Localisation of C-terminal gastrin immunoreactivity in gastrinoma cells. An immunoelectron microscopy study on conventionally processed tissue.

Localisation of C-terminal gastrin immunoreactivity has been studied, using the immunogold staining procedure, on ultrathin sections of 6 human gastrinomas conventionally processed for electron microscopy. The specific labelling, whose density depended on the mean diameter of the gold marker, was restricted to endocrine secretory granules. However, in poorly differentiated cells from malignant tumours, a number of granules remained unreactive. The labelling pattern depended also on the functional state of each cell. The immunoreactive granules showed various morphological features. A moderate number of gold particles was demonstrated over the floccular content of the infrequent diagnostic G-type granules. Non-diagnostic round granules of varying size and electron density were prevalent in most cells; their usually strong immunostaining allowed immediate recognition of cell specificity. Dense granules which were large in size and angular in shape and present in one case, were also intensely labelled. In the same tumour, unequal labelling occurred over polymorphous, often elongated granules, of varying size. Granules of different types, including intermediate forms, could be found in the same cell, indicating a spectrum of granule maturation towards well-defined types of the fetal or adult normal tissues. The present methodology would help to identify gastrin-producing cells in prospective or retrospective electron microscopy studies of multi-hormonal endocrine tumours.

[Immunodetection of gastrin-like peptides in routine electron microscopy by the "immunogold" method]. / Immunodétection des peptides de type gastrine en microscopie électronique de routine par la méthode "immunogold".

The present immunoelectron microscopic study was carried out in order to evaluate the influence of several technical parameters of the immunogold staining procedure on the labelling intensity of gastrin-storing cells in normal and pathological states. Ultra-thin sections of double-fixed Epon embedded human pyloric mucosa were immunolabelled using a C-terminal directed antigastrin serum, followed by colloidal gold linked immunoglobulins. When the labelling density occurring over the endocrine secretory granules was quantitatively evaluated, the following conclusions could be drawn: the density increased dramatically when particles of decreasing diameter (40 nm, 20 nm, 10 nm) were comparatively used, the dilution of the primary antiserum had to be adjusted in order to obtain an intense specific labelling together with a negligible background (non specific) labelling. However, a long incubation time of the primary antiserum, as well as the use of etching procedures, resulted in an increase of the labelling density when a highly diluted antiserum was applied. Subsequent attempts to immunolabel ultrathin sections of gastrin-secreting malignant tumours showed that the goal could be achieved on tissues routinely processed for electron microscopy, provide the previous parameters were carefully selected according to the control G cell model.

Light and electron microscope localization of G-17- and G-34-like immunoreactivities of human gastrinomas.

Gastrin- 17 (G-17) and gastrin-34-like immunoreactivities of human gastrinoma cells were investigated at light and electron microscope level using N-terminally directed antisera. The procedure includes (a) the 24 hr/immunoperoxidase staining of Bouin-fixed paraffin embedded tissues, (b) the immunoelectron microscopic labelling of aldehyde-fixed Epon-embedded tissues according to the immunogold technique. On light microscopy, a variable number of tumor cells stained for G-34. In contrast, G-17 immunoreactivity was very low or undetectable in the tumor material, although it was easily detected in endocrine cells of similarly processed human pyloric mucosa. On electron microscopy, most of the tumor cell granules belonging to the round compact or dense-cored type exhibited a variable labelling for G-34, whereas the vacuolar/floccular type remained unlabelled. In contrast, the labelling for G-17 occurred over most of the tumor cell granules, whether compact or floccular. Dense granules of varying size and shape, previously shown to store C-terminal gastrin immunoreactivity, were only faintly labelled by the two antisera. When compared to the labelling pattern of human pyloric G-cells, the predominance of round dense granules with G-34 and G-17 immunoreactivity in gastrinoma cells suggests an incomplete or defective post-translational processing of the precursor peptide.

Bronchial carcinoid with fibrillary inclusions related to cytokeratins: an immunohistochemical and ultrastructural study with subsequent investigation of 12 foregut APUDomas.

A bronchial P cell carcinoid, which was negative for all hormones immunocyto chemically tested, showed a globular intracytoplasmic inclusion in almost every cell. The inclusions were not clearly distinguishable using the haematoxylin- eosin- safran procedure; they were best demonstrated with the Masson trichrome stain and the Grimelius technique and were easily detected in 1 micron thick Epon sections as target-like structures. On electron microscopy, they were found to be composed of filamentous aggregates entrapping a few endosecretory granules, which showed degenerative changes. The filaments, 8-10 nm in diameter, lacked any periodicity; they were randomly dispersed in the central area and arranged in broken concentric swirls at the periphery of the inclusions. The globules lacked the tinctorial properties of amyloid, but showed a strong immunostaining for keratin-like proteins. A systematic investigation of 12 APUDomas of bronchial or duodenopancreatic origin, using both light and electron microscopy, identified a few filamentous bodies in one case, a somatostatin cell tumour of ampulla of Vater. In both cases, the structures appeared similar to those previously reported in growth hormone cell pituitary adenomas as well as in a few bronchial or gut carcinoids. Whatever their nature, morphological data suggest that they are related to abnormalities in the secretory function, involving the Golgi apparatus, the endosecretory granules and the microtubular microfilamentous system.

Placental site trophoblastic tumor of the uterus: an ultrastructural and immunohistochemical study.

