Biomedical subjects
Publications and source records attributed to E Horvath.
This review summarizes some aspects of pituitary adenoma pathology. A new embracing pituitary adenoma classification has been developed which correlates morphologic findings with endocrine activity. It is based on hormone content, histologic, immunohistochemical and ultrastructural features, cellular composition and cytogenesis, and separates pituitary adenomas into 7 distinct entities.
The present review deals with the use of electron microscopy in the identification of pituitary cell types as well as the assessment of their functional state, in rat and man. Application of immunoelectron microscopy, especially immunogold techniques, utilizing multiple labeling in establishing differentiation and hormone content of cell types, is emphasized. Recent evidence of plurihormonality in various pituitary cell types indicates that the once axiomatic one cell-one hormone theory is untenable and that the present perception of pituitary cell types and their function requires modification. Detection of hormonal and nonhormonal substances in pituitary cell types, not associated with their known endocrine function, suggests that hypophysial cells may have yet unknown roles, possibly in the realm of paracrine and autocrine regulation.
Isolated CNS myelin membranes were extracted with Triton X-100 under conditions previously established for the isolation of cytoskeletal proteins. Treated myelin retained much of its characteristic lamellar structure despite the removal of most of the major myelin basic protein (18.5 kDa) and the proteolipid protein, which together normally constitute 60% of the total myelin protein. The SDS-PAGE profile of this extract residue demonstrated an enrichment in proteins of Mr 30 to 60 kilodaltons (the Wolfgram group). The major myelin proteins were identified by antibodies on Western immunoblots, as were the 2'3'-cyclic nucleotide 3'-phosphodiesterase (CNP), actin, tubulin, myelin-associated glycoprotein (MGP) and the 21.5 kDA MBP. The overall behavior of CNP, the 21.5 kDa MBP, MGP and tubulin towards Triton extraction is reminiscent of the behavior of other membrane-skeletal complexes, supporting the idea that these and other minor myelin proteins might be part of heteromolecular complexes with interactions spanning several lamellae of the myelin sheath.
Primary hypothyroidism accompanies more than 50% of clinically significant pituitary thyrotroph adenomas. Hypothyroidism may also produce pituitary enlargement secondary to thyrotroph hyperplasia and present with a sellar mass and hyperprolactinemia. Three hypothyroid patients who presented with presumed prolactin-producing adenomas are reported. Although laboratory and radiologic abnormalities of pituitary enlargement may resolve after corrective thyroid therapy, our patients showed no such response and underwent operation. Histologic examination revealed no adenomas, but thyrotroph and lactotroph hyperplasia were present. Thyrotroph hyperplasia probably results from lack of negative feedback of thyroid hormone upon the anterior pituitary. Whether this is due to hypothalamic release of thyrotropin-releasing hormone (TRH) or another mechanism is unclear. Lactotroph hyperplasia may result from excess TRH, which stimulates lactotrophs with resultant hyperprolactinemia, or from reduced hypothalamic dopamine, thereby facilitating prolactin secretion. This study suggest that (a) hyperprolactinemia in hypothyroidism is not necessarily due to the "stalk section effect" secondary to pituitary enlargement, and (b) patients with primary hypothyroidism and sellar mass should initially be managed medically so that potentially reversible pituitary hyperplasia is not mistaken for adenoma.
The morphologic study of a large number of human pituitary tissues provided conclusive evidence that hyperplasia of various adenohypophysial cell types exists. These rare lesions may serve as the underlying cause of pituitary hypersecretory syndromes. The non-neoplastic proliferation of adenohypophysial cell types may be secondary to orthotopic or ectopic oversecretion of hypothalamic regulatory peptides, such as GRH, CRH, TRH and GnRH, disturbances of dopaminergic regulation or iatrogenic factors. Histologically, adenohypophysial hyperplasia may be diffuse or nodular. The former type does not result in major alterations in pituitary architecture and may be impossible to recognize in small surgical specimens. The latter type is associated with enlargement of pituitary acini. Examination of the reticulin network, using silver staining techniques, represents useful tools in the differential diagnosis. The possible emergence of adenoma on the grounds of hyperplasia is discussed.
The ultrastructural features and immunohistochemical characteristics of nontumorous adenohypophysial cells in the human pituitary are briefly reviewed and the usefulness of electron microscopy and immunoelectron microscopy is stressed in cell identification. Recent immunoelectron microscopic studies demonstrate the presence of plurihormonal cells containing more than one hormone, not only in pituitary adenomas but also in nontumorous adenohypophyses. These results lead to the conclusion that the one cell- one hormone theory, which dominated pituitary cytophysiology and cytopathology for several decades, is untenable.
