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Spontaneous mammary carcinoma of AKR mice and Xray-induced mammary carcinoma of C58 mice, both known as low-cancer strains, were investigated by electron microscopy.In AKR mammary carcinoma, intracytoplasmic type A and extracellular type B particles were present. In C58 mammary carcinoma, extracellular type C particles were observed. In both tumors, mature virus particles, i.e. type B and type C particles, were formed by the process of budding of cytoplasmic membrane of carcinoma cells and present in the lumina of the milk ducts. Aberrant form of type C particles, which contained a filamentous structure instead of a nucleoid was also seen in C58 mammary carcinoma.In serial in vivo transplantation of those carcinoma cells to the same strain mice, these virus particles were always present and formed from carcinoma cells, although the number of virus particles reduced almost successively with transplantations in AKR mammary carcinoma.In tissue culture of C58 mammary carcinoma cells, type C particles similar to those in the original tumor were observed extracellularly. On the contraty to the presence of type B particles in the original AKR mammary tumor, however, two tissue culture strains derived from in vivo transplanted AKR carcinoma cells contained type C particles. These type C particles were conceivable to be the leukemia virus that naturally harbored in AKR mice and happened to have been tranferred to the tissue culture probably through its latent infection in carcinoma cells.

惡性腫瘍の治療にかんして,直接癌細胞を対象とした従来の方法とは異なり,癌の間質成分を抑制することにより,二次的に癌実質を障害せしめんとする,全く独自の新しい理念に基づいて,線維芽細胞抑制剤であるクロロキンを用い基礎的臨床的研究を行なつた.その結果動物の移植腫瘍に対して諸種の面から抑制効果を認め,組織学的に壊死の増大と間質結合織の抑制像を認めた.ヒト癌においては多くの例に自覚症の改善を見,腫瘍の縮小を認めることもあり,また全身症状のかなり改善されることがしばしば見受けられた.本剤の作用機序については間質成分の抑制による二次的な実質の障害の他に,本剤の有する抗炎症作用,あるいは宿主に対する全身的な影響が考えられるところである.現在の段階では本療法は手術不能例あるいは手術前後にその適応が考えられる.なお間葉性腫瘍である惡性リンパ性腫瘍の治療に本剤を用い,目下腫大リンパ節あるいは脾腫の縮小傾向を認めている.

An electron microscope study was performed on the ultrastructure and developmental process of the MUKAI strain of Japanese B encephalitis virus propagated in vitro on porcine kidney stable cells.Tne virus particle of Japanese B encephalitis is hexagonal in sections and approximaely 40mμ in the longest diameter, composed of the outer mambran, 20A thick, viroplasm, 30A thick and an electron-dense nucleoid, 25mμ in diameter. The virus particles develop by budding process on the wall of the cytoplasmic vacuole. Thereafter, virus particles are densely packed in the vacuole usually in random arrangement and rarely in crystalline arrays. The vacuole containing virus particles gradually moves toward the cell surface and liberates the virus particles to the exterior of the cells through a narrow canaliculus.A structure suggestive of incomplete virus particles was also observed.

ABS>The incidence of leukemia in RF mice exposed to a single whole-body irradiation of 350 r x-rays was 77.4%, in 41 out of 53 animals. In all, 20 developed the thymic and 20 the non-thymic lymphocytic leukemia, while only one developed myelogenous leukemia. The incidence of leukemia by 20- methylcholanthrene painting was 88% in 22 out of 25 animals. Of them, 12 developed lymphocytic leukemia and 10 myelogenous leukemia. Cell-free filtrates from the leukemic mice, which were induced by a single whole-body x irradiation, were inoculated to the newborn mice of the same strain less than 16 hours old. Two (18.7%) of the twelve inoculated animals developed leukemia on the 357th and 374th days, respectively, after the inoculation. Attempts to inoculate cell-free filtrate from 20-methylcholanthrene-induced leukemic RF mice to newborn mice of the same strain were successful. Cell-free filtrate from the myelogenous leukemic mouse caused lymphocytic leukemia in one of three inoculated mice. This is suggestive of the possibility of transformation of the type of leukemia during cell-free transmission. Virus-like particles were demonstrated by electron microscope in the lymph nodes of 20-methylcholanthrene-induced leukemic mice. The results suggest that x-ray and chemical carcinogens may act to induce leukemia or mammary carcinomamore » by the activation of a latent virus naturally resident in the RF mouse. (P.C.H.)« less

