A 66-year-old man with tetraplegia developed hyperkalemia. Hypo-reninemic hypoaldosteronism was disclosed on the basis of a lack of response of plasma renin activity to furosemide administration or tilting with marked hypotension and a subnormal response of aldosterone to furosemide stimulation, tilting, angiotensin II infusion and ACTH administration, as well as increased vascular responsiveness to angiotensin II infusion. Of interest was the finding that urinary excretion of epinephrine and norepinephrine was markedly reduced, indicating that hyporeninemia may possibly be due to a chronic lack of sympathetic nervous stimuli. The patient was treated with sodium polystyrene sulfonate resin and/or 9-alpha-fluorohydrocortisone, and wheelchair rehabili-tation. However, even after stopping 8-month-mineralcorticoid replacement, normokalemia was maintained. Reexamination of the renin-angiotensin-aldosterone system revealed a normalized response to tilting or ACTH administration along with the normal catecholamine excretion. One more point to be noted is that ACTH admini-stration resulted in a rise in the plasma levels of cortisol, corticosterone and 18-OH-corticosterone, but not aldosterone. This may be attributed to ACTH-stimulated 18-OH-corticosterone derived from the zona fasciculata or alternatively to a partial defect of corticosterone methyl oxidase type II (18-dehydrogenase) in the adrenal glomerulosa cells. These results suggested that hyporeninemic hypoaldosteronism may have been attributable to a decrease in systemic nervous stimuli and that such abnormalities were reversible.

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We investigated the bioactivities of the recombinant DNA-derived methionyl 20K h GH (20K-Met-h GH) and methionyl h GH (22K-Met-h GH). The growthpromoting activities in Nb2 cells of 20K-Met-h GH and 22K-Met-h GH were 10.7% and 93.5% of pituitary h GH (P-h GH), respectively. In the IM-9 lymphocyte assay, the binding activities of 20K-Met-h GH and 22K-Met-h GH h GH receptor were 29.0% and 87.1% of P-h GH, respectively. Our data demonstrate that 20K-Met-h GH may have weaker biological potency than P-h GH.

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The effects of bile salts on the release of immunoreactive vasoactive intestinal polypeptide (IR-VIP) were investigated in men using a specific radioimmunoassay. Plasma IR-VIP was determined after extraction by the acidacetone method (recovery 75±5%). Oral administration of 400 mg sodium taurocholate caused a rise in plasma IR-VIP from 18.5±1.3pmol/l to 31.1±2.1 pmol/l after 30min and 39.0±1.7pmol/l after 60min and return to the initial value after 120min. Oral administration of chenodeoxycholic acid (CDCA) also increased plasma IR-VIP from a basal level of 14.5±1.5 pmol/l to 36.3±1.2 pmol/l after 60min. Oral administration of ursodeoxycholic acid (UDCA) increased plasma IR-VIP from 11.9±1.1 pmol/l to 25.6±1.8 pmol/l after 30min. Perifusion of 1m M taurocholate stimulated release of IR-VIP from human duodenal mucosa into the perifusate.
These results suggest that bile salts may participate, at least in part, in the release of IR-VIP from the gut.

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We measured the intra-platelet content of human epidermal growth factor (hEGF) and beta-thromboglobulin (β-TG) and the quantities of these released from platelets during in vitro aggregation. The intra-platelet amounts of hEGF and β-TG in 10⁸ platelets were 104.9±18.9 (Mean±SEM) pg and 2920.9±149.9 ng, respectively. During platelet aggregation elicited by 9, 11-epithio-11, 12-methano-thromboxane A2, a stable thromboxane A2 agonist, hEGF and β-TG were released in amounts about 50% and 40% of the respective content in platelets. Also during arachidonate-induced aggregation, hEGF and β-TG were released at about 60% and 50%, respectively. Various concentrations of thromboxane A2 antagonist, (9, 11), (11, 12)-di-deoxa-9, 11-dimethyl-methano-11, 12-methano-13, 14-dihydro-13-aza-14-oxo-l5-cyclopentyl-16, 17, 18, 19, 20-pentanor-15-epi-thromboxane A2, suppressed both aggregation and release reactions in a dose-dependent manner. There were good correlations between the platelet aggregation rate and released β-TG (r=0.9368, p<0.01) or hEGF (r=0.8931, p<0.01) and between released β-TG and hEGF (r=0.9385, p<0.01). These results suggest that hEGF is released from platelets in a similar fashion to β-TG in vitro.

