Inhibin and activin were initially isolated as regulators of pituitary or gonadal hormone and are now known to be growth factors belonging to the TGF-β family with diverse influences on the differentiation and proliferation of various tissues. To investigate the role of inhibin and activin in human brain tumors, the expression of inhibin α, and β_A mRNA as well as activin type II receptor (ACTR II) mRNA were studied in various human brain tumors. The tumors were divided into the following 4 groups: 3 Rathke's cleft cysts and 2 craniopharyngiomas (group 1), 8 meningiomas (group 2), 8 malignant gliomas (group 3), and various other tumors including 1 each of germinoma, astrocytoma, hemangioblastoma, and osteochondroma as well as 2 malignant lymphomas and 2 metastatic squamous cell carcinomas (group 4). Immediately after resection, tumor tissues were homogenized in guanidine thyiocyanate to extract total RNA. PCR was then performed with reverse-transcribed cDNA and the respective amplification primers. DNA bands were obtained by agarose gel electrophoresis. Messenger RNA for the inhibin β_A subunit was demonstrated in all of the tissues studied. In contrast, inhibin α subunit mRNA was expressed in 60%, 50%, 75%, and 75% of the tumors in groups 1, 2, 3 and 4, respectively, whereas ACTR II mRNA was demonstrated in 20%, 37.5%, 62.5% and 50% of the tumors in each group. Coexpression of mRNAs for the inhibin α, and β_A subunits and ACTR II occurred in some brain tumors. The levels of inhibin a and ACTR II mRNA tended to be higher in the tumors with a higher grade of malignancy. These results indicate that some human brain tumors express receptors for activin, and that inhibin and activin may play an autocrine or paracrine role in brain tumor tissue.

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We examined the protective effects of medical castration by means of gonadotropin-releasing hormone analogue (GnRH_A) on the toxic effects of cisplatin in rats. Twelve days after a s.c. injection of a slowly-releasable form of leuprolide acetate (GnRH_ASR), rats were injected i.p. with cisplatin daily (3 mg/kg body weight (BW) for males and 4mg/kg BW for females) for four days and sacrificed 24h after the last injection. The doses caused acute tubular necrosis and gastrointestinal (GI) symptoms, i.e., diarrhea and fluid retention and bleeding in GI tract. GnRH_ASR pretreatment reduced serum urea nitrogen (SUN) and serum creatinine (SCre) increase and the incidence of GI symptoms. Histological analysis showed that rats pretreated with GnRH_ASR had noticeably less kidney damage. GnRH_A thus demonstrated its ability to protect the kidneys and GI tract against cisplatin toxicity in both male and female rats. This finding suggests a potential clinical application of GnRH_A in antineoplastic chemotherapy.

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Prostaglandin E2 (PGE2) has a potent bone resorbing activity in vitro, but some recent studies have shown that PGE2 stimulates bone formation in vivo. The effects of PGE2 on the bone are therefore still controversial. We attempted to reveal the effects of PGE2 on bone in vivo more directly; we injected PGE2 continuously into the bone marrow and onto the periosteum and examined the local effects of PGE2 histologically or by bone densitometry. Following PGE2 infusion into the bone marrow, new bone was formed in the bone marrow around the infused site and following PGE2 infusion onto the periosteum, extensive periosteal bone formation was observed. Bone mineral content was also increased significantly in the PGE2 infused bones. The administration of cyclic AMP did not mimic the effects of PGE2. In contrast to in vitro experiments, the in vivo effect of PGE2 is predominantly to produce bone.