Ultrastructure of nonenzymatically glycated mesangial matrix in diabetic nephropathy. Advanced protein glycation has been proposed as a major factor in the development of diabetic nephropathy. Advanced glycation end products (AGEs) have altered the structure of extracellular matrix component and impaired self association in vitro . To elucidate the role of AGEs in the progression of diabetic nephropathy, the present study was undertaken to localize glomerular AGEs immunohistochemically. Ultrastructural changes of the mesangial matrix were analyzed with high resolution scanning electron microscopy. No glomerular AGEs staining was noted in normal control kidney specimens, or in tissue from glomerulonephritis patients without diabetes mellitus. The mesangium showed a positive AGEs staining in advanced stages of diabetic nephropathy, and the most characteristic finding was the strong AGEs staining in nodular lesions. By high resolution scanning electron microscopy, control and diabetic mesangial matrices revealed a meshwork structure composed of fine fibrils (10 nm in width) and numerous pores (12 to 13 nm in diameter). In the nodular lesions, however, loosening of the meshwork was significant, and the diameter of the pores was enlarged (approximately 24 nm). This study provides the first immunohistochemical evidence that AGEs are localized in diabetic glomeruli, most notably to nodular lesions. Advanced glycation might play a role in the progression of diabetic nephropathy through impairment of the assembly of matrix proteins in vivo .

The effect of nifedipine on CRF-41- and AVP-induced ACTH release was examined using monolayer cultured rat anterior pituitary cells and pituitary halves. Nifedipine inhibited ACTH release induced by synthetic rat CRF-41 in two systems. In pituitary halves, CRF-41 significantly stimulated both ACTH release and cyclic AMP accumulation. Nifedipine inhibited CRF-41-induced ACTH release and the inhibitory effect of nifedipine on CRF-41-induced ACTH release was accompanied by parallel decrease of cyclic AMP levels in pituitary halves. Nifedipine did not inhibit AVP-induced ACTH release in pituitary halves, and AVP did not significantly affect cyclic AMP accumulation in pituitary halves. These results suggest that CRF-41 stimulates ACTH release through the intracellular cyclic AMP system and calcium-calmodulin system which are accelerated by the influx of extracellular calcium ions. Moreover, it is suggested that the calcium required for AVP-induced ACTH release is derived primarily from intracellular rather than extracellular sources.

An intravenous administration of (D-ala2, met5)-enkephalinamide (DALA) caused a significant elevation of plasma ACTH and corticosterone at 10 to 20 min after injection in unanesthetized freely moving rats. An intraperitoneal administration of cyproheptadine tended to reduce plasma ACTH and corticosterone levels at 60 min after injection, but it did not attenuate the DALA-induced ACTH and corticosterone elevation. A large dose of naloxone (1-10 mg/kg body weight) caused a significant elevation in plasma corticosterone, but naloxone at 10 mg/kg body weight reduced the basal ACTH level and DALA-induced ACTH elevation. When both DALA and naloxone were injected, the steroidogenic effect was attenuated. Neither DALA nor naloxone affected the basal ACTH release and CRF-induced ACTH stimulation in rat anterior pituitary cell cultures. These results suggest that DALA acts at the extra-hypophyseal level to stimulate ACTH and corticosterone and that the naloxone stimulatory effect on steroidogenesis acts on the adrenal gland or is mediated by stimulating corticosterone stimulating factors other than ACTH.

Plasma adrenocorticotropic hormone (ACTH) has been measured after an intra-third ventricular administration of noradrenaline, an adrenergic agonist or an adrenergic antagonist. Centrally administered noradrenaline caused a significant increase in ACTH secretion. The alpha-agonist phenylephrine also increased the ACTH level. However, neither the alpha-antagonist phentolamine nor beta-agonist isoproterenol affected the ACTH level. The beta-antagonist propranolol evoked a significant elevation in ACTH. Passive immunoneutralization was examined with anti-rat corticotropin-releasing factor (CRF) rabbit serum, anti-arginine vasopressin (AVP) rabbit serum and normal rabbit serum (NRS) on the intra-third ventricular noradrenaline-induced ACTH secretion to study the involvement of endogenous CRF. An intra-third ventricular administration of noradrenaline caused a significant increase of ACTH levels in NRS-injected rats and anti-AVP-injected rats, whereas an i.v. anti-rat CRF injection significantly reduced the intra-third ventricular noradrenaline-induced ACTH secretion. These results suggest that central catecholamine stimulated ACTH secretion via the alpha-adrenergic mechanism and that endogenous CRF is at least partly involved in the noradrenaline-induced ACTH secretion.

B型肝炎ウイルス(HBV)関連膜性糸球体腎炎とHBV関連膜性増殖性糸球体腎炎の2例を経験し,おのおのの腎糸球体に, HBe抗原の存在を証明した.これまで, HBV腎症においてはseroconversionと相前後して腎症状の改善が見られることは知られているが,積極的にseroconversionを誘導して治療を行った報告は少ない.そこで,今回の2症例に対して, HBe抗原抗体系のseroconversionを自的に,副腎皮質ステロイド中断療法,インターフェロン療法を行ったところ,血中HBe抗原の消失, HBe抗体の出現と共に,尿蛋白の減少,消失が認められ,良好な経過をとった.以上の経験により, HBV腎症に対して積極的なseroconversionの誘導が有効と考え報告した.