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Pregnant female rats were given daily injections of a potent aromatase inhibitor, 4-hydroxy-Δ4-androstenedione (4OHA), throughout the latter half of the pregnancy (days 11 to 22; the day of insemination was designated as Day O) and male fetuses and pups were obtained. Control animals were male offspring of mothers treated with oil vehicle. When measured by the tritiumwater method, significant reductions in the aromatase activity were detected in the hypothalamic and preoptic continuum (HPOA) of the male fetuses and pups, over a period from day 16 of the pregnancy until a day after birth. All parturitions in the experimental as well as control animals occurred on day 22 of the pregnancy. Behavioral and anatomical consequences of the prenatal treatment were examined in adulthood. When mounted by stud male rats, the male litters of the 4OHA-treated mother showed lordosis at a significantly higher frequency both in terms of the number of positive test sessions (each consisted of a 20-min period in which the subject showed lordosis at least once) and the lordosis quotient (percent lordosis occurrence per 10 mounts). When placed with receptive female rats, the experimental animals were no less active in mounting or ejaculating than the control. No significant difference existed between the experimental and control animals in the weight of the testes or the accessorial genitalia, or the serum testosterone levels. Partial but significant intervention of behavioral defeminization of the brain was associated with decreased HPOA aromatase activity during the prenatal period.

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We reported a case of sporadic multiple endocrine neoplasia type 1, with multiple insulinoma, parathyroid adenoma, and pituitary tumor.
Measurement of hormone contents and immunohistochemical studies of the pancreatic tumors showed that the tumors contained insulin, glucagon, somatostatin, and pancreatic polypeptide. Furthermore, the concentrations of these hormones were different in each tumor.
Insulin extracted from the pancreatic tumors analyzed by reversed-phase high performance liquid chromatography revealed no structural abnormalities. On the other hand, in gel filtration evaluation of the extract of the parathyroid adenoma, it was found that the tumor extract contained a macromolecular parathyroid hormone (molecular weight 20, 000 to 25, 000).

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The acid saline extract (ASE) of rat submaxillary gland exerts a powerful degrading effect on ¹²⁵I-glucagon. In order to study the degradation of other ¹²⁵I-peptides by ASE and the effects of their inhibitors, ¹²⁵I-pancreatic polypeptide (PP) and ¹²⁵I-insulin were used together with ¹²⁵I-glucagon. The degradation studies were done by the trichloroacetic acid (TCA) method or gel filtration. Besides ¹²⁵I-glucagon, ¹²⁵I-PP was found to be destroyed by ASE in the ordinary immunoassay system using the TCA method, but ¹²⁵I-insulin was intact in the presence of ASE. Leupeptin, and to a lesser extent pchloromercuriphenyl-sulfonic acid (PCMS) and N-ethylmaleimide, inhibited the destruction of ¹²⁵I-glucagon or -PP under the TCA method. PCMS was especially protective at high concentrations, for example 16mM. These findings were confirmed by gel filtration of the assay mixture. In the presence of leupeptin (0.4mM) and PCMS (16mM), no shift in the peak of labelled glucagon or PP occurred. Thus ASE degrades not only ¹²⁵I-glucagon but-PP, and thiol proteinase inhibitors have a strong inhibitory action on them.

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To investigate the pathophysiology of patients with autoimmune thyroid diseases, we measured serum thyroid stimulating antibody (TSAb) activity and thyroid stimulation blocking antibody (TSBAb) activity by determining the radioiodine (¹²⁵I) uptake into FRTL-5 cells. FRTL-5 cells were pre-incubated for seven days with 5H medium and then incubated for 48 hours with patients' crude IgG prepared by polyethylene glycol precipitation. In order to measure TSBAb, 10μU/ml TSH was also added. ¹²⁵I was added one hour before the end of the 48 hour incubation period. After the incubation, the medium was aspirated, and the radioactivity in the cells was counted. In patients with untreated hyperthyroid Graves' disease, TSAb was detectable in 18 of 20 patients, the detectability being 90%, and activity showed a statistically significant positive correlation with TSAb activity determined by c-AMP accumulation. Out of 41 patients with hypothyroidism, TSBAb determined by ¹²⁵I uptake was positive in six cases, the detectability being 14.6%. The inhibition of ¹²⁵I uptake by one of these six IgGs was suggested to be at the TSH receptor level because it inhibited TSH induced c-AMP accumulation and showed positive thyrotropin binding inhibitor immunoglobulin (TBI I) activity, but did not inhibit the forskolin- and (Bu) ₂cAMP-induced ¹²⁵I uptake. Inhibition of another IgG was suggested at the post-receptor level because it did not inhibit TSH induced cAMP accumulation and showed negative TBI I activity, but inhibited forskolin- and (Bu) ₂cAMP-induced 19 uptake. Inhibition of four other IgGs was at both receptor and post-receptor levels because they inhibited TSH induced cAMP accumulation, showed positive TBI I activity, and inhibited forskolin-induced ¹²⁵I uptake. In conclusion, the present assays for TSAb and TSBAb are sensitive, practical and make it possible to evaluate the process after c-AMP accumulation and the heterogeneity of TSBAb is suggested.

