The change in the levels of free thyroid hormones and the pathophysiology of the hypothalamo-pituitary-thyroid axis of patients with nonthyroidal illness (NTI) have not been clearly elucidated so far. Therefore, it was thought of interest to investigate this problem by determining free thyroid hormones and TSH in serum and the response of TSH to TRH in these patients. The subjects employed in this study were 71 cases with hemodialysis, 40 cases with diabetes mellitus, 24 cases with liver cirrhosis, 12 cases with various cancers, 10 cases with anorexia nervosa and 110 normal subjects as controls. The serum total protein, albumin, free T₄, free T₃ TSH and other parameters of throid function were determined, and the TRH test was performed on about 10 patients of each group. Serum TSH was not only determined by a conventional assay system, but with a highly sensitive method, and the data were compared with one another. It was found that the serum free T₃ levels were significantly low in all the groups investigated, but the serum free T₄ levels were significantly low only in the groups with hemodialysis, decompensated liver cirrhosis, cancers and anorexia nervosa. No significant lowering of serum free T₄ was observed in the patients with diabetes mellitus, acute hepatitis and compensated liver cirrhosis. However, serum TSH levels tended to be higher in all the groups studied, though they were not significant. The response of TSH to TRH was low or delayed in about 20-50% of patients with hemodialysis, diabetes mellitus, liver cirrhosis, cancers and anorexia nervosa. It was observed that the serum rT₃ concentration was significantly high in the patients with diabetes mellitus and anorexia nervosa but significantly low in the patients on hemodialysis. In the rest of the groups, there were found many cases who showed high levels of serum rT₃ although they were not statistically significant. These results indicate that low concentrations of serum free T₃ observed in the majority of the patients with severe NTI were, at least in part, due to the decrease in the peripheral conversion of T₄ to T₃ and the lowered sensitivity of the anterior pituitary to thyroid hormones and TRH.

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Monoclonal antibody 3A4 to islet cell surface antigen has been previously established in our laboratory, using hybridization of spleen lymphocytes from non-obese diabetic (NOD) mice transferred into immunologically incompetent recipient mice.
In the present study, monoclonal islet cell surface antibody 5C12 could be newly obtained in the 10 : 1 ratio of NOD mice spleen cells and mouse myeloma cells (SP2/0) without any modifications.
Protein A radioligand assay and indirect immnofluorescence on living cells showed that 5C12 antibody reacted to normal rat islet cells and cultured rat insulinoma cells (RIN-r), but not to cultured lymphocytes (Bri-7, IM-9) and Chang-liver cells. Analysis of ¹²⁵I-labeled antibody binding revealed that unlabeled 5C12 effectively inhibited subsequent ¹²⁵I-5C12 binding to RIN-r cells, whereas unlabeled 3A4 did not. The scatchard plot from these data showed the curvilinearity, and about 150,000 binding sites to antibody per RIN-r cell were counted.
The treatment of RIN-r cells with papain and neuraminidase reduced the binding of 5C12 to RIN-r cells, whereas the effect of trypsin was not observed.
Immunoprecipitation of ¹²⁵I-labeled insulinoma cell lysates followed by SDS-PAGE and autoradiography indicated that 5C12 recognized 105K dalton cell surface protein in RIN-r cells. Immunoblotting also showed that 5C12 antibody recognized 105Kdalton cell surface protein in RIN-r cells.
These results demonstrated that 5C12 was an important tool for clarifying the immunoresponse against certain antigenic determinants on pancreatic B cells.
Furthermore, 5C12 has not only qualitatively and quantitatively improved diagnostic methodology, but it may also provide new reagents useful to the treatment and prevention of type 1 diabetes.

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The concentrations of plasma transferrin (Tf), which has been described as possessing growth promoting activity in vitro, were determined in patients with various endocrine diseases before and after treatment. Plasma Tf levels in 74 healthy subjects were 269 ± 3 (mean ± SE) mg/dl. In 11 patients with active acromegaly, they were elevated to 353 ± 11 mg/dl (p<0.001), while they were reduced to 168 ± 14 mg/dl in 8 patients with hypopituitarism (p<0.001). They were normalized after treatment. These data indicate that plasma Tf varies according to the endocrine conditions with good correlation of the status of plasma somatomedin C, and therefore its measurement may be useful clinically for the evaluation of the status of growth factors. However, the values should be assessed carefully in cases with proper Tf abnormalities, such as hematological, hepatic, or renal disorders.

