Sexual differentiation of the rodent brain is recognized to involve transcriptional activation of multiple genes induced by gonadal steroids at developmental stages. To identify the genes differing in expression level between sexes, we analyzed gene expression in male and female rat hypothalami at postnatal day 5 by means of a cDNA microarray consisting of 2352 genes. By comparing the expression pattern between sexes, we identified 12 male-enriched genes and 20 female-enriched genes. Among them, the expression pattern of 1 male-enriched gene, jagged homolog 1, and those of 2 female-enriched genes, p27Kip1 and p130, were confirmed to be consistent with microarray data by RT-PCR. Investigation of these genes should help to elucidate the molecular and cellular mechanisms underlying sexual differentiation of the rodent central nervous system.

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To determine the effect of octreotide acetate on urinary excretion of uric acid and plasma concentration of uridine, we subcutaneously administered octreotide acetate (1 μg/kg of body weight) to 5 healthy subjects. Ninety minutes after administration, octreotide acetate increased the plasma concentration of uridine by 15% and decreased the plasma concentration of glucagon by 24% and that of insulin to below the detection limits. In addition, octreotide acetate decreased the urinary excretion of uric acid, sodium, and chloride by 60%, 40%, and 38%, respectively, at 1 hour after administration. However, octreotide acetate did not affect the concentrations of hypoxanthine, xanthine, uric acid, cyclic AMP in plasma, lactic acid and pyruvic acid in blood, urinary excretion of hypoxanthine and xanthine, or creatinine clearance. From these results, we speculated that octreotide acetate decreases the urinary excretion of uric acid by decreasing the concentration of glucagon and/or urinary excretion of sodium, and increases the plasma concentration of uridine via decreased concentrations of glucagon and insulin.

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The present study was conducted in order to characterize the secretory pattern of GH in the Shiba goat, a native Japanese miniature goat, and to examine the effects of castration and KP102, a GH secretagogue, on this pattern. Blood samples were taken from an indwelling jugular catheter every 15 min for 24 h, and plasma GH was measured by radioimmunoassay. In intact males, GH was secreted in a pulsatile manner with very regular 5-h periodicity, which consisted of a distinctive GH pulse and a trough of virtually no GH secretion. Postpubertal castration increased the height and decreased the width of GH pulses, though it did not affect the interpulse interval and area under the curve. Modification of the shape of each GH pulse by testicular androgen might play a role in the expression of GH action in the male. KP102 (10 μg/kg, i.v.) immediately induced a robust GH pulse, which was followed by a spontaneous GH pulse of normal characteristics at regular intervals, suggesting that the clock generating GH pulses was reset by KP102. From these observations, we concluded that the Shiba goat is a very suitable experimental model for elucidating the mechanisms underlying GH pulse generation, and in particular, the involvement of androgen and GH secretagogues.

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A 61-year-old woman who had a nonfunctioning pituitary tumor removed was later found to have a pituitary carcinoma with metastasis to the liver that produced adrenocorticotropic hormone (ACTH), causing Cushing's disease. Despite the second removal of the pituitary tumor, Cushing's disease persisted. Endocrinologic examination demonstrated marked elevation of plasma ACTH, β-endorphin, and serum cortisol concentrations, which were not suppressed by low- or high-dose dexamethasone administration. A computed tomographic scan of the abdomen revealed a solitary hypodense area in the liver consistent with a metastatic lesion. Immediately after resection of the liver tumor, plasma ACTH concentrations fell to normal range. Such a decline in ACTH secretion was not observed even after the pituitary tumor was removed. Histologic characteristics of both the pituitary and liver tumors were similar. The immunohistochemical study using anti-ACTH antibody showed similar ACTH-positive cells in the pituitary and liver tumors. Based on these findings, the patient was diagnosed as having liver metastasis of ACTH-secreting pituitary carcinoma. This is the first case report in which a metastatic liver tumor of a pituitary carcinoma caused ACTH.