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The present studies were designed to clarify the contribution of the liver to the development of hyperglycemia in Wistar fatty rats. The hepatic activities of insulin-inducible enzymes involved in glycolysis (glucokinase GK and pyruvate kinase) and lipogenesis (glucose-6-phosphate dehydrogenase), were higher in fatty rats than in lean rats at 4 and 8 weeks of age because of the higher insulin levels in the former. Thereafter, the GK activities of fatty rats decreased slightly in spite of severe hyperinsulinemia, and did not differ from those of lean rats. In addition, fatty rats had higher levels of insulinsuppressible gluconeogenic enzymes, glucose-6-phsphatase (G6Pase) and fructose-1, 6-diphosphatase. These findings indicate that the hepatic enzymes of fatty rats are resistant to insulin. This postulation was supported by the fact that the hepatic enzyme activities of fatty rats showed a lower response to changes in plasma insulin levels produced by fasting and refeeding, The G6Pase/GK ratio, which indicates net glucose handling in the liver, increased in fatty rats and decreased in lean rats with advancing age, suggesting that hepatic glucose production in fatty rats becomes dominant with advancing age. The changes in hepatic glycolytic intermediates supported this suggestion: the glycolytic steps both from glucose to glucose-6-phosphate and from phosphoenolpyruvate to pyruvate in fatty rats were accelerated at 5 weeks of age, but suppressed at 12 weeks of age. These results indicate that insulin resistance in the hepatic enzyme regulation may contribute to the development of hyperglycemia in Wistar fatty rats.

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Human pancreatic cancer cells (FA-6) producing bone resorbing factor were established in culture. A biopsied lymphnode from a patient with pancreatic cancer associated with humoral hypercalcemia of malignancy (HHM) was transplanted to nude mice, and the cells producing high parathyroid hormone (PTH)-like activity were selected by a limited dilution from outgrowth of the xenografts of the tumor grown in nude mice. The conditioned media contained an activity to stimulate the resorption of mouse calvaria in vitro which was indomethacin-insensitive. The conditioned media had both alphatype and beta-type transforming growth factor (TGF) activity but no interleukin-1 activity. TGF-alpha activity was co-eluted with PTH-like activity from gel-chromatography at around 15 kDa. The FA-6 cells now established are the first cells of pancreatic cancer associated with HHM producing both PTH-like and TGF-alpha activities along with bone resorbing activity.

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To study neuronal mechanism through which the hypothalamus exerts its influence on growth hormone (GH) synthesis, anterolateral hypothalamic knife cuts (ALHD) were used. After ALHD, GH messenger RNA (mRNA) content in the pituitary was reduced to 47% of the control value. Neither the prolactin mRNA level nor the total DNA content showed any significant change. In such animals, the serum GH level was significantly higher (184%) and pituitary GH content was lower (43%) than the control values. These results suggest that neuronal factor (s) outside the mediobasal hypothalamus plays an important role in the regulation of GH synthesis.

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Analysis of urinary metabolites of [1, 2-³H]-3α, 5β-tetrahydroaldosterone was performed in male guinea-pigs. Separation of urinary metabolites was carried out by means of DEAE-Sephadex A-25 column chromatography and four fractions (fraction A to fraction D) Were detected. 46-50% of fraction A was extractable with ethyl acetate. When ethyl acetate extract was submitted to reversed phase high pressure liquid chromatography, a number of peaks were present and one of these peaks cochromatographed with 3α, 5β-tetrahydroaldosterone. Fraction B, fraction C and fraction D were incubated with enzyme and the free steroid released was identified on the basis of retention time on reversed phase high pressure liquid chromatography. Thus, fraction B contained monoglucosiduronate of 3α, 5β-tetrahydroaldosterone, whereas identification of fraction C was not successful. Fraction D was characterized as a monosulphate 3α, 5β-tetrahydroaldosterone, 3α, 5β-tetrahydroaldosterone and 3β, 5α-tetrahydroaldosterone were identified as aglycones.
It is probable that 3β, 5α-tetrahydroaldosterone is converted from other tetrahydroisomers by intestinal bacteria of the guinea-pig, since mammalian tissues do not contain enzymes that introduce a double bond into ring A.