A 24 year old female presented with signs and symptoms of postganglionic cholinergic autonomic dysfunction manifested by impaired lachrymation and salivation, mydriasis of the pupil, decreased gastrointestinal motility, atony of the bladder, and sweating and taste disturbance. Clinical and pharmacological studies confirmed that the abnormalities were restricted mainly to the postganglionic cholinergic autonomic systems. The titre of serum complement was low, antinuclear antibodies revealed a diffuse pattern, and anti SS-A antibody was positive, suggesting that this disorder was caused by an immune mechanism.

Brain corticotropin-releasing hormone (CRH) concentration and pituitary adreno-cortical responses were examined in chronically stressed rats: body restraint stress (6 h/day) for 4 or 5 weeks. Stressed rats showed a reduction in weight gain. CRH concentration in the median eminence and the rest of the hypothalamus were not different between control and chronically immobilized rats. The anterior pituitary adenocorticotropic hormone (ACTH) concentration was elevated in chronically stressed rats, whereas plasma ACTH and corticosterone levels did not differ from the control values. The median eminence CRH concentration was reduced to the same extent at 5 min after onset of ether exposure (1 min) in chronically immobilized rats and controls. However, plasma ACTH and corticosterone showed greater responses to ether stress in chronically immobilized rats than in control rats. Plasma ACTH and corticosterone responses to exogenous CRH were not different between control and chronically immobilized rats, while the response to arginine vasopressin (AVP) was significantly greater in chronically immobilized rats. These results suggest that chronic stress caused an increase in the ACTH-secreting mechanism and that pituitary hypersensitivity to vasopressin might at least be partly responsible for this.

Using a cellular digestion technique combined with scanning electron microscopy, we examined the three-dimensional ultrastructural changes of the glomerular basement membrane (GBM) in various types of human glomerulonephritis (GN). Gaps of the GBM, which were not found by routine transmission electron microscopy, were observed in a patient with IgA nephropathy manifesting microscopic hematuria. The dense material of the GBM in dense deposit disease (DDD) was not affected by this cellular digestion technique, while the deposits in other immune complex diseases were almost removed. Thus the dense material in DDD differs in composition from the immune complex deposits in other GN. In membranous GN and lupus nephritis, the morphological changes of the urinary surface of the GBM varied from pinhole to craters and reticula, according to the severity of the disease stage. This technique is useful for examining ultrastructural changes of the GBM in various human GN.

Characterization of the cells responsible for spontaneous DNA synthesis and/or IgG secretion in systemic lupus erythematosus (SLE) was undertaken by fractionation of the peripheral blood mononuclear cells (PBM). Each fraction was analyzed for its capacity to incorporate (/sup 3/H)thymidine ((/sup 3/H)TdR) and secrete IgG without mitogen. The non-E rosette-forming cell (non-ERRC) fraction, which consisted of the surface immunoglobulin-positive (sIg(+)) cells and null cells, revealed a markedly increased spontaneous DNA synthesis (620.0 +/- 586.9 cpm) during the first hour of culture and an elevated spontaneous IgG secretion (8639 +/- 2630 ng/ml) during 9 days of culture. Of particular interest was the finding that both increased responses were conducted by the null cells; the null cell population had approximately a fourfold relative increase of (/sup 3/H)TdR incorporation and a 60-fold relative increase of IgG secretion compared with the sIg(+) cell population. These results suggest that SLE patients have a small population of preactivated B-cell lineage cells, which lack sIg or have a very low density of sIg.

An intra-third ventricular administration of (D-Ala 2 ,Met 5 )-enkephalinamide (DALA) did not elevate plasma ACTH and corticosterone levels in unanesthetized freely moving rats, but intra-third ventricular administration of DALA and methionine (Met)-enkephalin potentiated a mild stress (hanging for 10 or 30 sec)-induced plasma ACTH and corticosterone elevations in unanesthetized freely moving rats. DALA and Met-enkephalin seemed to stimulate CRF release from the median eminence to increase plasma ACTH, as the CRF concentration in the median eminence area was reduced after injection in these stressed rats. When hypothalamic tissues were perifused in vitro, DALA (1–100 ng/ml) reduced the release of CRF. These results suggest that the opiates seem to have a dual effect on the CRF-ACTH system depending on which action overrides the other.

