Okayama University Medical School

IgE-mediated histamine release from whole blood was analyzed in 44 patients with bronchial asthma by observing maximum present release and dose-response curves of histamine release induced by anti-IgE and house dust extract. The maximum histamine release from whole blood induced by anti-IgE correlated with total serum IgE levels. There was a close correlation between allergen-induced release from whole blood and the serum levels of specific IgE antibodies. In the maximum histamine release from whole blood induced by both anti-IgE and allergen, the interaction with a serum factor was not clearly recognized. Effect of a serum factor was shown in the dose-response curves of anti-IgE-induced histamine release, but not in those of allergen-induced histamine release. The dose-response curves caused by anti-IgE showed that basophils from cases with a high serum IgE level require much more anti-IgE to produce maximum histamine release than basophils from cases with a low serum IgE level. The results showed that IgE molecules contained in the serum participate in anti-IgE-induced histamine release from whole blood.

Rats were fed a choline-free low protein diet for 12 or 26 weeks. In the 12-week group, the brain tyrosine concentration did not change. Dopamine levels were low in both the cerebral cortex and striatum. Norepinephrine level was low in the diencephalon. In the 26-week group, the tyrosine concentration was high in the brain. However, the dopamine and norepinephrine levels did not change in the cerebral cortex, striatum and hypothalamus. Furthermore, in another group of rats which were intraperitoneally injected with tyrosine, the brain tyrosine concentration was high, whereas the dopamine and norepinephrine levels in the hypothalamus were not significantly different from control levels.

The genetic polymorphism of factor B (Bf) was investigated in Okayama Prefecture, Japan. Cellogel immunofixation electrophoresis was employed according to Martin and Ziegler (1981) with minor modifications. In 316 non-blood related Japanese, the Bf was: Bf S, 70.6%; Bf FS, 27.8%; and Bf F, 1.6%. No rare variants were observed. The gene frequencies of Bfs and BfF were 0.845 and 0.155, respectively. The gene frequencies in Okayama Prefecture were quite similar to those in other districts of Japan. Considering the phenotype distribution in Japan, the Bf system might be a useful marker for personal identification and in disputed paternity cases.

The effects of angiotensin II, catecholamines and glucocorticoid on CRF-induced ACTH release were examined using rat anterior pituitary cells in monolayer culture. Synthetic ovine CRF induced a significant ACTH release in this system. Angiotensin II produced an additive effect on CRF-induced ACTH release. The ACTH releasing activity of CRF was potentiated by epinephrine and norepinephrine. Dopamine itself at 0.03-30 ng/ml did not show any significant effect on ACTH release, but it inhibited CRF-induced ACTH release. Corticosterone at 10(-7) and 10(-6)M inhibited CRF-induced ACTH release. These results indicate that angiotensin II, catecholamines and glucocorticoid modulate ACTH release at the pituitary level.

Simian virus 40 (SV40) large T antigen was partially purified from small amounts of SV40-infected and SV40-transformed cells by immunoaffinity chromatography with high recovery. T antigen, in both crude and partially purified states, was detected rapidly by a sensitive and quantitative enzyme-linked immunosorbent assay (ELISA). Stability of the partially purified T antigen was found to increase by addition of 0.01% bovine serum albumin (BSA).

The incidence of hepatitis A (HA), hepatitis B (HB), and non-A, non-B hepatitis (NANBH) was 27%, 30% and 43% among 73 patients with sporadic hepatitis. Epidemiological data (geographical distribution, seasonal variation, age, sex, and occupation) were not distinguishing of the type of hepatitis. Neither intrafamilial infection nor previous contact with viral hepatitis patients could be demonstrated in the NANBH cases. Fever and jaundice were less frequent in NANBH than in HA. Maximum levels of SGPT, serum bilirubin, ZTT, and gamma-globulin were significantly lower in NANBH than in HA and HB. Ten of 29 NANBH patients (35%) presented abnormal SGPT activities for more than 6 months, and four (14%) more than 12 months. In the ten patients with prolonged courses, jaundice was more frequent and maximum levels of SGPT were higher than in patients with transient courses. Histopathologic findings were not markedly different from those of HA and HB. Bile duct damage, fatty deposition, and giant multi-nucleated cells were recognized in 6, 12, and 2 NANBH patients, respectively. There were no characteristic ultrastructural changes in NANBH.

