Immunoreactivity for both myosin and myoglobin was detected in a large number of these cells, irrespective of their polygonal or spindle morphology, whereas no expression of desmin was observed. These findings support the embryological identity of mesothelium and mesoderm, the former being, in the post-embryonic stage, potentially capable of differentiation along the same lineages which the latter normally displays during embryogenesis. In the light of this concept it can be assumed that primary malignancies arising from the mesothelium have the competence to express the pluripotent nature of embryonic mesoderm, and hence the term mesodermoma is appropriate for this group of tumours, including mesotheliomas in a classical sense. A postulated mechanism for the phenotypic change of mesothelial cells is also outlined, involving atrazine conversion to 5-aza-chloro-cytidine, a probable DNA hypomethylating and gene activating agent, like its analogue 5-azacytidine.

Ultrastructural morphometry of gastric endocrine cells before and after omeprazole. A study in the oxyntic mucosa of duodenal ulcer patients.

Long-term toxicological experiments with inhibitors of acid secretion were found to induce hyperplasia and eventually carcinoid tumors of the enterochromaffin-like cells of the oxyntic mucosa. To evaluate the effects of 6 months' treatment with omeprazole in humans, the oxyntic endocrine cells were morphometrically investigated at the ultrastructural level in five patients with active duodenal ulcer. No omeprazole-induced changes were found in the volume density of the total endocrine cell population and specific cell types (including the enterochromaffin-like cell) as well as in the other cytological parameters investigated (number of cell profiles per unit area, mean cross-sectional area of cell profiles, nuclear-cytoplasmic ratio, and density of cytoplasmic secretory granules). Both pretreatment and post-treatment values in our patients with duodenal ulcer significantly differed from those of a previous investigation of healthy volunteers with regard to the volume density of enterochromaffin-like cells and non-granulated cells, which increased, and of D cells, which markedly decreased. The latter result may provide a cellular basis for impairment in the paracrine release of fundic somatostatin in peptic ulcer disease. Finally, morphometric data on endocrine cell volume density provided by electron microscopy were found to correlate with those obtained in the same patients using light microscopy techniques (Grimelius silver impregnation and chromogranin A immunostaining). It is concluded that 6 months' treatment with pharmacological doses of omeprazole is devoid of appreciable trophic effect on endocrine cells of human oxyntic mucosa.

A one-year carcinogenicity study with 2,6-dichlorobenzonitrile (Dichlobenil) in male Swiss mice: preliminary note.

2,6-Dichlorobenzonitrile (Dichlobenil) is the active principle of a commercial herbicide that was previously shown to be carcinogenic for animals. Dichlobenil was administered to male Swiss albino mice in order to assess the possible oncogenic properties of the active principle alone. The substance at 2 ppm concentration was injected (0.0005 mg/injection) either by subcutaneous or intraperitoneal route to 2 randomized groups of 30 animals each every third day for 13 times. Dichlobenil induced a significant increase of malignant tumors (lymphoma, mesothelioma, hepatocellular carcinoma, and pulmonary alveologenic carcinoma) with respect to the controls (p less than 0.01 in both treated groups). As for tumor histotypes, only lymphoma incidence was significantly increased in the intraperitoneally treated animals (p less than 0.05). Dichlobenil can be suspected of being a possible carcinogen in Swiss mice but the mechanism of its action is not precisely known.

A morphometrically-based classification rule for the diagnosis of primary mesothelial lesions.

A computer-aided morphometrical study was performed on histological specimens of reactive hyperplastic (n = 10) and malignant (n = 17) mesothelium. For each cell, seven nuclear features were measured and 13 parameters computed. Using stepwise variable selection, discriminant analysis chose the nuclear contour index, the standard deviation of the nuclear area, and the mean of the nuclear perimeter as discriminating features between hyperplastic and malignant mesothelium. The coefficients of these variables were included in a discriminant function which gave perfect discrimination between the two groups of lesions. When the function was assessed on a test set of hyperplastic (n = 10) and malignant (n = 17) mesothelial lesions treated as 'unknown', complete separation between these two diagnostic categories was achieved. This classification rule may help to increase the level of confidence with which a histological diagnosis of mesothelioma can be established.

