Dendritic cells infected with adenovirus expressing the thyrotrophin receptor induce Graves' hyperthyroidism in BALB/c mice.

Dendritic cells (DCs) are the most potent antigen-presenting cells and a prerequisite for the initiation of primary immune response. This study was performed to investigate the contribution of DCs to the initiation of Graves' hyperthyroidism, an organ-specific autoimmune disease in which the thyrotrophin receptor (TSHR) is the major autoantigen. DCs were prepared from bone marrow precursor cells of BALB/c mice by culturing with granulocyte macrophage-colony stimulating factor and interleukin-4. Subcutaneous injections of DCs infected with recombinant adenovirus expressing the TSHR (but not beta-galactosidase) in syngeneic female mice induced Graves'-like hyperthyroidism (8 and 35% of mice after two and three injections, respectively) characterized by stimulating TSHR antibodies, elevated serum thyroxine levels and diffuse hyperplasitc goiter. TSHR antibodies determined by ELISA were of both IgG1 (Th2-type) and IgG2a (Th1-type) subclasses, and splenocytes from immunized mice secreted interferon-gamma (a Th1 cytokine), not interleukin-4 (a Th2 cytokine), in response to TSHR antigen. Surprisingly, IFN-gamma secretion, and induction of antibodies and disease were almost completely suppressed by co-administration of alum/pertussis toxin, a Th2-dominant adjuvant, whereas polyriboinosinic polyribocytidylic acid, a Th1-inducer, enhanced splenocyte secretion of IFN-gamma without changing disease incidence. These observations demonstrate that DCs efficiently present the TSHR to naive T cells to induce TSHR antibodies and Graves'-like hyperthyroidism in mice. In addition, our results challenge the previous concept of Th2 dominance in Graves' hyperthyroidism and provide support for the role of Th1 immune response in disease pathogenesis.

[Methods of sleep induction for EEG recordings: comparison between three hypnotics and natural sleep].

We compared the effects of three oral hypnotics, monosodium trichloroethyl phosphate (MTP), chloral hydrate (CH), and pentobarbital calcium (PTB), to those of non-medication on wake-sleep states and sleep activation of epileptic seizure discharges. The subjects consisted of 410 epileptics and 171 non-epileptic outpatients (mean age: 12.5 years) of the Department of Pediatrics, Nagoya City University Hospital. Complete EEG records including awake and sleep states were obtained in 230/241 (95%) of patients with MTP, 20/22 (91%) of those with CH, 72/85 (85%) of those with PTB, and 225/233 (97%) of those without any hypnotics. There were no statistically significant differences in the effect of sleep induction among the four groups. Sleep activation effects were observed in 25% of patients with natural sleep and 35% of those with induced sleep. There was no statistical difference (p > 0.05). These results suggested MTP, CH, and PTB are useful hypnotics for sleep EEG recordings.

Carcinoma of the thyroid manifested as hyperthyroidism caused by functional bone metastasis.

A rare case of hyperthyroidism in the presence of a functioning bone metastasis secondary to an occult thyroid cancer is reported. A 59-year-old woman's pelvic bone metastasis was much too extensive and hypervascular to permit resection. An I-131 scan showed striking activity in the pelvic metastasis, which reflected ectopic excessive production of thyroid hormone by a functioning metastatic thyroid carcinoma. After total thyroidectomy, the patient received I-131 ablation and transcutaneous intra-arterial embolization therapy but her metastasis progressively enlarged. Microscopically, concomitant follicular and papillary cancer was found in close proximity to the thyroid. The patient now has vertebral metastases, and her hyperthyroid state still requires methymazole to prevent thyrotoxicosis.

[A case of nonresectable gastric cancer treated by sequential methotrexate and 5-fluorouracil].

An advanced gastric cancer patient with multiple retroperitoneal lymph node metastases and bone metastases was treated with sequential MTX and 5-FU. Complete response was obtained against both gastric primary lesion and retroperitoneal lymph nodes observed endoscopically and by computed tomography. Partial response was obtained against bone metastases observed by bone scintigraphy. Side effects of the chemotherapy were not observed.

