We studied the relation between body size and bone mineral density in elderly females. The study included a total of 93 ambulatory females aged over 60 years. They were divided into 3 groups according to their body mass index (BMI; kg/m2): slender group with BMI<20 (n=28), normal group with BMI of 20 to 24.9 (n=43) and obese group with BMI≥25 (n=22). Fracture incidence, bone mineral density, calcium regulating hormones and steroid hormones were studied in an intergroup comparative manner. The incidence of vertebral fracture was found to be negatively correlated with BMI (the incidences of vertebral fracture in slender, normal and obese were 78.6, 48.8 and 22.7%, respectively) and bone mineral density was also BMI-related (0.390±0.016, 0.456±0.015 and 0.493±0.018g/cm², respectively: p<0.01 in ANOVA; mean±SE). The number of years after menopause was shorter in patients with a higher BMI. There was no intergroup difference in serum levels of PTH, vitamin D and estrogens. On the other hand, serum levels of calcitonin, DHEA, DHEAS, delta-4 androstenedione and testosterone were found to be higher in subjects with a higher BMI. From the present results, it seems that bone mineral density is supported not only by weight-bearing stress upon bone, but also by serum levels of calcitonin and androgens in obese females.

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A 63-year-old housewife with a history of partial thyroidectomy was referred to our hospital because of a neck mass and abdominal tumor. Aspiration biopsy of the neck tumor revealed the recurrence of papillary thyroid carcinoma. Magnetic resonance imaging (MRI) of the abdomen and urinary and plasma catecholamine levels indicated that the tumor beside the abdominal aorta was an extraadrenal pheochromocytoma. Two tumors were excised and histologic studies confirmed the diagnosis. So far two cases of extraadrenal pheochromocytoma with papillary thyroid carcinoma have been reported. The present case indicates that the presence of papillary thyroid carcinoma should be considered in patients with extraadrenal pheochromocytoma.

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The effects of human recombinant interleukin-1β and -6 and tumor necrosis factor-α(TNF-α) on the releases of PRL and dopamine were examined using monolayer cultures of rat pituitary cells and hypothalamic cells. The release of PRL from rat pituitary cells in 30 min was increased about 2-fold (p<0.05) by 10⁵ U/l interleukin-1β, 10⁵ U/l interleukin-6 or 100μg/l TNF-α. TNF-α at 100μg/l significantly increased PRL release within 5 min incubation and this effect continued throughout the next 30 min of incubation. Incubation for 5 min with TNF-α caused dose-dependent stimulation of PRL release. These cytokines did not modulate [³H]-dopamine release from primary cultures of hypothalamic cells. These results suggest that these cytokines stimulate PRL release directly at the pituitary gland, without modifying the release of dopamine from the hypothalamus.

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We recently reported that the red blood cell (RBC) carbonic anhydrase I (CAI) concentration in patients with hyperthyroidism is reduced and reflects the patient's mean thyroid hormone level over the preceding months. In this study, RBC CAI concentrations were measured in patients with thyroid nodules who were receiving suppressive doses of thyroxine (group 1) and compared with those obtained in patients with primary hypothyroidism receiving replacement doses of thyroxine (group 2). Of the 17 patients in group 1, 16 (94%) had elevated plasma free T₄ levels, but all 17 had normal free T₃ levels. Of the 17 patients in group 2, 16 (94%) had normal free T₄ levels and all 17 had normal free T₃ levels. Plasma TSH concentrations in group 1 were all below the lower limit of sensitivity of 0.04 mU/l. In group 2, 11 had normal and 6 had slightly elevated plasma TSH concentrations. The mean (±SD) RBC CAI concentration in group 1 (300 ± 53 nmol/g Hb) was significantly lower than that in group 2 (340± 57nmol/g Hb). The RBC CAI concentration was significantly correlated with both the concentration of plasma free T₄ and free T₃. These observations indicate that in patients receiving suppressive doses of thyroxine a slight increase in the plasma free T₄ concentration produces a slight but significant decrease in RBC CAI levels.