The effects of clinofibrate on serum lipoprotein concentrations, lecithin cholesterol acyl transferase activity and atherogenic index were studied in 10 diabetes mellitus patients. The patients comprised five with well-controlled non-insulin-dependent diabetes, and five with poorly controlled insulin-dependent diabetes; six non-insulin-dependent diabetics acted as placebo controls. No adverse side-effects were reported and there were no significant changes in total cholesterol, triglyceride or high-density lipoprotein 3-cholesterol concentrations following 600 mg/kg clinofibrate treatment for 4 weeks in either insulin-dependent or non-insulin-dependent diabetics. High-density lipoprotein-cholesterol concentrations and lecithin cholesterol acyl transferase activity were significantly ( P

To elucidate the role of atrial natriuretic peptide (ANP) and vasopressin (VP) in a hypertensive state, ANP and VP receptor bindings in spontaneously hypertensive rat (SHR) kidney were analyzed using the radiolabeled receptor assay (RRA) technique. Systolic blood pressure of SHR aged 12 weeks was statistically higher than that of age-matched Wistar Kyoto (WKY) rats. Maximum binding capacity (Bmax) of [125I]-ANP binding to the SHR kidney membrane preparations was statistically lower than that of WKY rats, but dissociation constant (Kd) was not significantly different. On the other hand, Bmax of [3H]-VP binding to the SHR kidney membrane preparations was statistically higher than that of WKY rats, but Kd were similar. Since the physiological action of ANP is natriuresis and VP is the most important antidiuretic hormone in mammalia, these opposite changes of ANP and VP receptor bindings in SHR kidney suggested that these peptides may play an important role in the pathophysiology of the hypertensive state, although it has not been confirmed as yet.

A 51-yr-old male patient with Cushing's syndrome due to huge nodular adrenocortical hyperplasia is described. Urinary 17-OHCS was not suppressed by a high dose of (8 mg) dexamethasone and showed rather a tendency to paradoxical response. There was no response to metyrapone. Plasma cortisol showed a hyperresponse to insulin-induced hypoglycemia and a rapid response to corticotropin releasing hormone-lysine vasopressin (CRH-LVP) administration without an obvious ACTH response. Plasma cortisol responded to synthetic ACTH. Urinary 17-OHCS did not show parallel changes with plasma cortisol. These results and computerized tomography data suggested huge multiple nodular adrenocortical hyperplasia, which was confirmed later by surgery. The left and right adrenal glands weighed 105 and 45 g, respectively. Hyper-reaction of the adrenal gland to a small change in plasma ACTH or "unknown factors" may cause not only the discrepancy between cortisol and ACTH response but also the development of autonomous nodules in the adrenal gland.

Masugi nephritis was induced in Sprague-Dawley rats by an intravenous injection of rabbit anti-rat kidney serum. In the autologous phase of the disease, three of 18 rats manifested continuous hematuria. Ultrastructural examination of renal glomeruli by transmission and scanning microscopy revealed gaps in the basement membranes, and the transcapillary passage of red blood cells through the discontinuous regions in the hematuric rats. Control animals revealed no gaps in the glomerular basement membranes regardless of the method used in their preparation for electron microscopy. These data support the hypothesis that hematuria is a result of the passage of red blood cells through gaps in the glomerular basement membrane in Masugi nephritis.

A male patient with corticotropin-releasing factor (CRF) and adrenocorticotropin (ACTH)-producing syndrome is described. Soon after being referred to us the patient developed pneumonia, anaemia, oedema and respiratory distress, and died on the 24th day after admission. Autopsy and histology revealed that he had a rare type of multiple endocrine neoplasia (type 1 + paraganglioma) with a mediastinal paraganglioma, parathyroidal hyperplasia, pancreatic islet cell adenoma, duodenal multiple carcinoid tumours and adrenocortical nodular hyperplasia. It was not possible to examine the pituitary. The paraganglioma contained a large amount of immunoreactive (IR)-CRF (606 ng/g wet weight), IR-ACTH (59.4 ng/g wet weight), IR-human proopiomelanocortin n-terminal (1–76) peptide (hNT, 156.8 ng/g wet weight) and IR-β-lipotropin (β-LPH, 146.9 ng/g wet weight). The major IR-ACTH, β-LPH and IR-hNT were eluted at ACTH-(1–39), β-LPH and hNT marker positions, respectively. Big ACTH was not detected. IR-CRF eluted at the human CRF marker position on Sephadex G-75 chromatography and high performance liquid chromatography (HPLC). The IR-CRF fraction from the HPLC showed CRF bioactivity which paralleled that of synthetic human CRF in monolayer cultured rat anterior pituitary cells. Our results suggest that not only ACTH but CRF produced by the paraganglioma was responsible for the patient's Cushing's syndrome.