This review summarizes current knowledge on pathology of proliferative lesions of the human pituitary. The morphologic classification of pituitary adenomas--based on histology, immunohistochemistry and electron microscopy--has now been firmly established. It has been conclusively proven that all presently recognized adenohypophysial cell types give rise to adenoma and all known pituitary hormones may be secreted in excess. Evidence is accumulating that hyperplasia of various adenohypophysial cell types can lead to hypersecretory syndromes similar to those associated with the corresponding adenomas. Owing to the rarity of studies on pituitary hyperplasia, the condition is still incompletely defined. The difficulties regarding morphologic diagnosis of pituitary hyperplasia are discussed. Despite major advances in the field of pituitary pathology, several problems concerning structure-function relationship, as well as pathogenesis of proliferative lesions are still unresolved. There is strong circumstantial evidence suggesting that the cytological mapping of the pituitary is incomplete and there are still cell types waiting to be discovered.
The pituitary glands were removed from 63 human fetuses from 5 weeks of gestation to term and studied by electron microscopy and ultrastructural immunocytochemistry to document the development of cell differentiation and hormone production in the adenohypophysis. At 5 weeks of gestation, Rathke's cleft was lined by columnar epithelium with abundant cytoplasmic glycogen and occasional secretory granules. By 6 weeks of gestation, cells resembling corticotrophs were identified; in 8-week-old fetuses, type I microfilaments were found in those cells. Well-differentiated somatotrophs were seen in adenohypophyses of 8- to 9-week-old fetuses. Although secretory granules were numerous, the Golgi complex was inconspicuous in early fetal glands. After 10 weeks of gestation, there was a change with morphologic evidence of active hormone secretion; large Golgi regions and sparsely granulated cells were found. Some somatotrophs at this stage contained aggregates of type II microfilaments which resembled the fibrous bodies of sparsely granulated somatotroph adenomas. Densely granulated mammosomatotrophs containing growth hormone and prolactin were identified at 12 weeks of gestation. Cells with characteristics of the glycoprotein hormone cell line were seen in pituitaries at 12 weeks of gestation; thyrotrophs and gonadotrophs were identified after 15 weeks. Typical lactotrophs were not recognized before 23 weeks, but were numerous in pituitaries of fetuses older than 35 weeks. This study documents for the first time the existence of a bihormonal mammosomatotroph in the human fetal pituitary and confirms that somatotrophs and lactotrophs, the two acidophil cell types, are embryologically related.(ABSTRACT TRUNCATED AT 250 WORDS)
Twenty patients with a novel, frequently aggressive type of pituitary adenoma, termed silent subtype 3 adenoma on the basis of fine structural criteria, are reported. The surgically removed tumors were studied by morphological techniques, and the findings were correlated with clinical and biochemical data. All tumors were macroadenomas, often with parasellar extension. The histologically diffuse tumors frequently exhibited focal immunopositivity for one or more adenohypophysial hormones, although the majority of adenoma cells were negative. The tumors had characteristic electron microscopic features, assuring specific diagnosis and delineating this tumor type as a distinct ultrastructural entity. The tumors were removed from 9 women and 11 men (median ages, 27 and 41 yr, respectively). In all women, mild to moderate hyperprolactinemia and its sequelae were present from the early phase of the disease, leading to the erroneous diagnosis of prolactinoma. Bromocriptine therapy (3 patients) reduced serum PRL levels to normal, but failed to halt tumor growth. In men, most adenomas were nonfunctioning; 4 men had mild to moderate hyperprolactinemia. Three men had elevated serum GH levels and acromegaly, suggestive of multidirectional differentiation. Although the putative cell type giving rise to silent subtype 3 adenomas is not known, the tumor should be recognized to avoid erroneous diagnosis and inappropriate treatment.
Injection of concentrated influenza B/Lee/40 virus into 4-week-old Balb C mice resulted in 60% inhibition of 14C-palmitate oxidation in isolated hepatic mitochondria. Oral feeding of carnitine to infected mice prevented the inhibition of fatty acid oxidation. High concentrations of salicylic acid given orally also inhibited 14C-palmitate oxidation. Serum free fatty acid concentrations of infected mice and of those fed salicylic acid were significantly higher than in control mice. A combination of low-dose virus and low-dose salicylic acid inhibited palmitate oxidation, suggesting an additive effect on the metabolic derangement when the two agents were present simultaneously.