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The objective of this study was to compare, in infertile women suffering from severe hypogonadotropic amenorrhea, the therapeutic utility and the incidence of complications arising from fertility treatment by the conventional human menopausal gonadotropin/human chorionic gonadotropin (hMG-hCG) method, the hMG step-down method, the sequential hMG/gonadotropin-releasing hormone (GnRH) method and a new, modified hMG-GnRH method that has been developed by us. In the step-down method, the daily dose of hMG was decreased from 150 IU to 75 IU when the follicle diameter reached 11-13 mm. In the sequential hMG-GnRH, hMG injection was switched to pulsatile GnRH administration (20 μg/120 min SC), when the follicle diameter reached 11-13 mm. In our new modified hMG-GnRH, pulsatile GnRH was injected together with hMG. Daily hMG was stopped and the GnRH dosage was changed from 10 μg to 20 μg when the follicle diameter reached 11-13 mm. Initially, the three established methods were applied randomly to treat 34 cycles in 20 women; and subsequently, five patients who failed to conceive following treatment by sequential hMG-GnRH were then treated by the modified hMG-GnRH method. More than eight growing follicles and multiple pregnancies were observed during treatment by the conventional method. The incidence of ovarian hyperstimulation syndrome (OHSS) was 25.7% with the conventional method, 20.0% with the step-down method and 0% with the sequential hMG-GnRH method; however, the rate of ovulation was only 50% with the sequential hMG-GnRH method. By contrast, with the modified hMG-GnRH method, less than three growing follicles occurred in 81.8% of patients, there was a 100% rate of ovulation, and neither OHSS nor multiple pregnancies were observed. Moreover, the modified hMG-GnRH method induced pregnancy in 3 out of 5 patients. These data indicate that this new method is favorable for the treatment of severe hypogonadotropic amenorrhea.

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Characteristic findings of the pituitary stalk on magnetic resonance (MR) imaging, which suggest a damming-up phenomenon of neurosecretory granules, were reported. Neurosecretory granules containing vasopressin influence the signal intensity on MR T1-weighted image (T1WI). The normal posterior lobe of the pituitary gland appears as a bright signal on T1WI. The bright signal of the posterior lobe represents the normal content of neurosecretory granules and disappears in patients with central diabetes insipidus. The normal pituitary stalk appears as a low-intermediate intensity signal on sagittal and coronal T1WIs with 3 mm-slice thickness. The pituitary stalk appeared as a bright signal in 20 patients; 13 with pituitary adenoma, 4 with an intrasellar cystic lesion, one with cavernous sinus mass, and 2 with no abnormal MR findings. The pituitary stalk was not severed in any of the cases. The normal bright signal of the posterior lobe disappeared in 17 patients. No patients suffered from symptoms of central diabetes insipidus when the bright pituitary stalk appeared. It is suggested that the origin of the bright signal in the pituitary stalk is the damming up and accumulation of neurosecretory granules in the nerve fibers of the hypothalamohypophyseal tract obstructed by adenoma, postoperative scarring, cystic mass and so on. Probably, the damming-up phenomenon on MR imaging represents the functional integrity of the hypothalamo-neurohypophyseal system, and should be distinguished from an ectopic posterior lobe formation which is caused by stalk transection.

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Northern blot analysis of human TSH receptor (hTSHR) messenger ribonucleic acid (mRNA) expression has previously demonstrated multiple species of transcripts in the thyroid gland, suggesting the presence of multiple transcription initiation sites, alternatively spliced forms or alternate polyadenylation (poly (A)) sites. The first two have already been reported elsewhere. To clarify alternate poly (A) sites in the hTSHR gene, the present study was designed to characterize three full-length hTSHR cDNAs with distinct poly (A) signals that we have previously cloned. The comparison of the nucleotide sequencing data on the 3'UTR of these three clones to the Draft Human Genome in NCBI database revealed that the 3' segment of exon 10 of hTSHR gene contains three tandem repeats of the poly (A) sites, from which are expressed three full-length TSHR mRNAs with distinct 3'UTR length. The longest one appears to be a predominant transcript. From these data, together with (i) the previously reported organization of hTSHR genome and (ii) use of the Draft Human Genome to localize the unidentified sequence in the alternatively spliced form of truncated hTSHR, we propose the complete structure of hTSHR gene. Rather than 10 exons, our analysis suggests that hTSHR gene seems to contain 13 exons and 12 introns. At least three full-length TSHR mRNAs with distinct poly (A) sites and five alternatively spliced forms of TSHR mRNAs are expressed from the single hTSHR gene.

