Hideo Tsuchiyama

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.

Coexistent carcinoma in congenital dilatation of the bile duct and anomalous arrangement of the pancreatico-bile duct. Carcinogenesis of coexistent gall bladder carcinoma.

From 1972 to 1985, 40 cases of congenital dilatation of the bile duct (CDBD) were experienced in the department of the authors. Those consisted of 19 cases of anomalous arrangement of the pancreaticobile duct (P‐B anomaly), five of bile duct carcinoma, and two of gall bladder carcinoma. In addition, four patients who showed no dilatation of the bile duct in spite of the presence of a P‐B anomaly were experienced, and coexistent gall bladder carcinoma was present in three of four cases. The incidence of bile duct carcinoma associated with CDBD is very high. However, coexistent gall bladder carcinoma in CDBD is a new topic, and coexistent intrahepatic bile duct carcinoma in CDBD is extremely rare. This report presents interesting and rare cases of coexistent carcinomas in these anomalies and investigates their carcinogenesis, particularly that of gall bladder 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

Receptor autoradiographic evidence of specific brain natriuretic peptide binding sites in the porcine subfornical organ

Specific binding sites of brain natriuretic peptide (BNP), a newly discovered peptide in the subfornical organ (SFO) of porcine brain were investigated, following incubation of related tissue sections with 125I-BNP, then using autoradiography and an image analysis coupled with computer-assisted microdensitometry. Specific 125I-BNP binding sites were found to be localized in the SFO, an area densely labeled by 125I-alpha-rat atrial natriuretic peptide and 125I-(Sar1,Ile8)-angiotensin II. Specific 125I-BNP binding to the SFO was displaced by unlabeled BNP, with a high affinity, and was calculated to be Ka = 0.385 x 10(-9) M and Bmax = 40.1 fmol/mg using a LIGAND computer program. Acquisition of these present findings enhances our knowledge of the physiology of BNP, atrial natriuretic peptides and angiotensin II system in the SFO.

MORPHOLOGICAL STUDIES OF HUMAN ADRENAL CORTEX UNDER PATHOLOGIC CONDITIONS

The human adrenal cortex of 203 cases, surgically resected and obtained at autopsy, were investigated morphologically. As controls, 30 biopsy specimens were procured from the patients at operations for renal stones, tuberculosis, tumors and wandering kidneys.

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.

Specific [125I]brain natriuretic peptide-26 binding sites in rat and pig kidneys

Specific binding sites for porcine brain natriuretic peptide-26 (BNP-26), a member of the atrial natriuretic peptide family (ANPs), were investigated in the kidney by using receptor autoradiographic and membrane binding techniques with [125I]BNP-26. The binding sites were discretely localized in rat and porcine kidney areas corresponding anatomically to the glomeruli and inner medulla. There were no differences between the localization of [125I]BNP-26 and [125I]alpha-rat ANP binding sites in the kidney. [125I]BNP-26 binding to solubilized membranes from isolated glomeruli of the rat kidney was saturable, and a single class of high-affinity sites was labeled with a KD of 372 pM. The radioligand bound to two sites in solubilized inner medullary membranes of the rat, a low-affinity site with a KD of 30 nM, and a high-affinity site with a KD of 33 pM. The rank order of potency to inhibit binding was BNP-26 = alpha-rat ANP-(1-28) greater than atriopeptin III (ANP-(103-126)) much greater than atriopeptin I (ANP-(103-123)) greater than des-Cys105,Cys121- ANP-(104-126). Thus, [125I]BNP-26 presumably recognizes ANP receptors in the kidney. The possibility that BNP-26 regulates, as a circulating hormone, kidney functions by binding to ANP receptors would have to be considered.

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.