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The clinical and genetic features of a 43-year-old male patient with multiple endocrine neoplasia type 1 were reported. He developed hyperparathyroidism, a GHRH-producing pancreatic tumor, and acromegaly between 1980 and 1983. Because his pituitary gland increased in size even after resecting the GHRH-producing pancreatic tumor, transsphenoidal hypophysectomy was performed six years later. The pituitary contained two histologically-different adenomas composed of somatotroph cells and null cells. Genetic analyses revealed loss of heterozygosity on chromosome 11 in common in the pituitary adenomas, the pancreatic endocrine tumors, and a parathyroid hyperplasia. On the other hand, mutations of ras, p53, Gsα, and Gi2α genes were not found in these tumors. The loss of the tumor suppressor gene on chromosome 11q12-13 was involved in the formation of two pituitary adenomas, two pancreatic endocrine functioning tumors, and a parathyroid hyperplasia in this patient, but the tumorigenic factors in the specific endocrine organs remain to be studied.

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A mouse epithelioid hemangioendothelioma (mEHE) cell line, passagable in vivo without TSH stimulation, was established from a thyroid tumor. Before establishment of the cell line, the primary thyroid tumor which was co-transplanted with a TSH-producing pituitary adenoma showed signs of hyperplasia and then transformed during several in vivo transplantable passages. The cell line which was established from an in vitro culture, was deficient in thyroid-specific differentiation function and had characteristics of endothelial cell origin such as vascular cavity formation and factor VIII expression. The cell growth of mEHE/CH1-5 cell lines was independent of TSH but was inhibited by c-AMP and vitamin D₃. c-Myc proto-oncogene expression was also suppressed by vitamin D₃ treatment. Both serum depletion and heparin treatment induced morphological changes in mEHE/CH 5 from an epithelial cell-like to an endothelial cell-like shape. Immunohistochemical analysis showed that epithelial membrane antigen expression was decreased and factor VIII expression was increased in relation to the morphological changes. In a further in vivo transplantation experiment, the histology of the mEHE/CH5 cell tumor had an angiosarcoma-like structure. These results indicate that this cell line established from a thyroid tumor possesses both epithelial and endothelial characteristics. The mEHE/CH5 cells might provide a good model for analyzing thyroid tumorigenesis and allow functional characterization of endothelial cells.

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PIT1 abnormality is defined as a genetic abnormality in the PIT1 gene that encodes a pituitary specific transcription factor, Pit-1/GHF-1. PIT1 abnormality indicates combined deficiency of thyrotropin (TSH), growth hormone (GH) and prolactin (PRL), and has been reported in several cases. We studied the PIT1 gene in a patient with combined deficiency of TSH, GH and PRL. A novel mutation substituting a termination codon for Glutamate at 250th codon (E250X) was identified in the homozygous state in the patient. Both of the healthy parents harbored this mutation in the heterozygous state. This nonsense mutation results in complete loss of helix 3 of the POU homeodomain of Pit-1/GHF-1. As helix 3 of the homeodomain is involved directly in DNA binding, the mutant Pit-1/GHF-1 may lose the DNA binding activity of the POU homeodomain and lose its transcriptional activation. The E250X mutation is therefore considered to be the cause of the combined deficiency of TSH, GH and PRL in this patient.

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The responsiveness in vivo to dopamine of prolactin (PRL) secretion in patients with prolactinoma was compared with that in vitro of single cells obtained from the same prolactinomas by surgical operations. Six patients with prolactinoma showing various degrees of hyperprolactinemia were challenged by bromocriptine suppression test (2.5mg, peroral) before operation. Bromocriptine administration caused a decrease in the serum PRL concentration ranging 24-95% and there was no correlation between the basal PRL level and bromocriptine-induced inhibition. Monodispersed pituitary cells obtained from the prolactinomas by operation were subjected to a reverse hemolytic plaque assay for PRL to determine PRL secretion at the single cell level under basal conditions as well as in response to dopamine. The percentage of plaque-forming cells under basal conditions ranged 15-55% among the prolactinomas. The percentage of plaque-forming cells and plaque area were decreased in a dose-dependent manner by 10^-7-10^-5M dopamine for the pituitary cells obtained from some adenomas but not for those from other adenomas. When the inhibition rates in vitro due to 10^-5M dopamine in these two parameters were compared with the inhibition rate in vivo in the serum PRL concentration due to bromocriptine, it was found that there was a significant correlation between them. These results show that the reverse hemolytic plaque assay can be used to determine in vitro responsiveness to dopamine of PRL secretion from single prolactinoma cells. We conclude that 1) the relative abundance of PRL-secreting cells varies among prolactinomas; and 2) there is a correlation between in vivo and in vitro responsiveness of prolactinoma cells to dopaminergic inhibition.