Two pituitary gonadotroph adenomas were studied in vitro to characterize structure-function correlations. Both tumors were from men, aged 63 and 69 years, who had elevated blood levels of follicle-stimulating hormone (FSH) and normal blood luteinizing hormone (LH) and testosterone values. The surgically resected adenomas contained diffuse immunohistochemical positivity for beta-FSH, beta-LH, and alpha-subunit of glycoprotein hormones; by electron microscopy they were composed of well-differentiated gonadotrophs. In vitro, both tumors released FSH, LH, and alpha-subunit. Morphometric studies were performed on surgically resected and cultured adenoma cells. Compared with the surgical specimens, the cultured cells had decreased cytoplasmic volume densities of endoplasmic reticulum and Golgi apparatus and slightly increased cytoplasmic volume densities of secretory granules. Incubation with gonadotropin-releasing hormone (GnRH) for 2 and 24 hours increased FSH, LH, and alpha-subunit release by both tumors; morphometry after 2 consecutive days of exposure confirmed significant increases in cytoplasmic volume densities of endoplasmic reticulum and Golgi regions and marked decreases in that of secretory granules. There was no significant change in cell size, nuclear/cytoplasmic ratio, or secretory granule diameter. The two tumors differed in their response to gonadal steroids. Estradiol, testosterone, and progesterone stimulated release of FSH, LH, and alpha-subunit by one tumor and the morphologic changes paralleled the biochemical response; addition of testosterone suppressed the secretory and morphologic response to GnRH. The other tumor showed no significant response to estradiol or testosterone and addition of these steroids did not alter the response to GnRH. The results are consistent with the interpretation that GnRH stimulates not only release but also synthesis of gonadotropins by gonadotroph adenomas of men. The data also indicate variable sensitivity of these tumors to gonadal steroids with paradoxical stimulation alone and inhibition of response to GnRH. The structural changes correlate with the hormone release response in vitro.
An enzyme-linked immunosorbent assay (ELISA) was developed for detection of antibodies against the officially recognized bovine adenoviruses (BAV). Soluble antigens of adenoviruses were extracted from bovine kidney (BAV-1 and BAV-3) or bovine testicle (BAV-4, -6, and -7) cell cultures, respectively. Presence of the major hexon antigen, known to be the most essential compound for serological cross-reactivity, was demonstrated on a discontinuous sodium-dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The relatively crude polypeptide preparations were coated onto microelisa plates and tested for specific reactivity with available type-specific antisera of calves. Plates coated with BAV-1 showed reactivity with antibodies against types 1 and 3, but did not react with hyperimmune serum against BAV-4. Soluble antigens of BAV-4 react to a high extent with homologous antiserum but did not cross-react with a hyperimmune serum against BAV-1. Both these hyperimmune sera had been prepared in colostrum-free calves. Several calf sera produced by cooperating researches in colostrum-free calves were tested for homologous and cross-reactivity. In addition, ELISA plates coated with BAV-4 were tested on 33 sera of conventionally reared calves, earlier infected experimentally with homologous BAV-4. ELISA titres correlated well in 28 of them with seroneutralization titres; in 5 other sera correlation was poor, but in no case to a degree which would have resulted in a dissenting diagnosis. The use of BAV types 1 and 4 as representatives of bovine adenovirus subgroup I and II will allow easy and economical surveillance of these frequently occurring antibodies in diagnostic laboratories.
To study the effect of hyperprolactinemia on nontumorous prolactin cells, their morphology was investigated by transmission electron microscopy and morphometry in the nontumorous portion of seven human pituitaries containing prolactin-producing adenoma. For cell identification, immunoelectron microscopy was applied using the immunogold labelling method. In five cases, nontumorous prolactin cells demonstrated features of inactivity, whereas in two cases they showed morphological features of active hormone secretion. Our findings are consistent with the view that prolactin regulates its own release from prolactin cells in the anterior pituitary via the short loop negative feedback mechanism. Hyperprolactinemia presumably activates the short loop feedback mechanism resulting in inhibition of nontumorous prolactin cell. However, in two cases, hyperactivity of nontumorous prolactin cells was observed suggesting a dysfunction of the feedback mechanism.
Intracranial extension of rhabdomyosarcoma from the face, nasopharynx or middle ear is rare. A 16-year-old boy presented with deterioration of vision and headache. CT scan revealed a soft tissue mass occupying the sphenoid and ethmoid sinuses, extending to the suprasellar fossa and impinging on the optic chiasm. The tumour, mimicking pituitary carcinoma, was removed by transsphenoidal craniotomy. Morphologic studies, including immunohistochemistry and electron microscopy, revealed that the tumour was a rhabdomyosarcoma. This case stresses the value of immunohistochemical and ultrastructural studies in the diagnosis of tumours occurring in the region of the sella turcica. The origin of this tumour was thought to be the sphenoid or ethmoid sinus. The pituitary gland appeared intact.
