Kioko Kawai

Immunohistochemical expression of epidermal growth factor receptors in human adrenocortical carcinoma

Epidermal growth factor (EGF) receptors were examined immunohistochemically in 64 adrenocortical carcinomas obtained at autopsy, and in 23 adrenocortical adenomas and seven pheochromocytomas obtained during surgery. In the nonneoplastic adrenal gland, EGF receptors were scattered to the zona glomerulosa, zona fasciculata, and zona reticularis. Adrenocortical carcinomas (63 of 64), more than adrenocortical adenomas (10 of 23) or pheochromocytomas (four of seven), stained positively for EGF receptors (P less than .01). The immunoreactivity was limited to the cytoplasm, cell membrane, and chromatin. When the antibody was immunoabsorbed with an excess of immunogen peptide, there was no evidence of immunostaining. The adrenocortical carcinomas could be classified into 16 cases of the well-differentiated type, 33 cases of the moderately differentiated type, and 15 cases of the poorly differentiated type. There was no relationship between histologic grading and staining intensity of the EGF receptors. On the other hand, more than 80% of the cases of adrenocortical carcinomas revealed a moderate to high intensity for EGF receptors. In 62 of the 64 patients, there was already metastases to other organs. We conclude that the expression of EGF receptors is associated with tumor growth and/or metastatic potential in adrenocortical carcinoma.

Receptor autoradiographic localization of insulin-like growth factor-I (IGF-I) binding sites in human fetal and adult adrenal glands

We report here the first evidence of insulin-like growth factor-I (IGF-I) binding sites in human fetal and adult adrenal glands, obtained at autopsy. Sections of tissue were incubated with 0.1 nM [125I]IGF-I and analyzed using [3H]Ultrofilm autoradiography with image analysis coupled to computerized microdensitometry. Specific binding sites of [125I]IGF-I were found to be localized in the definitive zone, fetal zone, and fetal medulla of the fetal adrenal glands. In the adult adrenal glands, the entire cortex and medulla were specifically labeled with [125I]IGF-I. Specific binding obtained at a concentration of 0.1 nM [125I]IGF-I to areas in the fetal and adult human adrenal glands was competitively displaced by unlabeled IGF-I, with an IC50 value of 0.34-2.54 nM, and 0.38-0.73 nM, respectively, whereas insulin was much less potent in displacing the binding. Acquisition of this knowledge will aid in studies on cell growth and steroid-catecholamines biosynthesis of the human adrenal gland.

AN AUTOPSY CASE OF CREUTZFELDT‐JAKOB DISEASE WITH KURU‐LIKE NEUROPATHOLOGICAL CHANGES

An autopsy case of Creutzfeldt‐Jakob disease with Kuru‐llke neuropatho‐logical changes which revealed clinically extrapyramidal, pyramidal and psychic symptoms is presented in this report. On microscopic examination, status spongiosus, neuronal degeneration, proliferation of hypertrophic astrocytes and numerous plaques were observed in the cerebrum and cerebellum accompanied with widespread demyelinization. These plaques which suggested Kuru plaques measuring 10 to 60 μ were strongly PAS positive and had a dense central core surrounded by a halo of fine radially arranged fibrils. As for the relationship between Creutzfeldt‐Jakob disease and Kuru, the significance of these morphological changes is discussed

Adrenocorticotropic hormone-independent bilateral macronodular adrenocortical hyperplasia associated with Cushing's syndrome

A case of adrenocorticotropic hormone independent bilateral adrenocortical macronodular hyperplasia (AIMAH) is reported. A 59 year old male was admitted to hospital because of hypertension. Subsequently, hypercortisolism, low plasma adrenocorticotropic hormone (ACTH), loss of diurnal rhythm of ACTH, lack of suppression with high dose dexa‐methasone were found and bilateral adrenal enlargement was detected by abdominal computerized tomography and adrenal scintlgraphy. Bilateral total adrenalectomy was performed under a diagnosis of bilateral adrenal hyperplasia associated with Cushings syndrome. Both adrenal glands were enlarged in size and weight. Bulging nodules were found at the cut section. Microscopically, a variegated histologic pattern including trabecular, adenoid and zona glomerulosa‐like (ZG‐like) structures was revealed in the nodules. lmnunohistochemical examination disclosed positive staining of cytochrome P‐450 1701, negative of 3p‐HSD in the ZG‐like structure. Ultrastructurally, the cells composing the ZG‐like structure were similar to those of the ZG in normal adrenal cortex. The authors agree that AIMAH is one of the entiiies causing Cushings syndrome, and advise patholo‐gists to keep this disorder in mind when they examine the adrenals in Cushings syndrome.