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Adhesion of lymphocytes to glomerular endothelial cells might be a critical step in the development of acute and chronic glomerulonephritis. The protective effect of anti-CD11a and anti-intercellular adhesion molecule (ICAM-1) antibodies on the cytotoxity elicited by phorbol 12-myristate 13-acetate (PMA)-stimulated lymphocytes was investigated by using cultured bovine glomerular endothelial cells (GEN). Both anti-CD11a and CD18 antibodies inhibited GEN injury induced by the activated lymphocytes in a dose-dependent manner. In addition, both antibodies also inhibited the adhesion of the activated lymphocytes to the GEN monolayers. These results suggest that it is important for activated lymphocytes to bind to GEN via adhesion molecules for cytotoxity to be produced by the activated lymphocytes.

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The association of familial hypogonadism with progressive cerebellar ataxia is only rarely encountered, and the exact link between the symptoms remains unknown. We report here two sisters presenting with Holmes type cerebellar ataxia, hypogonadotropic hypogonadism and retinochoroidal degeneration recently diagnosed as Boucher-Neuhauser syndrome. There was consanguinity between the parents of the affected individuals and the condition seemed to be inherited as an autosomal recessive defect. On endocrinological examinations, in both cases, the responses of LH and FSH to LH-RH (100μg) were impaired even after repetitive stimulation with LH-RH (400μg, 7 days), suggesting that the hypogonadism was due to a primary pituitary disturbance. Impaired GH responses to GRF (100 μg) and insulin-induced hypoglycemia (0.1U/kg) were also noted. The two sisters shared an almost identical clinical and endocrinological picture. Their karyotypes were 46, XX. They had been treated for primary and secondary amenorrhea at the age of 20 years and neurological problems had started at the age of 30 years. This unique family displays clinical evidence of a possible common mechanism responsible for a progressive hypothalamo-pituitary and cerebellar impairment of late onset.

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The present study was undertaken to determine whether improvement of hyperglycemia alters calcium and phosphorus handling, parathyroid hormone (PTH) secretion and bone turnover in patients with non-insulin-dependent diabetes mellitus (NIDDM). We measured serum and urinary mineral levels, serum intact PTH and osteocalcin on admission and at discharge (38±3 days later, Means±SEM) in 28 patients with poorly-controlled NIDDM (63±2 years old, 13 males and 15 females). During the hospitalization period, glycemic control was markedly improved. Serum calcium levels remained unchanged, but serum phosphorus increased. Urinary calcium and phosphorus excretion decreased. Serum intact PTH decreased from mid-normal (30.0±2.2ng/l) to low normal values (24.0± 1.3ng/l) (P<0.01, normal values: 10-65ng/l). Serum osteocalcin increased from 4.14±0.35 to 4.92± 0.40μg/l (P<0.01, normal values: 2.5-13μg/l). On admission, urinary calcium and phosphorus excretion showed a positive correlation with urinary glucose excretion. Serum calcium levels showed a negative correlation with serum intact PTH (r=-0.46, P<0.05). Moreover, the change in serum calcium during the hospitalization was negatively correlated to the change in serum intact-PTH (r=-0.45, P<0.05). Serum phosphorus concentrations showed a positive correlation with the renal threshold for phosphorus excretion on admission (r=0.86, P<0.01). These results indicate that hyperglycemia causes excess urinary calcium and phosphorus excretion in patients with NIDDM. In response to urinary calcium loss, PTH secretion is mildly stimulated. Bone formation seems to be suppressed in the hyperglycemic state in spite of increased PTH secretion.