Effects of percutaneous transluminal mitral valvuloplasty on plasma catecholamine levels during exercise.

Elevation of plasma catecholamine levels during exercise in patients with mitral stenosis correlated with the severity of the disease. We investigated the plasma norepinephrine changes in six patients before and after percutaneous transluminal mitral valvuloplasty (PTMV) during continuously graded ergometer exercise. Peak exercise intensity was increased from 65.8 W to 87.5 W after PTMV. Plasma norepinephrine level at 60 W workload intensity was decreased from 2308 +/- 864 pg/ml to 841 +/- 233 pg/ml after PTMV (p < 0.05). We concluded that PTMV decreased the plasma norepinephrine level during exercise in the patients with mitral stenosis. Percutaneous transluminal mitral valvuloplasty is a novel procedure for the improvement of symptoms in patients with mitral stenosis.

[A case of solitary plasmacytoma that originated in a rib].

A 52-year-old woman was admitted to our hospital with the chief complaint of left anterior chest pain. A chest X-ray showed a tumor shadow in the left 4th rib. No abnormalities were detected in peripheral blood and aspirated bone marrow samples. The tumor was surgically resected with the involved bone, the intercostal muscle and the parietal pleura. The resected specimen contained a 3 x 6 x 3 cm lesion, and histological investigation of the tumor revealed a plasmacytoma of the rib. It relapsed as a multiple myeloma within two months after surgery. Chemotherapy was administered and the bone lesions were observed to disappear. We believe that the best way to treat solitary plasmacytoma is to first resect the solitary tumor, and then to add cobalt irradiation and chemotherapy after surgery.

Total gastrectomy for gastric cancer associated with situs inversus totalis. A report of 2 cases.

A 69-year-old man and a 54-year-old woman with situs inversus totalis were admitted to our hospital for surgery for gastric malignant disease. The diagnosis was made radiologically and successful total gastrectomies were performed.

Responses of atrium and ventricle to sustained sympathetic nerve stimulation.

To determine whether chronotropic and atrial inotropic responses to sympathetic nerve stimulation are maintained longer than ventricular inotropic response, the present study was performed with control and acute reserpinized dogs. We stimulated the right stellate ganglion of both groups supramaximally for 60 min and compared right atrial responses (chronotropism and inotropism) with left ventricular (LV) dP/dtmax. In the control group, heart rate (HR) immediately increased and was only slightly attenuated with 60 min of stimulation, and right atrial (RA) inotropic response was less attenuated than was LV response (7% in HR, 33% in RA dP/dtmax, 50% in LV dP/dtmax, P less than 0.01, from the peak value of each response). RA and LV norepinephrine (NE) content was decreased by the stimulation but remained higher than the LV control value. In the reserpinized group, NE content in the RA was low before the stimulation and was further decreased by the stimulation. In this group, HR response was attenuated (27% in HR, P less than 0.01) as was LV dP/dtmax, and the difference in contractile responsiveness between atrium and ventricle disappeared (58% in RA dP/dtmax vs. 61% in LV dP/dtmax, NS). The results indicate that the chronotropic response was only slightly attenuated and that the atrial contractile response was attenuated less than the ventricular response, with sustained sympathetic nerve stimulation in the normal heart. This can be ascribed to the much higher NE content in the RA than that in the LV.

[A solitary metastatic osteosarcoma of the lung with thin walled cavity: report of a case].

A 13-year-old girl had a history of wide resection of the left fibula under the diagnosis of osteosarcoma three years before. Postoperatively, a thin walled cystic lesion in the right upper lung field, which enlarged gradually with thickening of the wall was found on the chest roentgenogram. The right upper lobectomy was performed after clinical evaluation. The pathologic examination confirmed that the tumor was the metastatic osteosarcoma of the lung. Although metastatic osteosarcoma is common in the lung, very rare cases demonstrate thin walled cavity formation in chest roentgenogram. We reported herein a case of a solitary metastatic osteosarcoma of the lung with thin walled cavity considering with pathogenesis of the lung.