Hypercalcemia and production of parathyroid hormon.e-like protein in adult T-cell leukemia-lymphoma

To the editor: Human T-lymphotropic virus type1 (HTLV-1) infection causes adult T-cell leukemia-lymphoma (ATLL) and a tropic spastic paraparesis/myelopathy (1-2). ATLL is geographically limitated to patients born in southwestern Japan and the Caribbean region, which are ATLL endemic areas. As our university is located in the southwestern part of Japan, ATLL cases are frequently autopsied. Hypercalcemia has been reported to be a frequent complication of ATLL, which is very difficult to treat and is the direct cause of death in 22% of patients with ATLL. Dazai and co-workers reported 2 cases of ATLL associated with acute pancreatitis due to hypercalcemia (3). We have also published I such case (4) and investigated the incidence of the same complication (5). Metastatic calcification due to hypercalcemia in ATLL was commonly seen in alveolar septae of the lungs, in renal tubules, and in myocardium. Hypercalcemia in patients with hematological cancers and other tumors without metastasis is probably due to the production of bone resorptionstimulating factors by the tumor cells (6-9). We investigated production of parathyroid hormone-like protein (PTHLP) in ATLL patients associated with hypercalcemia in the tissues. Three cases of severe metastatic calcification due to hypercalcemia in ATLL associated with osteolytic changes were studied by immunohistochemistry. The material was obtained at Nagasaki University Hospital from lymph nodes of autopsy cases. These specimens were fixed 10% formalin and were routinely embedded in paraffin. Paraffinembedded 4-micron sections were stained with PTHLP (Polyclonal antibody; Oncogene Science, Inc., New York; Lot. 93900202); this antiserum reacts with human PTHLP on immunoblots and formalin-fixed paraffin-embedded tissue sections, but shows no cross-reactivity with human parathyroid hormone. The antibody was diluted 1:lO with phosphate-buffered saline (PBS) pH 7.4. The indirect method was performed for the staining of PTHLP using Dako universal kit for polyclonal antibody (Dako PAP kit 548; Lot. 090-2). of in in adult

Analysis of mRNA expression for steroidogenic enzymes in the remaining adrenal cortices attached to adrenocortical adenomas.

DESIGN AND METHODS We have recently demonstrated that the adrenal cortices attached to aldosterone-producing adenoma (APA) contained microscopic subcapsular micronodules suggestive of active aldosterone production. In this study, we used in situ hybridization to investigate the mRNA expression of steroidogenic enzymes in the adrenal cortices attached to cortisol-producing adenoma (CPA) and clinically silent adenoma (non-functioning adenoma; NFA), in addition to APA. RESULTS Microscopic subcapsular micronodules, which were several hundreds of micrometers in size and spheroid in shape, were observed in the cortices attached to CPA and NFA, as well as APA, at high frequency. Most of the cortical nodules in zona fasciculata to zona reticularis showed a suppressed steroidogenesis in the cortices attached to adenoma, but some expressed intensely all necessary steroidogenic enzyme mRNAs for cortisol synthesis. CONCLUSIONS It is thus necessary to keep in mind, on the occasion of subtotal adrenalectomy, that lesions with the potential to later develop into functional adrenocortical nodules may be present in other parts of the ipsilateral or contralateral adrenal cortices.