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The effect of leuprolide acetate (D-Leu⁶-[des-Gly¹⁰-NH₂]-LH-RH ethylamide acetate) for depot suspension (TAP-144-SR), a synthetic analog of luteinizing hormone-releasing hormone, was examined in three doses in 36 patients (34 girls, 2 boys) with central precocious puberty. TAP-144-SR was injected subcutaneously every four weeks for twelve weeks, and clinical symptoms and plasma and urinary levels of various hormones were followed every four weeks. Eleven girls given 10μg/kg showed a significant decrease in peak plasma LH and FSH responses to LH-RH test, but basal plasma LH and FSH did not change significantly. In 13 patients (11 girls and 2 boys) given 30μg/kg and 12 girls given 90 μg/kg, both basal and peak LH and FSH were significantly suppressed.Urinary excretion of LH decreased significantly in all groups except in the 10μg/kg group. Urinary excretion of FSH did not change significantly in the 10 and 30μg/kg groups, but it decreased significantly in the 90μg/kg group. In girls, plasma and urinary estradiol also fell greatly, but the difference was insignificant except in the 90μg/kg group. Regression of sexual characteristics was observed in almost half of the patients at the 12th week of the treatment. Side effects were minimal. A dose of more than 30μg/kg of TAP-144-SR is effective in suppressing gonadotropins and causing improvement of clinical symptoms, and appears to be useful in treating children with central precocious puberty.

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An attempt was made to develop immunologic probes directed against follistatin/activinbinding protein (ABP), for use in investigating the distribution of ABP in various tissues. Five oligopeptides corresponding to different regions of the predicted ABP amino acid sequence (peptides 1-12, 28-43, 123-134, 270-281 and 300-315) were synthesized chemically, and coupled to Limulus polyphemus hemolymph hemocyanin. The peptide-hemocyanin conjugates were then used to immunize rabbits, and the immunoresponses were monitored by enzyme-linked immunosorbent assay. Reactivity of the antipeptide antisera with ABP was determined by SDS-polyacrylamide gel electrophoresis and immunoblotting analysis. All of the peptides produced immune responses. The antiserum to peptide 123-134 recognized all forms of ABP, whereas the antiserum to peptide 300-315 reacted specifically and sensitively with the long forms of ABP. These two antisera exhibited only a limited cross-reaction with other proteins or none at all. Therefore, they will be useful for studying the distribution of ABP in various ussues.

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We have used immunohistochemistry to localize follistatin/activin-binding protein in adult male and female rats. A polyclonal antibody directed against a follistatin peptide (residues 123-134) was used as a specific immunologic probe. Intense and specific follistatin immunoreactivity was evident in spermatogenic cells of seminiferous tubules in the testis. The predominant staining was in nuclei of spermatocytes and spermatids, but no immune reaction was observed in spermatogonia or spermatozoa. Moderate immunoreactivity was detected in Leydig cells. Sertoli cells were follistatin-negative. Significant immunoreactivity was evident in ovarian granulosa cells. The intensity of the staining changed with follicle development: no immunoreactivity was observed in granulosa cells of primordial to primary follicles, but the cells of secondary to Graafian follicles displayed moderate to strong staining and finally luteal cells of the corpus luteum became negative. The epithelial lining of the oviduct and the smooth muscle of the myometrium of the uterus were intensely immunoreactive. Immunoreactive follistatin staining was present in the pituitary: a group of round-shaped cells were specifically stained. Immunostainable follistatin was visible in the epithelial layers of renal tubules with moderate to strong staining reactivity. Hepatic cells in the liver demonstrated homogeneous immunoreactivity from moderate to strong. The cortex of the adrenal gland, white pulp of the spleen and the brain cortex were also stained weakly but distinctly with the antiserum. In conclusion, immunoreactive follistatin is widespread in rat tissues, suggesting that follistatin/activin-binding protein is a ubiquitous protein, regulating a wide variety of activin actions.

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Gonadotropin secreting pituitary adenomas have been reported with increasing frequency in men, but they are still rarely recognized in women. We report a 52-year-old postmenopausal woman with LH- and FSH-secreting pituitary adenoma. She had increased LH (37.0±13.7 IU/l)(mean±SD) and FSH (109.9±26.7 IU/l) but these concentrations were within normal ranges in 80 postmenopausal women (LH: 29.7±18.3 IU/l, FSH: 104.0±43.9 IU/l). The administration of GnRH and conjugated estrogen resulted in normal response of LH and FSH. No abnormal response of gonadotropin to TRH and bromocriptine was observed. After transsphenoidal adenomectomy both LH and FSH decreased (LH: 11.1±4.2 IU/l, FSH: 37.0±9.6 IU/l). An immunocytochemical. study revealed that the adenoma cells synthesize both LH and FSH. The rarity of gonadotropin secreting pituitary adenomas in women could be the result of greater difficulty in recognition due to an increase in serum gonadotropin in postmenopausal women.