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Cushing's syndrome (CS) may be sustained by a nodular adrenocortical pathology (NAP) in addition to hyperplastic or neoplastic lesions of adrenal glands. NAP, in turn, may be represented by macronodular (MACRO) or micronodular (MICRO) manifestations. There is debate as to whether the MACRO-NAP and MICRO-NAP represent an expression of the same disorder or relate to distinguishable anatomo-clinical entities. A case of CS sustained by a “giant” MACRO-NAP forced us to review the literature and to analyze the morpho-clinical findings from a statistical viewpoint. Testing procedures were able to significantly dissect some clinical, pathological and hormonal characteristics. The statistical probation clearly indicated that MACRO-NAP and MICRO-NAP causing CS are nosologic entities that can be clinically differentiated via their phenotypic symptomatology.

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A 54-yr-old woman with the symptoms of primary reninism, i. e. hypertension, metabolic alkalosis and elevated levels of plasma renin activity (PRA) and aldosterone, is described. She had an ileal cancer secreting active and inactive renin. The symptoms markedly improved after resection of the tumor. the tumor active and inactive renin were proved to be present by an assay angiotnesin I formation in the presence and absence of renin antibody, and renin immunoreactivity was found immunohistochemically. The mRNA coding for the renin precursor was identified in the RNA-rich extract of the tumor blot hybridization analysis with the human renin cDNA as a probe. The mRNA from the tumor was shown to be identical in molecular size to that from the human kidney by agarose gel electrophoresis. This is the first description of an ectopic renin-producing ileal carcinoma and the first demonstration of renin mRNA in the tumor tissue.

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A 47-year-old man with Graves' disease suffered from a feeling of hunger and sweating in the night, polyarthralgia and fever one month after the start of treatment with methimazole. The above symptoms were ascribed to the side effects of methimazole; insulin autoimmune syndrome and lupus-like syndrome. The change in the antithyroid drug to propylthiouracil caused an amelioration of the symptoms. In addition to an anti-insulin antibody with a high binding capacity, hyperglucagonemia (260pg/ml with a plasma glucose level of 61mg/dl) was observed, which returned to normal in parallel with the decrease in the insulin binding capacity of the plasma one month after beginning the treatment with propylthyouracil. A normal decrease in the plasma glucagon level due to exogenous insulin (2mU/kg/min) was observed with the euglycemic clamp. However, the plasma glucagon level was not suppressed by the oral glucose loading and elicited a poor response to the arginine infusion. Taking previous reports into account, this basal hyperglucagonemia seems to be a characteristic finding in the insulin autoimmune syndrome, while a sluggish response of glucagon to oral glucose or arginine infusion might be ascribed to hyperthyroidism.
This is the first case report concerning a kinetical study of the glucagon secretion in insulin autoimmune syndrome with Graves' disease.

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Functioning parathyroid lipoadenoma (hamartoma) composed of abundant adipose or myxomatous stroma and epithelial cell nests is an unusual cause of primary hyperparathyroidism. We report herein four new cases. None of them belongs in the category of multiple endocrine neoplasia or familial hyperparathyroidism. The clinical manifestations and the laboratory findings are indistinguishable from those of the usual forms of primary hyperparathyroidism. Ultrasonography of the neck demonstrated an enlarged parathyroid gland as a hyperechoic mass in the two patients tested. At operation in each case, a single enlarged gland was found and resected, the weight being 3, 0.3, 0.45 and 1g, respectively. The patients are normocalcemic 1 to 10 years after surgery. Pathological examination disclosed that the lesions were consistent with lipoadenoma or its variants. On reviewing 20 cases of functioning lipoadenoma which were reported in the literature, including the present cases, we found that the size of the tumors varied and a functioning lipoadenoma is hence by no means unusually large as previously reported. Without knowledge of this specific clinicopathologic entity, the lesion may be overlooked at the preoperative localization study and misdiagnosed as a normal or hyperplastic parathyroid.