MALIGNANT GERM CELL TUMOR OF THE ANTERIOR MEDIASTINUM WITH LEUKEMIA‐LIKE INFILTRATION

We present here a rare autopsy case of malignant germ cell tumor with leukemia‐like infiltration of the anterior mediastinum in a 35‐year‐old male. Chest X‐rays revealed an abnormal mediastinum, which was diagnosed as thymoma. During the course of treatment, huge abnormal cells 40–50 μ in size were found in the peripheral blood smear and disseminated bone metastasis of the malignant thymoma was suspected. The tumor was resistant to both chemotherapy and radiotherapy. The patient died of respiratory failure. The autopsy disclosed a huge tumor measuring 24x13x10 cm in the anterior mediastinum. Histological findings of the tumor revealed cells which had spread to almost all organs, indicating leukemia‐like infiltration. This pattern of metastasis has been reported in the cancer of non‐hematologic origin under the term “carcinocythemia (CCA)”. To our knowledge, the present case is the first report of a leukemia‐like infiltration in case of malignant germ cell tumor. Careful serial section revealed no primary foci in either testis. ACTA PATH‐OL. JPN. 35: 1561–1570, 1985.

ADRENAL MEDULLARY HYPERPLASIA : HYPERPLASIA-PHEOCHROMOCYTOMA SEQUENCE

We present a case of unilateral adrenal medullary hyperplasia in a 63‐year‐old woman with‐clinical signs and symptoms of pheochromocytoma unassociated with multiple endocrine neoplasia. The surgically removed adrenal gland revealed diffuse medullary hyperplasia with multiple micronodules measuring up to 2 mm. The micronodules were composed of enlarged chromaffin cells with atypia, histologically similar to those of pheochromocytoma, forming small solid alveolar patterns separated by a fibrovascular stroma. Removal of the hyperplastic adrenal gland resulted in disappearance of paroxysmal nocturnal hypertension and palpitation. These results suggest that diffuse and nodular medullary hyperplasia is the precursor of pheochromocytoma.

FUNCTIONAL PATHOLOGY OF ALDOSTERONE‐PRODUCING ADENOMA

In addition to the morphological examination, the measurement of the content of corticosteroids was done in aldosterone‐producing adenoma. Histologically, the adenoma consists of four types of cells. The major component of the adenoma was clear‐type cells. In this type of cells, the activities of 3β‐hydroxysteroid dehydrogenase and glucose‐6‐phosphate dehydrogenase showed weakly positive. The fine structure was characterized by numerous lipid vacuoles and poor organellae. On the contrary, intermediate‐ and compact‐type cells revealed higher than moderate activities of these enzymes. Moreover, marked development of smooth endoplasmic reticulum and mitochondria was noted. Zona glomerulosa‐type cells were observed only in a small part. The content of aldosterone and corticosterone in aldosterone‐producing adenoma was significantly larger than those of the other type of adenoma. A tendency of positive correlation between the increase of compact‐type cells and content of aldosterone was found. The cell origin of this aldosterone‐producing adenoma and functional role of clear‐ and compact‐type cells were also discussed.

NON‐FUNCTIONING ADRENOCORTICAL ADENOMA IN CULTURE

This report describes the morphological responses of unstimulated and stimulated non‐functioning adrenocortical adenoma in culture. The removed adrenocortical adenoma was composed mainly of clear‐type cells and partially had a small area of cholesterol granuloma. These adenoma cells had many lipid droplets and round to long rod‐shaped mitochondria with tubular or tubulo‐lamellar cristae which were similar to those in Cushings adenoma. The non‐functioning adrenocortical adenoma cells which were incubated in vitro under ACTH (10 mlU/ml) and angiotensin II (10–6 M/ml) stimulation, were examined by phase contrast microscopy, transmission and scanning electron microscopy, and the content of Cortisol and aldosterone in the culture medium was measured by radioimmunoassay. As a result of exposure of ACTH, the cultured cells revealed the retraction response and production of Cortisol and aldosterone. After administration of ACTH for many days, the cultured cells showed characteristic changes in sER and mitochondria. The sER were markedly developed and packed tightly into a network of dilated tubules. Mitochondria were larger and more numerous than in the unstimulated cells. The mitochondria appeared to be entwined by the tubules of the sER. Lipid droplets decreased in number. ACTA PATHOL. JPN. 35 : 871–884, 1985.