The concentrations and alpha-methyl-p-tyrosine (alpha-MPT) induced disappearance of catecholamines, adrenaline, noradrenaline and dopamine, were measured in selected areas of the brainstem and hypothalamus of spontaneously hypertensive rats (SHR) and deoxycorticosterone acetate (DOCA)-salt hypertensive rats. The catecholamine levels were measured by a sensitive radioenzymatic assay method combined with microdissection of the rat brain. The adrenaline concentration was higher in the area A1 of young SHR, but not in adult SHR, than in age-matched control rats. Noradrenaline concentrations and the alpha-MPT induced noradrenaline disappearance were less in the rostral part of the nucleus tractus solitarii (NTS) and the nucleus hypothalamic anterior of young SHR, and in the rostral part of the NTS of adult SHR. On the other hand in DOCA-salt hypertensive rats, the concentrations of adrenaline and noradrenaline were the same as in control rats in the examined areas. The alpha-MPT induced noradrenaline disappearance was less in the rostral part of the NTS of DOCA-salt hypertensive rats. Dopamine concentrations and the alpha-MPT induced dopamine disappearance were the same in the examined areas of SHR and DOCA-salt hypertensive rats. The results suggest that SHR have a change in adrenergic neural activity in the brainstem and a decrease in noradrenergic neural activity in the brainstem and hypothalamus while DOCA-salt hypertensive rats have a decrease in noradrenergic neural activity in the brainstem. Such changes in brain catecholaminergic neurons may have played an important role in the development of hypertension in these rats.

<p>A combination of agarose gel electrophoresis and a newly developed technique of electro-affinity transfer was applied to the detection of circulating immune complexes of human alpha-fetoprotein (AFP) and anti-AFP. After electrophoretic transfer to nitrocellulose membrane, to which affinity-purified polyclonal horse antibodies to human AFP were bound, the membranes were treated with or without rabbit immunoglobulins to human AFP, followed by overlaying with horseradish peroxidase-labeled goat anti-rabbit IgG for color development. Artificial complexes formed in vitro from human AFP and rabbit anti-AFP were clearly separated from free AFP by the agarose electrophoresis. The complexes were stained 20-40% as dark as the equivalent amount of free AFP by treatment with rabbit anti-AFP, and 10-20% as dark without the antibody treatment over a wide range of antigen-antibody ratios.</p>

Transaminative metabolism of L-cysteine was investigated using homogenates of guinea pig liver and kidney. L-Cysteine was transaminated in the presence of 2-oxoglutarate and the homogenate of either liver or kidney. S-(2-Hydroxy-2-carboxyethylthio)cysteine (HCETC) (3-mercaptolactate-cysteine disulfide) was formed by liver homogenate, but the amount was very small. On the other hand, a relatively large amount of HCETC was formed in the presence of kidney homogenate. Transamination between 3-mercaptopyruvate and certain amino acids was catalyzed actively by both liver and kidney homogenates in the presence of L-glutamate. However, more half-cysteine was formed by liver than kidney, and more HCETC was produced by kidney than liver. L-Glutamate was the most potent amino donor, and L-aspartate strongly inhibited the reaction. Results indicate that L-cysteine can be transaminated both in liver and kidney of the guinea pig, and that kidney is more active than liver. 2-Oxoglutarate is the most active 2-oxo acid for cysteine transamination. Oxaloacetate (and aspartate in the reverse reaction) is inhibitory to the reaction. These results are in agreement with the previous conclusion that cysteine aminotransferase is identical with aspartate aminotransferase.