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The anti-obesity and anti-diabetic actions of BRL 26830A, β₃-adrenoceptor agonist, (2mg/kg administered intramuscularly daily for 2 weeks) were evaluated in obese diabetic Yellow KK mice and C57B 1 control mice. The following parameters were compared in the treated vs. control animals: brown adipose tissue (BAT) thermogenesis, resting metabolic rate (RMR), insulin receptors in adipocytes, and blood glucose and serum insulin levels during a glucose overloading test. BRL 26830A significantly increased BAT thermogenesis and RMR but it decreased the amount of white adipose tissue without affecting food intake. Those actions contributed to the mitigation of obesity in Yellow KK mice. BRL 26830A also increased the concentration of insulin receptors and decreased the levels of serum insulin and blood glucose during the glucose overloading test in Yellow KK mice. In the glucose overloading test performed one hour after BRL 26830A injection, insulin secretion was significantly increased and the blood glucose level was markedly decreased in both groups. These observations suggest that BRL 26830A possesses anti-obesity and anti-diabetic actions and consequently may be useful for treating obesity as well as non-insulin-dependent diabetes mellitus with obesity.

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To investigate a possible effect of triiodothyronine (T₃) on the regulation of estrogen receptor, estrogen dependent rat pituitary tumor, MtT/F84, was studied in rats which received surgical thyroidectomy (Tx) or were given propylthiouracil (PTU) and were supplemented with T₃. In T₃ loaded rats, the grafted tumor showed high estrogen receptor levels (160-200% of control), whereas low estrogen receptor levels (20-35% of control) were observed in the tumors grown in Tx and PTU treated rats. A single injection of T₃ to Tx rats with MtT/F84 increased the estrogen receptor level in a time dependent manner and reached the maximal level at 6 h. In primary culture of MtT/F84 cells, T₃ increased the specific ³H-estrogen binding to tumor cells in a dose dependent manner (165% of control by 10^-7M T₃) and also in a time dependnet (maximum at 12 h) manner. These data suggest that T₃ directly up-regulates the estrogen receptor level in MtT/F84 cells.

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In order to clarify the mechanism of rapid growth of anaplastic thyroid carcinoma, growth stimulating activity produced by the cancer cells in culture was studied. A cell line (HTh 7) established from a biopsy specimen of anaplastic thyroid carcinoma was used throughout the study. Growth stimulating activity was determined as an activity to increase ³H-thymidine incorporation in rat thyroid cell line (FRTL 5). Conditioned medium of HTh7 cells contained significant growth stimulating activity for FRTL 5 cells. The activity was separated into two fractions with heparin agarose gel: heparin-binding and heparin-non-binding. In the medium, the heparin-non-binding activity was much greater than the heparin-binding one. The heparin-non-binding activity was acid stable. It was partially purified with gel filtration in an acidic condition followed by reverse phase HPLC. In gel filtration with a Sephacryi S-200 column, the activity was eluted later than the elution volume of cytochrome c (MW 12400) as several separated peaks. In reverse phase HPLC, however, the activity in these peaks was eluted as a single peak. The retention time of the active peak was almost the same as that of recombinant IGF-I. When measured by specific RIAs, the conditioned media concentrated 20 times contained both 0.35ng/ml of IGF-I and 5.21 ng/ml of IGF-II. As for the heparin-binding mitogenic activity, when applied to heparin affinity HPLC column and eluted with a linear gradient of NaCl, the activity came out as one major peak with approximately 1.0M NaCl. Our present study has revealed that anaplastic thyroid carcinoma cells in culture produce growth stimulating activity and secrete it into the culture media. It is suggested that the activity consists of more than two peptide growth factors with distinctive charcteristics.