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A patient with intrapericardial pheochromocytoma is presented and the literature on this subject is reviewed.
The patient was a 45-year-old housewife who was first referred to Keio University hospital in February 1981 for a 6-year-history of headaches, palpitations and excessive sweating. An episode of hypertensive crisis was observed during surgery for myoma uteri 3 months before her admission. On physical examination, the patient was a normal-appearing, pleasant woman. Supine blood pressure was 130/80mmHg and standing blood pressure was 124/72 mmHg. Results of routine laboratory studies (including fasting serum glucose and calcium determinations and thyroid function tests) were normal. A 24-hour-urine collection showed VMA level of 10.5 mg and noradrenaline of 809μg. Also repeated measurements of plasma noradrenaline on supine position revealed averages of 2.45 ng/ml. These high levels of catecholamines, clinical manifestations, and provocative tests including a metoclopramide test were strongly suggestive of pheochromacytoma, and further studies for determination of the localization were performed. Results of computed tomography (CT), adrenal scintigraphy using ¹³¹I-Adosterol and selective venous sampling for catecholamines were equivocal. Then she was followed in an outpatient clinic and treated with α-and β-bl oc ker In October 1985, she was readmitted to our hospital because of paroxysms which had gradually increased in frequency and severity. Extensive venous sampling in order to determine the localization of the tumor was performed after demonstration of an increased uptake area in the anterior thorax in ¹³¹I-metaiodobenzylguanidine scintigraphy. Further, an anterior mediastinal mass was identified in the CT scan of the chest with contrast material. Finally, a thoracic angiogram revealed that the tumor was highly vascular and was fed by the branch of the left internal mammary artery. In January 1986, the patient underwent middle sternotomy with cardiopulmonary bypass. A 4.5 by 3.5 by 3.0cm soft, dark brown tumor was found on the anterior surface of the heart and excised without difficulty. The tumor seemed to arise from the root of the aorta and main pulmonary artery, and extended inferiorly over the anterior surface of the right ventricle. It was also fed by branches of the right coronary artery. Histologically, the resected tumor was compatible with pheochromocytoma. The tumor content of noradrenaline was 3.34mg/g wet tissue; however, adrenaline and dopamine were not detected. The postoperative course was uneventful.
Intrapericardial pheochromacytoma is extremely rare and to our knowledge only 13 cases have been reported in the literature. Of 14 patients (including the present case) involved, 4 were males and 10 females; the age range was 18-76 years. Nine of the lesions were attached to the posterior aspect of the left or right atrium. Two of the lesions were located in the interatrial septum, and the remainder were found on the left lateral surface of the heart. It is assumed that a pheochromocytoma at this site may arise from chromaffin cells which are distributed adjacent to sympathetic fibers innervating the heart. Compared to the previously reported cases, the location of the described case herein is unusual, because it was found on the anterior surface of the heart.
Our case will support the concept that all paraganglion-bearing tissue is capable of giving rise to functioning paragangliomas.

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It has been widely reported that dysfunctions of pancreatic A-cell occur in diabetics. Since these pancreatic A-cell dysfunctions are not normalized by conventional insulin injection treatment, they were thought to be a primary defect of diabetes mellitus. Recently it was found that paradoxic glucagon secretion to oral glucose and excessive glucagon response to i.v. arginine could be perfectly normalized if strict blood glucose regulations were achieved with appropriate insulin treatment. However, there has been no report on the perfect normalization of glucagon secretion in response to insulin-induced hypoglycemia in diabetics. In this report, to elucidate the precise significance of A-cell function in hypoglycemia in diabetics, the effect of long-term strict glycemic regulations and the importance of intact autonomic nerve function on hypoglycemia-induced glucagon secretion were studied.
In experiments on hypoglycemia-induced glucagon secretion in diabetics, 0.2 to 0.3U/kg of regular insulin injection were usually employed to overcome the hyperglycemia and insulin resistance. However, hyperinsulinemia has been demonstrated to suppress A-cell function in experiments using the euglycemic clamp technique. Therefore, the effect of plasma insulin concentrations after insulin injections was first studied in 7 healthy volunteers by injecting insulin at doses of 0.1U/kg and 0.3U/kg. In this experiment with 0.3U/kg of insulin, the rate of fall in glycemia and the nadir of blood glucose were made similar to that with 0.1U/kg of insulin by using glucose clamp technique with artificial endocrine pancreas. The plasma glucagon response after 0.3U/kg of insulin was significantly suppressed as compared to that after 0.1U/kg of insulin. From these experiments, it was concluded that not only hypoglycemic stimuli but also plasma insulin concentrations are important factors for demonstrating significant glucagon secretion in response to insulin-induced hypoglycemia.
Second, the effects of strict glycemic control and autonomic nerve function on hypoglycemia-induced glucagon secretion were studied. Regular insulin at a dose of 0.1U/kg was injected in an i.v. bolus form into 21 insulin-dependent (IDDM) and 22 noninsulin-dependent (NIDDM) diabetics before and one to three months after strict glycemic control with multiple insulin injection therapy or continuous subcutaneous insulin infusion therapy. To reduce fasting blood glucose level and to obtain the same hypoglycemic stimuli, overnight insulin infusion at a basal dose was undertaken in IDDM who showed hyperglycemia before strict glycemic regulations. In patients with Shy-Drager syndrome who had no glucose intolerance but had severe autonomic nerve dysfunction, insulin-induced hypoglycemia was also performed. The evaluation of autonomic nerve function was carried out by measuring coefficients of variation (C.V.) of 100 R-R intervals on ECG.
In IDDM with or without autonomic neuropathy and NIDDM with autonomic neuropathy before strict glycemic regulations, no significant rise in plasma glucagon concentrations could be observed during the insulin-induced hypoglycemia. In NIDDM without autonomic neuropathy, a rise in plasma glucagon concentrations was observed, though the response was delayed. After strict glycemic regulations, in 16 IDDM and 17 NIDDM without autonomic neuropathy, the glucagon response to hypoglycemia improved remarkably in IDDM and normalized in NIDDM. On the other hand, in 5 IDDM and 5 NIDDM with autonomic neuropathy whose C.Vs were less than 2.5%, the rise in glucagon secretion could not be observed at all, even after strict glycemic regulations. During one to three months strict glycemic regulations, autonomic dysfunction expressing as C.V. did not improve significantly in either IDDM and NIDDM.
In 5 patients with Shy-Drager syndrome whose mean C.V. was 0.7%, hypoglycemiainduced glucagon response was not demonstrated.
In summary,