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Eight patients with active acromegaly due to GH-producing pituitary adenoma were studied. GH secretory dynamics in vitro was evaluated by adding GRF, CRF, or a somatostatin analog, SMS 201-995 to the perifusate of dispersed cells from tumors. A comparison was made between the data obtained in preoperative tests for GH secretion and those obtained in experiments in vitro. Before operation, the GRF test (100μg, iv) resulted in no GH response in three of six patients examined. The CRF test (100μg, iv) resulted in a paradoxical GH increase in two of the same six patients. in vitro studies performed on adenoma cells revealed that exposure to GRF (100ng/ml) elicited an increase in GH in seven of eight patients examined. Exposure to CRF (100ng/ml) caused an enhanced GH secretion in four of the same eight patients. There were cases in which GH response to these hypothalamic hormones was observed in vitro but not in vivo, whereas there was only one case in which CRF caused an increase in GH in vivo but not in vitro. Thus, GH secretory dynamics was not always the same in vivo and in vitro. The discrepancy could be ascribed to the different secretory status of hypothalamic hormone (e. g., GRF or somatostatin) in vivo in each acromegalic patient.

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The specificity of a “two-site” immunoradiometric assay (IRMA) has been reevaluated by examining its ability to detect heterogeneous adrenocorticotrophin-like immunoreactivity (ACTH-LI) separated by gel column chromatography. Plasma samples from patients with Addison's disease, Nelson's syndrome and ectopic ACTH syndrome and tissue extract of human anterior pituitary were subjected to ACTH-IRMA and the levels of ACTH-LI were compared with those measured by conventional ACTH-radioimmunoassay (RIA). The level of ACTH-LI measured by IRMA was considerably lower than that measured by RIA in the plasma of a case of ectopic ACTH syndrome and the ACTH-LI did not show a dilution curve parallel with that of the standard. Gel exclusion chromatography revealed that the plasma contained a relatively large quantity of “big ACTH” which was found to be poorly detected by the IRMA. In the plasma of Addison's disease or the extract of pituitary gland in which “big ACTH” constituted a small portion, whole ACTH-LI was apparently diluted in parallel with the ACTH standard, although the “big ACTH” also did not show full parallelism with the ACTH standard in the IRMA. These data suggest that “big ACTH” derived not only from an ectopic ACTHproducing tumour but also from a normal human pituitary gland cannot be detected as well as authentic ACTH by the ACTH-IRMA system. Therefore, samples which contain a relatively large proportion of “big ACTH” in the total ACTH-LI should be carefully evaluated by ACTH-IRMA.

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The central mechanism controlling food intake in response to the change in environmental temperature has been little examined. The GABA agonist, muscimol, was injected into the lateral ventricle of rats which were acclimated to temperate (26°C) and hot (33°C) environments. Muscimol obviously stimulated the feeding behavior of rats in both environments. However, when muscimol was administered at doses of 100 and 250ng, the food intake at 26°C was greater than that at 33°C. In addition, the stimulating effect of muscimol (250ng) on food intake at 26°C lasted longer than that at 33°C. These findings suggested that there might be a difference in muscimol metabolism at the two temperatures.

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A 22-yr-old female suffering from hypertension, hypokalemic alkalosis and suppressed plasma renin activity was studied. The plasma aldosterone concentration (PAC) ranged between subnormal and normal levels while the other adrenal mineralocorticoids were normal. Examinations through computed. tomography and ultrasonography showed no abnormal findings. For diferential diagnosis, dexamethasone, spironolactone and triamterene were administered. Triamterene alone corrected the abnormalities in this case, and the therapeutic effect was further enhanced by sodium restriction. Therefore, the present case is strongly suggested to be one of Liddle's syndrome, which is characterized by a primary defect in renal tubular sodium handling and can be corrected with triamterene. However, the patient in our study is different from the first reported case in which aldosterone secretion was estimated to be low. Analysis of the changes in PAC has shown that PAC is parallel with the level of plasma progesterone in accordance with the rhythm of the menstrual cycle and, in the follicular phase, PAC is rather low. It is concluded that the patient was suffering from Liddle's syndrome, and it is assumed that PAC is not always subnormal and, as same as in normal females, PAC may change in accordance with the rhythm of the menstrual cycle in a female case of Liddle's syndrome.

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