Structure-function correlations of growth hormone or/and prolactin-producing pituitary adenomas: an in vitro study with the reverse hemolytic plaque assay.

The purpose of this study was to detect in vitro growth hormone (GH) and prolactin (PRL) secretion from adenomas clinically associated with GH or PRL hypersecretion. The reverse hemolytic plaque assay (RHPA) was applied in order to reveal possible differences among various morphologic adenoma types, and to examine the inhibitory effects of octreotide on GH release as well. The 20 surgically resected pituitary adenomas studied included 15 from acromegalic patients and 5 from patients with hyperprolactinemia. All adenomas were diagnosed by histology, immunocytochemistry and electron microscopy. Among tumors associated with acromegaly, 5 were densely granulated (DG), 5 were sparsely granulated (SG) somatotroph (SM) adenomas, 2 were mammosomatotroph (MSM) and 3 mixed somatotroph-lactotroph cell (mixed SM-LT) adenomas; tumors causing hyperprolactinemia included 4 lactotroph (LT) adenomas and 1 mixed SM-LT adenoma. GH release assessed by the RHPA corresponded to in vivo hormone secretion and to tissue immunoreactivity. Statistical analysis showed significant differences among all morphologic types of SM adenomas, exclusive of SG-SM adenomas compared to mixed SM-LT adenomas. The mean plaque size in DG-SM and MSM adenomas was significantly greater than that of SG-SM and mixed SM-LT adenomas, indicating higher GH secretion by the former two types during the same incubation time. PRL secretion was documented in 2 mixed SM-LT adenomas. Plaques for PRL, but not for GH were formed in all LT adenomas. In all SM and LT adenomas, cells producing large plaques represented a minority of the plaque-forming cell population, however, they accounted for the largest part of the total plaque area, thus the largest part of hormone secretion. Octreotide effects on GH release were studied in 6 adenomas by the RHPA. Octreotide treatment induced a rapid and significant reduction in GH secretion by SM cells in vitro, with a selective effect on high-secreting cells.

[Cutaneous hyalinosis: 3 cases]. / Hyalinose cutanéomuqueuse: trois cas.

INTRODUCTION: The authors report three cases of "Hyalinosis cutis et mucosae"; the first and the second case concern two siblings children (brother and sister) while the third is reported on a 42-year-old woman. PRESENTATION OF CASES: The case history and the personal anamnesis, the morphology of the skin lesions, the symptoms indicating laryngeal mucous affection as well as the histopathological picture, are compatible with the "Urbach-Wiethe disease". Furthermore, in the third case, the electron microscopic findings confirm the diagnosis. DISCUSSION: The particularities of the present cases are discussed in connection with the corresponding bibliographical data.

DNA measurement in pituitary adenomas assessed on imprints by image analysis.

OBJECTIVE: To determine the DNA content and S-phase fraction (SPF) of pituitary adenomas by image analysis and to correlate them with clinical and morphologic parameters. STUDY DESIGN: The study group consisted of 26 prospectively collected cases of operated pituitary adenomas (3 microadenomas and 23 macroadenomas). The tumors were classified by histology, immunocytochemistry and electron microscopy. DNA measurement was performed on imprints from fresh pituitary tissue. Samples of nontumorous adenohypophysial parenchyma served as normal controls. RESULTS: Overall, 31% of adenomas, all but one functioning one, were aneuploid. The remaining nonfunctioning aneuploid tumor was a null cell adenoma with glycoprotein differentiation. All aneuploid tumors were macroadenomas, mostly at advanced stages, III and IV. Dural invasion, although frequent in macroadenomas (78%), was not correlated with DNA ploidy and SPF. An increased number of hyperpentaploid aneuploid cells was noted primarily in aneuploid tumors. The mean SPF was < 2.50%, with a statistically significant difference between aneuploid and diploid adenomas (3.60% vs. 1.70%). CONCLUSION: The results suggest that quantitative assessment of DNA content may provide important information, particularly in functioning adenomas. In addition, fresh tissue imprints represent excellent material for optimum cytometric measurements by image analysis systems, even for microadenomas.

Diagnosis of pituitary adenomas on touch preparations assisted by immunocytochemistry.