In the presence of extracellular Ca2+, 6,7-dihydro-6,8,8, 10-tetramethyl-8H-pyrano [3, 2-g] chromone-2-carboxylic acid (EAA) had an inhibitory effect on the substance P-induced histamine release from rat peritoneal mast cells. Not only Ca2+ but also Mg2+, Sr2+ and Ba2+ were effective in enhancing the activity of EAA. Marked tachyphylaxis to EAA developed irrespective of the presence or absence of extracellular Ca2+. Cross-tachyphylaxis was observed between EAA and disodium cromoglycate (DSCG). These results indicate that the mode of action of EAA is similar, but not identical, with that of DSCG.

1. Absorption maxima of hydrochloric biliverdins derived from the natural indirect bilirubin existed at 680 mμ and 375 mμ, but the maxima of biliverdins purified on the column of silica gel existed at 640 mμ and 390 mμ. 2. The natural salt-form bilirubin was oxidized by hydrochloric acid to biliverdin, of which absorption maxima existed at 685 mμ and 370 mμ in a methanolic solution as well as in 5% hydrochloric methanol, but the purified biliverdin in chloroform solution showed the maxima at 640 mμ and 390 mμ. 3. The natural ester-form bilirubin could be transformed into biliverdin by oxidation of its alcoholic solution in the presence of hydrochloric acid. The crude biliverdin had absorption maxima at 645 to 655 mμ, 600 mμ and 320 mμ, and the crude hydrochloric biliverdin had the maxima at 665 to 675 mμ, 620 mμ and in the near ultra-violet range, while the purified biliverdin in chloroform solution had the maxima at 640 mμ and 380 mμ. 4. The biliverdins derived from the indirect, salt-form and ester-form bilirubin had quite similar absorption maxima after purifications by adsorption chromatography.

Diacetyl can serve as an acetyl donor for the forlnation of citrate and the acetylation of sulfanilamide in the dog heart homogenate. Diphosphothiamine and coenzyme A are essential for these reactions.

Separation of both forms of the direct bilirubin were carried out from the dog's gallbladder bile, and further isolations of them were also done. 1. The natural salt-form bilirubin was isolated after separation on the column of aluminium oxide with a n-propanolic aqueous solution. 2. The natural salt-form bilirubin was obtained in amorphous yellow powders which were strongly hygroscopic and easily soluble in water and methanol but not in chloroform or carbon tetrachloride. An aqueous solution of these powders showed both the direct diazo and Gmelin reaction, but neither Ehrlich's aldehyde nor Schlesinger reaction. The salt-form bilirubin was transferred into chloroform only when some quantities of hydrochloric acid were added to a mixture of chloroform and an aqueous solution of it. 3. The absorption maxima of the natural salt-form bilirubin existed at 420 to 430 mμ in a methanolic solution and at 425 or 435mμ in 50% or 10% n-propanol. 4. The natural ester-form bilirubin was isolated after separating on the column of silica gel with a chloroformethanolic mixture. 5. The natural ester-form bilirubin was obtained in amorphous greenish yellow powders. It was further hygroscopic and easily soluble in water and methanol but not in chloroform or carbon tetrachloride. An aqueous solution of it showed the direct diazo and Gmelin reaction, but neither Ehrlich's aldehyde nor Schlesinger's reaction. No pigment was transferred into chloroform even if some quantities of hydrochloric acid were added to a mixture of chloroform and an aqueous solution of it, but did by saponification with 5% methanolic potash. 6. The absorption maxima of the natural ester-form bilirubin existed at 415 mμ in both methanolic and aqueous solutions.

Descriptions are carried on the method how to separate the indirect bilirubin from the chloroform extracts of the dried dog's gallbeadder bile by adsorption chromatography. 1. The optimal concentrations of the bilirubin content were 2 to 4 mg/100 ml when 1 ml of the sample was adsorbed on the Tswett tube of about 10 mm diameter. 2. Though several zones of the indirect bilirubin were separated on the column of silica gel when developed with various solvents, these zones were proved to be mingled with some oxidized or other intermediate products and the separation like this was thought to owe to the activity of the adsorbents. 3. The chromatogram of the crystalline bilirubin resembled to the one formed by the indirect bilirubin in the chloroform extracts. 4. The chromatogram of the chromatographically separated indirect bilirubin was similar to the former. 5. The absorption maxima of a chloroform solution of the natural indirect bilirubin existed at 450 mμ in the visible range, and it was the same as the maxima of the crystalline bilirubins.