Ultrastructural and immunohistochemical studies in a case of placental site trophoblastic tumor (PSTT) of the uterus were carried out in order to define the nature of the abnormal tissue. By electron microscopy, the large cells, whether mononuclear or syncytial, showed numerous ribosomes, prominent Golgi elements, and abundant rough endoplasmic reticulum (RER) filled with granular material. Pseudopods and microvilli were found on the cell surfaces. By immunofluorescence, the well-developed filamentous cytoskeleton proved to be actin-rich. beta-HCG (human chorionic gonadotropin) and SP1 (beta 1-specific pregnancy glycoprotein) were detected in only a few tumor cells, whereas most of them stained for HPL (human placental lactogen). The present results show the secretory nature of most of the tumor cells, which resemble the intermediate trophoblast of the placental bed. Together with previous studies, they suggest that a varying spectrum of syncytiotrophoblastic differentiation exists in PSTT. Decidual, myometrial, or histiocytic cells do not seem involved in the histogenesis of the tumor tissue.

Endometrial carcinoma of the intestinal type. A first case report.

A primary endometrial adenocarcinoma is reported that showed abundant foci suggestive of pathologically differentiated intestinal epithelium. The tumor epithelium was composed of four main cell types. Columnar cells resembled absorptive intestinal cells and displayed glycocalyceal carcinoembryonic antigen immunostaining; mucin-producing cells and a few Paneth-like lysozyme-rich cells were irregularly distributed; a massive quantity of argyrophil cells including a few amphicrine (muco-argyrophil) ones, were detected by Grimelius-Alcian blue method. Immunocytochemical evidence was obtained for the storage of serotonin, somatostatin and gastrin/cholecystokinin in some of the endocrine cells. These findings suggest that the tumor arose from a pluripotential stem cell of the glandular epithelium.

[Histochemical study of estradiol receptors and morphologic parameters in breast cancer]. / Etude histochemique des récepteurs d'estradiol et paramètres morphologiques dans les cancers du sein.

We present our experience with the use of estradiol - bovine serum albumin (BSA) - fluorescein isothiocyanate (FITC) employed by LEE to localize estradiol receptors on frozen sections of breast carcinomas. 41 cases were studied on parallel sections with both conventional staining and a histochemical procedure. Heterogeneity of the cell population was usually found; however the histochemical result was influenced by morphological parameters. As far as histopathological types were concerned, invasive lobular carcinomas (3 cases) manifested a bright cytoplasmic fluorescence, while medullary carcinomas (3 cases) were poorly or non stained. Among infiltrating ductal carcinomas, well differentiated cases were composed of strongly fluorescent tubules, while poorly differentiated ones classified in BLOOM'S grade III were poorly stained. In most of the cases, the amount of stroma, mucus and carcinomatous or hyperplastic ductal components has to be taken in account to evaluate the fluorescence pattern of each microscopic field.

[Amphicrine, mucoid and argyrophile tumors of the breast. Histochemical and ultrastructural study of 2 cases]. / Tumeurs amphicrines, mucoïdes et argyrophiles, du sein. Etude histochimique et ultrastructurale de deux observations.

The authors report two cases of mucoargyrophilic carcinomas of the breast, occurring in 73 and 81 year old women. Both tumors consist of a central mucoid carcinoma surrounded by solid areas with lobular or endocrinoid features. Most cells showed an abundant eosinophilic granular cytoplasm or a mucinous appearance. The histochemical reactions (Grimelius-Alcian Blue, Grimelius-Mucicarmine) and the ultrastructural study of the peripheral areas disclosed several cell types: poorly differentiated or mucinous cells were present in both cases; but most of the cells were argyrophilic and contained in their hyaloplasm numerous dense core "neurosecretory" granules; such cells revealed many microfilaments arranged in bundles or in whorls. Amphicrine cells were easily demonstrated by histochemical staining and their cytoplasm showed both endocrine granules and mucinous vesicles. These features suggest a possible relationship to other argyrophilic tumors of the breast. The histogenesis of these tumors is still little known.

[Tubal epithelium. Scanning electron microscopy in the menstrual cycle, in pregnancy and in the menopause (author's transl)]. / L'épithélium tubaire. Microscopie électronique à balayage au cours du cycle menstruel, de la grossesse et de la ménopause.

The Fallopian tubes of 18 women who were either in reproductive age or were menopausal were studied by scanning electron microscopy. There were different cells seen in the epithelium. These were ciliated and non-ciliated cells, and cells, with a single cilium which was central and cells with short cilia. These elements undergo cyclical changes: secretory phenomena are at their maximum during the ovulatory phase. There is no sudden desquamation in tubal epithelium, nor is there massive deciliation. All the same, there are images that look like marguerites (described in the text) which are evidence of cellular regeneration. The appearance suggests a state of rest in precocious menopausal women and in pregnancy. The phenomena seen during the secretory phases disappear and the distribution of the cells is modified. Deciliation is found 20 to 30 years after the menopause. The physiological significance of these phenomena is still uncertain and their role in pathology has still to be defined.

[Duodenal ulcer and pyloric micro-gastrinoma. A report of one case (author's transl)]. / Ulcère duodénal et micro-gastrinome pylorique. A propos d'une nouvelle observation.

We report a case of micro-gastrinoma associated with a duodenal ulcer. This ulcer was severe and a gastrectomy (Finsterer) was performed. Pathologic examination showed a "carcinoid islet cell-tumor" with argyrophil granules in the cells' cytoplasme and ultrastructural studies confirmed neurosecretory granules with dense central core. Using the immunofluorescence and immunoperoxydase technic with several anti sera against polypeptide hormones we were able to show it was a pure gastrinoma. We presume this association is a new entity in gastrin-cells pathology.