Touch preparations from surgically removed pituitary adenomas were studied by both routine staining and immunocytochemistry for all anterior pituitary hormones. The results were correlated with the histologic, histochemical, immunohistochemical and ultrastructural findings. Several features were recognized, some of them applicable in diagnosing various adenoma types: high cellularity, cell monomorphism, nuclear irregularities and heterogeneity of immunoreactivity, all major diagnostic criteria of pituitary adenoma. The presence of numerous fibrous bodies associated with nuclear pleomorphism, multinucleation and peripheral displacement of nuclei, in association with growth hormone (GH) immunopositivity, represent diagnostic criteria for sparsely granulated GH cell adenoma. The "Golgi" pattern of prolactin (PRL) immunopositivity in conjunction with the small size of cell and nucleus and the presence of microcalcifications represents diagnostic features of PRL-secreting adenoma. Cytoplasmic accumulation of hyaline material in association with peripheral distribution of periodic acid-Schiff stain and adrenocorticotropic hormone immunoreactivity is characteristic of sparsely granulated corticotroph cell adenoma. Knowledge and application of the described features may contribute significantly to the diagnostic approach to pituitary adenomas.

A cytologic, ultrastructural and immunocytochemical comparison of tumor cells and cell cultures originating in invasive bladder carcinoma.

Tumor cells and urine-voided cells from patients with invasive bladder carcinoma as well as from healthy patients were examined cytologically, ultrastructurally and immunocytochemically. The ultrastructure of tumor cells showed an abundant, dilated, rough endoplasmic reticulum in the form of membrane-bound vacuoles full of granular to fibrillar material located perinuclearly and/or paranuclearly. Some cells exhibited enlarged modified lysosomes containing sparce flocculent and particulate precipitate. Papanicolaou staining of these cells showed two basophilic cytoplasmic textures, one green glossy-patchy, perinuclearly and/or paranuclearly, well segregated from the other texture of peripheral hematoxylinophilic foamy cytoplasm, comparable to the cytologic features of cell cultures originating in invasive bladder carcinoma. PAS diastase showed double distribution and texture of the perinuclear glycosaminoglycans, a glossy accumulated mass and large granules. Glycosaminoglycan sacs similar to those of cell cultures were also present in tumor-dispersed cells. There was a nonspecific binding of antisera against lysozyme, human chorionic gonadotropin and alpha 1-trypsin in normal and tumor cells. Tumor cells and tissues were positive for alpha 1-chymotrypsin distributed perinuclearly and in large spheres. Normal cells lacked the above characteristics. The results indicate that it is feasible to use the aforementioned characteristics in conjunction with the existing bladder-cytologic criteria for malignancy as markers in urothelial cancer with regard to prognosis of superficial tumors with high malignant potential.

Common characteristics of primary cell cultures originated from invasive urothelial carcinoma.

Ten primary cultures from invasive urothelial carcinomas were studied by light and electron microscopy. In all cultures, cells with accumulations of glycosaminoglycans were observed. Long membrane extensions with accumulations of glycosaminoglycans invaded neighboring cells and formed an extracellular matrix as well. The extracellular matrix inhibited cell proliferation and enhanced tumor nodule formation.

Morphological and immunohistochemical characteristics of a cell line originated from noninvasive human bladder transitional-cell carcinoma.

A cell line (IG) was derived from a noninvasive transitional cell carcinoma grade II. The cells were propagated in vitro for 9 months, had a low proliferative capacity, formed tumor nodules, and showed four characteristic morphological stages. Primary epithelial-like cells were characterized as stage I; fibroblast-like cells with loss of contact inhibition of growth as stage II; fibroblast-like cells with long cytoplasmic projections, massive glycosaminoglycan content and tumor nodule formation as stage III; and, as stage IV, granulated stellate cells. The cell culture progressed cytologically from well-differentiated stage I to less-differentiated stage IV. The four stages were confirmed and examined by electron microscopy. Immunoreactivity for fibronectin, keratin, epithelial membrane antigen and carcinoembryonic antigen shown by the cell line and the tissue indicates that the cell line represents the original cell population in the tumor. Glycosaminoglycan production by these cells was found to be responsible for the formation of tumor nodules which appeared in culture at the same time as the patient had developed an invasive bladder tumor. Glycosaminoglycans were observed as a massive accumulation intracellularly and extracellularly, as rods or thick slices in layers of adherent vesicles and granules covering the cell culture. Normal bladder epithelial cells and fibroblasts grew in monolayer, showed contact inhibition of growth, were devoid of intracellularly accumulated glycosaminoglycans and produced uniform extracellular matrix different from that of IG cells.