<P>The effect of P³² in the experimental Japanese B Encephalitis infection was investigated. A remarkable delay of infection was observed, when a therapeutic dose of P³² was administered intraperitoneally to the mouse which had been inoculated intracerebrally with the virus suspension at the concentration of about LD50. Almost equal results were obtained by intraperitoneal administrations to the mice when they had deen inoculated intravenously or intraperitoneally.

1. Sinomenine and Irgapyrin, the two antirheumatics known to be capable of releasing histamine, caused a marked gastric secretion in the unanesthetized dog. 2. The facial edema and itching associated with histamine release by sinomenine was almost completely eliminated by NeoAntergan, but the gastric secretion was not suppressed, or rather increased - an observation also reported by Paton and Schachter with Compound 48/80. This indicates that the histamine release cannot be markedly prevented by antihistamine agents in this animal. 3. The gastric secretion induced by Irgapyrin was not suppressed by Neo-Antergan but Irgapyrin originally never caused other symptoms associated with histamine release. This is probably due to the antihistamine action inherent in this compound itself. 4. No such histamine-releasing activity, as determined by gastric secretion, could be observed in aminopyrine or butazolidine sodium, the components of Irgapyrin. 5. Sinomenine, differing from Irgapyrin and Compound 48/80, was ineffective by intramuscular injection.

Our methods of tissue culture of the bone marrow, lymphnodes, and peripheral blood were described. Furthermore, for the purpose of promoting wide clinical application of bone marrow tissue culture, our simple vital inspection method was also stated.

In decerebrated cats the impulse discharges were detected by means of an unipolar microelectrode from a single inspiratory neuron in the respiratory centers, and the change in discharge produced by administration of stimulating, as well as depressant agents, was studied. The results were summarized as follows. 1) Inhalation of the air containing all. excess of carbon dioxide, as well as the air deficient of oxygen and the intravenous administration of a small dose of caffeine, aminocordine and lobeline produced a) a remarkable increase of frequency of impulses in the stage of constant frequency of the volley, b) a rapid increase of frequency of impulses in the stage of crescent frequency of the volley and c) a shortening of the duration of the volleys, as well as of silent periods. 2) In narcosis by a moderate dose of morphine, as well as in the recovery stage from apnea produced by over-ventilation, there were observed the phenomena which were exactly opposite to those described in 1). 3) It was concluded that a most essential sign by which one can discern whether the activity of the respiratory Genters is raised or depressed, is the changes of the frequency of impulses produced from an inspiratory neuron. The expense of this research was defrayed from the grant in aid of the Ministry of Education.

1) As the boundary of the outgrowth zone of the bone marrow culture is sharply defined from the surrounding area and the cellular density becomes very high in leukemia, diagnosis can be made with ease and certainty. 2) The differential diagnosis between aleukemic leukemia and aplastic anemia becomes easy by application of the tissue cuIture method. 3) The vital observation of blood cells were used in classification of leukemia by cellular systems, and it was discovered that there were a lot of patients with monocytic leukemia and relatively many with acute lymphatic leukemia. 4) Because the cellular growth in the bone marrow culture of aplastic anemia declines extremely and abundant fat cells are observed, the diagnosis of this disease is easily made by employment of this method.

(1) The movement of the blood cells in the bone marrow was classified into 9 types. (2) The characteristics of moving types are so distinct according to the kinds of blood cells, that the differential diagnosis of the cells by moving types is easily and certainly made. In this way, (by the kind of blood cells), we have classified leukemia, as is described in our other articles. (3) The phagocytosis and vital staining of the blood cells in the bone marrow is different in degree and mode, according to the kind of blood cells, and thus becomes valuable ground for the differential diagnosis of the cells.