Starvation-induced changes in CRF concentration in major brain regions and abnormalities in the pituitary-adrenal axis were examined in rats using rat CRF radioimmunoassay. The CRF concentrations in the hypothalamus and cerebellum were significantly reduced in the completely starved rats, while those in the midbrain, thalamus and neurointermediate lobe of the pituitary were significantly increased in the semi-starved or completely starved rats. No significant changes in the CRF concentrations were found in the pons, medulla oblongata and cebral cortex. In the completely starved rats, the serum ACTH level was significantly reduced, whereas the serum corticosterone level was markedly elevated. These observations suggest that starvation may stimulate the CRF-ACTH-corticosterone system and that not only hypothalamic CRF but also extrahypothalamic CRF may be discretely related to feeding behavior or starvation. The reduced serum ACTH level in starved rats may be ascribed to the negative feedback effect of the elevated serum corticosterone.

Abnormalities in the hypothalamo-pituitary-adrenal axis in spontaneously hypertensive rats (SHR) during development of hypertension were investigated using in vivo and in vitro methods. Plasma ACTH responses to hemorrhage and ether stress were significantly smaller in 7-week-old SHR than in age-matched Wistar-Kyoto rats (WKY), while plasma corticosterone baseline levels and its response to stress were greater in SHR than in WKY. There was no significant difference in the plasma ACTH response to ether stress between bilaterally adrenalectomized SHR and WKY replaced with a 25% corticosterone pellet for 6 days. Adrenalectomy prevented the development of hypertension in SHR; however, corticosterone replacement restored hypertension. Plasma ACTH showed a smaller response to iv CRH injection in SHR than in WKY, while the ACTH response to arginine vasopressin was not different between SHR and WKY. CRH concentrations in the median eminence, posterior pituitary, and cerebral cortex were lower in SHR than in WKY, while the CRH concentration in the median eminence was not different in SHR and WKY when they were adrenalectomized with or without corticosterone replacement. Basal in vitro CRH release from hypothalamic tissue was reduced in SHR, while CRH release in response to 56 mM KCl was not different in SHR and WKY. These results suggest that adrenocortical function is enhanced in young SHR, that reduced ACTH response to stress and exogenous CRH in SHR may be ascribed to higher plasma corticosterone levels, and that corticosterone is essential for the development of hypertension in SHR.

Rat tubuli were isolated both by the method of Krisko and by a modified method of Cook and Pickering; renal tubular basement membrane (TBM) was isolated by sonic disruption and by the method of Carlson. Using electron microscopy after negative staining, TBM of rat kidney isolated under these different conditions was shown to be a fine meshwork. Strands of the meshwork were interwoven, apparently enclosing pores to make up whole basement membrane. These findings are compared with our previous observation that glomerular and alveolar basement membranes were made up of a similarly fine meshwork.

We report a case of IgG multiple myeloma with selective complete Clq deficiency. The patient was a 75-year-old Japanese woman who exhibited urticaria on the arm and an absence of serum hemolytic complement activity (CH50). Further studies revealed no vasculitis in the urticarial lesion but showed selective complete deficiency of Clq without low molecular weight Clq precipitin. Addition of highly purified Clq restored the CH50 level of the patient's serum to normal. It is suggested that this abnormality was a primary Clq deficiency. We discussed a relationship between the Clq deficiency and myeloma and reviewed the literature.

The effect of synthetic atrial natriuretic peptide (ANP) was examined on the in vivo and in vitro release of ACTH. Intravenous ANP (4 micrograms/kg body weight) administration did not affect the corticotropin releasing factor (CRF, 4 micrograms/kg body weight)-, arginine vasopressin (AVP, 2 micrograms/kg body weight)- and angiotensin II (A II, 4 micrograms/kg body weight)-induced ACTH release in unanesthetized freely moving rats. ANP did not inhibit the basal, CRF- and AVP-induced release of ACTH in pituitary cell cultures. ANP did not affect the CRF- and AVP-induced plasma corticosterone elevation, while it attenuated the AVP-induced corticosterone elevation. These results indicate that ANP does not affect the ACTH release at the pituitary level in vivo and in vitro.

Effect of rat atrial natriuretic peptide (rANP) on acetylcholine-induced release of corticotropin-releasing factor (CRF) from the rat hypothalamus was studied in vitro using perifusion method. Perifused acetylcholine at 100 and 1000 ng/ml evoked significant CRF release, whereas norepinephrine at 10, 100 and 1000 ng/ml did not show a definite effect on CRF release. Continuous administration of alpha-rANP(1-28) (20ng/ml) inhibited the acetylcholine (100ng/ml)-induced CRF release. It is likely that ANP is involved in the regulation of CRF release.

症例は58才,女性. 43才より慢性関節リウマチ(RA), 53才で強皮症(PSS)と多発性筋炎(PM)を併発し加療中,昭和62年蛋白尿と浮腫が出現した.ネフローゼ症候群と腎機能低下を認め,腎糸球体と血管壁にアミロイド(AAtype)が沈着していた.ステロイド薬とdimethylsulfoxide (DMSO)を併用し,炎症反応は改善し蛋白尿と腎機能は約半年間は増悪しなかった. RA, PSS, PMのoverlap症候群に伴う続発性amyloidosisは従来報告がないので報告する.