Quantitative ultrastructure of endocrine cells of oxyntic mucosa in Zollinger-Ellison syndrome. Correspondence with light microscopic findings.

The endocrine cells of the oxyntic mucosa of five patients with longstanding Zollinger-Ellison syndrome were quantitatively investigated with electron microscopy and two light microscopic methods (Grimelius and immunostaining for chromogranin A). Ultrastructurally, the volume density of endocrine cells was 3.2% +/- 1.1% of the mucosal epithelial component, a 168% increase (P less than 0.001) over the value found in normal subjects. Of the six endocrine cell types of human oxyntic mucosa, only enterochromaffinlike cells increased in cell density (65% +/- 15% of the total endocrine cell mass), size, and number of cell profiles per unit area. The enterochromaffinlike cells also underwent morphological changes of secretory granules with a decrease in vacuolated forms, increase in elongated profiles, and appearance of granules with a punctate structure of the core. The latter variety of granules was previously observed only in carcinoid tumors of the oxyntic mucosa and is possibly related to the enterochromaffinlike cell hyperplasia-neoplasia sequence seen in hypergastrinemic patients. A positive relationship was found between endocrine cell densities evaluated ultrastructurally and with chromogranin A immunostaining. It is concluded that in Zollinger-Ellison syndrome, the trophic effects induced by longstanding hypergastrinemia are strictly selective for enterochromaffinlike cells and are associated with ultrastructural features typical for enterochromaffinlike cell tumors.

[Carcinoma of the male breast in the province of Alessandria. Clinico-pathologic aspects of an 8-years' case load]. / Carcinoma della mammella maschile nella provincia di Alessandria. Aspetti clinico-patologici di una casistica di 8 anni.

The authors report their experience on male breast carcinoma based on a series of 16 cases diagnosed between 1981 and 1988 at the City Hospital of Alessandria. Main clinico-pathological findings are described and compared with those recorded in the relevant world literature. Emphasis is placed on the immunohistochemical evaluation of estrogen receptors and the frequency data of this uncommon cancer in hospital-based series are reviewed.

A new method of grading malignancy of prostate carcinoma using quantitative microscopic nuclear features.

A morphometrical assessment of nuclear features and a DNA study were performed on prostate tissue specimens from 33 patients with prostate carcinoma using an image analysing computer. Six nuclear geometric variables were measured and their mean, standard deviation (SD) and standard error (SE) were calculated for each case. The data on nuclear DNA content obtained by static cytometry were processed using an algorithm which provided a DNA grade of malignancy (DNA MG). Using the stepwise multiple regression, we found a significant correlation (p less than 0.01) between the DNA MG, chosen as the dependent variable in the statistical model, and the following nuclear features in decreasing order of importance: area SD, convex perimeter SE, and the mean of maximum diameter. From the correlation coefficients of the variables an equation was built up which provided a geometric nuclear grade of malignancy (GNMG) on a morphometrical basis more closely related to the clinical stage of the tumour (r = 0.75) than the visually assessed histological grade (r = 0.68) based on the Gleason score. This new method of grading malignancy allows an objective and quantitative evaluation to be made of the biological behaviour of the tumour, as measured by the patient's clinical stage.

Expression of epidermal growth factor receptor in human breast carcinoma and its correlation with morphological and biological features of tumour aggressiveness.

A series of 54 patients presenting with primary breast cancer were investigated for tumour expression of epidermal growth factor receptor (EGFR) by the indirect three-step immunoperoxidase technique which used the monoclonal antibody EGFR1. The percentage of malignant cells positive for EGFR was determined and scored on a four-point (0-3) scale in each case. EGFR was demonstrated in 35 (64.8%) tumours. EGFR expression did not correlate (p greater than 0.05) with growth fraction immunohistochemically visualized by means of the monoclonal antibody Ki-67, tumour size, axillary lymph node status and malignancy grade. In contrast, a significant inverse relationship (p less than 0.05) was found between EGFR and estrogen receptor (ER) patterns. Expression of EGFR per se does not appear to be relevant to the biological behaviour of breast cancer as revealed through an evaluation of proliferative activity, pathological stage and histological differentiation. Recognition that EGFR is negatively related to ER supports the present evidence that the control of malignant cell growth and differentiation depends on complex regulatory mechanisms in which several extracellular messenger molecules, including hormones and peptide growth factors together with their specific cellular receptors, are involved and inextricably interwoven.