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There have been several reports about the glucocorticoid receptor (GC-R) in human peripheral lymphocytes (HPL). However, little has been investigated about the validity of the assay method of GC-R in HPL. such as the effect of receptor occupancy by intrinsic cortisol or administered GC. In the present study, it was clarified that receptor-bound prednisolone or cortisol was almost completely dissociated from the receptor by washing lymphocytes three times with 30 min equilibration at 37°C between each centrifugation. The GC receptor was measured by whole cell binding assay using [³H] dexamethasone as a ligand. The receptor number and dissociation constant (Kd) of lymphocytes, which were preincubated with prednisolone (10^-7M) or cortisol (10^-6M) at 37°C for 1 h, were equal to those of lymphocytes preincubated without GC. These data indicated that the receptor number and Kd of HPL can be determined without the effects of receptor occupancy by this assay procedure.
The GC and GC-R complex in the nucleus are quite important for the action of GC. However, there have been few reports about the assay of nuclear GC-R in HPL. In this study, it was elucidated that nuclear receptor-bound [³H] dexamethasone could be measured by centrifugation (2000g, 10 min) of the nuclei of lymphocytes after being lysed in 1.5mM MgCl₂ (0°C).
By these assay procedures, the receptor number in whole lymphocytes of normal HPL (n=22) was determined to be 10.48 ± 2.40 (M ± SD) fmoles/10⁶ cells, and Kd was 5.8 ± 1.24nM, while the receptor number and Kd in the nuclei were 5.68 ± 1.53 and 6.29 ± 1.49, respectively.

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The effect of glucocorticoid (GC) on the GC receptor in human peripheral lymphocytes was studies in vivo and in vitro. Excluding the effect of receptor occupancy, the GC receptor was measured by whole cell binding assay using [³H] dexamethasone as a ligand. The numbers of GC receptor in lymphocytes (whole cell, 8.42 ± 1.88; nucleus, 3.56 ± 1.38 fmoles/ 10⁶ cells (M ± SD) of patients treated with prednisolone (n=26, mean : 33.7mg/day) were significantly reduced when compared with those of normal subjects (n=21; 10.48 ± 2.40, 5.68 ± 1.53) and those of patients without GC therapy (n=9 : 10.56 ± 2.10, 5.26 ± 1.07), respectively. The dissociation constants among these subjects were almost similar (5 -7nM). There were negative correlations between the receptor numbers (both whole cell and nucleus) of patients and daily dosage of prednisolone. The GC receptor numbers in both the whole cell and the nucleus were significantly decreased after 48h in vitro preincubation with active GC. When new protein synthesis was blocked by cycloheximide (1μg/ml), GC did not reduce its own receptor number in vitro. Thus, the new protein synthesis may be required for this reduction in receptor number by GC. These results strongly suggest that GC down-regulates its own receptor in human peripheral lymphocytes.

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Although the clinical response to glucocorticoid (GC) therapy varied considerably from patient to patient, it is not known how to predict the responsiveness of each patient. In the present study, it was evaluated whether the responsiveness to GC in an individual patient was predictable by the GC receptor number. GC receptor number in human peripheral lymphocytes was measured by whole cell binding assay using tritiated dexamethasone as a ligand. There was a significant negative correlation between receptor number and dose of GC being given to the patient (down-regulation). The relative receptor number was expressed as a percentage of the receptor number to the mean of the receptor number in the group of patients stratified by every 10mg/day of the dosage of prednisolone. This relative receptor number was constant in an individual subject regardless of GC dosage.
In 12 patients with lupus nephritis who had a high receptor number (more than 100%), 9 patients (83%) showed improvement in their urinary findings after GC treatment. On the other hand, only 3 of 11 lupus nephritis patients with a low receptor number (less than 100%) showed improvement after GC therapy. In patients with various collagen diseases treated with 40-60mg/day of prednisolone, there were significant correlations between receptor number and each of % creatinuria, total cholesterol in serum, and fasting urinary calcium excretion, which were known to be affected by GC administration. Thus, the relative receptor number in lymphocytes was well correlated with both therapeutic and metabolic effects of GC in vivo. These data suggested that the GC receptor number in human peripheral lymphocytes is useful as an indicator for the prediction of responsiveness to GC therapy.

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