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A rare case of von Hippel-Lindau (VHL) disease with bilateral pheochromocytomas, right renal cell carcinoma, right pelvic carcinoma, spinal hemangioblastoma and primary hyperparathyroidism is described. A 78-year-old woman had a history of hypertension from her forties. She suffered from headache and body weight loss. Abdominal CT revealed bilateral adrenal tumors and right external renal tumors enhanced in early stage. MIBG scintigraphy exhibited a high accumulation of tracer in both adrenal glands. On the basis of the radiographic findings and endocrinological results, the patient was diagnosed as having bilateral pheochromocytomas and right renal cell carcinoma. A bilateral adrenectomy was performed, followed by surgery for resection of the renal cell carcinoma. The other resected right kidney showed a clear cell subtype that was determined to be renal cell carcinoma, and proved that the pelvic tumor was transient cell carcinoma. Spinal MRI showed spinal hemangioblastoma. von Hippel-Lindau (VHL) gene mutation for the patient was found. We diagnosed the patient as VHL because of the existence of spinal hemangioma and a VHL disease gene. Parathyroid echo revealed a hypoechoic space on the back of the left lobe, and serum calcium and intact PTH to be elevated. The patient was diagnosed as primary hyperparathyroidism. We report the first case of a patient with VHL disease complicated with bilateral pheochromocytomas, right renal cell carcinoma, right renal pelvic carcinoma and primary hyperparathyroidism. The life expectancy of affected individuals has been less than 50 years. Since the prognosis may be improved by an early diagnosis, affected individuals with VHL complexes should undergo cranial, spinal MRI and abdomen CT. The families may benefit from presymptomatic detection of affected gene carriers and the exclusion of at-risk family members by negative test results.

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The apoptosis of lymphocytes, which occurs in autoimmune diseases, is usually induced by the Fas/Fas ligand system. As the assay of nucleosomes produced by apoptotic cells can be used to quantitate apoptosis, we evaluated nucleosome and soluble Fas ligand (sFasL) levels of cultured mononuclear cells to clarify the apoptosis of mononuclear cells in patients with autoimmune thyroid diseases by enzyme-linked immunosorbent assay. Nucleosome levels of cultured mononuclear cells in patients with untreated Graves' disease were significantly higher (3.27±2.90 U/ml) than those of control subjects (1.39±0.24 U/ml) and euthyroid patients with treated Graves' disease (1.53±0.33 U/ml). Nucleosome levels of cultured mononuclear cells were positively correlated with sFasL levels (r=0.544, p<0.01). It is therefore likely that increased sFasL levels elicit apoptosis of these cells in untreated Graves' disease.

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Interferon-tau (IFNτ), produced by the trophectoderm of ruminant ungulates, binds to the type I IFN receptor (IFNAR) located at the uterine endometrium in a paracrine manner. Since IFNτ attenuates the secretory pattern of an endometrial luteolysin, prostaglandin F_2α, IFNτ has been considered as a conceptus factor implicated in the process of maternal recognition of pregnancy. Here we report the presence of IFNAR subunit (IFNAR1) in ovine conceptuses during the period of peri-implantation development and demonstrate that ¹²⁵I-human (h) IFNα binds to membrane preparations from ovine corpus luteum and conceptus. Using an antibody against hIFNAR1, immunohistochemical analysis revealed that IFNAR1 protein was present in day 14 and 16 conceptuses (day 0=day of estrus) and luminal and glandular epithelia of the endometrium. Conceptus membrane proteins analyzed by western blot with the same antibody displayed immunoreactive bands at 95, 60 and 55 kDa while endometrial membrane proteins showed bands at 200, 95 and 55 kDa. Northern blot analysis revealed that IFNAR1 mRNA was present in days 15-19 conceptuses and day 18-19 allantoic membranes. Receptor binding studies indicated that ¹²⁵I-hIFNα binding to day 16, but not earlier, conceptus membrane proteins could be displaced with hIFNα or ovine IFNτ. Based on Scatchard analysis, day 16 conceptus membranes contained 28 fmol IFNAR/mg protein with a dissociation constant of 300 pM. Cross-linking experiments demonstrated that ¹²⁵I-hIFNα-receptor complex migrated at 120 kDa, indicating that the receptor component(s) was approximately 100 kDa. These data provide evidence that although the binding does not occur until day 16, ovine conceptuses possess IFNAR1 near or at the time of implantation, suggesting that IFNτ, a factor produced by the trophectoderm of ruminant ungulates, could act on the conceptus in an autocrine manner. In addition to functioning as an antiluteolytic factor, therefore, IFNτ may have a direct effect on conceptus development.