The involvement of prostaglandin E2 (PGE2), adenosine 3',5'-cyclic monophosphate (cAMP), and vasopressin in lithium-induced polyuria was investigated in rats. Administration of LiCl (4 mmol/kg body wt) for 7 days induced a marked polyuria with a significant excretion of urinary PGE2. Administration of indomethacin (IND, 5 mg/kg body wt) for 4 days to lithium-induced diabetes insipidus (LiDI) rats diminished urine volume by 80% and urinary PGE2 by 85%. The in vitro data of the intact rat kidney showed that lithium stimulated arginine vasopressin (AVP)-induced PGE2 production and suggested that PGE2 suppressed cAMP synthesis in rat renal medulla. The AVP-induced PGE2 synthesis was greater and the AVP-stimulated cAMP production lower in the LiDI rat kidney in vitro. Interference of the vasopressin-associated cAMP system and the increased PGE2 synthesis in the kidney may be involved in the development of LiDI. The reduced cAMP production in the LiDI rat kidney might be partly due to the increased PGE2 synthesis. In LiDI rats plasma vasopressin increased, whereas AVP concentration in the hypothalamus and the neurohypophysis significantly decreased. It is postulated that lithium stimulates vasopressin release from the central nervous system and that elevated plasma vasopressin potentiates PGE2 production in the kidney synergistically with lithium.

The effect of lithium on the hypothalamo-pituitary-adrenal axis was studied in vivo and in vitro. The levels of plasma vasopressin, ACTH and corticosterone increased after the administration of lithium (LiCl 4 mmol/kg BW, 11 days) in rats, while the tissue vasopressin concentration in the median eminence, the rest of the hypothalamus and the posterior pituitary was decreased. The CRF concentration in the posterior pituitary increased markedly, but it did not change significantly in the median eminence or the rest of the hypothalamus. The elevated plasma ACTH level might be at least partly due to the increased vasopression secretion. Lithium stimulated ACTH secretion per se and also enhanced vasopressin-induced ACTH secretion in cultured pituitary cells and in half pituitary incubations, while it did not affect CRF-induced ACTH secretion. Lithium inhibited CRF-induced cAMP accumulation in half pituitary incubations, while lithium and vasopressin did not affect cAMP accumulation per se or even when administered together. The results suggest that lithium-induced ACTH release is via a cAMP-independent mechanism. Thus, it is possible that lithium stimulates ACTH release by acting directly on the corticotroph, stimulating vasopressin release and potentiating vasopressin-induced ACTH release.

The effects of intraperitoneal and intra-third ventricular administration of morphine on the hypothalamic corticotropin-releasing factor (CRF) and the pituitary-adrenocortical activity were examined in unanesthetized, freely moving rats. Hypothalamic CRF was measured by rat CRF radioimmunoassay. Intraperitoneal or intra-third ventricular administration of morphine increased blood concentrations of ACTH and corticosterone while intraperitoneal administration tended to increase CRF concentration in the whole hypothalamus including the median eminence and intra-third ventricular administration increased CRF concentration in the hypothalamus excluding the median eminence. However, morphine seemed to inhibit the increase in CRF concentration in the hypothalamus induced by the ether-laparotomy stress. The main site of morphine action on the hypothalamo-pituitary-adrenocortical system seemed to be in the hypothalamic area.

An endogenous inhibitor of GABA receptor binding was prepared from synaptic membrane of rat brain with 0.05% Triton X-100. The endogenous inhibitor was competitive with GABA for GABA binding sites. The inhibition of GABA receptor binding by the endogenous inhibitor was blocked by the allosteric effect of diazepam. In the presence of diazepam, specific [3H]GABA binding was greater in a medium containing the endogenous inhibitor than in one containing an equal inhibitory potency of GABA, whereas there was no difference in the absence of diazepam. This indicated that the endogenous inhibitor was not GABA itself.

The effect of immobilization stress (IM-stress) on the muscarinic cholinergic (m-Ch) receptor binding was determined in 8 brain regions using [3H]quinuclidinyl benzilate (QNB). IM-stress produced an increase in specific QNB binding in the septum, striatum, hippocampus and pons + medulla oblongata. Scatchard analysis revealed that IM-stress produced an increase in the affinity of m-Ch receptors in the septum, hippocampus and pons + medulla oblongata, but did not alter the maximum number of binding sites (Bmax). In the striatum, an increase in specific QNB binding was due to both the increase in Bmax and reduction of the dissociation constant (Kd). The present study suggests that IM-stress induces supersensitivity of postsynaptic m-Ch receptors probably due to a decrease in presynaptic cholinergic activities in the septum, striatum, hippocampus and pons + medulla oblongata. As the m-Ch receptors in the striatum and pons + medulla oblongata are affected by IM-stress, further studies of the m-Ch neural system must be performed under stressful situations in these regions as well as in the septum, hippocampus and cerebral cortex.