The present study reports the effects of SMS 201-995, a long-acting somatostatin analogue, on blood GH levels, glucose tolerance and tumour morphology in a 36-year-old, previously untreated acromegalic woman. Treatment (50 micrograms s.c., 8-hourly) resulted in marked suppression of GH concentration and an improvement in glucose tolerance. After 10 d of treatment, the tumour was removed by transsphenoidal surgery and studied by histology, immunohistochemistry, transmission electron microscopy and morphometry. Histologically, the tumour was an acidophilic adenoma which contained immunoreactive GH in many adenoma cells. By electron microscopy, the tumour was composed of densely granulated somatotrophs containing numerous large secretory granules and many lysosomes showing crinophagy. No cell necrosis or vascular impairment were evident. Using morphometry, the tumour was compared with 10 densely granulated somatotroph adenomas, removed from acromegalic patients not treated with somatostatin. The nuclear and cytoplasmic areas of the adenoma subjected to SMS 201-995 treatment were smaller, and the lysosomes occupied more of the cytoplasmic volume than those of controls. The nuclear/cytoplasmic ratio, cytoplasmic volume densities of endoplasmic reticulum, Golgi apparatus, mitochondria, secretory granules and secretory granule diameters were within the range of control adenomas. In vitro, treated adenoma cells secreted GH and retained responsiveness to both GRH stimulation and somatostatin suppression. The morphologic findings after SMS 201-995 treatment, are consistent with suppression of GH release. There is no evidence that somatostatin has any direct cytotoxic or vasotoxic effects. It appears that SMS 201-995 represents a potent and promising drug in the medical treatment of acromegaly, however, more work is needed to elucidate the mechanism of somatostatin suppression and to provide evidence for adenoma shrinkage.
We report the first documentation of GHRH production by a tumour associated with proven multiple endocrine neoplasia (MEN). A 30-year-old woman had hypoglycaemia, hyperparathyroidism, and pituitary adenoma with hyperprolactinaemia. Serum growth hormone elevation was attributed to hypoglycaemia but plasma GHRH was elevated. Subtotal pancreatectomy revealed multiple endocrine tumours and nesidioblastosis. Immunohistochemistry demonstrated insulin, glucagon, and somatostatin in several tumours. GHRH was localized in the largest one and was released from that tumour in vitro. Post-operative plasma GH returned to normal. Excess secretion of humoural factors by one tumour may stimulate growth of other tumours in MEN syndromes. The prevalence of GHRH in MEN-I tumours remains to be established.
In a series of 1,500 pituitary adenomas surgically resected at Mayo Clinic, 41 (2.7%) occurred in the setting of multiple endocrine neoplasia, type I (MEN-I). Of the 40 patients (18 males, 22 females), 21 (52%) presented with clinical evidence of a pituitary neoplasm, 13 with hyperparathyroidism, and two with functional islet cell tumor. Of the 41 tumors, 11 (27%) were microadenomas, and 30 (73%) were macroadenomas. Immunocytochemical studies demonstrated the following reactivities: GH (4), GH/PRL (6), GH/PRL/glycoprotein (7), GH/ACTH/glycoprotein (1), PRL (16), PRL/TSH (1), ACTH (3), and null cell adenoma (3). We conclude that, in comparison with pituitary adenomas occurring in the general population, those occurring in association with MEN-I are (1) more often endocrinologically functional, (2) more frequently GH- or PRL-producing, and (3) clinicopathologically similar in terms of the subjects age and sex as well as of tumor size and invasiveness.
This article reviewed the classification, morphologic features, and pathogenesis of human hypophyseal adenomas and differences between hyperplasia and adenoma. Histologic, immunohistochemical, and electron microscopic investigation obtained a deeper insight into several aspects of pituitary cytopathology. Based on these studies, a meaningful classification has been developed that separates hypophyseal adenomas into distinct entities. It hinges on histology, hormone content, ultrastructural characteristics, cellular composition, and cytogenesis and correlates the morphologic features of adenoma cells with endocrine activity. Although progress has been spectacular during the last two decades in the better understanding of structural characteristics and hormonal function of pituitary adenomas, much more work is necessary to shed light on structure-function correlations and pathogenesis. Collaborative investigations between clinical investigators and basic scientists can further advance knowledge in this intriguing and important field.