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We used octreotide to treat a woman with acromegaly and observed pituitary adenoma shrinkage after 5 months. Diffuse scalp hair loss occurred after 5 months, resulting in the discontinuation of treatment. After the cessation of octreotide, the hair loss stopped and hair growth resumed. Since bromocriptine did not effectively decrease the GH level of the patient, we decided to perform transsphenoidal surgery. After resection of the pituitary adenoma, her GH and IGF-1 levels were normalized. Although octreotide-induced scalp hair loss has not been well recognized, we should pay more attention to this side effect.

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Granulocyte colony-stimulating factor (G-CSF) concentrations in serum were determined for the first time by a newly developed and highly sensitive chemiluminescent immunoassay (the limitation of detection, 0.5pg/ml) in ten patients with subacute thyroiditis, during treatment with glucocorticoid or indomethacin. Before therapy, circulating neurophil counts significantly increased to 5.15±2.07× 10³/μl compared with the convalescent phase (2.94±1.07×10³/μl), and the data were correlated with individual serum G-CSF levels (r=0.854, P<0.01). Serum concentrations of interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) were less than the detectable threshold of ELISA. During two weeks of glucocorticoid therapy, although the circulating neutrophil counts increased from 5.15±2.46×10³/μl to 7.73±1.64×10³/μl (P<0.01), serum G-CSF levels were depressed from 25.1±15.3pg/ml to 13.8±13.9pg/ml (P<0.01). These data indicate that G-CSF is one of the mediators of the increase of neutrophils in subacute thyroiditis, while it does no contribute to steroid-induced neutrophilia.

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The metabolic effect of recombinant human insulin-like growth factor I (IGF-I) was investigated by the glucose clamp technique in normal rats and streptozotocin-induced diabetic rats, a model of insulin-dependent diabetes mellitus (IDDM), and compared with that of insulin. Glucose uptake by peripheral tissues was stimulated by intravenous administration of IGF-I at rates of from 0.369 to 3.690nmol/kg/min in a dose dependent manner, with a potency of 1/52 that of insulin estimated on the basis of the ED₅₀ molar ratio in normal rats. In streptozotocin-induced diabetic rats, the maximum effects of IGF-I and insulin were reduced to 72% and 70% of those in normal rats, respectively, indicating the presence of both IGF-I and insulin resistance. Hepatic glucose output in normal rats was suppressed by IGF-I in a dose dependent manner with a weaker potency of 1/99 that of insulin assessed on the basis of the ED₅₀ values. In streptozotocin-induced diabetic rats, a dose-response curve of the suppressive effect of insulin on hepatic glucose output shifted to the right, indicating the presence of hepatic insulin resistance, but a leftward shifting of the suppressive effect of IGF-I on hepatic glucose output was observed. We concluded that the IGF-I effect on peripheral tissue was decreased but that on the liver was rather increased in streptozotocin-induced diabetic rats, in contrast to the resistance of both peripheral tissues and liver to insulin.

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We examined 13 patients with euthyroid Graves' disease suspected ophthalmologically, by comparing them with 20 patients with untreated Graves' disease and by following them up for 5 to 10 years. They had Graves' ophthalmopathy (NOSPECS class II-IV) without other ocular diseases, normal levels of serum thyroid hormones, and no previous history of Graves' disease. Proptosis in euthyroid Graves' disease was not significantly different from that in untreated Graves' disease. In 3 patients with euthyroid Graves' disease, TSH was suppressed. There was either no TSH response to TRH or it was low in 7 of 12 patients examined. The result of a T3-suppression test was abnormal in 8 of 11 patients examined. Titers of serum TGHA, MCHA, TSH-binding inhibitory immunoglobulin (TBII), and thyroid stimulating antibody (TSAb) were significantly lower in patients with euthyroid Graves' disease compared than in patients with untreated Graves' disease. TSAb, however, was positive in 12 of 13 (92%) patients. In spite of positive TSAb, 9 of 13 patients with euthyroid Graves' disease had normal radioactive iodine uptake (RAID). During the observation period, various abnormalities in thyroid function developed: persistent hyperthyroidism in 5 patients (38%), transient thyrotoxicosis in 2 (15%) and transient hypothyroidism in 1 (8%). We conclude that euthyroid Graves' disease is a subtype of Graves' disease that minimally develops thyrotoxicosis in spite of the existence of TSAb due to some mechanism inhibiting thyroid growth or stimulation, and that the measurement of TSAb provides a useful marker for the diagnosis of this disease.