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Changes in insulin-stimulated glucose metabolism were studied in young and aged subjects, subjects with impaired glucose tolerance, and patients with NIDDM by means of the glucose clamp technique. The diabetic group includes obese and non-obese patients treated without insulin and non-obese patients treated with insulin. The glucose disposal rate (GDR) was decreased in aged subjects (5.8±0.4mg/kg/min) compared with young controls (7.4±0.3mg/kg/min). In patients with IGT, it was further decreased to 3.6±0.5mg/kg/min, which was comparable to the rate in NIDDM without insulin treatment (3.3±0.4mg/kg/min). There were no differences in the GDR between obese (3.0±0.3 mg/kg/min) and non-obese (3.4-0.6mg/kg/min) diabetic patients. In insulin-treated diabetic patients, GDR ranged widely, but the mean value was partially normalized (5.2±0.9mg/kg/min). In the diabetic group, no correlation was observed between fasting blood glucose and GDR. These results suggest that in the course of developing NIDDM, a decrease in insulin-stimulated glucose uptake precedes a rise in fasting blood glucose. Thus, as previously reported for Caucasian NIDDM patients, resistance to insulin-stimulated glucose uptake may be one of the basic defects in Japanese patients with NIDDM. The degree of glycemia, however, is not directly related to the magnitude of the defect in insulin action.

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The outcome of ¹³¹I therapy for 109 patients with Gravest disease was analysed according to pretreatment laboratory data including thyrotropin receptor antibody (TRAb) activities. Forty-five percent of patients became euthyroid, and 13% of patients became hypothyroid within one year after ¹³¹I therapy. Forty-two percent of patients remained hyperthyroid one year after ¹³¹1 therapy. Pretreatment values for serum T₄, T₃, and the estimated weight of the thyroid were significantly higher in the hyperthyroid group. The mean for the TRAb index of the hyperthyroid group was significantly higher than that of the euthyroid group. Life table analysis revealed a significant effect of the TRAb index on the rate of hyperthyroidism after 3 months or later. These results appear to suggest that the TRAb index is one of the factors which influence the outcome of 131I therapy for Graves' disease.

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We examined the effect of endothelin-3 (ET-3) at a high dose (pressor dose) and a low dose (non-pressor dose) in rat perfused kidney (PK), since ET-3 has recently been reported to exert a vasodilator action especially at a low dose. Kidneys were perfused with Krebs-Henseleit buffer at a fixed flow rate (6ml/min) in situ. After collection of the renal venous effluent and urine for 20min, vehicle (saline; n=6), 10^-13M ET-3 (low dose; n=6) or 10^-8M ET-3 (high dose; n=6) was added to the perfusate, and sample collection was performed for the same period with each. The high dose of ET-3 significantly increased the perfusion pressure, fractional sodium excretion and synthesis of prostaglandins (PGs) consistently with a significant reduction in the glomerular filtration rate (GFR). On the other hand, the low dose of ET-3 significantly increased the GFR, urine volume and free-water clearance with no change in the perfusion pressure or synthesis of PGs. These findings suggest that a low dose of ET-3 can increase the glomerular capillary ultrafiltration coefficient and that ET-3 exerts an influence on sodium and water handing in the rat PK.

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The usefulness of spot determination of urinary cortisol in the screening of Cushing's syndrome was evaluated by measuring the cortisol concentration in randomly sampled urine in 68 normal subjects and in 9 patients with Cushing's syndrome. The urinary cortisol concentration in the morning was significantly higher in patients with Cushing's syndrome but some overlap existed between normal subjects and patients with Cushing's syndrome. In contrast, there was a clear discrimination between two groups when urinary cortisol was measured in the late evening: urinary cortisol was lower than 75μg per gram creatinine (μg/gCr) in normal subjects but higher than 150μg/gCr in patients with Cushing's syndrome. When 1 mg dexamethasone was administered at 2300 h in the evening, spot urinary cortisol the next morning was less than 80μg/gCr in normal subjects while it was above 100μg/gCr in patients with Cushing's syndrome. Dexamethasone-induced suppresslon ot urlnary cortlsol in normal subjects lasted until late in the afternoon, which allows sampling of urine at any time in the morning and possibly in the afternoon. These results suggest the usefulness of spot determination of urinary cortisol in the screening of Cushings' syndrome.

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Syndrome of inappropriate secretion of antidiuretic hormone (SIADH), hypothalamic hypogonadism and alopecia universalis occurred in a 31-year-old female with insulin-dependent diabetes mellitus (IDDM). Despite various clinical investigations and careful observation for 20 months, the cause and pathogenesis of SIADH and hypothalamic hypogonadism were not elucidated. The complex of these disorders had not been described. The presence of IDDM and alopecia universalis, in which an autoimmune process has been assumed to be involved, is interesting in considering the pathogenesis of the SIADH and hypothalamic hypogonadism.

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