Influence of reduced presynaptic myocardial norepinephrine stores on left ventricular contractility.

Many investigators have reported that myocardial norepinephrine content is decreased in congestive heart failure. However there have been no studies of how decrease in myocardial norepinephrine might influence myocardial contraction. To clarify whether decreased myocardial norepinephrine per se affects myocardial contraction, we observed the change in left ventricular contractility during 30 min of left stellate ganglion stimulation in control and acutely reserpinized dogs. We obtained left ventricular max dp/dt and left ventricular end-systolic pressure-segment length relationships as indicators of left ventricular contractility. Both parameters decreased after left stellate ganglion stimulation in reserpinized dogs (left ventricular max dp/dt: 2064 +/- 200 to 1608 +/- 168 mmHg/s, left ventricular end-systolic pressure-segment length slope 117 +/- 22 to 79 +/- 14 mmHg/mm, n = 8, P less than 0.05), while they did not change in controls. In reserpinized dogs, left ventricular norepinephrine content decreased to one-third that of controls before the stimulation, and further decreased after stimulation. These data indicate that lowered myocardial norepinephrine itself may be responsible for the negative effect on left ventricular contractility in congestive heart failure.

Androgenic actions of 5 alpha-androstane-3 alpha,17 beta-diol in rat submandibular gland.

To evaluate the action of 5 alpha-androstane-3 alpha,17 beta-diol(3 alpha-diol) in rat submandibular gland, 5 alpha-reductase, 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD) and oxidative 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSDO) activities, and trypsin-like protease activities, were assayed in control, castrated and 3 alpha-diol injected rats. 3 alpha-Diol (1 mg/kg) was injected subcutaneously in castrated male rats daily for 7 days. A 47% decrease of 5 alpha-reductase activity in the nuclei and a 30% decrease of 3 alpha-HSD(O) activities in the cytosol were shown after castration. 3 alpha-Diol restored the 5 alpha-reductase and 3 alpha-HSD(O) activities to 82 and 140% of the control submandibular gland, respectively. 3 alpha-Diol raised the trypsin-like protease activity to near control values in the submandibular gland of castrated rats. Morphological observations also revealed a distinct effect of 3 alpha-diol on the number of granules of granular duct cells. It is concluded that 3 alpha-diol has an androgenic action in the rat submandibular gland. It stimulates the 3 alpha-HSD(O). The 3 alpha-HSD(O) in its turn may be responsible for DHT accumulation in the cells.

3 alpha-hydroxysteroid dehydrogenase in the cytosol of rat submandibular gland.

We examined the in vitro shuttle metabolism between dihydrotestosterone (DHT) and 5 alpha-androstane-3 alpha, 17 beta-diol (3 alpha-diol) by 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD, E.C. 1.1.1.50) in rat submandibular gland (SMG) and ventral prostate (VP). The protein having molecular weight of 30 kDa, which was revealed by Sephacryl S-200 column chromatography, had 3 alpha-HSD activity to produce 3 alpha-diol from DHT, and also showed an oxidative 3 alpha-HSD (3 alpha-HSDO) ability to produce DHT from 3 alpha-diol. From the kinetic studies, the apparent Km and Vmax values of 3 alpha-HSD for DHT and NADPH were 6.4 microM, 1429 pmol/mg protein per min and 33.0 microM, 1205 pmol in SMG, and 9.3 microM, 377 pmol and 34.0 microM, 192 pmol in VP. The corresponding values of 3 alpha-HSDO for 3 alpha-diol and NADP+ were 18.0 microM, 714 pmol and 14.0 microM, 445 pmol in SMG, and 14.0 microM, 417 pmol and 36.0 microM, 77 pmol in VP. The affinities for DHT and 3 alpha-diol and the cosubstrate requirements of this enzyme in SMG were similar to those in VP. However, higher capacities of 3 alpha-HSD and 3 alpha-HSDO in SMG than in VP were shown. This suggests that there may be more 3 alpha-HSD in the SMG.