Bovine glomerular basement membrane (GBM) was isolated and purified according to a modification of Spiro’s method. Rat and bovine tubular basement membranes (TBM) were isolated and purified by sonic disruption or by the method of Carlson et al. Electron microscopic studies on the ultrastructure of GBM and TBM were performed after negative staining with 1% phosphotungstic acid solution, pH 7.3. When negatively stained, GBM and TBM were seen as fragments varying in size. The surface of the membranes showed a characteristic felt-like or spongy appearance. At higher magnification, GBM and TBM showed a fine meshwork composed of strands and pores which three-dimensionally resembled a crystal lattice. Pores were fairly uniform in size and shape. They were round, oval or polygonal in shape. Some of the pores were elongated to form short straight or bent channels. Strands were also uniform in diameter and surrounded a pore or channel. For an average of 50 pores, the long dimension was 3.1 ± 0.6 nm and the short dimension 2.5 ± 0.3 nm in bovine GBM, 3.8 ± 1.2 and 2.5 + 0.7 nm in bovine TBM, and 4.9 ± 1.5 and 2.8 ± 0.6 nm in rat TBM, respectively. The strand was 1.8 ± 0.3 nm in diameter in bovine GBM, 2.5 ± 0.6 nm in bovine TBM and 3.7 ± 0.7 nm in rat TBM for an average of 50 strands. The diameters of the pores were less than or close to the short axis of an albumin molecule. It was concluded that renal GBM and TBM were molecular sieves composed of pores and strands.

The characteristics of dopamine receptors, as well as the effects of denervation and dopamine infusion on dopamine receptors were studied using the radiolabeled receptor assay of [3H]-spiperone on rat kidney membrane preparations. The rat renal cortex was found to have a single class of [3H]-spiperone binding sites with a dissociation constant (Kd) of 13.5 +/- 2.2 nM. However, neither sulpiride nor serotonin strongly interacted with [3H]-spiperone binding, suggesting that DA1 receptors were predominant in the rat renal cortex. Denervation was performed by surgically stripping the nerves from the renal artery and coating them with 10% phenol. Chronic denervation had no significant effect on the affinity or maximum binding capacity of the renal dopamine receptors, although diuresis and the disappearance of catecholamine fluorophores in the denervated rats were observed. Chronic infusion of dopamine was performed using an osmotic minipump, resulting in a decrease in the number of rat kidney dopamine receptors. These results suggest that the dopamine receptor subtype in the renal cortex was mainly DA1, and that the major source of dopamine which affected the dopamine receptors in the rat kidney was not the nerve ending, but rather the circulation.

Painless (silent) thyroiditis (PT) occurred simultaneously in 1 male and 4 females aged 21 to 52 years working at a nursery school. Clinical symptoms did not include goiter, or pain or tenderness of the neck in any of the patients but were characterized by edema of the lower legs as well as palpitation and loss of body weight as observed in subacute thyroiditis. General blood analysis showed that all patients were negative for C-reactive protein (CRP) and 3 had mild impairment of liver function. Examination of thyroid function suggested transient thyrotoxicosis accompanied by a marked reduction in radioactive iodine uptake (RAIU), but antithyroglobulin hemagglutination antibody (TGHA) and antithyroid microsomal hemagglutination antibody (MCHA) were negative in all patients. Examination of various viral antibodies showed no significant changes in their titers. Thyrotoxicosis was transient and disappeared without treatment or by glucocorticoid administration. Our results suggested that PT observed in this study was caused by some environmental factor. The possibility of an unidentified virus as a factor cannot be ruled out.

The patient was admitted to our hospital at 19 and again at 22-yr of age for hirsutism and hypertension. Her baseline and ACTH-stimulated plasma 17-hydroxy pregnenolone, dehydroepiandrosterone and dehydroepiandrosterone sulfate were increased whereas plasma 17-hydroxy progesterone and androstenedione were normal and responded poorly to ACTH. Plasma deoxycorticosterone, corticosterone and cortisol baseline levels were normal, and they responded normally to ACTH. The plasma aldosterone concentration (PAC) was always high and responded well to ACTH, angiotensin III and furosemide-upright stimulation. However, plasma renin activity (PRA) was normal or slightly high, and responded normally to furosemide-upright stimulation and fluorohydrocortisone suppression. Dexamethasone (2 mg/day) for 1-2 weeks suppressed the androgens, cortisol and corticosterone levels. PRA and PAC were suppressed temporally, but PRA returned to normal and PAC to be a high level after 2 weeks of dexamethasone administration. Blood pressure was also reduced temporally but returned to a high level after 2 weeks of dexamethasone. These results indicate that primary aldosteronism and dexamethasone-suppressible hyperaldosteronism were not likely to be present, and unknown aldosterone stimulating factors which potentiated the action of endogenous angiotensin II or ACTH might be responsible for the hyperaldosteronism in this patient. We conclude that this patient had a mild and non-salt losing 3 beta-HSD deficiency in the zona reticularis with normal fasciculata and high glomerulosa function.