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Cabergoline (CG) is a dopamine agonist that inhibits secretion of prolactin (PRL) and growth hormone. The purpose of this study was to investigate the PRL-lowering effect and antitumor effect of CG on estradiol-induced rat pituitary tumors in vitro and to elucidate these mechanisms. We compared the effects of CG with those of bromocriptine (BC) in terms of the inhibition of hormone secretion as well as antitumor effects on rat pituitary tumors. Primary cultures of dissociated pituitary tumor cells were used in these studies. A significant inhibition of prolactin (PRL) secretion was observed for both drugs within 12h after treatment, and the inhibitory effects of CG and BC were antagonized by sulpiride or haloperidol. Inhibitory effect on PRL secretion after 12-h BC or CG pretreatment was more pronounced with CG than BC treatment at all time points. PRL secretion in group pretreated with CG was significantly suppressed at 72h when compared to that of vehicle. Inhibition of de novo PRL synthesis was better demonstrated in the CG group. These findings suggest that CG has a higher affinity for the D₂ receptor of pituitary cells as compared to BC and may preferentially inhibit PRL secretion rather than PRL production. An antitumor effect of CG has been confirmed at a lower dosage than that of BC.

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Studies of anterior pituitary function in patients treated for nonfunctioning pituitary adenomas have been limited by their short duration. The purpose of the present study was to examine pituitary function longitudinally among three types of patients: those with complete tumor removal (group A); those with subtotal or partial adenomectomy (group B); and those in group B who underwent additional radiation therapy (group C). The subjects were 33 patients whose anterior pituitary function was evaluated by provocative tests such as insulin induced hypoglycemia, thyrotropin releasing hormone administration test and luteinizing hormone releasing hormone administration test. They underwent preoperative evaluation and postoperative reevaluations at 2 weeks, 3 months, 6 months and annually thereafter, for over 10 years. Anterior pituitary function was restored within a year after surgery, if at all. No additional function was restored after one year from treatment. In group A, no one developed impairment of anterior pituitary function after one year from surgery. In group B, however, new impairment was noted at intervals, due to tumor regrowth. In Group C, deficiencies developed after one year, irrespective of tumor regrowth. In conclusion, lifelong endocrinological follow-up is recommended for patients receiving postoperative irradiation, and for patients with potential for tumor regrowth. On the other hand, patients with total adenomectomy may be exempted from periodic endocrinological follow-up if they do not need postoperative hormonal replacement therapy.

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The significance of serum insulin-like growth factor I (IGF-I) and insulin-like growth factor binding protein 3 (IGFBP-3) levels in uremia is still controversial. In this study we measured serum IGF-I by immunoradiometric assay (IRMA) and IGFBP-3 levels by specific radioimmunoassay (RIA) in 28 children (aged 3-16 years) with end-stage (n=14, on hemodialysis) and pre-terminal renal failure (n=14) and in 15 age-matched healthy children. Thyroid function of the patients was also investigated and GH-stimulation tests with L-Dopa and insulin were performed. Neither IGF-I nor IGFBP-3 levels significantly correlated with mean height SDS for bone age or for chronological age in either non dialysis patients or pubertal (n=10) or prepubertal patients (n=18). These data was consistent with the concept that growth in CRF was not related to abnormalities in serum IGF-I or IGFBP-3 levels.