Morphological and biochemical study on the constituent cells and estrogen level of the rat submandibular gland.

The constituent cells of the submandibular gland of immature rats were compared with those of adult animals as well as the estrogen and estrogen binding protein level. The submandibular gland of immature and that of adult animals were entirely different in structure. High levels of estrogen binding protein are present in the nuclei in the young male and female submandibular gland. Nuclear estradiol binding protein, however, was not detected in the gland of adults in both sexes. These results suggest that the submandibular gland of the immature rat is influenced by estrogen.

Very well differentiated adenocarcinoma of the colon: a case report.

A 47-year-old woman with chronic colitis was found to have a sessile tumor at the hepatic flexure of the colon. An endoscopic biopsy revealed no aytpia, but surgery was performed since, in view of the tumor's size, malignancy could not be ruled out. A tumor measuring 3 x 2 x 1.5 cm was found in the resected specimen. The tumor exhibited quite unusual histologic features, showing invasive, moderately atypical glands forming mucin-filled cysts, very similar to those seen in colitis cystica profunda, under pseudopolypoid mucosa covered by non-atypical glandular epithelial cells. The extension of glandular elements into the pericolonic fatty tissue, and the intermingling of anaplastic microglands led to a final diagnosis of adenocarcinoma. In a case such as this, it is very difficult to evaluate cancerous tissue by endoscopic biopsy, even with an amount of tissue adequate for histologic examination.

Thermal salivation in rats, anesthetized with barbiturates, chloralose, urethane and ketamine.

1. The relationship between thermal salivation (TS) and thermoregulation was studied in anesthetized rats. 2. Of the 6 anesthetics used, ketamine-anesthetized rats secreted the largest amount of saliva. Salivation, however, was thermal and not induced by ketamine itself. 3. Ketamine-anesthetized rats readily secreted saliva at core temperatures less than 40 degrees C but TS was remarkably enhanced by hyperthermia of 40-42.5 degrees C. 4. The equilibrium phase in the triphasic heat response of core temperature was a consequence of equilibrium between heat gain and heat loss by salivation.

[Biochemical characteristics of nuclear 5 alpha-reductase and cytosol 3 alpha-hydroxysteroid dehydrogenase in rat submandibular gland].

Biochemical studies of testosterone 5 alpha-reductase and 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD) in rat submandibular gland (SMG) were performed. 14C-labeled testosterone or dihydrotestosterone (DHT) was incubated with subcellular fractions from rat SMG in the presence of 0.2 mM NADPH at 37 degrees C for 20 min in an atmosphere of 95% O2 and 5% CO2. Among the subcellular fractions, the high 5 alpha-reductase activity was detected in the nuclear fraction and 3 alpha-HSD in cytosol. Nuclear 5 alpha-reductase was efficiently solubilized in 2 mg digitonin per mg protein and 0.3 M KCl solution at 4 degrees C for 30 min. The maximum velocities (Vmax) of nuclear and solubilized 5 alpha-reductase activity for testosterone were 71.4 pmol/mg protein per min and 25.4 pmol/mg protein per min. Apparent Michaelis constant (Km) of nuclear and solubilized enzymes for testosterone were calculated as 11.1 microM and 16.7 microM by the Lineweaver Burk plot, respectively. The activity of solubilized 5 alpha-reductase from nuclei was stable by NADPH and KCl, and the molecular weight of the enzyme was estimated as 158 K.D approximately 200 K.D by Bio-Gel A-1.5 m column chromatography. The column chromatography also showed a peak of 3 alpha-HSD activity in cytosol, revealing the molecular weight of approximately 50 K.D. However, the elution peak of the 3 alpha-HSD was effectively decreased by KCl in Tris-HCl buffer. The molecular weight of 5 alpha-reductase and 3 alpha-HSD in SMG were similar to those in prostate. A stable and extractable 5 alpha-reductase was demonstrated in nuclei of rat SMG with possessing a considerable affinity for testosterone and also high 3 alpha-HSD activity for DHT was revealed in cytosol of the tissue.