[Peritoneal splenosis. Report of a case and pathogenetic considerations]. / Splenosi peritoneale. Segnalazione di un caso e considerazioni patogenetiche.

A clinical case of splenosis recently observed in a young man subjected at the age of 8 to splenectomy for rupture of the spleen and operated at 19 for intestinal occlusion is reported. The pathogenetic aspects of the condition, characterised by the presence of nodules of splenic tissue in the abdominal cavity, are examined through a review of the literature.

Triazine herbicides and ovarian epithelial neoplasms.

The possible role of a class of herbicides, triazines, in ovarian carcinogenesis has been evaluated in a population-based case-referent study. The trade names reported by the study subjects, and the type of cultivation in which they worked, were used for the assessment of exposure. Women previously exposed to triazines showed a significant relative risk of 2.7 for ovarian neoplasms. Although none of the doses could be quantified for the study subjects, two risk trends in favor of the plausibility of the association were found: the first by duration and the second by probability of exposure. The population representativity of the study and the comparability of information between the cases and referents suggest the lack of any major bias in the results. Triazine-related risk remained consistent when the analysis was restricted to farmers and when the exposure to other herbicides and to other types of cultivation were considered. Unexposed farmers had the same risk as unexposed nonfarmers.

Putative mesothelial cell growth-promoting activity of a cytoplasmic protein expressed by the mesothelial cell. A preliminary report.

Mesothelioma cells produce a cytoplasmic protein unique to primary tumours of the mesothelium which induces the in vitro proliferation of human mesothelial cells in a dose-dependent fashion. When a polyclonal antibody to this protein was added to cultures of human mesothelioma cells, inhibition of their growth occurred. These results provide evidence for a growth-factor-like role of this mesothelial protein that may act through an autocrine mechanism.

Correlation of proliferative activity with pathological features in breast carcinoma.

A presently undefined nuclear antigen recognized by the monoclonal antibody Ki-67, and the transferrin receptor (TR), both expressed by proliferating cells, were visualized in cryostat sections of 40 consecutive cases of primary breast cancer using a three-step immunoperoxidase technique. The percentages of Ki-67 and TR positive cells were determined. A strong positive correlation was observed between these two indices of proliferation (p less than 0.01). Moreover, each of them was positively related to the histological tumour grade (p less than 0.01), although the scatter in the number of proliferating cells within each grade was large. No significant relation (p greater than 0.05) was found between the percentages of Ki-67 and TR positive cells and tumor size and between these values and axillary node status. These correlations are similar to those recently reported in the relevant literature and suggest that immunohistochemical assessment of proliferative activity may prove to be an objective indicator of biological behaviour and therefore be of clinical and therapeutical importance.

[Benign cystic mesothelioma of the peritoneum]. / Mesotelioma cistico benigno del peritoneo.

A case of peritoneal cystic mesothelioma in a 40-yr-old woman is reported. The patient was operated on for signs of acute appendicitis. The lesion was made up of small, multiple, thin-walled cysts and both visceral and parietal layers of the peritoneum in the appendix region were involved. The lesion must be differentiated from other cystic lesions of the abdominal cavity and in particular from the multilocular lymphangioma. The diagnostic role of special techniques, such as immunohistochemistry and electron microscopy, in difficult cases is emphasized.

The influence of perimenopausal age on estrogen receptor status and estradiol and DNA content in human breast cancer.

In 25 cases of breast cancer occurring in perimenopausal age, estrogen receptor (ER) content was determined by the dextran-coated charcoal (DCC) assay and both endogenous-bound estradiol and nuclear DNA concentrations were measured by computerized quantitative analysis on formalin-fixed paraffin-embedded tissue samples. No statistically significant relationship (p greater than 0.05) was found within these parameters. The high incidence (64%) of ER-negative cases in this menopausal age was mainly due (62.5%) to interference of high levels of endogenous estradiol occupying the receptor sites in-vivo. The prevalence of hormone-insensitive and aneuploid cell clones accounted for the remaining true ER-negative tumours (37.5%).