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To investigate the role of nitric oxide (NO) in pituitary ACTH secretion, the effect of a nitric oxide synthase (NOS) inhibitor, Nω-Nitro-L-arginine (Nitro-Arg), on ACTH secretion induced by interleukin (IL)-1/β, corticotropin releasing hormone (CRH), arginine vasopressin (AVP) and phorbol myristate acetate (PMA) was examined in rat anterior pituitary cell cultures. Nitro-Arg did not affect IL-1β-induced ACTH release in pituitary cell cutures incubated with Dulbecco modified Eagle's medium, Basal Medium Eagle or Krebs Ringer bicarbonate-glucose buffer. It did not affect CRH-, AVP- or PMA-induced ACTH release either. These results suggest that NO does not play an important role in ACTH release at the pituitary level.

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Nine patients with Graves' ophthalmopathy (GO) were treated with intravenous methylprednisolone pulse therapy and followed up by ophthalmological assessment, magnetic resonance imaging, and thyroid-associated autoantibody (thyroid stimulating antibody (TSAb), TSH binding inhibitor immunoglobulins (TBII), and anti-eye muscle antibody (EMAb)). Ophthalmological assessment was performed by the ophthalmopathy index (OI) which was made on the basis of the system recommended by the American Thyroid Association Committee. EMAb was expressed as the ratio of density of the 64kDa band of eye muscle membrane to that of 92kDa non-specific band found with all normal sera when assessed by western blotting. Five patients with mild ophthalmopathy (OI<4) did not show progressive improvement in OI. Three of 4 patients with severe eye disease (OI>4) showed a progressive and distinct improvement in OI. These 3 patients had high TSAb levels before methylprednisolone pulse therapy. One patient with severe ophthalmopathy did not respond to this pulse therapy; this patient's TSAb was negative. A significant positive correlation was observed between the activity of TSAb before treatment and the improvement in OI (ΔOI) (r=0.86, P<0.01, n=9). The relationship between ΔOI and EMAb did not reach significance. These results suggest that TSAb in sera of GO patients can be a useful marker for predicting the efficacy of methylprednisolone pulse therapy.

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We developed a new method for measuring an unidentified ketosteroid glucuronide (US-G) detected by the method of Iwata et al. for measuring 17-ketosteroid glucuronides by reversed phase HPLC on a Capcell-Pak C₈ column with three kinds of mobile phase solutions (Iwata method; Clin Chem 35: 795-799, 1989). The Iwata method inadequately separated US-G and two hydroxy 17-ketosteroides, 11β-hydroxyetiocholanolone and 11β-hydroxyandrosterone, and it exhibits insufficient sensitivity for measuring traces of US-G in the urine of healthy subjects. We solved these problems by developing a new method which measures US-G in urine, as a free type by hydrolyzing the glucuronide type enzymatically, by normal phase HPLC on a Capcell-Pak Silica column with one kind of mobile phase solution. By this method, the levels of US excreted as a glucuronide in the urine of healthy subjects and of patients with Cushing's syndrome were determined as proportions of the levels of 11β-hydroxyandrosterone. The average daily urinary excretion of US was 971μg (125-4, 995μg) in patients with Cushing's syndrome (n=22: two males and 20 females aged 26 to 65 years), and 34μg (0-141μg) in healthy subjects (n=63: 49 males, and 14 females aged 21 to 54 years), and the differences were clearly significant. However, there were no differences between the urinary US levels of patients with pituitary adenoma and patients with adrenal adenoma. Furthermore, no US was detected in the urine of patients with aldosteronism (two males and eight females aged 34 to 61 years). The daily level of urinary US excretion in two of the patients with polycystic ovary syndrome was 159 and 142μg, but no US was detected in the other two patients.

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