乳糜胸水を伴った40才の女性が来院,胸部CTでの多発性小嚢胞,肺生検によるごく軽度肺胞壁の線維化,胸管の通過障害の存在などから,びまん性過誤腫性肺脈管筋腫症と診断した.本疾患に乳糜胸水を伴うのは欧米と比べて本邦では少なく,本症例で3例目である.乳糜胸水貯留阻止を目的として,絶食と中心静脈栄養法に加え,胸腔内にOK-432,ドキシサイクリンを投与し,乳糜胸水貯留を阻止できたので報告する.

Synthetic ovine corticotropin releasing factor (CRF) at 0.03–30 ng/ml evoked a significant linear release of ACTH in pituitary monolayer cell cultures. Arginine vasopressin (AVP) at 3–30 ng/ml induced a slight ACTH release in the same culture system. CRF at 100 ng/ml greatly increased the medium ACTH with intracellular ACTH remaining relatively constant. In perifused pituitary quarters, both AVP and CRF evoked significant release of ACTH. When AVP was added in combination with synthetic ovine CRF, an additive or synergistic effect was observed on the CRF-induced ACTH release in both in vitro systems. Preincubation of cultured pituitary cells with synthetic ovine CRF for 1 h did not have a significant effect on the subsequent AVP-induced ACTH release. However, when cells were preincubated for 6 h with CRF at 1 and 100 ng/ml, the AVP-induced ACTH release was significantly increased, but preincubation with AVP suppressed both the basal and AVP-induced ACTH release. These results suggest that synthetic ovine CRF stimulates not only the release of ACTH but also the synthesis of ACTH and that AVP can evoke significant ACTH release from corticotrophs which have enough releasable ACTH or substances needed for ACTH release under the influence of CRF.

CRF and vasopressin concentrations in major brain regions after bilateral adrenalectomy and their involvement in adrenalectomy-induced ACTH secretion were investigated. At 5, 14 and 28 days after bilateral adrenalectomy, the plasma ACTH level was greatly elevated, whereas hypothalamic CRF content was reduced at 5 days and was not changed at 14 and 28 days after adrenalectomy. The CRF concentration in the medulla oblongata was reduced at 2-4 weeks after adrenalectomy. On the other hand, the arginine vasopressin (AVP) concentration was significantly elevated 2-4 weeks after adrenalectomy. An intrajugular administration of anti-ovine or anti-rat CRF serum significantly suppressed the elevated plasma ACTH level in adrenalectomized, freely moving rats, whereas anti-AVP serum or antipressor AVP antagonist, dpTyr(Me)AVP did not suppress the ACTH level. These results indicate that CRF played an important role in the adrenalectomy-induced ACTH elevation but that vasopressin was not involved.

白血病の治療を行なうに当つては,各種抗白血病剤の治療効果判定基準並びに治療目標の適確なものを得ることが極めて重要な臨床上の問題であると考え,臨床骨髄組織培養法によつてこの研究を行なつた.主として,白血病に特有なる増生様式が治療による緩解と共に如何に変貌するかを観察した.その結果,その増生様式の変化が最も端的に抗白血病剤の治療効果を反映するものであることを知つた.更に生体染色によつてこれら骨髄細胞の細胞学的変化を追求したところ,治療後の緩解時にも白血病はその細胞学的特徴を保持していることが明らかとなつた.又,骨髄巨核球の機能をも併せ検討することにより,白血病の治療に当つては急性白血病の場合は増生様式が正常型変化を,慢性白血病の場合は低形成型変化を目標として治療を行ない,慢性白血病においては低形成型に至るや直ちに投薬を中止して,経過を観察するのが最も良い方法であるとの結論を得た.

An electron microscope study was performed on the ultrastructure and developmental process of the Mukai strain of Japanese B encephalitis virus propagated in vitro on porcine kidney stable cells. On the basis of electron microscopic observations, it is suggested that the virus particles develop by “budding” from the membrane of the cytoplasmic vacuole. The particles then collect in the vacuole, usually in random arrangement, but occasionally in crystalline arrays. The densely packed vacuole migrates to the cell surface and there liberates the virus particles. It is also suggested that the JE virus particle is hexagonal in shape and approximately 40 mμ in the longest diameter; it is composed of an outer membrane 20 A thick, viroplasm 30 A thick, and an electron dense nucleoid 25 mμ in diameter.