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Although subacute thyroiditis (de Quervain's thyroiditis) is presumed to be caused by a viral infection, only 2 familial occurrences of subacute thyroiditis have been reported in the literature. Typical and severe subacute thyroiditis was diagnosed in an older sister who was hospitalized for 8 days. During this period her younger sister nursed her. Three weeks after, the same clinical picture occurred in the younger sister. We postulate that subacute thyroiditis might occur by transmission of possible viral infection in genetically predisposed individuals.

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Many clinical studies have shown that osteoporosis is associated with atherosclerosis and cardiovascular death. Although both high plasma levels of low density lipoprotein cholesterol (LDL-C) and low plasma levels of high density lipoprotein cholesterol (HDL-C) are known to be risk factors for atherosclerosis, it is unclear whether such lipid derangements are also associated with the pathogenesis of osteoporosis. In this study, we evaluated the relationships between plasma levels of total C, LDL-C, HDL-C, or triglyceride (TG) versus bone mineral density (BMD) at the lumbar spine, femoral neck, radius, or total body as well as the presence of vertebral fractures in 214 Japanese postmenopausal women (age range, 47-86 years, mean 62.7). Multiple regression analysis was performed between BMD at each skeletal site versus each lipid level adjusted for age, years after menopause, body mass index (BMI), and %fat. Plasma LDL-C levels were significantly and inversely correlated with the absolute values of both one-third radial (1/3R) and distal radial (UDR) BMD (p<0.01), and tended to be inversely correlated with the absolute values of L-BMD (p=0.051). In contrast, plasma HDL-C levels were significantly and positively correlated with the absolute values of L, 1/3R and UDR BMD (p<0.05). On the other hand, plasma TG levels were significantly lower in women with vertebral fractures than in those without fractures (97.0±36.5 vs. 126.4±65.8 mg/dl, mean±SD, p<0.05). When multivariate logistic regression analysis was performed with the presence of vertebral fractures as a dependent variable and each lipid level adjusted for age, years after menopause, BMI, and %fat as independent variables, TG alone was selected as an index affecting the presence of vertebral fractures (odds ratio: 0.51, 95% confidential interval: 0.29-0.89 per SD increase, p<0.05). Our study showed that plasma LDL-C and HDL-C levels were inversely and positively correlated with both R- and L-BMD values, respectively, while low plasma TG levels were associated with the presence of vertebral fractures in postmenopausal women. Thus, plasma lipids might be related to bone mass and bone fragility, and might be the common factor underlying both osteoporosis and atherosclerosis.

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Nitric oxide synthase (NOS) and nitric oxide (NO) play important roles in ovulation. The purpose of this study was to investigate the changes of intrafollicular nitrate/nitrite concentration and NOS mRNA expression in preovulatory follicles during equine CG (eCG) and human CG (hCG) induced ovulation in immature rats. Immature Sprague-Dawley rats received 15 IU eCG and then 15 IU hCG 48 h later. Rats were killed immediately before, 5 h after or 10 h after hCG injection, and their preovulatory follicles were dissected. Follicular fluid, granulosa cell, and theca cell layers were collected from preovulatory follicles and assayed for NO or NOS mRNA or for in vitro incubation study. Nitrate/nitrite concentration in the follicular fluid decreased significantly 5 and 10 h after hCG injection. Inducible NOS (iNOS) mRNA expression, which was greater in granulosa cell than in the theca cell layer, decreased significantly 5 and 10 h after hCG injection. However, endothelial NOS (eNOS) mRNA expression was detected mainly in the theca cell layer and further increased 5 and 10 h after hCG injection but remained low in granulosa cells. In vitro treatment of granulosa cells with 10^-4 or 5×10^-4 M S-nitroso-L-acetyl penicillamine (NO donor) decreased progesterone production and increased DNA fragmentation. We concluded that the decrease in nitrate/nitrite concentration in preovulatory follicles after hCG injection was due mainly to decreased iNOS expression in granulosa cells. These changes in nitrate/nitrite concentration may prevent apoptosis in preovulatory follicles.

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We report ACTH-independent bilateral macronodular adrenal hyperplasia (AIMAH) seen in a 48-year-old male with a history of rectal and skin cancer. Bilateral multiple adrenal nodular lesions on abdominal CT scans, elevated early morning plasma cortisol levels with undetectable plasma ACTH, increased 24-hour urinary free cortisol excretion, and loss of the normal circadian rhythm in cortisol secretion established the diagnosis of AIMAH. Subtotal resection of the adrenal glands preserving lower one third of the left side was performed, considering the unfavorable effects of total adrenalectomy on future treatment of his malignancies. He has been doing well for four years with normal plasma ACTH and cortisol levels. This case suggests that subtotal resection of the adrenal glands may be applicable to selected patients.

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ACTH deficiency gradually develops in patients with growth hormone deficiency (GHD) who have abnormalities of the pituitary stalk on magnetic resonance imaging (MRI) following perinatal complications. We report here a rare case of GHD manifesting ACTH deficiency in middle age. A 48-year-old male patient was admitted to our hospital due to fever and hyponatremia. He was diagnosed as GHD and hypothyroidism at the age of 9, and had received lysine treatment until age 20, which was then replaced by thyroid hormone. He was not mentally retarded, but was the shortest in his class throughout his schooldays, reaching a final height of 148 cm. Hormonal examination revealed the presence of hypoadrenalism as indicated by poor responses of plasma cortisol to intravenous administration of corticotropin-releasing hormone (CRH) and insulin-induced hypoglycemia. Plasma ACTH responded well to CRH, but not to insulin-induced hypoglycemia, indicating that his hypoadrenalism was of hypothalamic origin. MRI showed an invisible pituitary stalk and relatively small pituitary gland. Since he had a perinatal abnormality, the damage around the pituitary and GHD could have originated from birth. In the literature, around 60% of GHD patients with pituitary stalk abnormalities develop hypoadrenalism due to ACTH deficiency, and more than 90% of such cases are diagnosed by age 30. We suggest that the appearance of hypoadrenalism should be carefully monitored in GHD patients with pituitary stalk abnormalities even after they reach middle age.

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The presence of TSH receptor antibody (TRAb) is rarely responsible for hyperthyroidism due to metastatic lesions of thyroid carcinoma. A 70-year-old woman was incidentally found to be thyrotoxic around the time that external irradiation was performed for multiple bone metastases 9 years after subtotal thyroidectomy for follicular carcinoma. Hyperthyroidism persisted after oral administration of thiamazole. Relevant laboratory data were as follows: FT₄ 9.6 ng/L, FT₃ 7.3 ng/L, TSH <0.19 mU/L, TBII 70, TSAb 735, and Tg 32,000 μg/L. ¹³¹I-total body scan showed ¹³¹I accumulation in the occipital bone, cervical vertebra, thoracic vertebra, ilium, and residual thyroid gland. Since the ilium uptake (11.6) was markedly higher compared to the residual thyroid gland uptake (0.14), four subsequent ¹³¹I therapies were performed. The patient became hypothyroid, and TBII became negative. TSAb became negative after the first ¹³¹I-therapy but has increased again to 204 at present. Tg was 1,962 μg/L despite high TSH levels. ¹³¹I accumulation in the residual thyroid, cervical vertebra, and thoracic vertebra disappeared. Also ¹³¹I accumulation in the ilium has gradually decreased, but the image in the occipital bone has become markedly distinctive. This is a rare case characterized by TRAb-positive hyperthyroidism, by T₃-predominant thyrotoxicosis, and by stronger accumulation of ¹³¹I in the metastatic tumor than in the residual thyroid gland. Thus, the response to TRAb and ¹³¹I-therapy is different among metastatic thyroid tissues.

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